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JZL184, as a monoacylglycerol lipase inhibitor, down-regulates inflammation in a cannabinoid pathway dependent manner
Biomedicine & Pharmacotherapy 103 (2018) 1720–1726
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JZL184, as a monoacylglycerol lipase inhibitor, down-regulates inflammation in a cannabinoid pathway dependent manner
T
Mohammad-Reza Rahmania,b, Ali Shamsizadeha,c, Amir Moghadam-Ahmadia,d, ⁎ Gholamreza Bazmandegana,c, Mohammad Allahtavakolia,c, a
Physiology-Pharmacology Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran Student Research Committee, Rafsanjan University of Medical Sciences, Rafsanjan, Iran c Department of Physiology and Pharmacology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran d Department of Neurology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran b
A R T I C LE I N FO
A B S T R A C T
Keywords: Stroke PMCAO JZL-184 AM251 Cannabinoids
Introduction: Stroke is a prevalent disorder which is associated with several complications including inflammation. JZL-184 (JZL) inhibits arachidonic acid (AA) production and consequently results in two-arachidonoylglycerol (2-AG) accumulation. Both reduced production of AA metabolic products and increased 2-AG, the agonist of type 1 cannabinoid receptor (CB1), can result in reduced inflammation. In this study, we investigated the mechanisms of JZL in the improvement of stroke complications in mouse permanent cerebral ischemia (PPMCAO) model using AM251, the antagonist of CB1. Material and methods: PMCAO mice were divided into six groups including intact, controls, vehicle, JZL, AM251 and JZL plus AM251 administrated groups. Brain infarction and edema, brain levels of matrix metalloperoteinase-9 (MMP9), interleukin (IL)-10 and tumor necrosis factor-α (TNF-α) and behavioral functions have been examined in all groups. Results: The results showed that JZL lowered brain infarction, neurological disorders, TNF-α and MMP9 more effectively than JZL plus AM251. JZL and JZL plus AM251 reduced brain edema and increased brain IL-10. JZL, AM251 and JZL plus AM251 improve behavioral functions. Discussion: JZL reduces brain infarction and brain pro-inflammatory molecules in CB1 pathway dependent manner. JZL also reduces brain edema and increased IL-10 in CB1 pathways or decreased AA metabolites. Further, AM251 improves behavioral functions via unknown mechanisms.
1. Introduction Cannabinoids are a set of molecules which have several effects including anti-nausea, anti-cancer, anti-seizure, anti-oxidation, anti-inflammation and neuroprotection [1]. Cannabinoids perform their functions via interactions with two series of receptors, type 1 and 2 cannabinoid receptor (CB1 and 2). CB1is expressed in either brain or peripheral tissues, while CB2 is expressed in the peripheral tissues only [2]. Therefore, the altered behaviors following administration of cannabinoids are related to the interaction of the molecules with CB1 [3]. It has been indicated that two series of molecules including anandamide (N-arachidonoylethanolamine) and two-arachidonoylglycerol (2-AG) are the endogenous ligands for CB1 which are entitled endocannabinoids [4]. Thus, interactions of these ligands with CB1 mimic the effects of cannabinoids such as anti-inflammatory effects and neuron protection [5]. Many investigators indicate that, the protective ⁎
properties of CB1 receptors and using its agonists in ischemic brain injuries have been proved by investigators [6,7]. Accordingly, Nagayama et al., revealed that R(+)-WIN 55212-2, a synthetic cannabinoid agonist, reduced hippocampal neuronal damages and also brain infarctions following induction of MCAO in rat animal models [7]. Interestingly, some investigators reported that using CB1 antagonists results in blocking neuroprotection of CB1 agonist [6,7]. Therefore, therapeutic strategies which increased the brain endocannabinoids may hamper brain pro-inflammatory based disorders such as stroke [8]. JZL-184 (JZL) is a potential irreversible inhibitor of monoacylglycerol lipase (MAGL). MAGL is an important enzyme which facilitates the catabolism of 2-AG to produce arachidonic acid (AA) [9,10]. Prostaglandins (PGs), leukotrienes (LKs) and platelet activator factor (PAF), the important pro-inflammatory factors, are produced from AA, hence, we hypothesized that administration of JZL might decrease inflammation via two distinct pathways, increased levels of 2-
Corresponding author at: Physiology- Pharmacology Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran. E-mail address: [email protected] (M. Allahtavakoli).
https://doi.org/10.1016/j.biopha.2018.05.001 Received 27 September 2017; Received in revised form 1 May 2018; Accepted 2 May 2018 0753-3322/ © 2018 Published by Elsevier Masson SAS.
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(3 mg/kg), respectively. Finally, the sixth group, JZL plus AM251, contains the ischemic mice which have been treated with both JZL (4 mg/kg) and AM251 (3 mg/kg) intra-peritoneal. Accordingly, the drugs were administrated immediately after induction of PMCAO and the experiments were performed 48 h after single dose of the drug injections.
AG and consequently activation of CB1 as well as decreased levels of PGs, LKs and PAF [10,11]. Brain stroke is a human disorder which is associated with several mortalities and disabilities [12]. Following brain stroke, the central cells within the region of brain ischemia are damaged at once, and the penumbra region cells are alive, though they do not have normal function and are at risk of necrosis and consequently induction of the stroke complications [13,14]. Thus, several investigations are designed to find new strategies to protect the neurons in the penumbra area [15,16]. Ischemia and its related complications including brain inflammation and edema, is one of the most important inducers of penumbra region cell damage [12,17,18]. Tumor necrosis factor-alpha (TNF-α) and matrix metalloproteinase-9 (MMP9), as the pro-inflammatory molecules, and interleukin-10 (IL-10), as the anti-inflammatory cytokines, play crucial roles in the pathogenesis of stroke [19]. Therefore, expression of these cytokines and also the examination of brain edema and stroke infarctions might be remarkable determinants to check stroke treatment procedures. The previous study reported that JZL at doses of 4, 8 and 16 mg/kg has similar effects on 2-AG in animal model [20], Therefore, in order to reduce the side effects, the minimum effective dose should be used [21,22]. Thus, this project was designed to evaluate the effect of JZL (4 mg/kg) on the improvement of PMCAO behavioral functions and also inflammatory markers. Infarct area, edema cerebral inflammatory molecules TNF and MMP9 [23,24], as well as IL-10, as anti-inflammatory cytokine [25] were measured in the mouse permanent cerebral ischemia (PMCAO) model using JZL at dose of 4 mg/kg [26]. Additionally, another investigation by our research team revealed that MMP9 has pro-inflammatory effects during stroke [27]. Another study by Montaner et al., proved that MMP-9 is the main reason for brain hemorrhage and edema in the animal stroke model [28]. The main mechanisms used by JZL to hamper inflammation in the PMCAO model have yet to be clarified. Thus, this study was aimed to evaluate the effects of JZL, at 4 mg/kg, on the CB1/endocannabinoid pathway in reduction of inflammation, via evaluation of MMP9, TNF-α and IL-10 as pro/anti-inflammatory markers, brain edema/infarction and improvement of behavioral functions in PMCAO stroke model using the CB1 antagonist, AM251. AM251 is an antagonist of CB1 which is used in this project to block CB1 pathway to evaluate JZL mechanisms used to improve stroke secondary side effects.
2.3. Mouse permanent model of middle cerebral artery occlusion (PMCAO) establishment PMCAO model was induced in the experimental mice under anesthesia (ketamine 90 mg/kg plus xylazine 4/5 mg/kg) as described previously [32,33]. Briefly, after producing a small vertical incision on the midline of right ear and eye, temporalis muscle was retracted. Then, the bone was laterally cut and moved on the temporal area of the skull to make an approximately 1mm2 in diameter hole, just above the MCA. Accordingly, the dura was carefully removed, and then the root of MCA was cauterized immediately. Eventually, the mouse muscle was replaced and the skin was closed carefully.
2.4. Infarct volume and brain edema measurement Infarct volume was evaluated using the Zhang and Iadecola protocol [32], briefly, the mice were sacrificed 48 h after PMCAO induction and the brains were removed. After that, coronal slices, 1-mm-thick were made. Then the slices were stained using 2,3,5-triphenyltetrazolium chloride 1% (Sigma Chemical Co., St. Louis, MO, USA). The protocol results in staining and no red staining of the non-infarcted and infarcted brain tissue, respectively. The infarcted tissues were analyzed using the Image J software (NIH Image, version1.61, Bethesda, Maryland, USA) after demarcation. In order to calculate the total infarction, all the infarct zones were summed and multiplied in the thickness of the brain sections according to the Zhang and Iadecola protocol [32]. The corrected infarct volume was calculated by the following formula: infarct area × (1- [(ipsilateral hemisphere area-contralateral hemisphere area)/contralateral hemisphere]) [34]. Brain edema was calculated and reported as percentage using the formula of O’Donnell and colleagues [35] as follows: (volume of left hemisphere - volume of right hemisphere)/volume of right hemisphere.
2. Material and method
2.5. MMP-9, IL-10 and TNF-α brain levels
2.1. Animals
MMP-9 (R and D system), IL-10 and TNF-α (eBiosciences) brain levels were assessed using commercial ELISA kits based on the companies` instructions. In brief, the ischemic brain tissue samples were homogenized mechanically and then added to the anti-IL-10, antiMMP-9, and anti-TNF-α capture antibody pre-coated ELISA plates. After incubation for 2 h in the dark (at room temperature), the ELISA plates were washed 3 times with washing buffer and incubated in the dark at room temperature for an hour with conjugated HRP detection secondary antibodies. Then, HRP substrate (3, 3′, 5, 5′Tetramethylbenzidine (TMB) + H2O2) was added to the ELISA plates and they were incubated in dark place for 15 min and the chemical reaction was stopped via sulfuric acid (2 N). The optical densities (OD) were evaluated at 450 nm by a microplate ELISA reader (Bio-Rad, USA).
