菖蒲郁金汤对抽动秽语综合征大鼠突触胞吞相关蛋白表达的影响

Effects of Changpu Yujin Decoction on the expression of synaptic endocytosis related proteins in a rat model of Tourette syndrome

目的研究菖蒲郁金汤对抽动秽语综合征模型大鼠神经元胞吞相关蛋白表达的影响。方法大鼠随机分为空白组、模型组、硫必利组及菖蒲郁金汤组。采用腹腔注射亚氨基二丙腈制备抽动秽语综合征模型,连续灌胃给予相应药物4周,在第0、7、14、21、28天,采用刻板行为及运动行为评分法评定行为学变化,Percoll密度梯度离心法制备突触体,RT-qPCR和Western blot法检测突触体中clathrin、AP-2、dynamin-1 mRNA及蛋白表达,免疫组织化学法检测纹状体clathrin、AP-2、dynamin-1蛋白表达。结果与模型组比较,硫必利组和菖蒲郁金汤组行为学评分随时间延长逐渐降低(P<0.05,P<0.01);在给药第28天,菖蒲郁金汤组刻板行为和运动行为评分均低于硫必利组(P<0.01)。与空白组比较,模型组各观察点突触体/纹状体clathrin、AP-2、dynamin-1 mRNA及蛋白表达均升高(P<0.01);与模型组比较,硫必利组和菖蒲郁金汤组在给药期间突触体/纹状体clathrin、AP-2、dynamin-1 mRNA及蛋白表达随时间延长逐渐降低(P<0.05,P<0.01);在给药第28天,菖蒲郁金汤组突触体clathrin、dynamin-1 mRNA和clathrin蛋白表达低于硫必利组(P<0.05),2组纹状体clathrin、AP-2、dynamin-1蛋白表达差异无统计学意义(P>0.05)。结论菖蒲郁金汤抗抽动的作用可能是通过调控clathrin、AP-2、dynamin-1的表达来实现。

AIM To observe the effects of Changpu Yujin Decoction on the expression of neuronal endocytosis-related proteins in rat models of Tourette syndrome(TS).METHODS The rats were randomly divided into the blank group,the model group,the tiapride group and Changpu Yujin Decoction group.The TS rat models were induced by intraperitoneal injection of imindipropionitrile followed by the corresponding drug administration by gavage for 4 weeks.On the 0th,7th,14th,21st and 28th days,the rats had their behavioral changes evaluated by scoring on stereotyped behavior and motor behavior;their synaptosomes prepared by Percoll density gradient centrifugation for detection of the mRNA and protein expressions of clathrin,AP-2 and dynamin-1 by RT-qPCR and Western blot;and their protein expressions of clathrin,AP-2 and Dynamin-1 in striatum determined by immunohistochemical method.RESULTS Compared with the blank group,the model group was found to be with increased mRNA and protein expressions of clathrin,AP-2,dynamin-1 in synaptosomes/striatum(P<0.01).Compared with the model group,both groups intervened with tiapride and Changpu Yujin Decoction displayed decreased behavioral scores(P<0.05,P<0.01);reduced mRNA and protein expressions of clathrin,AP-2,dynamin-1 in synaptosomes/striatum(P<0.05,P<0.01)in line with the length of the intervention time.On the 28th day after administration,Changpu Yujin Decoction group demonstrated lower scores of rigid behavior and motor behavior(P<0.01);reduced expressions of synaptosome clathrin,dynamin-1 mRNA and clathrin protein(P<0.05),in contrast to the tiapride group.Tiapride and Changpu Yujin Decoction produced no significant difference in the expressions of clathrin,AP-2 and dynamin-1 protein in the striatum(P>0.05).CONCLUSION Changpu Yujin Decoction may achieve the anti-tic effects by regulating the expressions of clathrin,AP-2 and dynamin-1.