Pharmacodynamic Study And Mechanism Discussion Of Compound Anti-anxiety Capsules ?

Anxiety disorder,a common mental illness,is an expected response to inevitable vicious incidents which are known beforehand and will happen soon.It is characterized by mental disorders such as fear,anxiety,and being nervous,etc.,and it is often accompanied by vegetative nerve function disturbances such as heart palpitations,hidrosis,respiratory distress,and cold hands and feet,etc.Anxiety has become a common disease and frequently-occurring disease in our country,which seriously affects the quality of people's lives.At present,there lacks effective Chinese patent medicines in domestic mental disease treatment.There is only one kind of pure TCM preparation-Jiuwei Zhenxin Particle(JW)-approved by the State Food and Drug Administration(SFDA)to treat anxiety disorders.Anxiety is related with a variety of behavioral responses(such as convulsion,impulsivity,aggression and drug dependence),which may be regulated by multiple genes.But now its pathological mechanism has not been completely elucidated.Therefore,it is very necessary to research on the development of Chinese patent medicine against anxiety and on its mechanism.Compound Anti-anxiety Capsule Component(CZZX)is the compound of saruma henryi,a clinical effective anti-anxiety prescription,which is composed of four herbs,including saruma henryi(as known as Rhizoma Valeriana Jatamansi),Cortex Albizia,the parched Semen Ziziphi Spinosae and Medulla Junci.The therapeutic principles are mainly soothing liver-qi stagnation and nourishing blood for tranquillization.It has significant effects against anxiety disorders with stagnation of liver qi and uneasiness.The research group has developed research on the pharmacodynamics and mechanism of CZZX with diazepam as the positive control medicine at an earlier stage.The results show that it has stable anti-anxiety effects.CZZX can sooth liver-qi stagnation and nourish blood for tranquillization,which is suitable for the anxiety disorders with stagnation of liver qi and uneasiness.Its mechanism is related with the regulatory effect on GABA transduction,regulatory effect on Cdk5/p35 pathway expression,inhibition of monoamine neurotransmitter release(5-HT,NE)in the brain tissue,and the regulatory effect on immune dysfunction.JW is mainly suitable for anxiety disorder with insufficiency of heart and spleen.Then,how is the anti-anxiety effect of CZZX compared with JW?What signaling pathways are associated with the action mechanism of CZZX in addition to the above mentioned?These questions are worthy of our study.The purpose of this study was to determine whether CZZX and JW exerts anxiolytic-like effects in mice and rats using several behavioral assays.Determine BDNF expression in cortex and hippocampus on acute stress rat,The effect of ERK-CREB pathways,The c-fos expression in hypothalamus.Through above study,we has laid a good foundation for treatment of anxiety disorders via traditional Chinese medicine.The archieved results are as follow:1 the pharmacological research1.1 pharmacodynamics experiment in miceAccording to the results of the elevate plus maze test(EPM)in mice,CMT?? groups(4.8g/kg/d)could significantly increase the open entries percent and open time percent,the result of EPM in mice verified that CMTX had a stable anxiolytic effect without increasing the activity of the animals.JW only increased The results of the open time percent.Light/dark box test suggested that CMT?? groups(4.8g/kg/d)significantly increased both time spent in the light as well as the number of transitions between compartments.comparing with the control group(P<0.05).In open field test,CMTX ? groups(2.4g/kg/d)could significantly significantly increased the number of center entries.Data of the locomotor activity test indicated an anxiolytic-like profile of action for CZZX and JW without sedative side effects.1.2 pharmacodynamics experiment in ratsAccording to the results of the elevate plus maze test(EPM)in rat,CMTX ?,?groups(1.5g/kg/d,3 g/kg/d)and JW could significantly increase the open entries percent and open time percent,the result of EPM in rats verified that CMTX and JW had a stable anxiolytic effect without increasing the activity of the animals.The results of the Vogel's conflict test suggested that CMTX ?-? groups(0.75g/kg/d,1.5g/kg/d,3g/kg/d)significantly increased the number of licks during the punished period comparing with the control group.In the open field test,rats treated with CMTX ?(1.5g/kg/d)and JW significantly increased the time spent in center area,rats treated with CMT?? significantly increased the number of center entries.Animals subjected to restraint showed an excessive anxiety in the EPM,since they exhibited a reduction in the percentage of time spent and entries on open arms.However,CMTX could prevent the emergence of stress-induced excessive anxiety.2 Mechanism studies2.1 Effects of CMTX on the CREB/ERK pathway by stress exposureThe protein levels of ERK1/2,CREB,phosphorylated ERK1/2(p-ERK1/2),and phosphorylated CREB(p-CREB)in the hippocampus(HP)and prefrontal cortex(PFC)were examined by Western blot analysis.Our results showed that the rats exposure to stress resulted in the increse of p-ERK1/2 and p-CREB in the HP and PFC.In addition,CMT??decreased the ratio of p-ERK1/2 to ERK1/2 and the ratio of p-CREB to CREB in the HP and PFC.Our study suggested that one potential way,by which CMTX had anti-anxiety-like efect,might be mediated by activating the ERK-CREB pathway in the brain.2.2 Effects of CMTX on the BDNF protein expression by stress exposureThe protein levels of BDNF in the hippocampus(HP)and prefrontal cortex(PFC)were examined by Western blot analysis.Our results showed that the rats exposure to stress resulted in the decrese of BDNF in the HP and PFC.In addition,CMTXIII increased the ratio of BDNF in the HP and PFC.Our study suggested that one potential way,by which CMTX had anti-anxiety-like efect,might be mediated by activating the BDNF in the brain.2.3 Effects of CMTX on the expression of c-fos geneUse the immunohistochemical method to analyze the effects on expression of immo-bilization stress in rats.compared with control,the c-fos levels were increased,the number of the c-fos positive cells was increased and their color became darker in mode.DZP and CMTX could decrease c-fos levels and positive cells,and light the color of the cells in cortex and hippocampus.3 conclusionWith the adoption of the elevated plus-maze test(EPM),light/dark box(LDB),open field test(OFT)and locomotor activity test on mice,and the EPM,Vogel conflict drinking,OFT and restraint stress test on rats,the paper makes a comparative evaluation on anti-anxiety effects of CZZX and JW in aspects of the unconditioned reflex model and the punitive conditional reflex model,etc.,increasing the comprehensiveness and reliability of pharmacodynamic evaluation.Researches on the mechanism of CZZX show:its anti-anxiety role is mainly realized by regulating ERK/CREB pathway,affecting BDNF expression,and decreasing the expression of the third messenger c-fos in brain tissues.Researches on this topic lay a certain experimental foundation for subsequent "clinical research on new drug" on CZZX,and provide a train of thought for the systematic in-depth research on the anti-anxiety efficacy and mechanism of TCM compound preparations.