This experimental study has been performed on 78 male mice between 25–35 g and 8–10 weeks-old in the standard conditions (freely available food and water and animals kept at 37 °C and 12:12-hr light/ dark cycles). Local Ethical Committee has approved the experimental procedures (Code: IR.RUMS.REC.1394.88). Accordingly, all the mice handling and the experiments were done using the methods to minimize suffering. 2.2. Experimental groups 78 male mice were randomly divided into the six experimental groups including intact, control, vehicle, (JZL) 4 mg/kg [9,29], JZL plus AM251 and AM251 3 mg/kg groups [30]. Accordingly, 13 mice were placed in each group (8 animals were used to assess edema, infarct volume and behavioral experiments and five animals were used to determine cerebral levels of TNF-α, IL-10 or MMP-9). The control group contained the ischemic mice that were not treated with any drug, while vehicle group was treated with 300 mg/kg dimethyl sulfoxide (DMSO 10%) intra-peritoneal [31]. JZL and AM251 groups contain the ischemic animals which were treated with JZL (4 mg/kg) and AM251
2.6. Sticky testing Adhesive removal (sticky tape) test was applied to explore the sensorimotor functions, based on the Whishaw method protocol [36]. The sensorimotor behavior was determined before and 24 or 48 h after PMCAO insult, the averaged latency to remove sticky tape during 3 trials was recorded [37] 1721
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2.7. Hanging wire test In order to evaluate muscle strength of the PMCAO models, the animal was hung by forepaw from horizontal steel wire in a triplicate format. Using a stopwatch, the fall was recorded from the time the animal grasped the wire. 2.8. Neurological disorder assessment Rating neurological disorders was recorded using Bederson grading system [18] at 2, 24 and 48 h after PMCAO induction. Neurological disorders were rated using the score scales: 0, no observable sign; 1, forelimb flexion; 2, forelimb flexion plus reduced resistance to lateral push; 3, unidirectional circling; 4, unidirectional circling plus reduced levels of consciousness; and 5, death.
Fig. 2. The effects of JZL-184 and AM251 on brain edema in MCAO model. The figure shows that JZL (*) significantly reduced brain edema when compared to control and vehicle groups. AM251 alone and JZL plus AM251 could not reduce brain edema.
2.9. Statistical analysis
3.2. Brain edema
Data were presented as mean ± standard error. Differences in brain levels of MMP9, IL-10 and TNF-α, infarct volume, paresis and sensorimotor disorder were analyzed using Two Way ANOVA followed by TUKEY test. Neurological disorder was also analyzed using nonparametric tests including Kruskal-Wallis and Mann-Whitney tests and, hence, the results were reported as medians. A P value of less than 5% was considered to be significant.
Brain edema was assessed using O'Donnell et al. method as a marker of stroke damage to explain the effects of drugs on reduction of stroke secondary side effects. The brain edema, 48 h after PMCAO, was significantly decreased in JZL (p = 0.002) in comparison to vehicle group (Fig. 2), while it was not significant for AM251 plus JZL group (p = 0.188) and AM251 (p = 0.765). The brain edema in JZL treated group was significantly decreased when compared to AM251 (p < 0.001) and control (p < 0.001) groups, but was not different when compared to AM251 plus JZL group (p = 0.367, Fig. 2).
3. Results 3.1. Brain infarction
3.3. TNF-a, IL-10, and MMP9 levels
Brain infarction was evaluated to show the size of neurons damages by stroke and also the efficacy of the drugs to reduce the stroke effects. Fig. 1 presents the raw data regarding the brain infarct volumes 48 h after PMCAO induction. The infarct volumes in the JZL group were decreased significantly in comparison to control (p = 0.002), vehicle (p = 0.009) and AM251 (p = 0.008) groups. Additionally, there were not significant differences between JZL and JZL plus AM251 (p = 0.365). While there were not significant differences between AM251 and control (p = 0.979), vehicle (p > 1.00), as well as JZL plus AM251 (p = 0.360) groups. JZL plus AM251 group also did not have significant differences with vehicle group.
TNF-α, IL-10, and MMP9 brain levels were evaluated using ELISA method to determine inflammation scales. Data analysis demonstrated that the following 48 h’ cerebral levels of TNF-α were significantly hampered after treatment with JZL (p = 0.014), while it was not changed after treatment with AM251 (p = 0.216) and JZL plus AM251 (p = 0.100) when compared to vehicle. There were no significant differences between the following groups regarding brain levels of TNF-α: Vehicle and controls (p = 0.1), JZL and intact (p = 0.920), JZL and AM251 (p = 0.651), JZL and JZL plus AM251 (p = 0.876) as well as AM251 and JZL plus AM251 (p = 0.992) groups. Fig. 3 illustrates the raw data regarding brain levels of TNF-α. Treatment of the PMCAO animals with JZL (p < 0.001), but not JZL plus AM251 (p = 0.106) and AM251 (p = 0.744), led to increased expression of IL-10 in comparison to vehicle group. Interestingly, JZL increased IL-10 brain levels when compared to intact group (p < 0.001). Although there was a significant difference between JZL and AM251 groups (p = 0.008), the difference between JZL and JZL plus AM251 groups was not significant (p = 0.138, Fig. 3). Administration of JZL (p = 0.011), but not AM251 (p = 0.524) and JZL plus AM251 (p = 0.099), led to decreased expression of MMP9 in comparison to vehicle group. There were not significant differences between JZL and AM251 (p = 0.255), JZL and JZL plus AM251 (p = 0.832) and JZL with intact (p = 0.170) groups (Fig. 3). 3.4. Sticky test
Fig. 1. The effects of JZL, AM251 and JZL plus AM251 on infarct volumes in MCAO model. Infarct volumes are presented as a percentage of affected ipsilateral hemispheres. Data are expressed as a mean ± SEM. (*) JZL 4 mg/kg significantly reduced the infarct volumes when compared to control and vehicle group.Control and vehicle groups had not significant difference.AM251 was unable to reduce infract volume as well as JZL. AM251 also neutralized the protective roles of JZL in the group which treated with JZL and AM251 simultaneously.
Sticky test was used to evaluate the sensorimotor function, as a behavioral function using Whishaw method. Fig. 4 shows latency in removing the contralateral forepaw label 24 and 48 h after induction of PMCAO. JZL (p < 0.001 for 24 h and p = 0.004 for 48 h), AM251 (p = 0.003 for 24 h and p = 0.002 for 48 h) and JZL plus AM251 (p < 0.001 for 24 h and p = 0.001 for 48 h) improved the time of sticky test when compared to the vehicle group. There were significant 1722
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Fig. 3. The effects of AM251, JZL and JZL plus AM251 on response latency in adhesive removal test. JZL (*), AM251 (*) and JZL plus AM251 (*) administration were associated with improved response latency in adhesive removal test when compared to control and vehicle groups. There were not significant differences between AM251/JZL plus AM251 and JZL group (*). Control and vehicle groups also had not significant differences regarding hanging time.
PMCAO complications. Additionally, AM251 also neutralized the protective roles operated by JZL in the reduction of infarct volume. And while the figure illustrates a various range of infarctions between groups, the statistical analysis showed that the differences were not significant. Interestingly, the results obtained from brain edema also proved the neuroprotective roles operated by JZL and revealed that JZL, but not AM251 and JZL plus AM251, reduced the cerebral edema in the PMCAO animal models. According to the results and based on the mechanisms used by JZL regarding inhibition of production of arachidonic acid metabolites and elevation of CB1 agonist, 2-AG, it appears that JZL may be considered to decrease inflammation in the infarcted brain tissues. However, due to the fact that the differences between JZL and JZL plus AM251treated groups were not significant, hence, it may be hypothesized that JZL reduces brain infarction and also edema via down-regulation of AA metabolites which need to be explored by additional investigations. Baba et al., reported that AA metabolites are the most important molecules that participate in the induction of contraction in endothelial cell and consequently blood brain barrier (BBB) leak which is damaged during brain edema [38]. Thus, it may be supposed that JZL uses down-regulation of AA metabolites to suppress infarction and edema. The results also showed that although JZL decreased brain levels of MMP9 and TNF-α, treatment of the PMCAO mice with a combination of JZL and AM251 had no effects on the brain levels of these inflammatory molecules. Additionally, JZL decreased brain levels of the studied mice to reach to the intact group. The use of JZL is associated with 2-AG accumulation, the molecule is the ligand for CB1 and according to the results which showed that CB1 antagonist, AM251, neutralized the antiinflammatory and anti-infarct properties of JZL, it may be hypothesized that JZL may reduce infarct volumes and brain edema via down-regulation of pro-inflammatory molecules, MMP9 and TNF-α. Although the data directed the conclusion to the involvement of CB1 pathway in reduction of inflammation in the stroke region of the brain, AM251 has not affected the inflammation when compared to JZL treated group. Thus, again, it seems that down-regulation of AA metabolites may be considered as the main pathway which is used by JZL to reduce inflammation in the brain stroke. Pihlaja et al., also showed that JZL plays important neuroprotective roles in an Alzheimer's disease animal model through suppression of inflammation [39]. Furthermore, based on the mentioned results, it may also be demonstrated that reduction in production of AA and its pro-inflammatory metabolites, PGs, LKs and PAFs,
differences between JZL group at 24 h (p = 0.002) and 48 h (p < 0.001) with the intact group. There were no significant differences between JZL and AM251 groups (p = 0.698 for 24 h and p = 0.992 for 48 h), JZL and JZL plus AM251 groups (p = 0.978 for 24 h and p = 0.982 for 48 h) and also between AM251 and JZL plus AM251 groups (p = 0.953 for 24 h and p = 0.999 for 48 h). 3.5. Hanging wire test Hanging test is a test to evaluate the muscle performance which is categorized as behavioral function. Hanging wire test in both 24 and 48 h had similar results. Accordingly, the results revealed that treatment with JZL (p = 0.006 for 24 and p < 0.001 for 48 h), AM251 (p = 0.029 for 24 and p < 0.001 for 48 h) and JZL plus AM251 (p = 0.018 for 24 h and p < 0.001 for 48 h) significantly increased endurance time when compared to vehicle group (Fig. 5). However, JZL after both 24 and 48 h, was unable to improve hanging test to reach to the intact group (p < 0.001). There were no significant differences between two untreated groups including control and vehicle groups (p = 0.782 for 24 h and p = 0.966 for 48 h) and also between JZL and AM251 (p = 0.973 for 24 h and p = 0.927 for 48 h) and JZL plus AM251 (p = 0.993 for 24 h and p = 0.994 for 48 h) groups (Fig. 5). 3.6. Bederson test Rating neurological disorders was recorded using Bederson grading system. The test shows that JZL significantly improved the neurologic damages in the JZL group only (p < 0.001 for both 24 and 48 h). Neurological deficits were not changed after administration of AM251 (p > 0.05 for both 24 and 48 h) and JZL plus AM251 groups (p > 0.05 for both 24 and 48 h, Table1). 4. Discussion JZL component plays as an irreversible MAGL inhibitor, the enzyme responsible for the catabolism of 2-AG to produce arachidonic acid. Thus, JZL not only reduces production of arachidonic acid metabolites, it increases the 2-AG as ligands for CB1 receptors [10,20,37]. Therefore, JZL administration may be associated with hampering inflammation in stimulation of CB1 and suppression of AA metabolites behavior [10,11]. The results demonstrated that, although JZL reduced infarct volume in 4 mg/kg concentration, AM251 was unable to protect the mice from
Fig. 4. The effects of AM251, JZL and JZL plus AM251 on hanging time. JZL (*), AM251 (*) and JZL plus AM251 (*) administration were associated with improved behavioral functions when compared to control and vehicle groups. There were not significant differences between AM251/JZL plus AM251 and JZL group. Control and vehicle groups also had not significant differences regarding hanging time.
