- Email: [email protected]
Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 Neurochemistry International xxx (2014) xxx–xxx 1
Contents lists available at ScienceDirect
Neurochemistry International journal homepage: www.elsevier.com/locate/nci 5 6
Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice
3 4 7
Q1
a
8 9
b
10 1 4 2Q 2 13 14 15 16 17 18 19 20 21 22 23
Qing-Qiu Mao a,b, Zhen Huang b, Xiao-Ming Zhong b, Yan-Fang Xian a, Siu-Po Ip a,⇑ School of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong College of Pharmacy, Zhejiang Chinese Medicine University, Hangzhou 310053, Zhejiang, China
a r t i c l e
i n f o
a b s t r a c t
2 Article history: Received 23 January 2014 Received in revised form 15 April 2014 Accepted 29 April 2014 Available online xxxx Keywords: Antidepressant Brain-derived neurotrophic factor Corticosterone Piperine
A mouse model of depression has been recently developed by exogenous corticosterone administration. The present study aimed to examine the antidepressant-like effect and the possible mechanisms of piperine, a major alkaloid of black pepper (Piper nigrum Linn.) and long pepper (Piper longum Linn.), in corticosterone-induced depression in mice. The results showed that 3-weeks corticosterone injections caused depression-like behavior in mice, as indicated by the significant decrease in sucrose consumption and increase in immobility time in the forced swim test and tail suspension test. Moreover, it was found that brain-derived neurotrophic factor protein and mRNA levels in the hippocampus were significantly decreased in corticosterone-treated mice. Treating the animals with piperine significantly suppressed behavioral and biochemical changes induced by corticosterone. The results suggest that piperine produces an antidepressant-like effect in corticosterone-treated mice, which is possibly mediated by increasing brain-derived neurotrophic factor expression in the hippocampus. Ó 2014 Published by Elsevier Ltd.
25 26 27 28 29 30 31 32 33 34 35 36 37 38
39 40
1. Introduction
41
Depression is a commonly occurring, debilitating, and lifethreatening psychiatric disorder characterized by a pervasive low mood, loss of interest or pleasure in daily activities, low self-esteem, and suicidal tendencies (Hankin, 2006; Perahia et al., 2009). Although the pathophysiology of depression has not been fully understood, it is believed than it may involve the malfunction of monoaminergic system, hyperactivation of hypothalamic–pituitary–adrenal (HPA) axis, increased oxidative stress and decreased contents of neurotrophins (Krishnan and Nestler, 2008; Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009). At present, there are several types of antidepressants available for clinical use; they include tricyclic antidepressants, monoamine oxidase inhibitors, selective serotonin reuptake inhibitors, noradrenergic reuptake inhibitors, as well as serotonin and noradrenaline reuptake inhibitors (Bouvier et al., 2003). However, due to the multiple pathogenic factors involved in depression, many antidepressant drugs show low response rates and may cause adverse side-effects such as cardiotoxicity, hypertensive crisis, sexual dysfunction, and sleep disorder (Park et al., 2007). Therefore, it is desirable to seek new antidepressants. Piperine, a major alkaloid of black pepper
42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
⇑ Corresponding author. Tel.: +852 3163 4457; fax: +852 3163 4459.
(Piper nigrum Linn.) and long pepper (Piper longum Linn.), has been used extensively as condiment and flavoring for all types of savory dishes (Li et al., 2007a,b). In recent years, pharmacological studies have shown that piperine possesses analgesic, anti-inflammatory, anti-convulsant, anti-oxidant and cognitive enhancing effects (Gupta et al., 2000; Selvendiran et al., 2003; Chonpathompikunlert et al., 2010). Moreover, piperine is reported to inhibit monoamine oxidase activity, increase monoamine neurotransmitters levels and thus produce antidepressant-like activity in various mouse models of behavioral despair (Lee et al., 2005; Li et al., 2007a). It has also been shown that piperine could potentiate the protective effects of curcumin against chronic stress induced significant reduction in sucrose consumption in mice (Rinwa and Kumar, 2013). The Q3 antidepressive effect of piperine has also been demonstrated in mice exposed to chronic mild stress, which were related to up-regulation of hippocampal progenitor cell proliferation (Li et al., 2007b). Studies from our laboratory showed that treating mice intraperitoneally with piperine caused a significant reduction of immobility time in both forced swim and tail suspension tests, which was related to the serotonergic system (Mao et al., 2011a,b). However, the mechanisms by which piperine exerts its antidepressant-like effects are yet to be fully elucidated. On the other hand, high levels of corticosterone have been demonstrated to produce changes in behavior, neurochemistry and brain anatomy, which may be indicative of or consistent with
E-mail addresses: [email protected], [email protected] (S.-P. Ip). http://dx.doi.org/10.1016/j.neuint.2014.04.017 0197-0186/Ó 2014 Published by Elsevier Ltd.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 2 86 87 88 89 90 91 92 93 94 95 96
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
depressive-like symptoms (Dwivedi et al., 2006; Johnson et al., 2006; Murray et al., 2008; Lee et al., 2009; Zhao et al., 2009; Crupi et al., 2010), while antidepressants and acupuncture treatment could significantly reverse these changes (Lee et al., 2009; Crupi et al., 2010). These findings suggest that corticosterone-treated rodents may be a valid depression model to evaluate the efficacy of antidepressant candidates and explore the mechanism of action of antidepressants (Zhao et al., 2008; Iijima et al., 2010). Therefore, in this study, the antidepressant-like effect of piperine and its possible mechanisms was further evaluated in a mouse model of depression induced by corticosterone.
replaced with tap water for 24 h. After the adaptation, mice were deprived of water and food for 24 h. Sucrose preference test was conducted at 9:00 a.m. in which rats were housed in individual cages and were free to access to two bottles containing 100 ml of sucrose solution (1% w/v) and 100 ml of water, respectively. After 24 h, the volumes of consumed sucrose solution and water were recorded and the sucrose preference was calculated by the following formula:
132 133 134 135 136 137 138 139
140
sucrose consumption Sucrose preference ¼ water consumption þ sucrose consumption 100%:
142
97
2. Materials and methods
98
2.1. Animals
2.4. Forced swim test
143
99
The forced swim test was carried out 24 h after the sucrose preference test. The test was performed according to the method of Porsolt et al. (1977). Briefly, mice were forced to swim in a transparent glass vessel (25 cm high, 14 cm in diameter) filled with 10 cm of water at 24–26 °C. The total duration of immobility (seconds) was measured during the last 4 min of a single 6-min test session. Mice were considered immobile when they made no attempts to escape except the movements necessary to keep their heads above the water.
144
108
Male ICR mice weighing 20–25 g were obtained from the Laboratory Animal Services Center, The Chinese University of Hong Kong, Hong Kong. The animals were maintained on a 12-h light/ dark cycle under regulated temperature (22 ± 2 °C) and humidity (50 ± 10%) and fed with standard diet and water ad libitum. They were allowed to acclimate 3 days before use. The experiments on animals have been approved by the Animal Experimentation Ethics Committee of the Chinese University of Hong Kong and conformed to the guidelines of the ‘‘Principles of Laboratory Animal Care’’ (NIH publication No. 80–23, revised 1996).
2.5. Tail suspension test
153
109
2.2. Groups and drug treatment The tail suspension test, which was performed based on the method of Steru et al. (1985), was carried out 24 h after the forced swim test. Briefly, mice were suspended 5 cm above the floor by means of an adhesive tape, placed approximately 1 cm from the tip of the tail. The total duration of immobility (seconds) was quantified during a test period of 6 min. Mice were considered immobile when they were completely motionless.
154
2.6. Tissue sample collection
161
Twenty-four hours after the forced swim test, all mice were sacrificed by decapitation. Whole brains were rapidly removed from mice and chilled in ice-cold saline. The hippocampus was dissected on a cold plate and frozen in liquid nitrogen immediately. The tissue samples were then stored at 80 °C until assay.
