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Antidepressant- and anxiolytic-like effects of “biased” 5-HT1A receptor agonists F15599 and F13714 in rats
Abstracts – XIX Congress PTF / Pharmacological Reports 67S (2015) 2–45
Effect of prolonged stress on NO synthases and IL-1b levels in brain structures and HPA axis activity Anna Ga˛dek-Michalska, Joanna Tadeusz, Paulina Rachwalska, Jan Bugajski Department of Physiology, Institute of Pharmacology, Polish Academy of Sciences, Krako´w, Poland The aim of this study was to determine the effect of prior repeated restraint stress (RS) on a single restraint-induced changes in IL-1b and nitric oxide synthase (NOS) levels in prefrontal cortex (PFC), hippocampus, hypothalamus and plasma and assess if these alterations may influence HPA axis responses. Experiments were performed on male Wistar rats. The first group consisted of control non-stressed animals. In the second group, animals were subjected to single restraint stress (RS) for 10 min. In the third group, the animals were restrained for 10 min twice a day repeatedly for 3, 7 and 14 consecutive days and decapitated 24 h after the last restraint. The fourth group consisted of prior repeatedly restrained rats (for 3, 7 and 14 days), which were subjected to 10 min restraint stress 24 h after the last restraint period and rapidly decapitated 0, 1, 2 and 3 h later. IL-1b, ACTH and corticosterone levels were determined in plasma using commercially available kits. IL-1b, neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) levels in the above mentioned brain structures samples were analyzed by western blot procedure. The present results indicate time-related similarities in the potent alterations in IL-1b levels in brain structures, and pituitary-adrenal hormones. Single restraint for 10 min significantly decreases iNOS levels in PFC and hypothalamus. Repeated RS for 3 days most strongly enhances iNOS level induced by single RS, which was gradually reduced by prior RS for 7 and 14 days. Parallel but much weaker changes appear with nNOS level. In brain structures, changes in NOS levels induced by single RS are enhanced or reduced by prior restraint, depending on brain structure and the period of prior stress. Changes in the rapid and strong ACTH and corticosterone secretion, induced by a single restraint or preceded by repeated RS, do not seem to be directly correlated with alterations of IL-1b and NOS levels in brain structures. This research was supported by grant: POIG 01.01.02-12-004/09-00 financed by European Regional Development Fund as well as by the statutory funds from the Institute of Pharmacology, Polish Academy of Sciences. http://dx.doi.org/10.1016/j.pharep.2015.06.070 P-glycoprotein inhibitors modulate levetiracetam effects in zebrafish epilepsy model Piotr Jakubowski 1, Michał Majewski 1, Piotr Podlasz 2, Anna Jakimiuk 3, Natalia Kasica 2 1
Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland 2 Department of Animal Anatomy, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland 3 Department of Pathophysiology, Forensic Veterinary and Administration, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland Epilepsy is one of the most common neurological diseases with 50–60 million people affected worldwide. Unfortunately, only 70% of patients are effectively treated with currently available antiepileptic drugs (AEDs). One of the leading hypothesis for refractory
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epilepsy is overexpression of p-glycoprotein (p-gp) in blood–brain barrier. This protein is responsible for efflux of numerous xenobiotics, including AEDs and thus may decrease concentration of this drugs in a brain and thus compromise therapy. Zebrafish, a vertebrate with complementary advantages to rodents, has been recently widely used in laboratories of both basic medical studies and clinical research. Combining the complexity of animal studies and in vitro high throughput screening, the zebrafish has proven to be a powerful tool for target identification, disease modelling and drug development. In our study, effects of p-gp inhibitors ritonavir and tariquidar on the efficacy of levetiracetam in PTZ epilepsy model in 7 dpf zebrafish larvae were investigated. After 24 h pretreatment period with levetiracetam and p-gp inhibitors larvae were deposited into multiwell plate and then treated with 10 mM pentylenetetrazole solution for epilepsy induction. Plate was then recorded by means of camcorder and fish movement