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Fig. 5. The effects of AM251, JZL-184 and a combination of JZL and AM251 on the brain levels of TNF-α, IL-10 and MMP9 in the MCAO mice. JZL (*), but not AM251 and AM251 plus JZL, significantly reduced brain levels of TNFα and MMP9 in MCAO mice. Both JZL and AM251 plus JZL were able to increase brain levels of IL-10. The differences between control and vehicle groups regarding brain levels of TNF-α, IL-10 and MMP9 were not significant.
inflammation [40]. However, there are several anti-inflammatory cytokines such as tumor growth factor-beta (TGF-β), IL-25 and so on, hence it may be hypothesized that other anti-inflammatory cytokines may also participate in the declination of inflammation using JZL which need to be explored by further investigations. Collectively, it seems that JZL may play important roles in the reduction of brain stroke complication via down-regulation of inflammation. Additionally, Bederson test also proved the roles of CB1 pathway in the neuron functions. Accordingly, using JZL alone, but not in combination with AM251, leads to improved neurological disorders. AM251 blocked the CB1 pathway, hence, this pathway may play pivotal roles in the prevention of neuron damages during stroke which is proved by several investigations [41,42]. However, JZL also blocked AA metabolite production; further investigations need to be performed to evaluate the measure of AA metabolites after administration of AM251. Previous investigations also proved the hypothesis and demonstrated that JZL improves neuron functions in Parkinson's disease animal models and down syndrome in cannabinoid receptor dependent manner [43,44]. The important roles played by JZL in modulation of neuron functions via CB1 pathway have also been evidenced by several investigators [1,4,45]. Collectively, it appears that JZL may inhibit penumbra region cells damages in the PMCAO animal model through reduction of infarction in CB1 and edema in both CB1 and AA metabolites dependent manners. Moreover, both endogenous and exogenous cannabinoids play neuroprotective roles during cerebral damages such as ischemia [43,46–48], therefore, it seems that JZL via up-regulation of endogenous cannabinoids activates CB1 pathway and inhibits AA metabolites production, reduces brain infarct and edema, protects penumbra cells and improves neuron functions. Therefore, it has been demonstrated that each factor which rescues the ischemic neurons from death and induces brain reperfusion can be considered as a
Table 1 Bederson test results in different groups at 2, 24 and 48 h after brain ischemia. Group
2h
24 h
48 h
Control Vehicle JZL AM251 JZL plus AM251
3 (2–3) 3 (3–3) 3 (2.75–4) 3.5 (3–4)** 3 (2–3)¶
3 3 2 3 3
4 (3–4) 3.5 (3–4) 1 (1–2)* 3 (3–4)# 3 (2.75–3.5)###
(3–3.25) (3–3.25) (1.75–2)* (3–3)# (2.75–4.25)##
Effect of JZl, AM251 and JZL plus AM251 on neurological deficits was assessed by a 5-score scales at 2, 24 and 48 h the following stroke. Data are presented as median, 25th and 75th percentiles (percentiles in the parentheses). * p < 0.001. ** p < 0.05 versus vehicle group. ¶ p < 0.05 versus AM251. # p < 0.001. ## p < 0.01 versus JZL.
were not associated with reduced infarct volumes because infarct volumes were not altered in the PMCAO group which was treated with JZL plus AM251. Thus, it seems that JZL may reduce infarct volume via decreased inflammation in both CB1 dependent manner and also downregulation of AA metabolites. In parallel with the results, JZL was able to up-regulate IL-10 in the brain of PMCAO mice and also increased its levels in the JZL group when compared to intact group which confirms its anti-inflammatory effects. IL-10 is the most powerful anti-inflammatory cytokine, thus, it can be concluded that increased brain expression of IL-10 following administration of JZL is associated with down-regulation of pro-inflammatory molecules, MMP9 and TNF-α. Interestingly, our previous investigation on the PMCAO model using transient receptor potential vanilloid-1 (TRPV1) antagonist (AMG9810) leads to decreased brain infarct and edema via down-regulation and upregulation of TNF-α and IL-10 which collectively leads to decreased 1724
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neuroprotective agent [49,50]. Thus, based on the outcomes obtained from JZL on the PMCAO models, it may be hypothesized that JZL can be considered as a neuroprotective drug. Although JZL and JZL plus AM251 were unable to improve the sticky and hanging tests in the PMCAO models to reach to the intact group, the outcomes revealed that hanging test and sticky removal test were improved in all of the three groups including JZL and JZL plus AM251 receiving mice (Figs. 3 and 4). However, due to the fact that AM251 suppresses CB1 pathway, it seems that JZL progresses the behavioral functions independent of CB1 pathway. Interestingly, administration of AM251 alone also improved the behavioral functions. Knowles et al., also proved the roles of AM251 in improvement of behavioral functions in stroke animal models [51]. Some investigations proved that there were not significant correlations between the behavioral tests, such as sticky tape and hanging test, with infarction volumes and edema percentage [52]. Moreover, in agreement with our results, it has been shown that AM251 can improve behavioral tests [53]. Thus, the results which are presented in this project, and also suppression of CB1 receptor are associated with increased 2-AG and also decreased AA metabolites, hence, it can perhaps be hypothesized that 2-AG may improve the behavioral functions via an unknown 2-AG dependent mechanism. Collectively, based on the results it seems that JZL is an important agent which might be considered as anti-stroke complication. However, in order to describe the main mechanisms used by JZL to reduce inflammation, brain infarcts and edema, authors of this article suggest different CB1R compound tools (agonist/antagonist) be used to study the link between MAGL inhibition by JZL184 and CB1 receptor signaling pathway.
[14] [15]
[16]
[17] [18]
[19] [20]
[21] [22] [23]
[24]
[25]
Acknowledgment [26]
This work is derived from Mr. Mohammad-Reza Rahmani’s PhD dissertation and was supported by grant no. 20/611 from the ViceChancellor for Research and Technology, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.
[27]
References
[28]
[1] F. Grotenhermen, Cannabinoids, Curr. Drug Target CNS Neurol. Disord. 4 (5) (2005) 507–530. [2] F. Chaperon, M.H. Thiebot, Behavioral effects of cannabinoid agents in animals, Crit. Rev. Neurobiol. 13 (3) (1999) 243–281. [3] D. Panikashvili, C. Simeonidou, S. Ben-Shabat, L. Hanus, A. Breuer, R. Mechoulam, E. Shohami, An endogenous cannabinoid (2-AG) is neuroprotective after brain injury, Nature 413 (6855) (2001) 527–531. [4] M.R. Pazos, E. Nunez, C. Benito, R.M. Tolon, J. Romero, Functional neuroanatomy of the endocannabinoid system, Pharmacol. Biochem. Behav. 81 (2) (2005) 239–247. [5] R. Gallily, A. Breuer, R. Mechoulam, 2-Arachidonylglycerol, an endogenous cannabinoid, inhibits tumor necrosis factor-alpha production in murine macrophages, and in mice, Eur. J. Pharmacol. 406 (1) (2000) R5–R7. [6] S. Parmentier-Batteur, K. Jin, X.O. Mao, L. Xie, D.A. Greenberg, Increased severity of stroke in CB1 cannabinoid receptor knock-out mice, J. Neurosci. 22 (22) (2002) 9771–9775. [7] T. Nagayama, A.D. Sinor, R.P. Simon, J. Chen, S.H. Graham, K. Jin, D.A. Greenberg, Cannabinoids and neuroprotection in global and focal cerebral ischemia and in neuronal cultures, J. Neurosci. 19 (8) (1999) 2987–2995. [8] J. Fernandez-Ruiz, C. Garcia, O. Sagredo, M. Gomez-Ruiz, E. de Lago, The endocannabinoid system as a target for the treatment of neuronal damage, Expert Opin. Ther. Targets 14 (4) (2010) 387–404. [9] J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavón, A.M. Serrano, D.E. Selley, L.H. Parsons, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem Biol. 5 (1) (2009) 37–44. [10] M.M. Mulvihill, D.K. Nomura, Therapeutic potential of monoacylglycerol lipase inhibitors, Life Sci. 92 (8-9) (2013) 492–497. [11] D.K. Nomura, B.E. Morrison, J.L. Blankman, J.Z. Long, S.G. Kinsey, M.C. Marcondes, A.M. Ward, Y.K. Hahn, A.H. Lichtman, B. Conti, B.F. Cravatt, Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation, Science (New York, N.Y.) 334 (6057) (2011) 809–813. [12] K.W. Muir, P. Tyrrell, N. Sattar, E. Warburton, Inflammation and ischaemic stroke, Curr. Opin. Neurol. 20 (3) (2007) 334–342. [13] P.U. Heuschmann, P.L. Kolominsky-Rabas, J. Roether, B. Misselwitz, K. Lowitzsch,
[29]
[30]
[31]
[32]
[33]
[34]
[35]
[36] [37]
[38]
1725