162
2.7. Measurement of brain-derived neurotrophic factor (BDNF) protein level
167
BDNF content in the hippocampus was measured using a commercially available enzyme-linked immunosorbent assay kit (Chemicon International, Temecula, CA) according to the manufacturer’s instructions. Briefly, six hippocampus samples were weighed and homogenized in a ten-fold volume of lysis buffer. The homogenate was centrifugated at 14,000g for 30 min at
169
100 101 102 103 104 105 106 107
125
The mice were divided into four groups of ten animals: vehicle control group, corticosterone plus vehicle group, corticosterone plus piperine (5 mg/kg) group and corticosterone plus piperine (10 mg/kg) group. The doses of piperine were selected on the basis of literature data and our previous studies (Lee et al., 2005; Li et al., 2007a,b; Mao et al., 2011a,b). Corticosterone (Sigma–Aldrich, St. Louis, MO, USA, 20 mg/kg, dissolved in saline containing 0.1% dimethyl sulfoxide and 0.1% Tween-80) was administrated subcutaneously in a volume of 5 ml/kg once daily for 21 days (Zhao et al., 2009). Piperine (Sigma–Aldrich, St. Louis, MO, USA, dissolved in saline containing 0.1% Tween-80) was administered intraperitoneally (i.p.) in a volume of 10 ml/kg 30 min prior to the corticosterone injection for 21 days. The behavioral tests were carried out 24 h after the last injection. One animal from each group was tested in sequence. The detailed experimental schedules are depicted in Fig. 1.
126
2.3. Sucrose preference test
127
Sucrose preference test was carried out 24 h after the last injection. The test was performed as described previously (Mao et al., 2012b). Briefly, 72 h before the test, mice were trained to adapt 1% sucrose solution (w/v): two bottles of 1% sucrose solution were placed in each cage, and 24 h later 1% sucrose in one bottle was
110 111 112 113 114 115 116 117 118 119 120 121 122 123 124
128 129 130 131
Corticosterone and drugs treatment
1
2 3
4
5 6
7 8
Sacrifice
9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Days Sucrose Behavioural preference despair test tests Fig. 1. Experimental design and schedule.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
145 146 147 148 149 150 151 152
155 156 157 158 159 160
163 164 165 166
168
170 171 172 173 174
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 3
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
186
4 °C, and the supernates were collected and stored at 80 °C until assay. All samples and standards were applied in duplicate into 96-well immunoplates precoated with rabbit anti-human BDNF antibodies, which were incubated overnight on a shaker at 4 °C. After washing four times, biotinylated mouse anti-BDNF antibodies were added and the immunoplates were incubated for 3 hours at room temperature. After washing, streptavidin–HRP conjugate solution was added and the immunoplates were incubated at room temperature for 1 h. TMB/E substrate was added and the immunoplates were incubated at room temperature for 15 min. The plates were immediately read using a microplate reader at 450 nm. The detection limit of the assay was 7.8 pg/ml.
187
2.8. Measurement of BDNF mRNA levels
188
208
BDNF mRNA levels in the hippocampus were measured using TaqMan real-time reverse transcription-PCR analysis. Briefly, six hippocampus samples were weighed, and total RNA from hippocampus was isolated with TRIzol Reagent (Gibco, Grand Island, NY) according to the manufacturer’s protocol. Concentrations of extracted RNA were calculated from the absorbance at 260 nm. Total RNA (1.5 lg) was transcribed using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Inc., FosterCity, CA) according to the manufacturer’s protocol. cDNA was stored at 80 °C until assay. Assays-on-Demand primers for BDNF (Mm00432069_m1) and b-actin (Mm02619580_g1) were purchased from Applied Biosystems, Inc (FosterCity, CA). Real-time quantitative PCR was performed with cDNA, using an StepOnePlus™ Real-Time PCR System (Applied Biosystems, Inc., FosterCity, CA) with the following reaction program: 2 min hold at 50 °C, 10 min hold at 95 °C, followed by 40 cycles of 15 s at 95 °C and 1 min at 60 °C. Sequence Detection Software 2.0 (Applied Biosystems, Inc., FosterCity, CA) was used for data analysis. The relative mRNA expression of BDNF was normalized to the amount of b-actin in the same cDNA using the relative quantification method (2DDCT method) described by the manufacturer.
209
2.9. Statistical analysis
210
213
Data are expressed as means ± SEMs. Multiple group comparisons were performed using one-way analysis of variance (ANOVA) followed by Dunnett’s test, to detect inter-group differences. A difference was considered statistically significant when p < 0.05.
214
3. Results
215
The effect of piperine on the percentage of sucrose consumption in corticosterone-treated mice are given in Fig. 2. One-way ANOVA showed a significant different on the percentage of sucrose
178 179 180 181 182 183 184 185
189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207
211 212
216 217
(A) Duration of immobility (s)
177
consumption among groups (F (3, 36) = 15.85, p < 0.01). The corticosterone injections resulted in a significant reduction in the percentage of sucrose consumption in the animals (p < 0.01) compared with the controls. Treatment with piperine at daily doses of 5 or 10 mg/kg significantly increased the percentage of sucrose consumption in the corticosterone-treated mice (p < 0.01 and p < 0.01, respectively) compared with the corticosterone-treated mice. The effect of piperine on the immobility time of corticosteronetreated mice in the forced swim test and tail suspension test are given in Fig. 3. One-way ANOVA showed a significant difference t on the immobility time among groups in the forced swim test (F (3, 36) = 6.35, p < 0.01) and tail suspension test (F (3, 36) = 7.66, p < 0.01). The corticosterone injections resulted in a significant increase in the immobility time of mice in the forced swim test (p < 0.01) and tail suspension test (p < 0.01) compared with the controls. Treatment with piperine at daily doses of 5 or 10 mg/kg significantly attenuated the increase in the immobility time of corticosterone-treated mice in the forced swim test (p < 0.05 and p < 0.01, respectively) and tail suspension test (p < 0.01 and p < 0.01, respectively) compared with the corticosterone-treated mice. The effect of piperine on BDNF protein and mRNA levels in the hippocampus of corticosterone-treated mice are given in Fig. 4. One-way ANOVA showed a significant different on BDNF protein (F (3, 20) = 17.19, p < 0.01) and mRNA (F (3, 20) = 19.37, p < 0.01) levels in the hippocampus among groups. Exposure to corticosterone significantly decreased BDNF protein (p < 0.01) and mRNA (p < 0.01) levels in the hippocampus compared with the controls. Treatment with at daily doses of 5 or 10 mg/kg significantly attenuated the decrease in protein (p < 0.05 and p < 0.01, respectively) and mRNA (p < 0.05 and p < 0.01, respectively) levels in the hippocampus of corticosterone-treated mice compared with the corticosterone-treated mice.
250 #
200 * 150
**
100 50 0
Control
5
10
(mg/kg)
Piperine Corticosterone
(B) 100
Duration of immobility (s)
176
Sucrose consumption (%)
175
** 80
**
60
#
40 20
250 #
200 150
Control
5
10
(mg/kg)
Piperine Corticosterone Fig. 2. Effect of piperine on the percentage of sucrose consumption in corticosterone-treated mice.
**
5
10
100 50 0
0
**
Control
(mg/kg)
Piperine C orticosterone Fig. 3. Effect of piperine on the immobility time of corticosterone-treated mice in the forced swim test (A) and tail suspension test (B). Values are given as mean ± SEM (n = 10). #p < 0.01 compared with the control group; ⁄p < 0.05, ⁄⁄ p < 0.01 compared with the corticosterone group.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 4
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
BDNF protein level (ng/g tissue)
(A) 10 8
**
* 6
#
4 2 0
Control
5
10
(mg/kg)
BDNF mRNA level (Fold of control)
Piperine C orticosterone
(B) 1.2 1.0
** **
0.8 #
0.6 0.4 0.2 0.0
Control
5
10(mg/kg)
Piperine Corticosterone
Fig. 4. Effect of piperine on BDNF protein (A) and mRNA (B) levels in the hippocampus of corticosterone-treated mice. Values are given as mean ± SEM (n = 6). #p < 0.01 compared with the control group; ⁄p < 0.05, ⁄⁄p < 0.01 compared with the corticosterone group.