was tracked and then locomotion parameters were analyzed. Results indicate that simultaneous use of levetiracetam and p-gp inhibitors significantly changes movement patterns in 7 dpf larvae suggesting possible use in epilepsy treatment. http://dx.doi.org/10.1016/j.pharep.2015.06.071 Antidepressant- and anxiolytic-like effects of ‘‘biased’’ 5-HT1A receptor agonists F15599 and F13714 in rats Magdalena Jastrze˛bska-Wie˛sek 1, Anna Partyka 1, Anna Wasik 1, Joanna S´niecikowska 2, Adrian Newman-Tancredi 3, Anna Wesołowska 1 1 Department of Clinical Pharmacy, Jagiellonian University Medical College, Krako´w, Poland 2 Department of Pharmaceutical Chemistry, Jagiellonian University Medical College, Krako´w, Poland 3 Neurolixis Inc, San Diego, CA, USA
The 5-HT1A receptors have drawn attention as a target for pharmacotherapy for variety CNS disorders. 5-HT1A receptors are expressed as both 5-HT1A autoreceptors, located in the raphe nucleus, and post-synaptic 5-HT1A heteroreceptors, located in various brain regions, i.e. cortex, hypothalamus and septum/ hippocampus. Activation of these serotonin receptors, in the different brain regions, exerts different effects. 5-HT1A receptor agonists have been investigated to function as antidepressants and anxiolytics. Furthermore, they are used as add-ons to atypical antipsychotics, and also to ameliorate female sexual dysfunction and Parkinson’s disease. Compounds F15599 and F13714 have been developed as two ‘‘biased’’ 5-HT1A receptor agonists. It has been demonstrated that F15599 preferentially activates postsynaptic 5-HT1A heteroreceptors over raphe-located autoreceptors, and F13714 exerts an opposite pharmacological profile with more pronounced activity at raphe-located autoreceptors, and only modest activation at post-synaptic heteroreceptors. The aim of this study was to examine antidepressant- and anxiolytic-like effects of F15599 and F13714, administered orally 30 min before tests to rats. The antidepressant-like effect was investigated in the forced swim test while anxiolytic-like activity was assessed in the conflict drinking Vogel and the elevated plus maze tests in rats. It would be interesting if any differences in pharmacological activity would be observed for these ‘‘biased’’ 5-HT1A receptor agonists. Supported by JU MC funds: K/ZDS/004121. http://dx.doi.org/10.1016/j.pharep.2015.06.072
Effect of prolonged stress on NO synthases and IL-1b levels in brain structures and HPA axis activity Anna Ga˛dek-Michalska, Joanna Tadeusz, Paulina Rachwalska, Jan Bugajski Department of Physiology, Institute of Pharmacology, Polish Academy of Sciences, Krako´w, Poland The aim of this study was to determine the effect of prior repeated restraint stress (RS) on a single restraint-induced changes in IL-1b and nitric oxide synthase (NOS) levels in prefrontal cortex (PFC), hippocampus, hypothalamus and plasma and assess if these alterations may influence HPA axis responses. Experiments were performed on male Wistar rats. The first group consisted of control non-stressed animals. In the second group, animals were subjected to single restraint stress (RS) for 10 min. In the third group, the animals were restrained for 10 min twice a day repeatedly for 3, 7 and 14 consecutive days and decapitated 24 h after the last restraint. The fourth group consisted of prior repeatedly restrained rats (for 3, 7 and 14 days), which were subjected to 10 min restraint stress 24 h after the last restraint period and rapidly decapitated 0, 1, 2 and 3 h later. IL-1b, ACTH and corticosterone levels were determined in plasma using commercially available kits. IL-1b, neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) levels in the above mentioned brain structures samples were analyzed by western blot procedure. The present results indicate time-related similarities in the potent alterations in IL-1b levels in brain structures, and pituitary-adrenal hormones. Single restraint for 10 min significantly decreases iNOS levels in PFC and hypothalamus. Repeated RS for 3 days most strongly enhances iNOS level induced by single RS, which was gradually reduced by prior RS for 7 and 14 days. Parallel but much weaker changes appear with nNOS level. In brain structures, changes in NOS levels induced by single RS are enhanced or reduced by prior restraint, depending on brain structure and the period of prior stress. Changes in the rapid and strong ACTH and corticosterone secretion, induced by a single restraint or preceded by repeated RS, do not seem to be directly correlated with alterations of IL-1b and NOS levels in brain structures. This research was supported by grant: POIG 01.01.02-12-004/09-00 financed by European Regional Development Fund as well as by the statutory funds from the Institute of Pharmacology, Polish Academy of Sciences. http://dx.doi.org/10.1016/j.pharep.2015.06.070 P-glycoprotein inhibitors modulate levetiracetam effects in zebrafish epilepsy model Piotr Jakubowski 1, Michał Majewski 1, Piotr Podlasz 2, Anna Jakimiuk 3, Natalia Kasica 2 1