J. Heidrich, P. Hermanek, C. Leffmann, M. Sitzer, M. Biegler, Predictors of inhospital mortality in patients with acute ischemic stroke treated with thrombolytic therapy, JAMA 292 (15) (2004) 1831–1838. S.E. Lakhan, A. Kirchgessner, M. Hofer, Inflammatory mechanisms in ischemic stroke: therapeutic approaches, J. Transl. Med. 7 (1) (2009) 97. M. Allahtavakoli, R. Moloudi, M.K. Arababadi, A. Shamsizadeh, K. Javanmardi, Delayed post ischemic treatment with rosiglitazone attenuates infarct volume, neurological deficits and neutrophilia after embolic stroke in rat, Brain Res. 1271 (2009) 121–127. W. Hacke, G. Albers, Y. Al-Rawi, J. Bogousslavsky, A. Davalos, M. Eliasziw, M. Fischer, A. Furlan, M. Kaste, K.R. Lees, The desmoteplase in acute ischemic stroke trial (DIAS) a phase II MRI-based 9-hour window acute stroke thrombolysis trial with intravenous desmoteplase, Stroke 36 (1) (2005) 66–73. U. Dirnagl, C. Iadecola, M.A. Moskowitz, Pathobiology of ischaemic stroke: an integrated view, Trends Neurosci. 22 (9) (1999) 391–397. J.B. Bederson, L.H. Pitts, M. Tsuji, M. Nishimura, R. Davis, H. Bartkowski, Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination, Stroke 17 (3) (1986) 472–476. R. Jin, G. Yang, G. Li, Inflammatory mechanisms in ischemic stroke: role of inflammatory cells, J. Leuk. Biol. 87 (5) (2010) 779–789. J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavon, A.M. Serrano, D.E. Selley, L.H. Parsons, A.H. Lichtman, B.F. Cravatt, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem. Biol. 5 (1) (2009) 37–44. R.L. Strawderman, Encyclopedia of biopharmaceutical statistics, J. Am. Stat. Assoc. 96 (455) (2001) 1141. L. Brunton, B. Knollman, R. Hilal-Dandan, McGraw Hill Professional, New York, Goodman and Gilman’s the Pharmacological Basis of Therapeutics, (2017). C.C. Lin, C.S. Pan, C.Y. Wang, S.W. Liu, L.D. Hsiao, C.M. Yang, Tumor necrosis factor-alpha induces VCAM-1-mediated inflammation via c-Src-dependent transactivation of EGF receptors in human cardiac fibroblasts, J. Biomed. Sci. 22 (53) (2015) 53. H.S. Rossi, N.M. Koho, M. Ilves, M.M. Rajamaki, A.K. Mykkanen, Expression of extracellular matrix metalloproteinase inducer and matrix metalloproteinase-2 and -9 in horses with chronic airway inflammation, Am. J. Vet. Res. 78 (11) (2017) 1329–1337. W. Mu, X. Ouyang, A. Agarwal, L. Zhang, D.A. Long, P.E. Cruz, C.A. Roncal, O.Y. Glushakova, V.A. Chiodo, M.A. Atkinson, W.W. Hauswirth, T.R. Flotte, B. Rodriguez-Iturbe, R.J. Johnson, IL-10 suppresses chemokines, inflammation, and fibrosis in a model of chronic renal disease, J. Am. Soc. Nephrol. 16 (12) (2005) 3651–3660. M.R. Rahmani, A. Shamsizadeh, A. Moghadam-Ahmadi, A. Kayedi, M. Allahtavakoli, Monoacylglycerol lipase inhibitor, JZL-184, confers neuroprotection in the mice middle cerebral artery occlusion model of stroke, Life Sci. 26 (18) (2018) 30092–30094. M. Allahtavakoli, F. Amin, A. Esmaeeli-Nadimi, A. Shamsizadeh, M. KazemiArababadi, D. Kennedy, Ascorbic acid reduces the adverse effects of delayed administration of tissue plasminogen activator in a rat stroke model, Basic Clin. Pharmacol. Toxicol. 117 (5) (2015) 335–339. J. Montaner, C.A. Molina, J. Monasterio, S. Abilleira, J.F. Arenillas, M. Ribo, M. Quintana, J. Alvarez-Sabin, Matrix metalloproteinase-9 pretreatment level predicts intracranial hemorrhagic complications after thrombolysis in human stroke, Circulation 107 (4) (2003) 598–603. J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavón, A.M. Serrano, D.E. Selley, L.H. Parsons, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem. Biol. 5 (1) (2009) 37–44. C. Berger, A. Stauder, F. Xia, C. Sommer, S. Schwab, Neuroprotection and glutamate attenuation by acetylsalicylic acid in temporary but not in permanent cerebral ischemia, Exp. Neurol. 210 (2) (2008) 543–548. D. Fernandez-Suarez, M. Celorrio, J.I. Riezu-Boj, A. Ugarte, R. Pacheco, H. Gonzalez, J. Oyarzabal, C.J. Hillard, R. Franco, M.S. Aymerich, Monoacylglycerol lipase inhibitor JZL184 is neuroprotective and alters glial cell phenotype in the chronic MPTP mouse model, Neurobiol. Aging 35 (11) (2014) 2603–2616. F. Zhang, C. Iadecola, Stimulation of the fastigial nucleus enhances EEG recovery and reduces tissue damage after focal cerebral ischemia, J. Cereb. Blood Flow Metab. 12 (6) (1992) 962–970. A. Tamura, D. Graham, J. McCulloch, G. Teasdale, Focal cerebral ischaemia in the rat: 1. Description of technique and early neuropathological consequences following middle cerebral artery occlusion, J. Cereb Blood Flow Metab. 1 (1) (1981) 53–60. S. Ashwal, D.J. Cole, T.N. Osborne, W.J. Pearce, Low dose L-NAME reduces infarct volume in the rat MCAO/reperfusion model, J. Neurosurg. Anesthesiol. 5 (4) (1993) 241–249. M.E. O’Donnell, Y.J. Chen, T.I. Lam, K.C. Taylor, J.H. Walton, S.E. Anderson, Intravenous HOE-642 reduces brain edema and Na uptake in the rat permanent middle cerebral artery occlusion model of stroke: evidence for participation of the blood-brain barrier Na/H exchanger, J. Cereb. Blood Flow Metab. 33 (2) (2013) 225–234. I.Q. Whishaw, The Behavior of the Laboratory Rat: A Handbook with Tests, Oxford University Press, 2004. S. Ghosh, L.E. Wise, Y. Chen, R. Gujjar, A. Mahadevan, B.F. Cravatt, A.H. Lichtman, The monoacylglycerol lipase inhibitor JZL184 suppresses inflammatory pain in the mouse carrageenan model, Life Sci. 92 (8-9) (2013) 498–505. T. Baba, K.L. Black, K. Ikezaki, K.N. Chen, D.P. Becker, Intracarotid infusion of
Biomedicine & Pharmacotherapy 103 (2018) 1720–1726
M.-R. Rahmani et al.
[39]
[40]
[41]
[42]
[43]
[44]
[45]
leukotriene C4 selectively increases blood-brain barrier permeability after focal ischemia in rats, J. Cereb. Blood Flow Metab. 11 (4) (1991) 638–643. R. Pihlaja, J. Takkinen, O. Eskola, J. Vasara, F.R. Lopez-Picon, M. Haaparanta-Solin, J.O. Rinne, Monoacylglycerol lipase inhibitor JZL184 reduces neuroinflammatory response in APdE9 mice and in adult mouse glial cells, J. Neuroinflamm. 12 (1) (2015) 81. E. Hakimizadeh, A. Shamsizadeh, A. Roohbakhsh, M.K. Arababadi, M.R. Hajizadeh, M. Shariati, M.R. Rahmani, M. Allahtavakoli, Inhibition of transient receptor potential vanilloid-1 confers neuroprotection, reduces tumor necrosis factor-alpha, and increases IL-10 in a rat stroke model, Fundam. Clin. Pharmacol. 31 (4) (2017) 420–428. E. Vendel, E.C. de Lange, Functions of the CB1 and CB 2 receptors in neuroprotection at the level of the blood-brain barrier, Neuromol. Med. 16 (3) (2014) 620–642. D. Panikashvili, R. Mechoulam, S.M. Beni, A. Alexandrovich, E. Shohami, CB1 cannabinoid receptors are involved in neuroprotection via NF-kappa B inhibition, J. Cereb. Blood Flow Metab. 25 (4) (2005) 477–484. M.S. Aymerich, E. Rojo-Bustamante, C. Molina, M. Celorrio, J.A. Sánchez-Arias, R. Franco, Neuroprotective effect of JZL184 in MPP+-treated SH-SY5Y cells through CB2 receptors, Mole Neurobiol. 53 (4) (2016) 2312–2319. L.V. Lysenko, J. Kim, C. Henry, A. Tyrtyshnaia, R.A. Kohnz, F. Madamba, G.M. Simon, N.E. Kleschevnikova, D.K. Nomura, R.A. Ezekowitz, A.M. Kleschevnikov, Monoacylglycerol lipase inhibitor JZL184 improves behavior and neural properties in Ts65Dn mice, a model of down syndrome, PloS One 9 (12) (2014) e114521. S.G. Kinsey, L.E. Wise, D. Ramesh, R. Abdullah, D.E. Selley, B.F. Cravatt, A.H. Lichtman, Repeated low-dose administration of the monoacylglycerol lipase
[46] [47]
[48] [49]
[50]
[51]
[52]
[53]
1726
inhibitor JZL184 retains cannabinoid receptor type 1-mediated antinociceptive and gastroprotective effects, J. Pharmacol. Exp. Ther. 345 (3) (2013) 492–501. R. Mechoulam, D. Panikashvili, E. Shohami, Cannabinoids and brain injury: therapeutic implications, Trends Mol. Med. 8 (2) (2002) 58–61. A. Franklin, S. Parmentier-Batteur, L. Walter, D.A. Greenberg, N. Stella, Palmitoylethanolamide increases after focal cerebral ischemia and potentiates microglial cell motility, J. Neurosci. 23 (21) (2003) 7767–7775. M. van der Stelt, V. Di Marzo, Cannabinoid receptors and their role in neuroprotection, Neuromol. Med. 7 (1-2) (2005) 37–50. M. Kotoda, T. Ishiyama, K. Mitsui, S. Hishiyama, T. Matsukawa, Neuroprotective effects of amiodarone in a mouse model of ischemic stroke, BMC Anesthesiol. 17 (1) (2017) 168. X.N. Mao, H.J. Zhou, X.J. Yang, L.X. Zhao, X. Kuang, C. Chen, D.L. Liu, J.R. Du, Neuroprotective effect of a novel gastrodin derivative against ischemic brain injury: involvement of peroxiredoxin and TLR4 signaling inhibition, Oncotarget 8 (53) (2017) 90979–90995. M.D. Knowles, P.B. de la Tremblaye, I. Azogu, H. Plamondon, Endocannabinoid CB1 receptor activation upon global ischemia adversely impact recovery of reward and stress signaling molecules, neuronal survival and behavioral impulsivity, Prog. Neuropsychopharmacol. Biol. Psychiatry 66 (2016) 8–21. L. Zhang, T. Schallert, Z.G. Zhang, Q. Jiang, P. Arniego, Q. Li, M. Lu, M. Chopp, A test for detecting long-term sensorimotor dysfunction in the mouse after focal cerebral ischemia, J. Neurosci. Methods 117 (2) (2002) 207–214. H. Abbassian, B.J. Whalley, V. Sheibani, M. Shabani, Cannabinoid type 1 receptor antagonism ameliorates harmaline-induced essential tremor in rat, Br. J. Pharmacol. 173 (22) (2016) 3196–3207.