252
4. Discussion
253
The sucrose preference test is an indicator of anhedonia-like behavioral change. Anhedonia, a core symptom of major depression among humans, is modeled by inducing a decrease in responsiveness to rewards, as reflected by the reduced consumption of and/or preference for sweetened solutions (Willner, 2005). In the present study, our data are in line with other findings showing that repeated corticosterone injections results in significant decreased in the percentage of sucrose consumption of animals (Gourley et al., 2008; Mao et al., 2012a,b). Piperine treatment significantly suppressed this behavioral change which suggested the antidepressant-like action of piperine. The forced swim test and tail suspension test are the widely used for assessing the effectiveness of candidate antidepressants (Porsolt et al., 1977; Steru et al., 1985). It has been reported that repeated corticosterone injections significantly increased the immobility time of mice and rats in the forced
254 255 256 257 258 259 260 261 262 263 264 265 266 267
Hyperactivation of hypothalamic-pituitary-adrenal
Oxidative stress
Decrease brain contents of neurotransmitters (serotonin, norepinephrine, dopamine, etc.)
swim test and tail suspension test (Murray et al., 2008; Zhao et al., 2008, 2009; Lee et al., 2009; Crupi et al., 2010), which could be reversed by antidepressants and acupuncture treatment (Lee et al., 2009; Crupi et al., 2010). Consistently, in this study, we found that 21-days corticosterone injections caused a significant increase in the immobility time of mice in the forced swim test and tail suspension test, and that piperine treatment could attenuate these changes. Taken together, the results obtained from the behavioral studies indicate that piperine treatment produced an antidepressant-like action in the corticosterone-treated mice. The pathophysiology of depression was still poorly understood. Monoaminergic hypothesis is an early milestone in the field of depression (Krishnan and Nestler, 2008). However, recent research on the pathophysiology of depression has revealed some serious gaps and limitations in the monoaminergic hypothesis. For example, drugs that target monoamines affect these neurotransmitter systems within hours of initial treatment, but antidepressant drugs exert their antidepressant action in patients after several weeks of daily treatment (Wong and Licinio, 2001). Recently, with the advance of neuroscience, it has been suggested that the hippocampus may play an important role in the neurobiological basis of depressive syndromes. Several clinical and laboratory experiments have demonstrated that depression was closely associated with hippocampal neuronal atrophy and destruction, while chronic antidepressant treatment produced neuroprotective effect against hippocampal neuronal damage (Manji and Duman, 2001; Fuchs et al., 2004). To date, the mechanisms of hippocampal neuronal damage in depression still remains to be uncovered. However, several lines of evidence have supported the involvement of hyperactivation of the hypothalamic–pituitary–adrenal (HPA) axis, increased oxidative stress and decreased neurotrophins contents (Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009) (Fig. 5). These factors induce hippocampal neuronal damage and eventually cause the development of depression (Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009). Previous studies have shown that piperine is able to inhibit monoamine oxidase activity and increase monoamine neurotransmitters levels (Lee et al., 2005; Li et al., 2007a). Our recently studies have also demonstrated the antidepressant-like effect of piperine which were mediated through the regulation of serotonergic system (Mao et al., 2011a, 2011b) and the inhibition of oxidative stress and HPA axis hyperactivation (Mao et al., 2012a). In this study, we focus on BDNF, one of the most prevalent neurotrophic factors in the central nervous system, as a target to study the mechanisms underlying the antidepressant-like effects of piperine (Fig. 5). The role of BDNF in the pathogenesis of depression and in the mechanism of action of antidepressants has been well appreciated. Clinical studies have found decreased BDNF levels in the blood of
Hippocampal neuronal atrophy and destruction
Decrease brain levels of neurotrophins (brain-derived neurotrophic factor, nerve growth factor etc.)
Depression
Fig. 5. Pathology of depression and the acting mechanisms of piperine. Actions of piperine: increase brain levels of neurotransmitters and neurotrophins; inhibit oxidative stress and hypothalamic–pituitary–adrenal axis hyperactivation.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
347
depressive patients (Aydemir et al., 2006, 2007), while antidepressant treatment seems to normalize BDNF levels (Basßterzi et al., 2009). As well as clinical studies, it has also been shown that BDNF expression was decreased in depressive animals, which could be reversed by long term antidepressant treatment (Li et al., 2007a,b; Mao et al., 2011a,b). Furthermore, several studies reported that treatment of the animals with exogenous corticosterone caused a significant decrease in BDNF expression in the hippocampus and frontal cortex (Jacobsen and Mørk, 2006; Dwivedi et al., 2006; Huang et al., 2011; Mao et al., 2012b). In vitro studies also showed that corticosterone treatment decreased BDNF expression in the primary culture of hippocampal neurons (Li et al., 2007b; Yu et al., 2004). Consistent with these findings, in the present study, 3-weeks corticosterone injections were found to significantly decrease BDNF protein and mRNA levels in the hippocampus of mice. Piperine treatment significantly reversed the corticosterone-induced changes in BDNF expression, suggesting BDNF may be involved in the antidepressant-like effect of piperine. In addition, our previous studies have demonstrated that piperine treatment could protect against corticosterone-induced neurotoxicity in rat pheochromocytoma (PC12) cells, which was likely associated with the inhibition of oxidative stress and the up-regulation of BDNF mRNA level (Mao et al., 2012a,b). These findings have shown that the antidepressive action of piperine may be mediated by the inhibition of oxidative stress and the up-regulation of BDNF expression, thereby leading to the neuroprotective effects. In conclusion, piperine treatment was found to alleviate corticosterone-induced depressive-like behavior. The mechanism underlying the antidepressant-like action of piperine may be mediated by increasing BDNF expression in the hippocampus.
348
5. Uncited references
317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346
349
Q4
Mao et al. (2010a,b).
350
Acknowledgment
351 352
This project was supported by the Zhejiang Provincial Natural Science Foundation of China (Y2110307).
353
References
354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383
Aydemir, C., Yalcin, E.S., Aksaray, S., Kisa, C., Yildirim, S.G., Uzbay, T., Goka, E., 2006. Brain-derived neurotrophic factor (BDNF) changes in the serum of depressed women. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 30, 1256–1260. Aydemir, O., Deveci, A., Taskin, O.E., Taneli, F., Esen-Danaci, A., 2007. Serum brainderived neurotrophic factor level in dysthymia: a comparative study with major depressive disorder. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 31, 1023– 1026. Basßterzi, A.D., Yazici, K., Aslan, E., Delialiog˘lu, N., Tasßdelen, B., Acar, S.T., Yazici, A., 2009. Effects of fluoxetine and venlafaxine on serum brain derived neurotrophic factor levels in depressed patients. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 33, 281–285. Bouvier, N., Trenque, T., Millart, H., 2003. Development of antidepressant drugs. Experience and prospects. Presse Med. 32, 519–522. Chonpathompikunlert, P., Wattanathorn, J., Muchimapura, S., 2010. Piperine, the main alkaloid of Thai black pepper, protects against neurodegeneration and cognitive impairment in animal model of cognitive deficit like condition of Alzheimer’s disease. Food Chem. Toxicol. 48, 798–802. Crupi, R., Mazzon, E., Marino, A., La Spada, G., Bramanti, P., Cuzzocrea, S., Spina, E., 2010. Melatonin treatment mimics the antidepressant action in chronic corticosterone-treated mice. J. Pineal Res. 49, 123–129. Drzyzga, Ł.R., Marcinowska, A., Obuchowicz, E., 2009. Antiapoptotic and neurotrophic effects of antidepressants: a review of clinical and experimental studies. Brain Res. Bull. 79, 248–257. Dwivedi, Y., Rizavi, H.S., Pandey, G.N., 2006. Antidepressants reverse corticosteronemediated decrease in brain-derived neurotrophic factor expression: differential regulation of specific exons by antidepressants and corticosterone. Neuroscience 139, 1017–1029. Fuchs, E., Czéh, B., Kole, M.H., Michaelis, T., Lucassen, P.J., 2004. Alterations of neuroplasticity in depression: the hippocampus and beyond. Eur. Neuropsychopharmacol. 5, S481–S490.