Department of Pharmacology and Toxicology, Faculty of Medical Sciences, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland 2 Department of Animal Anatomy, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland 3 Department of Pathophysiology, Forensic Veterinary and Administration, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland Epilepsy is one of the most common neurological diseases with 50–60 million people affected worldwide. Unfortunately, only 70% of patients are effectively treated with currently available antiepileptic drugs (AEDs). One of the leading hypothesis for refractory
23
epilepsy is overexpression of p-glycoprotein (p-gp) in blood–brain barrier. This protein is responsible for efflux of numerous xenobiotics, including AEDs and thus may decrease concentration of this drugs in a brain and thus compromise therapy. Zebrafish, a vertebrate with complementary advantages to rodents, has been recently widely used in laboratories of both basic medical studies and clinical research. Combining the complexity of animal studies and in vitro high throughput screening, the zebrafish has proven to be a powerful tool for target identification, disease modelling and drug development. In our study, effects of p-gp inhibitors ritonavir and tariquidar on the efficacy of levetiracetam in PTZ epilepsy model in 7 dpf zebrafish larvae were investigated. After 24 h pretreatment period with levetiracetam and p-gp inhibitors larvae were deposited into multiwell plate and then treated with 10 mM pentylenetetrazole solution for epilepsy induction. Plate was then recorded by means of camcorder and fish movement was tracked and then locomotion parameters were analyzed. Results indicate that simultaneous use of levetiracetam and p-gp inhibitors significantly changes movement patterns in 7 dpf larvae suggesting possible use in epilepsy treatment. http://dx.doi.org/10.1016/j.pharep.2015.06.071 Antidepressant- and anxiolytic-like effects of ‘‘biased’’ 5-HT1A receptor agonists F15599 and F13714 in rats Magdalena Jastrze˛bska-Wie˛sek 1, Anna Partyka 1, Anna Wasik 1, Joanna S´niecikowska 2, Adrian Newman-Tancredi 3, Anna Wesołowska 1 1 Department of Clinical Pharmacy, Jagiellonian University Medical College, Krako´w, Poland 2 Department of Pharmaceutical Chemistry, Jagiellonian University Medical College, Krako´w, Poland 3 Neurolixis Inc, San Diego, CA, USA
The 5-HT1A receptors have drawn attention as a target for pharmacotherapy for variety CNS disorders. 5-HT1A receptors are expressed as both 5-HT1A autoreceptors, located in the raphe nucleus, and post-synaptic 5-HT1A heteroreceptors, located in various brain regions, i.e. cortex, hypothalamus and septum/ hippocampus. Activation of these serotonin receptors, in the different brain regions, exerts different effects. 5-HT1A receptor agonists have been investigated to function as antidepressants and anxiolytics. Furthermore, they are used as add-ons to atypical antipsychotics, and also to ameliorate female sexual dysfunction and Parkinson’s disease. Compounds F15599 and F13714 have been developed as two ‘‘biased’’ 5-HT1A receptor agonists. It has been demonstrated that F15599 preferentially activates postsynaptic 5-HT1A heteroreceptors over raphe-located autoreceptors, and F13714 exerts an opposite pharmacological profile with more pronounced activity at raphe-located autoreceptors, and only modest activation at post-synaptic heteroreceptors. The aim of this study was to examine antidepressant- and anxiolytic-like effects of F15599 and F13714, administered orally 30 min before tests to rats. The antidepressant-like effect was investigated in the forced swim test while anxiolytic-like activity was assessed in the conflict drinking Vogel and the elevated plus maze tests in rats. It would be interesting if any differences in pharmacological activity would be observed for these ‘‘biased’’ 5-HT1A receptor agonists. Supported by JU MC funds: K/ZDS/004121. http://dx.doi.org/10.1016/j.pharep.2015.06.072