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JZL184, as a monoacylglycerol lipase inhibitor, down-regulates inflammation in a cannabinoid pathway dependent manner
T
Mohammad-Reza Rahmania,b, Ali Shamsizadeha,c, Amir Moghadam-Ahmadia,d, ⁎ Gholamreza Bazmandegana,c, Mohammad Allahtavakolia,c, a
Physiology-Pharmacology Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran Student Research Committee, Rafsanjan University of Medical Sciences, Rafsanjan, Iran c Department of Physiology and Pharmacology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran d Department of Neurology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran b
A R T I C LE I N FO
A B S T R A C T
Keywords: Stroke PMCAO JZL-184 AM251 Cannabinoids
Introduction: Stroke is a prevalent disorder which is associated with several complications including inflammation. JZL-184 (JZL) inhibits arachidonic acid (AA) production and consequently results in two-arachidonoylglycerol (2-AG) accumulation. Both reduced production of AA metabolic products and increased 2-AG, the agonist of type 1 cannabinoid receptor (CB1), can result in reduced inflammation. In this study, we investigated the mechanisms of JZL in the improvement of stroke complications in mouse permanent cerebral ischemia (PPMCAO) model using AM251, the antagonist of CB1. Material and methods: PMCAO mice were divided into six groups including intact, controls, vehicle, JZL, AM251 and JZL plus AM251 administrated groups. Brain infarction and edema, brain levels of matrix metalloperoteinase-9 (MMP9), interleukin (IL)-10 and tumor necrosis factor-α (TNF-α) and behavioral functions have been examined in all groups. Results: The results showed that JZL lowered brain infarction, neurological disorders, TNF-α and MMP9 more effectively than JZL plus AM251. JZL and JZL plus AM251 reduced brain edema and increased brain IL-10. JZL, AM251 and JZL plus AM251 improve behavioral functions. Discussion: JZL reduces brain infarction and brain pro-inflammatory molecules in CB1 pathway dependent manner. JZL also reduces brain edema and increased IL-10 in CB1 pathways or decreased AA metabolites. Further, AM251 improves behavioral functions via unknown mechanisms.
1. Introduction Cannabinoids are a set of molecules which have several effects including anti-nausea, anti-cancer, anti-seizure, anti-oxidation, anti-inflammation and neuroprotection [1]. Cannabinoids perform their functions via interactions with two series of receptors, type 1 and 2 cannabinoid receptor (CB1 and 2). CB1is expressed in either brain or peripheral tissues, while CB2 is expressed in the peripheral tissues only [2]. Therefore, the altered behaviors following administration of cannabinoids are related to the interaction of the molecules with CB1 [3]. It has been indicated that two series of molecules including anandamide (N-arachidonoylethanolamine) and two-arachidonoylglycerol (2-AG) are the endogenous ligands for CB1 which are entitled endocannabinoids [4]. Thus, interactions of these ligands with CB1 mimic the effects of cannabinoids such as anti-inflammatory effects and neuron protection [5]. Many investigators indicate that, the protective ⁎
properties of CB1 receptors and using its agonists in ischemic brain injuries have been proved by investigators [6,7]. Accordingly, Nagayama et al., revealed that R(+)-WIN 55212-2, a synthetic cannabinoid agonist, reduced hippocampal neuronal damages and also brain infarctions following induction of MCAO in rat animal models [7]. Interestingly, some investigators reported that using CB1 antagonists results in blocking neuroprotection of CB1 agonist [6,7]. Therefore, therapeutic strategies which increased the brain endocannabinoids may hamper brain pro-inflammatory based disorders such as stroke [8]. JZL-184 (JZL) is a potential irreversible inhibitor of monoacylglycerol lipase (MAGL). MAGL is an important enzyme which facilitates the catabolism of 2-AG to produce arachidonic acid (AA) [9,10]. Prostaglandins (PGs), leukotrienes (LKs) and platelet activator factor (PAF), the important pro-inflammatory factors, are produced from AA, hence, we hypothesized that administration of JZL might decrease inflammation via two distinct pathways, increased levels of 2-
Corresponding author at: Physiology- Pharmacology Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran. E-mail address: [email protected] (M. Allahtavakoli).
https://doi.org/10.1016/j.biopha.2018.05.001 Received 27 September 2017; Received in revised form 1 May 2018; Accepted 2 May 2018 0753-3322/ © 2018 Published by Elsevier Masson SAS.
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(3 mg/kg), respectively. Finally, the sixth group, JZL plus AM251, contains the ischemic mice which have been treated with both JZL (4 mg/kg) and AM251 (3 mg/kg) intra-peritoneal. Accordingly, the drugs were administrated immediately after induction of PMCAO and the experiments were performed 48 h after single dose of the drug injections.
AG and consequently activation of CB1 as well as decreased levels of PGs, LKs and PAF [10,11]. Brain stroke is a human disorder which is associated with several mortalities and disabilities [12]. Following brain stroke, the central cells within the region of brain ischemia are damaged at once, and the penumbra region cells are alive, though they do not have normal function and are at risk of necrosis and consequently induction of the stroke complications [13,14]. Thus, several investigations are designed to find new strategies to protect the neurons in the penumbra area [15,16]. Ischemia and its related complications including brain inflammation and edema, is one of the most important inducers of penumbra region cell damage [12,17,18]. Tumor necrosis factor-alpha (TNF-α) and matrix metalloproteinase-9 (MMP9), as the pro-inflammatory molecules, and interleukin-10 (IL-10), as the anti-inflammatory cytokines, play crucial roles in the pathogenesis of stroke [19]. Therefore, expression of these cytokines and also the examination of brain edema and stroke infarctions might be remarkable determinants to check stroke treatment procedures. The previous study reported that JZL at doses of 4, 8 and 16 mg/kg has similar effects on 2-AG in animal model [20], Therefore, in order to reduce the side effects, the minimum effective dose should be used [21,22]. Thus, this project was designed to evaluate the effect of JZL (4 mg/kg) on the improvement of PMCAO behavioral functions and also inflammatory markers. Infarct area, edema cerebral inflammatory molecules TNF and MMP9 [23,24], as well as IL-10, as anti-inflammatory cytokine [25] were measured in the mouse permanent cerebral ischemia (PMCAO) model using JZL at dose of 4 mg/kg [26]. Additionally, another investigation by our research team revealed that MMP9 has pro-inflammatory effects during stroke [27]. Another study by Montaner et al., proved that MMP-9 is the main reason for brain hemorrhage and edema in the animal stroke model [28]. The main mechanisms used by JZL to hamper inflammation in the PMCAO model have yet to be clarified. Thus, this study was aimed to evaluate the effects of JZL, at 4 mg/kg, on the CB1/endocannabinoid pathway in reduction of inflammation, via evaluation of MMP9, TNF-α and IL-10 as pro/anti-inflammatory markers, brain edema/infarction and improvement of behavioral functions in PMCAO stroke model using the CB1 antagonist, AM251. AM251 is an antagonist of CB1 which is used in this project to block CB1 pathway to evaluate JZL mechanisms used to improve stroke secondary side effects.
2.3. Mouse permanent model of middle cerebral artery occlusion (PMCAO) establishment PMCAO model was induced in the experimental mice under anesthesia (ketamine 90 mg/kg plus xylazine 4/5 mg/kg) as described previously [32,33]. Briefly, after producing a small vertical incision on the midline of right ear and eye, temporalis muscle was retracted. Then, the bone was laterally cut and moved on the temporal area of the skull to make an approximately 1mm2 in diameter hole, just above the MCA. Accordingly, the dura was carefully removed, and then the root of MCA was cauterized immediately. Eventually, the mouse muscle was replaced and the skin was closed carefully.
2.4. Infarct volume and brain edema measurement Infarct volume was evaluated using the Zhang and Iadecola protocol [32], briefly, the mice were sacrificed 48 h after PMCAO induction and the brains were removed. After that, coronal slices, 1-mm-thick were made. Then the slices were stained using 2,3,5-triphenyltetrazolium chloride 1% (Sigma Chemical Co., St. Louis, MO, USA). The protocol results in staining and no red staining of the non-infarcted and infarcted brain tissue, respectively. The infarcted tissues were analyzed using the Image J software (NIH Image, version1.61, Bethesda, Maryland, USA) after demarcation. In order to calculate the total infarction, all the infarct zones were summed and multiplied in the thickness of the brain sections according to the Zhang and Iadecola protocol [32]. The corrected infarct volume was calculated by the following formula: infarct area × (1- [(ipsilateral hemisphere area-contralateral hemisphere area)/contralateral hemisphere]) [34]. Brain edema was calculated and reported as percentage using the formula of O’Donnell and colleagues [35] as follows: (volume of left hemisphere - volume of right hemisphere)/volume of right hemisphere.
2. Material and method
2.5. MMP-9, IL-10 and TNF-α brain levels
2.1. Animals
MMP-9 (R and D system), IL-10 and TNF-α (eBiosciences) brain levels were assessed using commercial ELISA kits based on the companies` instructions. In brief, the ischemic brain tissue samples were homogenized mechanically and then added to the anti-IL-10, antiMMP-9, and anti-TNF-α capture antibody pre-coated ELISA plates. After incubation for 2 h in the dark (at room temperature), the ELISA plates were washed 3 times with washing buffer and incubated in the dark at room temperature for an hour with conjugated HRP detection secondary antibodies. Then, HRP substrate (3, 3′, 5, 5′Tetramethylbenzidine (TMB) + H2O2) was added to the ELISA plates and they were incubated in dark place for 15 min and the chemical reaction was stopped via sulfuric acid (2 N). The optical densities (OD) were evaluated at 450 nm by a microplate ELISA reader (Bio-Rad, USA).
This experimental study has been performed on 78 male mice between 25–35 g and 8–10 weeks-old in the standard conditions (freely available food and water and animals kept at 37 °C and 12:12-hr light/ dark cycles). Local Ethical Committee has approved the experimental procedures (Code: IR.RUMS.REC.1394.88). Accordingly, all the mice handling and the experiments were done using the methods to minimize suffering. 2.2. Experimental groups 78 male mice were randomly divided into the six experimental groups including intact, control, vehicle, (JZL) 4 mg/kg [9,29], JZL plus AM251 and AM251 3 mg/kg groups [30]. Accordingly, 13 mice were placed in each group (8 animals were used to assess edema, infarct volume and behavioral experiments and five animals were used to determine cerebral levels of TNF-α, IL-10 or MMP-9). The control group contained the ischemic mice that were not treated with any drug, while vehicle group was treated with 300 mg/kg dimethyl sulfoxide (DMSO 10%) intra-peritoneal [31]. JZL and AM251 groups contain the ischemic animals which were treated with JZL (4 mg/kg) and AM251
2.6. Sticky testing Adhesive removal (sticky tape) test was applied to explore the sensorimotor functions, based on the Whishaw method protocol [36]. The sensorimotor behavior was determined before and 24 or 48 h after PMCAO insult, the averaged latency to remove sticky tape during 3 trials was recorded [37] 1721
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2.7. Hanging wire test In order to evaluate muscle strength of the PMCAO models, the animal was hung by forepaw from horizontal steel wire in a triplicate format. Using a stopwatch, the fall was recorded from the time the animal grasped the wire. 2.8. Neurological disorder assessment Rating neurological disorders was recorded using Bederson grading system [18] at 2, 24 and 48 h after PMCAO induction. Neurological disorders were rated using the score scales: 0, no observable sign; 1, forelimb flexion; 2, forelimb flexion plus reduced resistance to lateral push; 3, unidirectional circling; 4, unidirectional circling plus reduced levels of consciousness; and 5, death.