5
Gourley, S.L., Wu, F.J., Taylor, J.R., 2008. Corticosterone regulates pERK1/2 map kinase in a chronic depression model. Ann. NY. Acad. Sci. 1148, 509–514. Gupta, S.K., Bansal, P., Bhardwaj, R.K., Velpandian, T., 2000. Comparative antinociceptive, anti-inflammatory and toxicity profile of nimesulide vs nimesulide and piperine combination. Pharmacol. Res. 41, 657–662. Hankin, B.L., 2006. Adolescent depression: description, causes, and interventions. Epilepsy Behav. 8, 102–114. Huang, Z., Zhong, X.M., Li, Z.Y., Feng, C.R., Pan, A.J., Mao, Q.Q., 2011. Curcumin reverses corticosterone-induced depressive-like behavior and decrease in brain BDNF levels in rats. Neurosci. Lett. 493, 145–148. Iijima, M., Ito, A., Kurosu, S., Chaki, S., 2010. Pharmacological characterization of repeated corticosterone injection-induced depression model in rats. Brain Res. 1359, 75–80. Jacobsen, J.P., Mørk, A., 2006. Chronic corticosterone decreases brain-derived neurotrophic factor (BDNF) mRNA and protein in the hippocampus, but not in the frontal cortex, of the rat. Brain Res. 1110, 221–225. Johnson, S.A., Fournier, N.M., Kalynchuk, L.E., 2006. Effect of different doses of corticosterone on depression-like behavior and HPA axis responses to a novel stressor. Behav. Brain Res. 168, 280–288. Krishnan, V., Nestler, E.J., 2008. The molecular neurobiology of depression. Nature 455, 894–902. Lee, S.A., Hong, S.S., Han, X.H., Hwang, J.S., Oh, G.J., Lee, K.S., Lee, M.K., Hwang, B.Y., Ro, J.S., 2005. Piperine from the fruits of Piper longum with inhibitory effect on monoamine oxidase and antidepressant-like activity. Chem. Pharm. Bull. 53, 832–835. Lee, B., Shim, I., Lee, H.J., Yang, Y., Hahm, D.H., 2009. Effects of acupuncture on chronic corticosterone-induced depression-like behavior and expression of neuropeptide Y in the rats. Neurosci. Lett. 453, 151–156. Li, S., Wang, C., Li, W., Koike, K., Nikaido, T., Wang, M.W., 2007a. Antidepressant-like effects of piperine and its derivative, antiepilepsirine. J. Asian Nat. Prod. Res. 9, 421–430. Li, S., Wang, C., Wang, M., Li, W., Matsumoto, K., Tang, Y., 2007b. Antidepressant like effects of piperine in chronic mild stress treated mice and its possible mechanisms. Life Sci. 80, 1373–1381. Manji, H.K., Duman, R.S., 2001. Impairments of neuroplasticity and cellular resilience in severe mood disorders: implications for the development of novel therapeutics. Psychopharmacol. Bull. 35, 45–49. Mao, Q.Q., Huang, Z., Zhong, X.M., Feng, C.R., Pan, A.J., Li, Z.Y., Ip, S.P., Che, C.T., 2010a. Effects of SYJN, a Chinese herbal formula, on chronic unpredictable stress-induced changes in behavior and brain BDNF in rats. J. Ethnopharmacol. 128, 336–341. Mao, Q.Q., Xian, Y.F., Ip, S.P., Tsai, S.H., Che, C.T., 2010b. Long-term treatment with peony glycosides reverses chronic unpredictable mild stress-induced depressive-like behavior via increasing expression of neurotrophins in rat brain. Behav. Brain Res. 210, 171–177. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2011a. Role of 5-HT(1A) and 5-HT(1B) receptors in the antidepressant-like effect of piperine in the forced swim test. Neurosci. Lett. 504, 181–184. Mao, Q.Q., Xian, Y.F., Ip, S.P., Che, C.T., 2011b. Involvement of serotonergic system in the antidepressant-like effect of piperine. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 35, 1144–1147. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2012a. Protective effects of piperine against corticosterone-induced neurotoxicity in PC12 cells. Cell. Mol. Neurobiol. 32, 531–537. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2012b. Peony glycosides reverse the effects of corticosterone on behavior and brain BDNF expression in rats. Behav. Brain Res. 227, 305–309. Murray, F., Smith, D.W., Hutson, P.H., 2008. Chronic low dose corticosterone exposure decreased hippocampal cell proliferation, volume and induced anxiety and depression like behaviours in mice. Eur. J. Pharmacol. 583, 115–127. Park, S.W., Kim, Y.K., Lee, J.G., Kim, S.H., Kim, J.M., Yoon, J.S., Park, Y.K., Lee, Y.K., Kim, Y.H., 2007. Antidepressant-like effects of the traditional Chinese medicine kami-shoyo-san in rats. Psychiatry Clin. Neurosci. 61, 401–406. Perahia, D.G., Quail, D., Desaiah, D., Montejo, A.L., Schatzberg, A.F., 2009. Switching to duloxetine in selective serotonin reuptake inhibitor non- and partialresponders: effects on painful physical symptoms of depression. J. Psychiatr. Res. 43, 512–518. Porsolt, R.D., Pichon, M.L.E., Jalfre, M., 1977. Behavioral despair in mice: a primary screening test for antidepressant. Arch. Int. Pharmacodyn. Ther. 229, 327–336. Rinwa, P., Kumar, A., 2013. Piperine potentiates the protective effects of curcumin against chronic unpredictable stress-induced cognitive impairment and oxidative damage in mice. Brain Res. 1488, 38–50. Selvendiran, K., Singh, J.P., Krishnan, K.B., Sakthisekaran, D., 2003. Cytoprotective effect of piperine against benzo[a]pyrene induced lung cancer with reference to lipid peroxidation and antioxidant system in Swiss albino mice. Fitoterapia 74, 109–115. Steru, L., Chermat, R., Thierry, B., Simon, P., 1985. The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology 85, 367–370. Watson, S., Mackin, P., 2007. HPA axis function in mood disorders. Psychiatry 5, 166–170. Willner, P., 2005. Chronic mild stress (CMS) revisited: consistency and behavioural– neurobiological concordance in the effects of CMS. Neuropsychobiology 52, 90–110.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 6 470 471 472 473 474 475 476
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
Wong, M.L., Licinio, J., 2001. Research and treatment approaches to depression. Nat. Rev. Neurosci. 2, 343–351. Yu, I.T., Lee, S.H., Lee, Y.S., Son, H., 2004. Differential effects of corticosterone and dexamethasone on hippocampal neurogenesis in vitro. Biochem. Biophys. Res. Commun. 317, 484–490. Zhang, Y.M., 2001. Stress, brain damage and depression. Foreign Med. Sci. Psychiatr. 28, 105–109.
