自拟菖蒲郁金汤对抽动秽语综合征大鼠突触超微结构及突触素、突触后致密物-95蛋白的影响

Effect of Self-Designed Changpu Yujin Decoction on Synapse Ultrastructure and Synapse-Related Proteins in Tourette Syndrome Rats

目的观察自拟菖蒲郁金汤(CPYJD)对抽动秽语综合征(TS)模型大鼠纹状体突触超微结构及突触素(SYN)、突触后致密物-95(PSD-95)蛋白表达的影响,探讨其抗抽动的作用机制。方法依据随机数字表法将140只SD大鼠分为空白组(35只)和造模组(105只),采用腹腔注射亚氨基二丙腈制备TS动物模型,随后将造模组大鼠随机分为模型组、硫必利组及CPYJD组,每组35只。从造模完成后次日开始灌胃给药,空白组与模型组给予生理盐水,其余各组给予相应药物,1次/d,连续给药4周。各组大鼠分别于治疗前、治疗7、14、21、28 d,随机抽取7只大鼠,行为学评定后麻醉剥离出纹状体,利用透射电镜观察突触超微结构,免疫组织化学法检测大鼠纹状体中SYN、PSD-95蛋白水平的变化。结果与空白组比较,模型组各时间刻板、运动行为评分、突触间隙升高(P<0.05),突触致密物厚度、SYN、PSD-95蛋白表达降低(P<0.05)。与本组治疗前比较,硫必利组和CPYJD组28 d刻板、运动行为评分、突触间隙降低,SYN、PSD-95蛋白升高(P<0.05),CPYJD组28 d突触致密物厚度增加(P<0.05)。与模型组比较,硫必利组及CPYJD组28 d刻板、运动行为评分、突触间隙降低,SYN、PSD-95蛋白表达升高(P<0.05),CPYJD组28d致密物厚度增加(P<0.05)。与硫必利组比较,CPYJD组14 d刻板行为及7 d运动行为评分升高(P<0.05),21、28 d刻板行为评分、突触间隙降低,28 d运动行为评分降低,28 d突触致密物厚度增大,7、14、21、28 d PSD-95蛋白表达升高(P<0.05)。结论CPYJD可改善TS大鼠刻板及运动行为,相较于硫必利起效时间晚,但起效后抗抽动效应稳定,其机制可能与改善突触结构及功能,提高SYN及PSD-95蛋白表达相关。

Objective To observe the effect of self-designed Changpu Yujin Decoction(CPYJD)on the ultrastructure of synaptic and the expression of synaptophysin(SYN)and postsynaptic density-95(PSD-95)protein in the striatum of Tourette syndrome(TS)model rats,and to explore the mechanism.Methods According to random number table,140 SD rats were divided into blank group(n=35)and building model group(n=105).The model was established by intraperitoneal injection of 3,3’-iminodipropionitrile.Then the TS rats were randomly divided into model group,tiapride group and CPYJD group,35 rats in each group.The intragastric administration was started from the next day after the TS model was established.The blank group and the model group were given normal saline,and the other groups were given corresponding drugs,once a day,for 4 consecutive weeks.At before treatment,7,14,21 and 28 d after treatment,7 rats were randomly selected from each group.After behavioral assessment,the striatum was strip out after anesthesia to strip out the striatum,the synapses ultrastructure were observed by transmission electron microscope,the expressions of SYN and PSD-95 in striatum were detected by immunohistochemistry.Results Compared with the blank group,the stereotyped behavior and motor behavior scores and the width of the synaptic cleft increased(P<0.05),the thickness of postsynaptic density(PSD)and the expression of SYN and PSD-95proteins decreased(P<0.05)in the model group at each observation point.Comparing with before treating in the same group,the stereotyped behavior and motor behavior scores,synaptic cleft decreased,the expression of SYN and PSD-95 increased in tiapride group and CPYJD group at 28 d(P<0.05),the thickness of PSD increased in CPYJD group at 28 d(P<0.05).Compared with the model group,the stereotyped behavior and motor behavior scores,synaptic cleft decreased,the expression of SYN and PSD-95 proteins increased in tiapride group and CPYJD group at 28 d(P<0.05).Compared with the tiapride group,the stereotypic behavior at score 14 d and motor behavior at score 7 d in CPYJD group increased(P<0.05),the stereotypical behavior score and synaptic cleft at 21,28 d were decreased,the motor behavior at score 28 d decreased,and the thickness of PSD at 28 d increased,expression of PSD-95 increased at 7,14,21 and 28 d in the CPYJD group(P<0.05).Conclusions CPYJD can improve the stereotypic and motor behavior of TS rats,compared with tiapride,the onset time is later,but the effect is stable,the mechanism may be related to the improvement of synaptic structure and function,and increase of SYN and PSD-95 protein expression.