Fig. 2. The effects of JZL-184 and AM251 on brain edema in MCAO model. The figure shows that JZL (*) significantly reduced brain edema when compared to control and vehicle groups. AM251 alone and JZL plus AM251 could not reduce brain edema.
2.9. Statistical analysis
3.2. Brain edema
Data were presented as mean ± standard error. Differences in brain levels of MMP9, IL-10 and TNF-α, infarct volume, paresis and sensorimotor disorder were analyzed using Two Way ANOVA followed by TUKEY test. Neurological disorder was also analyzed using nonparametric tests including Kruskal-Wallis and Mann-Whitney tests and, hence, the results were reported as medians. A P value of less than 5% was considered to be significant.
Brain edema was assessed using O'Donnell et al. method as a marker of stroke damage to explain the effects of drugs on reduction of stroke secondary side effects. The brain edema, 48 h after PMCAO, was significantly decreased in JZL (p = 0.002) in comparison to vehicle group (Fig. 2), while it was not significant for AM251 plus JZL group (p = 0.188) and AM251 (p = 0.765). The brain edema in JZL treated group was significantly decreased when compared to AM251 (p < 0.001) and control (p < 0.001) groups, but was not different when compared to AM251 plus JZL group (p = 0.367, Fig. 2).
3. Results 3.1. Brain infarction
3.3. TNF-a, IL-10, and MMP9 levels
Brain infarction was evaluated to show the size of neurons damages by stroke and also the efficacy of the drugs to reduce the stroke effects. Fig. 1 presents the raw data regarding the brain infarct volumes 48 h after PMCAO induction. The infarct volumes in the JZL group were decreased significantly in comparison to control (p = 0.002), vehicle (p = 0.009) and AM251 (p = 0.008) groups. Additionally, there were not significant differences between JZL and JZL plus AM251 (p = 0.365). While there were not significant differences between AM251 and control (p = 0.979), vehicle (p > 1.00), as well as JZL plus AM251 (p = 0.360) groups. JZL plus AM251 group also did not have significant differences with vehicle group.
TNF-α, IL-10, and MMP9 brain levels were evaluated using ELISA method to determine inflammation scales. Data analysis demonstrated that the following 48 h’ cerebral levels of TNF-α were significantly hampered after treatment with JZL (p = 0.014), while it was not changed after treatment with AM251 (p = 0.216) and JZL plus AM251 (p = 0.100) when compared to vehicle. There were no significant differences between the following groups regarding brain levels of TNF-α: Vehicle and controls (p = 0.1), JZL and intact (p = 0.920), JZL and AM251 (p = 0.651), JZL and JZL plus AM251 (p = 0.876) as well as AM251 and JZL plus AM251 (p = 0.992) groups. Fig. 3 illustrates the raw data regarding brain levels of TNF-α. Treatment of the PMCAO animals with JZL (p < 0.001), but not JZL plus AM251 (p = 0.106) and AM251 (p = 0.744), led to increased expression of IL-10 in comparison to vehicle group. Interestingly, JZL increased IL-10 brain levels when compared to intact group (p < 0.001). Although there was a significant difference between JZL and AM251 groups (p = 0.008), the difference between JZL and JZL plus AM251 groups was not significant (p = 0.138, Fig. 3). Administration of JZL (p = 0.011), but not AM251 (p = 0.524) and JZL plus AM251 (p = 0.099), led to decreased expression of MMP9 in comparison to vehicle group. There were not significant differences between JZL and AM251 (p = 0.255), JZL and JZL plus AM251 (p = 0.832) and JZL with intact (p = 0.170) groups (Fig. 3). 3.4. Sticky test
Fig. 1. The effects of JZL, AM251 and JZL plus AM251 on infarct volumes in MCAO model. Infarct volumes are presented as a percentage of affected ipsilateral hemispheres. Data are expressed as a mean ± SEM. (*) JZL 4 mg/kg significantly reduced the infarct volumes when compared to control and vehicle group.Control and vehicle groups had not significant difference.AM251 was unable to reduce infract volume as well as JZL. AM251 also neutralized the protective roles of JZL in the group which treated with JZL and AM251 simultaneously.
Sticky test was used to evaluate the sensorimotor function, as a behavioral function using Whishaw method. Fig. 4 shows latency in removing the contralateral forepaw label 24 and 48 h after induction of PMCAO. JZL (p < 0.001 for 24 h and p = 0.004 for 48 h), AM251 (p = 0.003 for 24 h and p = 0.002 for 48 h) and JZL plus AM251 (p < 0.001 for 24 h and p = 0.001 for 48 h) improved the time of sticky test when compared to the vehicle group. There were significant 1722
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Fig. 3. The effects of AM251, JZL and JZL plus AM251 on response latency in adhesive removal test. JZL (*), AM251 (*) and JZL plus AM251 (*) administration were associated with improved response latency in adhesive removal test when compared to control and vehicle groups. There were not significant differences between AM251/JZL plus AM251 and JZL group (*). Control and vehicle groups also had not significant differences regarding hanging time.
PMCAO complications. Additionally, AM251 also neutralized the protective roles operated by JZL in the reduction of infarct volume. And while the figure illustrates a various range of infarctions between groups, the statistical analysis showed that the differences were not significant. Interestingly, the results obtained from brain edema also proved the neuroprotective roles operated by JZL and revealed that JZL, but not AM251 and JZL plus AM251, reduced the cerebral edema in the PMCAO animal models. According to the results and based on the mechanisms used by JZL regarding inhibition of production of arachidonic acid metabolites and elevation of CB1 agonist, 2-AG, it appears that JZL may be considered to decrease inflammation in the infarcted brain tissues. However, due to the fact that the differences between JZL and JZL plus AM251treated groups were not significant, hence, it may be hypothesized that JZL reduces brain infarction and also edema via down-regulation of AA metabolites which need to be explored by additional investigations. Baba et al., reported that AA metabolites are the most important molecules that participate in the induction of contraction in endothelial cell and consequently blood brain barrier (BBB) leak which is damaged during brain edema [38]. Thus, it may be supposed that JZL uses down-regulation of AA metabolites to suppress infarction and edema. The results also showed that although JZL decreased brain levels of MMP9 and TNF-α, treatment of the PMCAO mice with a combination of JZL and AM251 had no effects on the brain levels of these inflammatory molecules. Additionally, JZL decreased brain levels of the studied mice to reach to the intact group. The use of JZL is associated with 2-AG accumulation, the molecule is the ligand for CB1 and according to the results which showed that CB1 antagonist, AM251, neutralized the antiinflammatory and anti-infarct properties of JZL, it may be hypothesized that JZL may reduce infarct volumes and brain edema via down-regulation of pro-inflammatory molecules, MMP9 and TNF-α. Although the data directed the conclusion to the involvement of CB1 pathway in reduction of inflammation in the stroke region of the brain, AM251 has not affected the inflammation when compared to JZL treated group. Thus, again, it seems that down-regulation of AA metabolites may be considered as the main pathway which is used by JZL to reduce inflammation in the brain stroke. Pihlaja et al., also showed that JZL plays important neuroprotective roles in an Alzheimer's disease animal model through suppression of inflammation [39]. Furthermore, based on the mentioned results, it may also be demonstrated that reduction in production of AA and its pro-inflammatory metabolites, PGs, LKs and PAFs,
differences between JZL group at 24 h (p = 0.002) and 48 h (p < 0.001) with the intact group. There were no significant differences between JZL and AM251 groups (p = 0.698 for 24 h and p = 0.992 for 48 h), JZL and JZL plus AM251 groups (p = 0.978 for 24 h and p = 0.982 for 48 h) and also between AM251 and JZL plus AM251 groups (p = 0.953 for 24 h and p = 0.999 for 48 h). 3.5. Hanging wire test Hanging test is a test to evaluate the muscle performance which is categorized as behavioral function. Hanging wire test in both 24 and 48 h had similar results. Accordingly, the results revealed that treatment with JZL (p = 0.006 for 24 and p < 0.001 for 48 h), AM251 (p = 0.029 for 24 and p < 0.001 for 48 h) and JZL plus AM251 (p = 0.018 for 24 h and p < 0.001 for 48 h) significantly increased endurance time when compared to vehicle group (Fig. 5). However, JZL after both 24 and 48 h, was unable to improve hanging test to reach to the intact group (p < 0.001). There were no significant differences between two untreated groups including control and vehicle groups (p = 0.782 for 24 h and p = 0.966 for 48 h) and also between JZL and AM251 (p = 0.973 for 24 h and p = 0.927 for 48 h) and JZL plus AM251 (p = 0.993 for 24 h and p = 0.994 for 48 h) groups (Fig. 5). 3.6. Bederson test Rating neurological disorders was recorded using Bederson grading system. The test shows that JZL significantly improved the neurologic damages in the JZL group only (p < 0.001 for both 24 and 48 h). Neurological deficits were not changed after administration of AM251 (p > 0.05 for both 24 and 48 h) and JZL plus AM251 groups (p > 0.05 for both 24 and 48 h, Table1). 4. Discussion JZL component plays as an irreversible MAGL inhibitor, the enzyme responsible for the catabolism of 2-AG to produce arachidonic acid. Thus, JZL not only reduces production of arachidonic acid metabolites, it increases the 2-AG as ligands for CB1 receptors [10,20,37]. Therefore, JZL administration may be associated with hampering inflammation in stimulation of CB1 and suppression of AA metabolites behavior [10,11]. The results demonstrated that, although JZL reduced infarct volume in 4 mg/kg concentration, AM251 was unable to protect the mice from
Fig. 4. The effects of AM251, JZL and JZL plus AM251 on hanging time. JZL (*), AM251 (*) and JZL plus AM251 (*) administration were associated with improved behavioral functions when compared to control and vehicle groups. There were not significant differences between AM251/JZL plus AM251 and JZL group. Control and vehicle groups also had not significant differences regarding hanging time.