Zhao, Y., Ma, R., Shen, J., Su, H., Xing, D., Du, L., 2008. A mouse model of depression induced by repeated corticosterone injections. Eur. J. Pharmacol. 581, 113–120. Zhao, Y., Xie, W., Dai, J., Wang, Z., Huang, Y., 2009. The varying effects of short-term and long-term corticosterone injections on depression-like behavior in mice. Brain Res. 1261, 82–90.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
477 478 479 480 481 482
No. of Pages 6, Model 5G
10 May 2014 Neurochemistry International xxx (2014) xxx–xxx 1
Contents lists available at ScienceDirect
Neurochemistry International journal homepage: www.elsevier.com/locate/nci 5 6
Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice
3 4 7
Q1
a
8 9
b
10 1 4 2Q 2 13 14 15 16 17 18 19 20 21 22 23
Qing-Qiu Mao a,b, Zhen Huang b, Xiao-Ming Zhong b, Yan-Fang Xian a, Siu-Po Ip a,⇑ School of Chinese Medicine, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong College of Pharmacy, Zhejiang Chinese Medicine University, Hangzhou 310053, Zhejiang, China
a r t i c l e
i n f o
a b s t r a c t
2 Article history: Received 23 January 2014 Received in revised form 15 April 2014 Accepted 29 April 2014 Available online xxxx Keywords: Antidepressant Brain-derived neurotrophic factor Corticosterone Piperine
A mouse model of depression has been recently developed by exogenous corticosterone administration. The present study aimed to examine the antidepressant-like effect and the possible mechanisms of piperine, a major alkaloid of black pepper (Piper nigrum Linn.) and long pepper (Piper longum Linn.), in corticosterone-induced depression in mice. The results showed that 3-weeks corticosterone injections caused depression-like behavior in mice, as indicated by the significant decrease in sucrose consumption and increase in immobility time in the forced swim test and tail suspension test. Moreover, it was found that brain-derived neurotrophic factor protein and mRNA levels in the hippocampus were significantly decreased in corticosterone-treated mice. Treating the animals with piperine significantly suppressed behavioral and biochemical changes induced by corticosterone. The results suggest that piperine produces an antidepressant-like effect in corticosterone-treated mice, which is possibly mediated by increasing brain-derived neurotrophic factor expression in the hippocampus. Ó 2014 Published by Elsevier Ltd.
25 26 27 28 29 30 31 32 33 34 35 36 37 38
39 40
1. Introduction
41
Depression is a commonly occurring, debilitating, and lifethreatening psychiatric disorder characterized by a pervasive low mood, loss of interest or pleasure in daily activities, low self-esteem, and suicidal tendencies (Hankin, 2006; Perahia et al., 2009). Although the pathophysiology of depression has not been fully understood, it is believed than it may involve the malfunction of monoaminergic system, hyperactivation of hypothalamic–pituitary–adrenal (HPA) axis, increased oxidative stress and decreased contents of neurotrophins (Krishnan and Nestler, 2008; Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009). At present, there are several types of antidepressants available for clinical use; they include tricyclic antidepressants, monoamine oxidase inhibitors, selective serotonin reuptake inhibitors, noradrenergic reuptake inhibitors, as well as serotonin and noradrenaline reuptake inhibitors (Bouvier et al., 2003). However, due to the multiple pathogenic factors involved in depression, many antidepressant drugs show low response rates and may cause adverse side-effects such as cardiotoxicity, hypertensive crisis, sexual dysfunction, and sleep disorder (Park et al., 2007). Therefore, it is desirable to seek new antidepressants. Piperine, a major alkaloid of black pepper
42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60
⇑ Corresponding author. Tel.: +852 3163 4457; fax: +852 3163 4459.
(Piper nigrum Linn.) and long pepper (Piper longum Linn.), has been used extensively as condiment and flavoring for all types of savory dishes (Li et al., 2007a,b). In recent years, pharmacological studies have shown that piperine possesses analgesic, anti-inflammatory, anti-convulsant, anti-oxidant and cognitive enhancing effects (Gupta et al., 2000; Selvendiran et al., 2003; Chonpathompikunlert et al., 2010). Moreover, piperine is reported to inhibit monoamine oxidase activity, increase monoamine neurotransmitters levels and thus produce antidepressant-like activity in various mouse models of behavioral despair (Lee et al., 2005; Li et al., 2007a). It has also been shown that piperine could potentiate the protective effects of curcumin against chronic stress induced significant reduction in sucrose consumption in mice (Rinwa and Kumar, 2013). The Q3 antidepressive effect of piperine has also been demonstrated in mice exposed to chronic mild stress, which were related to up-regulation of hippocampal progenitor cell proliferation (Li et al., 2007b). Studies from our laboratory showed that treating mice intraperitoneally with piperine caused a significant reduction of immobility time in both forced swim and tail suspension tests, which was related to the serotonergic system (Mao et al., 2011a,b). However, the mechanisms by which piperine exerts its antidepressant-like effects are yet to be fully elucidated. On the other hand, high levels of corticosterone have been demonstrated to produce changes in behavior, neurochemistry and brain anatomy, which may be indicative of or consistent with
E-mail addresses: [email protected], [email protected] (S.-P. Ip). http://dx.doi.org/10.1016/j.neuint.2014.04.017 0197-0186/Ó 2014 Published by Elsevier Ltd.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 2 86 87 88 89 90 91 92 93 94 95 96
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
depressive-like symptoms (Dwivedi et al., 2006; Johnson et al., 2006; Murray et al., 2008; Lee et al., 2009; Zhao et al., 2009; Crupi et al., 2010), while antidepressants and acupuncture treatment could significantly reverse these changes (Lee et al., 2009; Crupi et al., 2010). These findings suggest that corticosterone-treated rodents may be a valid depression model to evaluate the efficacy of antidepressant candidates and explore the mechanism of action of antidepressants (Zhao et al., 2008; Iijima et al., 2010). Therefore, in this study, the antidepressant-like effect of piperine and its possible mechanisms was further evaluated in a mouse model of depression induced by corticosterone.
replaced with tap water for 24 h. After the adaptation, mice were deprived of water and food for 24 h. Sucrose preference test was conducted at 9:00 a.m. in which rats were housed in individual cages and were free to access to two bottles containing 100 ml of sucrose solution (1% w/v) and 100 ml of water, respectively. After 24 h, the volumes of consumed sucrose solution and water were recorded and the sucrose preference was calculated by the following formula:
132 133 134 135 136 137 138 139
140
sucrose consumption Sucrose preference ¼ water consumption þ sucrose consumption 100%:
142
97
2. Materials and methods
98
2.1. Animals
2.4. Forced swim test
143
99
The forced swim test was carried out 24 h after the sucrose preference test. The test was performed according to the method of Porsolt et al. (1977). Briefly, mice were forced to swim in a transparent glass vessel (25 cm high, 14 cm in diameter) filled with 10 cm of water at 24–26 °C. The total duration of immobility (seconds) was measured during the last 4 min of a single 6-min test session. Mice were considered immobile when they made no attempts to escape except the movements necessary to keep their heads above the water.
144
108
Male ICR mice weighing 20–25 g were obtained from the Laboratory Animal Services Center, The Chinese University of Hong Kong, Hong Kong. The animals were maintained on a 12-h light/ dark cycle under regulated temperature (22 ± 2 °C) and humidity (50 ± 10%) and fed with standard diet and water ad libitum. They were allowed to acclimate 3 days before use. The experiments on animals have been approved by the Animal Experimentation Ethics Committee of the Chinese University of Hong Kong and conformed to the guidelines of the ‘‘Principles of Laboratory Animal Care’’ (NIH publication No. 80–23, revised 1996).
2.5. Tail suspension test
153
109
2.2. Groups and drug treatment The tail suspension test, which was performed based on the method of Steru et al. (1985), was carried out 24 h after the forced swim test. Briefly, mice were suspended 5 cm above the floor by means of an adhesive tape, placed approximately 1 cm from the tip of the tail. The total duration of immobility (seconds) was quantified during a test period of 6 min. Mice were considered immobile when they were completely motionless.
154
2.6. Tissue sample collection
161
Twenty-four hours after the forced swim test, all mice were sacrificed by decapitation. Whole brains were rapidly removed from mice and chilled in ice-cold saline. The hippocampus was dissected on a cold plate and frozen in liquid nitrogen immediately. The tissue samples were then stored at 80 °C until assay.
