Effect Of Jianpi Zhidong Decoction On 5-HT_1AR Expression In Hippocampus And Hypothalamus Of TS Model Rats With Chronic Restraint Stress

Objective:1.In the clinical aspect,the clinical efficacy of Jian-Pi-Zhi-Dong Decoction(JPZDD)was further confirmed by summarizing the clinical experience of tutor in treatment of Tourette Syndrome(TS)for many years,which laid a foundation for its further extension and application.2.In the experiment,build a rat model induced by chronic restraint stress with intraperitoneal injecting IDPN.Using chronic restraint stimulation to simulate emotional factors,then revealing the mechanism of stress factors how to induce and aggravate TS.Explore the relationship between 5-HT1AR,the emotional factors and the pathogenesis of TS by detecting the expression of 5-HT1AR in hippocampus and hypothalamus of ratsin each group,and revealing the partial anti-twitching mechanism of JPZDD.Methods:Twenty-four male SD rats were randomly divided into four groups according to body weight after adaptive feeding for one week,including blank control group,model group,Chinese herbal group and Tiapride group,6 rats in each group.The blank control group were intraperitoneally injected with normal saline once a day for 7 consecutive days.The model groups,the Chinese herbal group and the Tiapride group were intraperitoneally injected with IDPN,250mg/kg/d for 7 consecutive days,and chronic restraint stimulation was given at 8:00a.m,every day for 3 hours.Chronic restraint stimulation lasted until the 21st day,the stability of the model was tested and regrouped by behaviorally.After successful modeling,the blank control group and model group were gavaged with normal saline(0.9%)at 10ml/kg/d,the Chinese herbal group was gavaged with JPZDD at 16g/kg/d,and the Tiapride group was gavaged with tiapride suspension at 21 mg/kg/d,respectively for six weeks.Behavioral tests were carried out on the first day after model establishment and on the eighth,fifteenth and twenty-ninth days after dosing.After 4 weeks of administration,the head was severed and the brain was removed,detecting 5-HT1AR in hippocampus and hypothalamus of rats through Western Blot and RT-PCR.Results:The results of stereotyped behavior score in rats:Compared with blank control group,the stereotyped behavior score of TS rats in model group,the Chinese herbal group and the Tiapride group increased significantly at the end of 21 days after modeling(P<0.05);However,there was no statistical significance between the three groups(P>0.05),At the end of 4 weeks of intragastric administration,compared with the model group and the Tiapride group,the stereotyped behavior score decreased significantly in the Chinese herbal group(P<0.01).The results of open-field experiment:After 21 days of modeling,the total activity scores of rats in the model group,the Chinese herbal group and the Tiapride group increased compared to blank control group,but there was no statistical significance between these groups(P>0.05).After 4 weeks of gavage,the total activity score of the model group was significantly higher than the blank control group(P<0.01).Compared with the blank control group,there was no statistical significance in the total activity score of rats in the Chinese herbal group(P>0.05),The total activity scores of the rats in the Chinese herbal group were significantly lower than those in the model group and the Tiapride group(P<0.01).The results of the elevated plus-maze test:After 21 days of modeling,the OE/TE values of the model group,the Chinese herbal group were significantly lower than those of the blank control group(P<0.01).The OE/TE values of the Tiapride group was lower than those of the blank control group(P<0.05),But there was no statistical significance between these groups(P>0.05).After 4 weeks of gavage,the OE/TE values of model group significantly lower than the blank control group(P<0.05).Compared with the blank control group,the OE/TE values of Chinese herbal group not significantly different from that of blank control group(P>0.05).Compared with model group,OE/TE value of Chinese herbal group was higher than that of model group(P<0.05).There was no significant difference in OE/TE value between Tiapride group and model group(P>0.05).Results of the expression level of 5-HT1AR protein in the hippocampus of each group:The expression levels of 5-HT1AR protein in the hippocampus of rats in the model group,the Chinese herbal group and the Tiapride group were all lower than those in the blank control group,but there was no significant difference among the groups(P>0.05).Results of the expression level of 5-HT1AR protein in the hypothalamus of each group:The expression levels of 5-HT1AR protein in the hypothalamus of the model group,the Chinese herbal group and the Tiapride group were all higher than those of the blank control group,but there was no significant difference among the groups(P>0.05).Results of the expression level of 5-HT1AR gene in the hippocampus of each group:The expression levels of 5-HT1AR gene in the hippocampus of rats in the model group and the Tiapride group were lower than those in the blank control group,the expression of hippocampus 5-HT1A R gene in the Chinese herbal group was higher than those in the the blank control group,but there was no significant difference among the groups(P>0.05).Results of the expression level of 5-HT1AR gene in the hypothalamus of each group:The expression of hypothalamic 5-HT1AR gene in the model group was higher than those in the blank control group,there was statistical significance(P<0.05).The expression of hypothalamic 5-HT1AR gene in the Chinese herbal group was higher than those in the model group,there was no statistical significance(P>0.05).The expression of hypothalamic 5-HT1AR gene in the Tiapride group was lower than those in the model group,there was no statistical significance(P>0.05).Conclusion:JPZDD can resist the influence of emotional factors,effectively inhibit the excitability of the TS model rat’s central nervous system and can obviously decrease the stereotyped movement,improve autonomic behavior.Detecting 5-HT1AR through Western Blot and RT-PCR,the results suggest that the pathogenesis of TS is related to the abnormality of 5-HT system,but it cannot confirm whether 5-HT1ARis the target of JPZDD.