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Fig. 5. The effects of AM251, JZL-184 and a combination of JZL and AM251 on the brain levels of TNF-α, IL-10 and MMP9 in the MCAO mice. JZL (*), but not AM251 and AM251 plus JZL, significantly reduced brain levels of TNFα and MMP9 in MCAO mice. Both JZL and AM251 plus JZL were able to increase brain levels of IL-10. The differences between control and vehicle groups regarding brain levels of TNF-α, IL-10 and MMP9 were not significant.
inflammation [40]. However, there are several anti-inflammatory cytokines such as tumor growth factor-beta (TGF-β), IL-25 and so on, hence it may be hypothesized that other anti-inflammatory cytokines may also participate in the declination of inflammation using JZL which need to be explored by further investigations. Collectively, it seems that JZL may play important roles in the reduction of brain stroke complication via down-regulation of inflammation. Additionally, Bederson test also proved the roles of CB1 pathway in the neuron functions. Accordingly, using JZL alone, but not in combination with AM251, leads to improved neurological disorders. AM251 blocked the CB1 pathway, hence, this pathway may play pivotal roles in the prevention of neuron damages during stroke which is proved by several investigations [41,42]. However, JZL also blocked AA metabolite production; further investigations need to be performed to evaluate the measure of AA metabolites after administration of AM251. Previous investigations also proved the hypothesis and demonstrated that JZL improves neuron functions in Parkinson's disease animal models and down syndrome in cannabinoid receptor dependent manner [43,44]. The important roles played by JZL in modulation of neuron functions via CB1 pathway have also been evidenced by several investigators [1,4,45]. Collectively, it appears that JZL may inhibit penumbra region cells damages in the PMCAO animal model through reduction of infarction in CB1 and edema in both CB1 and AA metabolites dependent manners. Moreover, both endogenous and exogenous cannabinoids play neuroprotective roles during cerebral damages such as ischemia [43,46–48], therefore, it seems that JZL via up-regulation of endogenous cannabinoids activates CB1 pathway and inhibits AA metabolites production, reduces brain infarct and edema, protects penumbra cells and improves neuron functions. Therefore, it has been demonstrated that each factor which rescues the ischemic neurons from death and induces brain reperfusion can be considered as a
Table 1 Bederson test results in different groups at 2, 24 and 48 h after brain ischemia. Group
2h
24 h
48 h
Control Vehicle JZL AM251 JZL plus AM251
3 (2–3) 3 (3–3) 3 (2.75–4) 3.5 (3–4)** 3 (2–3)¶
3 3 2 3 3
4 (3–4) 3.5 (3–4) 1 (1–2)* 3 (3–4)# 3 (2.75–3.5)###
(3–3.25) (3–3.25) (1.75–2)* (3–3)# (2.75–4.25)##
Effect of JZl, AM251 and JZL plus AM251 on neurological deficits was assessed by a 5-score scales at 2, 24 and 48 h the following stroke. Data are presented as median, 25th and 75th percentiles (percentiles in the parentheses). * p < 0.001. ** p < 0.05 versus vehicle group. ¶ p < 0.05 versus AM251. # p < 0.001. ## p < 0.01 versus JZL.
were not associated with reduced infarct volumes because infarct volumes were not altered in the PMCAO group which was treated with JZL plus AM251. Thus, it seems that JZL may reduce infarct volume via decreased inflammation in both CB1 dependent manner and also downregulation of AA metabolites. In parallel with the results, JZL was able to up-regulate IL-10 in the brain of PMCAO mice and also increased its levels in the JZL group when compared to intact group which confirms its anti-inflammatory effects. IL-10 is the most powerful anti-inflammatory cytokine, thus, it can be concluded that increased brain expression of IL-10 following administration of JZL is associated with down-regulation of pro-inflammatory molecules, MMP9 and TNF-α. Interestingly, our previous investigation on the PMCAO model using transient receptor potential vanilloid-1 (TRPV1) antagonist (AMG9810) leads to decreased brain infarct and edema via down-regulation and upregulation of TNF-α and IL-10 which collectively leads to decreased 1724
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neuroprotective agent [49,50]. Thus, based on the outcomes obtained from JZL on the PMCAO models, it may be hypothesized that JZL can be considered as a neuroprotective drug. Although JZL and JZL plus AM251 were unable to improve the sticky and hanging tests in the PMCAO models to reach to the intact group, the outcomes revealed that hanging test and sticky removal test were improved in all of the three groups including JZL and JZL plus AM251 receiving mice (Figs. 3 and 4). However, due to the fact that AM251 suppresses CB1 pathway, it seems that JZL progresses the behavioral functions independent of CB1 pathway. Interestingly, administration of AM251 alone also improved the behavioral functions. Knowles et al., also proved the roles of AM251 in improvement of behavioral functions in stroke animal models [51]. Some investigations proved that there were not significant correlations between the behavioral tests, such as sticky tape and hanging test, with infarction volumes and edema percentage [52]. Moreover, in agreement with our results, it has been shown that AM251 can improve behavioral tests [53]. Thus, the results which are presented in this project, and also suppression of CB1 receptor are associated with increased 2-AG and also decreased AA metabolites, hence, it can perhaps be hypothesized that 2-AG may improve the behavioral functions via an unknown 2-AG dependent mechanism. Collectively, based on the results it seems that JZL is an important agent which might be considered as anti-stroke complication. However, in order to describe the main mechanisms used by JZL to reduce inflammation, brain infarcts and edema, authors of this article suggest different CB1R compound tools (agonist/antagonist) be used to study the link between MAGL inhibition by JZL184 and CB1 receptor signaling pathway.
[14] [15]
[16]
[17] [18]
[19] [20]
[21] [22] [23]
[24]
[25]
Acknowledgment [26]
This work is derived from Mr. Mohammad-Reza Rahmani’s PhD dissertation and was supported by grant no. 20/611 from the ViceChancellor for Research and Technology, Rafsanjan University of Medical Sciences, Rafsanjan, Iran.
[27]
References
[28]
[1] F. Grotenhermen, Cannabinoids, Curr. Drug Target CNS Neurol. Disord. 4 (5) (2005) 507–530. [2] F. Chaperon, M.H. Thiebot, Behavioral effects of cannabinoid agents in animals, Crit. Rev. Neurobiol. 13 (3) (1999) 243–281. [3] D. Panikashvili, C. Simeonidou, S. Ben-Shabat, L. Hanus, A. Breuer, R. Mechoulam, E. Shohami, An endogenous cannabinoid (2-AG) is neuroprotective after brain injury, Nature 413 (6855) (2001) 527–531. [4] M.R. Pazos, E. Nunez, C. Benito, R.M. Tolon, J. Romero, Functional neuroanatomy of the endocannabinoid system, Pharmacol. Biochem. Behav. 81 (2) (2005) 239–247. [5] R. Gallily, A. Breuer, R. Mechoulam, 2-Arachidonylglycerol, an endogenous cannabinoid, inhibits tumor necrosis factor-alpha production in murine macrophages, and in mice, Eur. J. Pharmacol. 406 (1) (2000) R5–R7. [6] S. Parmentier-Batteur, K. Jin, X.O. Mao, L. Xie, D.A. Greenberg, Increased severity of stroke in CB1 cannabinoid receptor knock-out mice, J. Neurosci. 22 (22) (2002) 9771–9775. [7] T. Nagayama, A.D. Sinor, R.P. Simon, J. Chen, S.H. Graham, K. Jin, D.A. Greenberg, Cannabinoids and neuroprotection in global and focal cerebral ischemia and in neuronal cultures, J. Neurosci. 19 (8) (1999) 2987–2995. [8] J. Fernandez-Ruiz, C. Garcia, O. Sagredo, M. Gomez-Ruiz, E. de Lago, The endocannabinoid system as a target for the treatment of neuronal damage, Expert Opin. Ther. Targets 14 (4) (2010) 387–404. [9] J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavón, A.M. Serrano, D.E. Selley, L.H. Parsons, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem Biol. 5 (1) (2009) 37–44. [10] M.M. Mulvihill, D.K. Nomura, Therapeutic potential of monoacylglycerol lipase inhibitors, Life Sci. 92 (8-9) (2013) 492–497. [11] D.K. Nomura, B.E. Morrison, J.L. Blankman, J.Z. Long, S.G. Kinsey, M.C. Marcondes, A.M. Ward, Y.K. Hahn, A.H. Lichtman, B. Conti, B.F. Cravatt, Endocannabinoid hydrolysis generates brain prostaglandins that promote neuroinflammation, Science (New York, N.Y.) 334 (6057) (2011) 809–813. [12] K.W. Muir, P. Tyrrell, N. Sattar, E. Warburton, Inflammation and ischaemic stroke, Curr. Opin. Neurol. 20 (3) (2007) 334–342. [13] P.U. Heuschmann, P.L. Kolominsky-Rabas, J. Roether, B. Misselwitz, K. Lowitzsch,
[29]
[30]
[31]
[32]
[33]
[34]
[35]
[36] [37]
[38]
1725