162
2.7. Measurement of brain-derived neurotrophic factor (BDNF) protein level
167
BDNF content in the hippocampus was measured using a commercially available enzyme-linked immunosorbent assay kit (Chemicon International, Temecula, CA) according to the manufacturer’s instructions. Briefly, six hippocampus samples were weighed and homogenized in a ten-fold volume of lysis buffer. The homogenate was centrifugated at 14,000g for 30 min at
169
100 101 102 103 104 105 106 107
125
The mice were divided into four groups of ten animals: vehicle control group, corticosterone plus vehicle group, corticosterone plus piperine (5 mg/kg) group and corticosterone plus piperine (10 mg/kg) group. The doses of piperine were selected on the basis of literature data and our previous studies (Lee et al., 2005; Li et al., 2007a,b; Mao et al., 2011a,b). Corticosterone (Sigma–Aldrich, St. Louis, MO, USA, 20 mg/kg, dissolved in saline containing 0.1% dimethyl sulfoxide and 0.1% Tween-80) was administrated subcutaneously in a volume of 5 ml/kg once daily for 21 days (Zhao et al., 2009). Piperine (Sigma–Aldrich, St. Louis, MO, USA, dissolved in saline containing 0.1% Tween-80) was administered intraperitoneally (i.p.) in a volume of 10 ml/kg 30 min prior to the corticosterone injection for 21 days. The behavioral tests were carried out 24 h after the last injection. One animal from each group was tested in sequence. The detailed experimental schedules are depicted in Fig. 1.
126
2.3. Sucrose preference test
127
Sucrose preference test was carried out 24 h after the last injection. The test was performed as described previously (Mao et al., 2012b). Briefly, 72 h before the test, mice were trained to adapt 1% sucrose solution (w/v): two bottles of 1% sucrose solution were placed in each cage, and 24 h later 1% sucrose in one bottle was
110 111 112 113 114 115 116 117 118 119 120 121 122 123 124
128 129 130 131
Corticosterone and drugs treatment
1
2 3
4
5 6
7 8
Sacrifice
9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26
Days Sucrose Behavioural preference despair test tests Fig. 1. Experimental design and schedule.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
145 146 147 148 149 150 151 152
155 156 157 158 159 160
163 164 165 166
168
170 171 172 173 174
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 3
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
186
4 °C, and the supernates were collected and stored at 80 °C until assay. All samples and standards were applied in duplicate into 96-well immunoplates precoated with rabbit anti-human BDNF antibodies, which were incubated overnight on a shaker at 4 °C. After washing four times, biotinylated mouse anti-BDNF antibodies were added and the immunoplates were incubated for 3 hours at room temperature. After washing, streptavidin–HRP conjugate solution was added and the immunoplates were incubated at room temperature for 1 h. TMB/E substrate was added and the immunoplates were incubated at room temperature for 15 min. The plates were immediately read using a microplate reader at 450 nm. The detection limit of the assay was 7.8 pg/ml.
187
2.8. Measurement of BDNF mRNA levels
188
208
BDNF mRNA levels in the hippocampus were measured using TaqMan real-time reverse transcription-PCR analysis. Briefly, six hippocampus samples were weighed, and total RNA from hippocampus was isolated with TRIzol Reagent (Gibco, Grand Island, NY) according to the manufacturer’s protocol. Concentrations of extracted RNA were calculated from the absorbance at 260 nm. Total RNA (1.5 lg) was transcribed using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Inc., FosterCity, CA) according to the manufacturer’s protocol. cDNA was stored at 80 °C until assay. Assays-on-Demand primers for BDNF (Mm00432069_m1) and b-actin (Mm02619580_g1) were purchased from Applied Biosystems, Inc (FosterCity, CA). Real-time quantitative PCR was performed with cDNA, using an StepOnePlus™ Real-Time PCR System (Applied Biosystems, Inc., FosterCity, CA) with the following reaction program: 2 min hold at 50 °C, 10 min hold at 95 °C, followed by 40 cycles of 15 s at 95 °C and 1 min at 60 °C. Sequence Detection Software 2.0 (Applied Biosystems, Inc., FosterCity, CA) was used for data analysis. The relative mRNA expression of BDNF was normalized to the amount of b-actin in the same cDNA using the relative quantification method (2DDCT method) described by the manufacturer.
209
2.9. Statistical analysis
210
213
Data are expressed as means ± SEMs. Multiple group comparisons were performed using one-way analysis of variance (ANOVA) followed by Dunnett’s test, to detect inter-group differences. A difference was considered statistically significant when p < 0.05.
214
3. Results
215
The effect of piperine on the percentage of sucrose consumption in corticosterone-treated mice are given in Fig. 2. One-way ANOVA showed a significant different on the percentage of sucrose
178 179 180 181 182 183 184 185
189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207
211 212
216 217
(A) Duration of immobility (s)
177
consumption among groups (F (3, 36) = 15.85, p < 0.01). The corticosterone injections resulted in a significant reduction in the percentage of sucrose consumption in the animals (p < 0.01) compared with the controls. Treatment with piperine at daily doses of 5 or 10 mg/kg significantly increased the percentage of sucrose consumption in the corticosterone-treated mice (p < 0.01 and p < 0.01, respectively) compared with the corticosterone-treated mice. The effect of piperine on the immobility time of corticosteronetreated mice in the forced swim test and tail suspension test are given in Fig. 3. One-way ANOVA showed a significant difference t on the immobility time among groups in the forced swim test (F (3, 36) = 6.35, p < 0.01) and tail suspension test (F (3, 36) = 7.66, p < 0.01). The corticosterone injections resulted in a significant increase in the immobility time of mice in the forced swim test (p < 0.01) and tail suspension test (p < 0.01) compared with the controls. Treatment with piperine at daily doses of 5 or 10 mg/kg significantly attenuated the increase in the immobility time of corticosterone-treated mice in the forced swim test (p < 0.05 and p < 0.01, respectively) and tail suspension test (p < 0.01 and p < 0.01, respectively) compared with the corticosterone-treated mice. The effect of piperine on BDNF protein and mRNA levels in the hippocampus of corticosterone-treated mice are given in Fig. 4. One-way ANOVA showed a significant different on BDNF protein (F (3, 20) = 17.19, p < 0.01) and mRNA (F (3, 20) = 19.37, p < 0.01) levels in the hippocampus among groups. Exposure to corticosterone significantly decreased BDNF protein (p < 0.01) and mRNA (p < 0.01) levels in the hippocampus compared with the controls. Treatment with at daily doses of 5 or 10 mg/kg significantly attenuated the decrease in protein (p < 0.05 and p < 0.01, respectively) and mRNA (p < 0.05 and p < 0.01, respectively) levels in the hippocampus of corticosterone-treated mice compared with the corticosterone-treated mice.
250 #
200 * 150
**
100 50 0
Control
5
10
(mg/kg)
Piperine Corticosterone
(B) 100
Duration of immobility (s)
176
Sucrose consumption (%)
175
** 80
**
60
#
40 20
250 #
200 150
Control
5
10
(mg/kg)
Piperine Corticosterone Fig. 2. Effect of piperine on the percentage of sucrose consumption in corticosterone-treated mice.
**
5
10
100 50 0
0
**
Control
(mg/kg)
Piperine C orticosterone Fig. 3. Effect of piperine on the immobility time of corticosterone-treated mice in the forced swim test (A) and tail suspension test (B). Values are given as mean ± SEM (n = 10). #p < 0.01 compared with the control group; ⁄p < 0.05, ⁄⁄ p < 0.01 compared with the corticosterone group.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 4
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
BDNF protein level (ng/g tissue)
(A) 10 8
**
* 6
#
4 2 0
Control
5
10
(mg/kg)
BDNF mRNA level (Fold of control)
Piperine C orticosterone
(B) 1.2 1.0
** **
0.8 #
0.6 0.4 0.2 0.0
Control
5
10(mg/kg)
Piperine Corticosterone
Fig. 4. Effect of piperine on BDNF protein (A) and mRNA (B) levels in the hippocampus of corticosterone-treated mice. Values are given as mean ± SEM (n = 6). #p < 0.01 compared with the control group; ⁄p < 0.05, ⁄⁄p < 0.01 compared with the corticosterone group.
