健脾止动汤对Tourette综合征模型大鼠自主活动及纹状体多巴胺突触囊泡蛋白表达的影响  

Effect of Jian-Pi-Zhi-Dong decoction on autonomous activity and dopamine synaptic vesicle protein expression in the striatum of Tourette syndrome model rats

作　　者：李钰萍 薛小娜[1] 陈自佳[1] 王道涵[1] 林绿萍 刘晓芳[1] Li Yuping;Xue Xiaona;Chen Zijia;Wang Daohan;Lin Lyuping;Liu Xiaofang(Department of Paediatrics,Dongfang Hospital of Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学东方医院儿科,北京100078

出　　处：《中华行为医学与脑科学杂志》2024年第6期481-486,共6页Chinese Journal of Behavioral Medicine and Brain Science

基　　金：国家自然科学基金(82004422,82205191)。

摘　　要：目的研究健脾止动汤对Tourette综合征(Tourette syndrome,TS)模型大鼠自主活动及纹状体内多巴胺(dopamine,DA)突触囊泡蛋白表达的影响。方法4周龄雄性SD大鼠,采用亚氨基二丙腈腹腔注射法建立TS模型;将造模成功的30只大鼠按照随机数字表法分为模型组、中药组、泰必利组,每组10只,另取体质量相匹配的10只大鼠作为对照组。中药组、泰必利组大鼠分别给予健脾止动汤(1.6 g/100 g)和硫酸泰必利混悬液(2.1 mg/100 g)灌胃干预,对照组和模型组大鼠给予等体积0.9%氯化钠溶液灌胃,1次/d,持续4周。采用自主活动程控仪监测大鼠的自主活动次数,采用ELISA法检测大鼠纹状体DA水平,Western blot技术检测纹状体多巴胺转运蛋白(dopamine transporter,DAT)、Ⅱ型囊泡单胺转运体(vesicular monoamine transporter-2,VMAT2)以及α突触核蛋白(α-synuclein,α-syn)的蛋白表达水平。采用SPSS 25.0软件进行统计分析,多组间比较采用单因素方差分析、非参数检验和重复测量方差分析。结果4组大鼠自主活动次数比较,时间和组别的交互作用显著(F=184.354,P<0.001)。灌胃1~4周,模型组大鼠的自主活动次数均高于对照组(均P<0.05),中药组和泰必利组大鼠自主活动次数均低于模型组(均P<0.05),且中药组和泰必利组在灌胃1~4周的自主活动次数均较造模后减少(均P<0.05)。Western blot结果显示,4组大鼠纹状体的α-syn(H=29.098)、DAT(F=54.632)及VMAT2(H=18.982)蛋白相对表达量均差异有统计学意义(均P<0.001)。中药组和泰必利组大鼠α-syn蛋白表达水平均低于模型组[0.39(0.36,0.51),0.39(0.36,0.50),0.62(0.50,0.70)](均P<0.05);中药组DAT蛋白表达水平高于模型组且低于泰必利组[(0.37±0.06),(0.26±0.07),(0.49±0.09)](均P<0.05),VMAT2蛋白表达水平与模型组比较差异无统计学意义(P>0.05)。ELISA结果显示,4组大鼠纹状体DA水平差异有统计学意义(F=75.370,P<0.001)。模型组DA水平高于对照组[(7.65�Objective To explore the effect of Jian-Pi-Zhi-Dong decoction on autonomous activity and dopamine(dopamine,DA)synaptic vesicle protein expression in the striatum of Tourette syndrome(TS)model rats.Methods The 4-week-old male SD rats were used to establish the TS model by intraperitoneal injection of N-aminodipropionate.Thirty successfully modeled rats were randomly divided into model group,Chinese medicine group,and tiberide group according to random number table method,with 10 rats in each group.And another 10 rats with matched body mass were selected as the control group.Rats in Chinese medicine group were given Jian-Pi-Zhi-Dong decoction solution(1.6 g/100 g)and the rats in tiapride group were given sulfate tiapride suspension(2.1 mg/100 g),while rats in control group and model group were given an equal volume of 0.9%sodium chloride solution,once a day for 4 weeks.The number of autonomous activities in rats was determined by autonomous activity programmer.ELISA was used to detect the level of DA in the striatum of rats and the expression of dopamine transporter(DAT).Vesicular monoamine transporter-2(VMAT2)andα-synuclein(α-syn)were measured by Western blot.SPSS 25.0 software was used for data analysis.Multiple group comparisons were performed using one-way ANOVA,non-parametric test and repeated measures ANOVA.Results Comparing the number of autonomous activities among the 4 groups,the interaction effect between time and group was significant(F=184.354,P<0.001).At the 1-4 weeks of gavaging,the numbers of autonomic activities in the model group were more than those in the control group(all P<0.05).While the numbers of autonomic activities in Chinese medicine group and tiapride group were less than those in the model group(all P<0.05).Moreover,the numbers of autonomic activities in Chinese medicine group and tiapride group from 1 to 4 weeks were less than those after model making(all P<0.05).The Western blot results showed significant differences in the relative expression ofα-syn(H=29.098),DAT(F=54.632)and VMA

关 键 词：TOURETTE综合征 Α突触核蛋白 Ⅱ型囊泡单胺转运体 多巴胺转运蛋白 健脾止动汤 

分 类 号：R285.5[医药卫生—中药学]