J. Heidrich, P. Hermanek, C. Leffmann, M. Sitzer, M. Biegler, Predictors of inhospital mortality in patients with acute ischemic stroke treated with thrombolytic therapy, JAMA 292 (15) (2004) 1831–1838. S.E. Lakhan, A. Kirchgessner, M. Hofer, Inflammatory mechanisms in ischemic stroke: therapeutic approaches, J. Transl. Med. 7 (1) (2009) 97. M. Allahtavakoli, R. Moloudi, M.K. Arababadi, A. Shamsizadeh, K. Javanmardi, Delayed post ischemic treatment with rosiglitazone attenuates infarct volume, neurological deficits and neutrophilia after embolic stroke in rat, Brain Res. 1271 (2009) 121–127. W. Hacke, G. Albers, Y. Al-Rawi, J. Bogousslavsky, A. Davalos, M. Eliasziw, M. Fischer, A. Furlan, M. Kaste, K.R. Lees, The desmoteplase in acute ischemic stroke trial (DIAS) a phase II MRI-based 9-hour window acute stroke thrombolysis trial with intravenous desmoteplase, Stroke 36 (1) (2005) 66–73. U. Dirnagl, C. Iadecola, M.A. Moskowitz, Pathobiology of ischaemic stroke: an integrated view, Trends Neurosci. 22 (9) (1999) 391–397. J.B. Bederson, L.H. Pitts, M. Tsuji, M. Nishimura, R. Davis, H. Bartkowski, Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination, Stroke 17 (3) (1986) 472–476. R. Jin, G. Yang, G. Li, Inflammatory mechanisms in ischemic stroke: role of inflammatory cells, J. Leuk. Biol. 87 (5) (2010) 779–789. J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavon, A.M. Serrano, D.E. Selley, L.H. Parsons, A.H. Lichtman, B.F. Cravatt, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem. Biol. 5 (1) (2009) 37–44. R.L. Strawderman, Encyclopedia of biopharmaceutical statistics, J. Am. Stat. Assoc. 96 (455) (2001) 1141. L. Brunton, B. Knollman, R. Hilal-Dandan, McGraw Hill Professional, New York, Goodman and Gilman’s the Pharmacological Basis of Therapeutics, (2017). C.C. Lin, C.S. Pan, C.Y. Wang, S.W. Liu, L.D. Hsiao, C.M. Yang, Tumor necrosis factor-alpha induces VCAM-1-mediated inflammation via c-Src-dependent transactivation of EGF receptors in human cardiac fibroblasts, J. Biomed. Sci. 22 (53) (2015) 53. H.S. Rossi, N.M. Koho, M. Ilves, M.M. Rajamaki, A.K. Mykkanen, Expression of extracellular matrix metalloproteinase inducer and matrix metalloproteinase-2 and -9 in horses with chronic airway inflammation, Am. J. Vet. Res. 78 (11) (2017) 1329–1337. W. Mu, X. Ouyang, A. Agarwal, L. Zhang, D.A. Long, P.E. Cruz, C.A. Roncal, O.Y. Glushakova, V.A. Chiodo, M.A. Atkinson, W.W. Hauswirth, T.R. Flotte, B. Rodriguez-Iturbe, R.J. Johnson, IL-10 suppresses chemokines, inflammation, and fibrosis in a model of chronic renal disease, J. Am. Soc. Nephrol. 16 (12) (2005) 3651–3660. M.R. Rahmani, A. Shamsizadeh, A. Moghadam-Ahmadi, A. Kayedi, M. Allahtavakoli, Monoacylglycerol lipase inhibitor, JZL-184, confers neuroprotection in the mice middle cerebral artery occlusion model of stroke, Life Sci. 26 (18) (2018) 30092–30094. M. Allahtavakoli, F. Amin, A. Esmaeeli-Nadimi, A. Shamsizadeh, M. KazemiArababadi, D. Kennedy, Ascorbic acid reduces the adverse effects of delayed administration of tissue plasminogen activator in a rat stroke model, Basic Clin. Pharmacol. Toxicol. 117 (5) (2015) 335–339. J. Montaner, C.A. Molina, J. Monasterio, S. Abilleira, J.F. Arenillas, M. Ribo, M. Quintana, J. Alvarez-Sabin, Matrix metalloproteinase-9 pretreatment level predicts intracranial hemorrhagic complications after thrombolysis in human stroke, Circulation 107 (4) (2003) 598–603. J.Z. Long, W. Li, L. Booker, J.J. Burston, S.G. Kinsey, J.E. Schlosburg, F.J. Pavón, A.M. Serrano, D.E. Selley, L.H. Parsons, Selective blockade of 2-arachidonoylglycerol hydrolysis produces cannabinoid behavioral effects, Nat. Chem. Biol. 5 (1) (2009) 37–44. C. Berger, A. Stauder, F. Xia, C. Sommer, S. Schwab, Neuroprotection and glutamate attenuation by acetylsalicylic acid in temporary but not in permanent cerebral ischemia, Exp. Neurol. 210 (2) (2008) 543–548. D. Fernandez-Suarez, M. Celorrio, J.I. Riezu-Boj, A. Ugarte, R. Pacheco, H. Gonzalez, J. Oyarzabal, C.J. Hillard, R. Franco, M.S. Aymerich, Monoacylglycerol lipase inhibitor JZL184 is neuroprotective and alters glial cell phenotype in the chronic MPTP mouse model, Neurobiol. Aging 35 (11) (2014) 2603–2616. F. Zhang, C. Iadecola, Stimulation of the fastigial nucleus enhances EEG recovery and reduces tissue damage after focal cerebral ischemia, J. Cereb. Blood Flow Metab. 12 (6) (1992) 962–970. A. Tamura, D. Graham, J. McCulloch, G. Teasdale, Focal cerebral ischaemia in the rat: 1. Description of technique and early neuropathological consequences following middle cerebral artery occlusion, J. Cereb Blood Flow Metab. 1 (1) (1981) 53–60. S. Ashwal, D.J. Cole, T.N. Osborne, W.J. Pearce, Low dose L-NAME reduces infarct volume in the rat MCAO/reperfusion model, J. Neurosurg. Anesthesiol. 5 (4) (1993) 241–249. M.E. O’Donnell, Y.J. Chen, T.I. Lam, K.C. Taylor, J.H. Walton, S.E. Anderson, Intravenous HOE-642 reduces brain edema and Na uptake in the rat permanent middle cerebral artery occlusion model of stroke: evidence for participation of the blood-brain barrier Na/H exchanger, J. Cereb. Blood Flow Metab. 33 (2) (2013) 225–234. I.Q. Whishaw, The Behavior of the Laboratory Rat: A Handbook with Tests, Oxford University Press, 2004. S. Ghosh, L.E. Wise, Y. Chen, R. Gujjar, A. Mahadevan, B.F. Cravatt, A.H. Lichtman, The monoacylglycerol lipase inhibitor JZL184 suppresses inflammatory pain in the mouse carrageenan model, Life Sci. 92 (8-9) (2013) 498–505. T. Baba, K.L. Black, K. Ikezaki, K.N. Chen, D.P. Becker, Intracarotid infusion of
Biomedicine & Pharmacotherapy 103 (2018) 1720–1726
M.-R. Rahmani et al.
[39]
[40]
[41]
[42]
[43]
[44]
[45]
leukotriene C4 selectively increases blood-brain barrier permeability after focal ischemia in rats, J. Cereb. Blood Flow Metab. 11 (4) (1991) 638–643. R. Pihlaja, J. Takkinen, O. Eskola, J. Vasara, F.R. Lopez-Picon, M. Haaparanta-Solin, J.O. Rinne, Monoacylglycerol lipase inhibitor JZL184 reduces neuroinflammatory response in APdE9 mice and in adult mouse glial cells, J. Neuroinflamm. 12 (1) (2015) 81. E. Hakimizadeh, A. Shamsizadeh, A. Roohbakhsh, M.K. Arababadi, M.R. Hajizadeh, M. Shariati, M.R. Rahmani, M. Allahtavakoli, Inhibition of transient receptor potential vanilloid-1 confers neuroprotection, reduces tumor necrosis factor-alpha, and increases IL-10 in a rat stroke model, Fundam. Clin. Pharmacol. 31 (4) (2017) 420–428. E. Vendel, E.C. de Lange, Functions of the CB1 and CB 2 receptors in neuroprotection at the level of the blood-brain barrier, Neuromol. Med. 16 (3) (2014) 620–642. D. Panikashvili, R. Mechoulam, S.M. Beni, A. Alexandrovich, E. Shohami, CB1 cannabinoid receptors are involved in neuroprotection via NF-kappa B inhibition, J. Cereb. Blood Flow Metab. 25 (4) (2005) 477–484. M.S. Aymerich, E. Rojo-Bustamante, C. Molina, M. Celorrio, J.A. Sánchez-Arias, R. Franco, Neuroprotective effect of JZL184 in MPP+-treated SH-SY5Y cells through CB2 receptors, Mole Neurobiol. 53 (4) (2016) 2312–2319. L.V. Lysenko, J. Kim, C. Henry, A. Tyrtyshnaia, R.A. Kohnz, F. Madamba, G.M. Simon, N.E. Kleschevnikova, D.K. Nomura, R.A. Ezekowitz, A.M. Kleschevnikov, Monoacylglycerol lipase inhibitor JZL184 improves behavior and neural properties in Ts65Dn mice, a model of down syndrome, PloS One 9 (12) (2014) e114521. S.G. Kinsey, L.E. Wise, D. Ramesh, R. Abdullah, D.E. Selley, B.F. Cravatt, A.H. Lichtman, Repeated low-dose administration of the monoacylglycerol lipase
[46] [47]
[48] [49]
[50]
[51]
[52]
[53]
1726
inhibitor JZL184 retains cannabinoid receptor type 1-mediated antinociceptive and gastroprotective effects, J. Pharmacol. Exp. Ther. 345 (3) (2013) 492–501. R. Mechoulam, D. Panikashvili, E. Shohami, Cannabinoids and brain injury: therapeutic implications, Trends Mol. Med. 8 (2) (2002) 58–61. A. Franklin, S. Parmentier-Batteur, L. Walter, D.A. Greenberg, N. Stella, Palmitoylethanolamide increases after focal cerebral ischemia and potentiates microglial cell motility, J. Neurosci. 23 (21) (2003) 7767–7775. M. van der Stelt, V. Di Marzo, Cannabinoid receptors and their role in neuroprotection, Neuromol. Med. 7 (1-2) (2005) 37–50. M. Kotoda, T. Ishiyama, K. Mitsui, S. Hishiyama, T. Matsukawa, Neuroprotective effects of amiodarone in a mouse model of ischemic stroke, BMC Anesthesiol. 17 (1) (2017) 168. X.N. Mao, H.J. Zhou, X.J. Yang, L.X. Zhao, X. Kuang, C. Chen, D.L. Liu, J.R. Du, Neuroprotective effect of a novel gastrodin derivative against ischemic brain injury: involvement of peroxiredoxin and TLR4 signaling inhibition, Oncotarget 8 (53) (2017) 90979–90995. M.D. Knowles, P.B. de la Tremblaye, I. Azogu, H. Plamondon, Endocannabinoid CB1 receptor activation upon global ischemia adversely impact recovery of reward and stress signaling molecules, neuronal survival and behavioral impulsivity, Prog. Neuropsychopharmacol. Biol. Psychiatry 66 (2016) 8–21. L. Zhang, T. Schallert, Z.G. Zhang, Q. Jiang, P. Arniego, Q. Li, M. Lu, M. Chopp, A test for detecting long-term sensorimotor dysfunction in the mouse after focal cerebral ischemia, J. Neurosci. Methods 117 (2) (2002) 207–214. H. Abbassian, B.J. Whalley, V. Sheibani, M. Shabani, Cannabinoid type 1 receptor antagonism ameliorates harmaline-induced essential tremor in rat, Br. J. Pharmacol. 173 (22) (2016) 3196–3207.