252
4. Discussion
253
The sucrose preference test is an indicator of anhedonia-like behavioral change. Anhedonia, a core symptom of major depression among humans, is modeled by inducing a decrease in responsiveness to rewards, as reflected by the reduced consumption of and/or preference for sweetened solutions (Willner, 2005). In the present study, our data are in line with other findings showing that repeated corticosterone injections results in significant decreased in the percentage of sucrose consumption of animals (Gourley et al., 2008; Mao et al., 2012a,b). Piperine treatment significantly suppressed this behavioral change which suggested the antidepressant-like action of piperine. The forced swim test and tail suspension test are the widely used for assessing the effectiveness of candidate antidepressants (Porsolt et al., 1977; Steru et al., 1985). It has been reported that repeated corticosterone injections significantly increased the immobility time of mice and rats in the forced
254 255 256 257 258 259 260 261 262 263 264 265 266 267
Hyperactivation of hypothalamic-pituitary-adrenal
Oxidative stress
Decrease brain contents of neurotransmitters (serotonin, norepinephrine, dopamine, etc.)
swim test and tail suspension test (Murray et al., 2008; Zhao et al., 2008, 2009; Lee et al., 2009; Crupi et al., 2010), which could be reversed by antidepressants and acupuncture treatment (Lee et al., 2009; Crupi et al., 2010). Consistently, in this study, we found that 21-days corticosterone injections caused a significant increase in the immobility time of mice in the forced swim test and tail suspension test, and that piperine treatment could attenuate these changes. Taken together, the results obtained from the behavioral studies indicate that piperine treatment produced an antidepressant-like action in the corticosterone-treated mice. The pathophysiology of depression was still poorly understood. Monoaminergic hypothesis is an early milestone in the field of depression (Krishnan and Nestler, 2008). However, recent research on the pathophysiology of depression has revealed some serious gaps and limitations in the monoaminergic hypothesis. For example, drugs that target monoamines affect these neurotransmitter systems within hours of initial treatment, but antidepressant drugs exert their antidepressant action in patients after several weeks of daily treatment (Wong and Licinio, 2001). Recently, with the advance of neuroscience, it has been suggested that the hippocampus may play an important role in the neurobiological basis of depressive syndromes. Several clinical and laboratory experiments have demonstrated that depression was closely associated with hippocampal neuronal atrophy and destruction, while chronic antidepressant treatment produced neuroprotective effect against hippocampal neuronal damage (Manji and Duman, 2001; Fuchs et al., 2004). To date, the mechanisms of hippocampal neuronal damage in depression still remains to be uncovered. However, several lines of evidence have supported the involvement of hyperactivation of the hypothalamic–pituitary–adrenal (HPA) axis, increased oxidative stress and decreased neurotrophins contents (Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009) (Fig. 5). These factors induce hippocampal neuronal damage and eventually cause the development of depression (Zhang, 2001; Watson and Mackin, 2007; Drzyzga et al., 2009). Previous studies have shown that piperine is able to inhibit monoamine oxidase activity and increase monoamine neurotransmitters levels (Lee et al., 2005; Li et al., 2007a). Our recently studies have also demonstrated the antidepressant-like effect of piperine which were mediated through the regulation of serotonergic system (Mao et al., 2011a, 2011b) and the inhibition of oxidative stress and HPA axis hyperactivation (Mao et al., 2012a). In this study, we focus on BDNF, one of the most prevalent neurotrophic factors in the central nervous system, as a target to study the mechanisms underlying the antidepressant-like effects of piperine (Fig. 5). The role of BDNF in the pathogenesis of depression and in the mechanism of action of antidepressants has been well appreciated. Clinical studies have found decreased BDNF levels in the blood of
Hippocampal neuronal atrophy and destruction
Decrease brain levels of neurotrophins (brain-derived neurotrophic factor, nerve growth factor etc.)
Depression
Fig. 5. Pathology of depression and the acting mechanisms of piperine. Actions of piperine: increase brain levels of neurotransmitters and neurotrophins; inhibit oxidative stress and hypothalamic–pituitary–adrenal axis hyperactivation.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
347
depressive patients (Aydemir et al., 2006, 2007), while antidepressant treatment seems to normalize BDNF levels (Basßterzi et al., 2009). As well as clinical studies, it has also been shown that BDNF expression was decreased in depressive animals, which could be reversed by long term antidepressant treatment (Li et al., 2007a,b; Mao et al., 2011a,b). Furthermore, several studies reported that treatment of the animals with exogenous corticosterone caused a significant decrease in BDNF expression in the hippocampus and frontal cortex (Jacobsen and Mørk, 2006; Dwivedi et al., 2006; Huang et al., 2011; Mao et al., 2012b). In vitro studies also showed that corticosterone treatment decreased BDNF expression in the primary culture of hippocampal neurons (Li et al., 2007b; Yu et al., 2004). Consistent with these findings, in the present study, 3-weeks corticosterone injections were found to significantly decrease BDNF protein and mRNA levels in the hippocampus of mice. Piperine treatment significantly reversed the corticosterone-induced changes in BDNF expression, suggesting BDNF may be involved in the antidepressant-like effect of piperine. In addition, our previous studies have demonstrated that piperine treatment could protect against corticosterone-induced neurotoxicity in rat pheochromocytoma (PC12) cells, which was likely associated with the inhibition of oxidative stress and the up-regulation of BDNF mRNA level (Mao et al., 2012a,b). These findings have shown that the antidepressive action of piperine may be mediated by the inhibition of oxidative stress and the up-regulation of BDNF expression, thereby leading to the neuroprotective effects. In conclusion, piperine treatment was found to alleviate corticosterone-induced depressive-like behavior. The mechanism underlying the antidepressant-like action of piperine may be mediated by increasing BDNF expression in the hippocampus.
348
5. Uncited references
317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346
349
Q4
Mao et al. (2010a,b).
350
Acknowledgment
351 352
This project was supported by the Zhejiang Provincial Natural Science Foundation of China (Y2110307).
353
References
354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383
Aydemir, C., Yalcin, E.S., Aksaray, S., Kisa, C., Yildirim, S.G., Uzbay, T., Goka, E., 2006. Brain-derived neurotrophic factor (BDNF) changes in the serum of depressed women. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 30, 1256–1260. Aydemir, O., Deveci, A., Taskin, O.E., Taneli, F., Esen-Danaci, A., 2007. Serum brainderived neurotrophic factor level in dysthymia: a comparative study with major depressive disorder. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 31, 1023– 1026. Basßterzi, A.D., Yazici, K., Aslan, E., Delialiog˘lu, N., Tasßdelen, B., Acar, S.T., Yazici, A., 2009. Effects of fluoxetine and venlafaxine on serum brain derived neurotrophic factor levels in depressed patients. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 33, 281–285. Bouvier, N., Trenque, T., Millart, H., 2003. Development of antidepressant drugs. Experience and prospects. Presse Med. 32, 519–522. Chonpathompikunlert, P., Wattanathorn, J., Muchimapura, S., 2010. Piperine, the main alkaloid of Thai black pepper, protects against neurodegeneration and cognitive impairment in animal model of cognitive deficit like condition of Alzheimer’s disease. Food Chem. Toxicol. 48, 798–802. Crupi, R., Mazzon, E., Marino, A., La Spada, G., Bramanti, P., Cuzzocrea, S., Spina, E., 2010. Melatonin treatment mimics the antidepressant action in chronic corticosterone-treated mice. J. Pineal Res. 49, 123–129. Drzyzga, Ł.R., Marcinowska, A., Obuchowicz, E., 2009. Antiapoptotic and neurotrophic effects of antidepressants: a review of clinical and experimental studies. Brain Res. Bull. 79, 248–257. Dwivedi, Y., Rizavi, H.S., Pandey, G.N., 2006. Antidepressants reverse corticosteronemediated decrease in brain-derived neurotrophic factor expression: differential regulation of specific exons by antidepressants and corticosterone. Neuroscience 139, 1017–1029. Fuchs, E., Czéh, B., Kole, M.H., Michaelis, T., Lucassen, P.J., 2004. Alterations of neuroplasticity in depression: the hippocampus and beyond. Eur. Neuropsychopharmacol. 5, S481–S490.
5
Gourley, S.L., Wu, F.J., Taylor, J.R., 2008. Corticosterone regulates pERK1/2 map kinase in a chronic depression model. Ann. NY. Acad. Sci. 1148, 509–514. Gupta, S.K., Bansal, P., Bhardwaj, R.K., Velpandian, T., 2000. Comparative antinociceptive, anti-inflammatory and toxicity profile of nimesulide vs nimesulide and piperine combination. Pharmacol. Res. 41, 657–662. Hankin, B.L., 2006. Adolescent depression: description, causes, and interventions. Epilepsy Behav. 8, 102–114. Huang, Z., Zhong, X.M., Li, Z.Y., Feng, C.R., Pan, A.J., Mao, Q.Q., 2011. Curcumin reverses corticosterone-induced depressive-like behavior and decrease in brain BDNF levels in rats. Neurosci. Lett. 493, 145–148. Iijima, M., Ito, A., Kurosu, S., Chaki, S., 2010. Pharmacological characterization of repeated corticosterone injection-induced depression model in rats. Brain Res. 1359, 75–80. Jacobsen, J.P., Mørk, A., 2006. Chronic corticosterone decreases brain-derived neurotrophic factor (BDNF) mRNA and protein in the hippocampus, but not in the frontal cortex, of the rat. Brain Res. 1110, 221–225. Johnson, S.A., Fournier, N.M., Kalynchuk, L.E., 2006. Effect of different doses of corticosterone on depression-like behavior and HPA axis responses to a novel stressor. Behav. Brain Res. 168, 280–288. Krishnan, V., Nestler, E.J., 2008. The molecular neurobiology of depression. Nature 455, 894–902. Lee, S.A., Hong, S.S., Han, X.H., Hwang, J.S., Oh, G.J., Lee, K.S., Lee, M.K., Hwang, B.Y., Ro, J.S., 2005. Piperine from the fruits of Piper longum with inhibitory effect on monoamine oxidase and antidepressant-like activity. Chem. Pharm. Bull. 53, 832–835. Lee, B., Shim, I., Lee, H.J., Yang, Y., Hahm, D.H., 2009. Effects of acupuncture on chronic corticosterone-induced depression-like behavior and expression of neuropeptide Y in the rats. Neurosci. Lett. 453, 151–156. Li, S., Wang, C., Li, W., Koike, K., Nikaido, T., Wang, M.W., 2007a. Antidepressant-like effects of piperine and its derivative, antiepilepsirine. J. Asian Nat. Prod. Res. 9, 421–430. Li, S., Wang, C., Wang, M., Li, W., Matsumoto, K., Tang, Y., 2007b. Antidepressant like effects of piperine in chronic mild stress treated mice and its possible mechanisms. Life Sci. 80, 1373–1381. Manji, H.K., Duman, R.S., 2001. Impairments of neuroplasticity and cellular resilience in severe mood disorders: implications for the development of novel therapeutics. Psychopharmacol. Bull. 35, 45–49. Mao, Q.Q., Huang, Z., Zhong, X.M., Feng, C.R., Pan, A.J., Li, Z.Y., Ip, S.P., Che, C.T., 2010a. Effects of SYJN, a Chinese herbal formula, on chronic unpredictable stress-induced changes in behavior and brain BDNF in rats. J. Ethnopharmacol. 128, 336–341. Mao, Q.Q., Xian, Y.F., Ip, S.P., Tsai, S.H., Che, C.T., 2010b. Long-term treatment with peony glycosides reverses chronic unpredictable mild stress-induced depressive-like behavior via increasing expression of neurotrophins in rat brain. Behav. Brain Res. 210, 171–177. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2011a. Role of 5-HT(1A) and 5-HT(1B) receptors in the antidepressant-like effect of piperine in the forced swim test. Neurosci. Lett. 504, 181–184. Mao, Q.Q., Xian, Y.F., Ip, S.P., Che, C.T., 2011b. Involvement of serotonergic system in the antidepressant-like effect of piperine. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 35, 1144–1147. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2012a. Protective effects of piperine against corticosterone-induced neurotoxicity in PC12 cells. Cell. Mol. Neurobiol. 32, 531–537. Mao, Q.Q., Huang, Z., Ip, S.P., Xian, Y.F., Che, C.T., 2012b. Peony glycosides reverse the effects of corticosterone on behavior and brain BDNF expression in rats. Behav. Brain Res. 227, 305–309. Murray, F., Smith, D.W., Hutson, P.H., 2008. Chronic low dose corticosterone exposure decreased hippocampal cell proliferation, volume and induced anxiety and depression like behaviours in mice. Eur. J. Pharmacol. 583, 115–127. Park, S.W., Kim, Y.K., Lee, J.G., Kim, S.H., Kim, J.M., Yoon, J.S., Park, Y.K., Lee, Y.K., Kim, Y.H., 2007. Antidepressant-like effects of the traditional Chinese medicine kami-shoyo-san in rats. Psychiatry Clin. Neurosci. 61, 401–406. Perahia, D.G., Quail, D., Desaiah, D., Montejo, A.L., Schatzberg, A.F., 2009. Switching to duloxetine in selective serotonin reuptake inhibitor non- and partialresponders: effects on painful physical symptoms of depression. J. Psychiatr. Res. 43, 512–518. Porsolt, R.D., Pichon, M.L.E., Jalfre, M., 1977. Behavioral despair in mice: a primary screening test for antidepressant. Arch. Int. Pharmacodyn. Ther. 229, 327–336. Rinwa, P., Kumar, A., 2013. Piperine potentiates the protective effects of curcumin against chronic unpredictable stress-induced cognitive impairment and oxidative damage in mice. Brain Res. 1488, 38–50. Selvendiran, K., Singh, J.P., Krishnan, K.B., Sakthisekaran, D., 2003. Cytoprotective effect of piperine against benzo[a]pyrene induced lung cancer with reference to lipid peroxidation and antioxidant system in Swiss albino mice. Fitoterapia 74, 109–115. Steru, L., Chermat, R., Thierry, B., Simon, P., 1985. The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology 85, 367–370. Watson, S., Mackin, P., 2007. HPA axis function in mood disorders. Psychiatry 5, 166–170. Willner, P., 2005. Chronic mild stress (CMS) revisited: consistency and behavioural– neurobiological concordance in the effects of CMS. Neuropsychobiology 52, 90–110.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469
NCI 3574
No. of Pages 6, Model 5G
10 May 2014 6 470 471 472 473 474 475 476
Q.-Q. Mao et al. / Neurochemistry International xxx (2014) xxx–xxx
Wong, M.L., Licinio, J., 2001. Research and treatment approaches to depression. Nat. Rev. Neurosci. 2, 343–351. Yu, I.T., Lee, S.H., Lee, Y.S., Son, H., 2004. Differential effects of corticosterone and dexamethasone on hippocampal neurogenesis in vitro. Biochem. Biophys. Res. Commun. 317, 484–490. Zhang, Y.M., 2001. Stress, brain damage and depression. Foreign Med. Sci. Psychiatr. 28, 105–109.
Zhao, Y., Ma, R., Shen, J., Su, H., Xing, D., Du, L., 2008. A mouse model of depression induced by repeated corticosterone injections. Eur. J. Pharmacol. 581, 113–120. Zhao, Y., Xie, W., Dai, J., Wang, Z., Huang, Y., 2009. The varying effects of short-term and long-term corticosterone injections on depression-like behavior in mice. Brain Res. 1261, 82–90.
Please cite this article in press as: Mao, Q.-Q., et al. Piperine reverses the effects of corticosterone on behavior and hippocampal BDNF expression in mice. Neurochem. Int. (2014), http://dx.doi.org/10.1016/j.neuint.2014.04.017
477 478 479 480 481 482