文献类型：期刊文献

中文题名：菖蒲郁金汤对抽动秽语综合征模型大鼠突触体SNARE蛋白复合体的影响研究

英文题名：Effect of Changpu Yujin Decoction on Synaptosome SNARE Protein Complex in Tourette Syndrome Model Rats

作者：冯鹏[1,2];孙治前[1];罗文珍[1];史正刚[1];李玉霞[3];吴丽萍[3];尚菁[1];田文霞[3];陈静[3]

第一作者：冯鹏

机构：[1]甘肃中医药大学中医临床学院,甘肃兰州730000;[2]河西学院医学院,甘肃张掖734000;[3]甘肃中医药大学附属医院,甘肃兰州730020

第一机构：甘肃中医药大学中医临床学院

年份：2021

卷号：37

期号：5

起止页码：709

中文期刊名：南京中医药大学学报

外文期刊名：Journal of Nanjing University of Traditional Chinese Medicine

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

基金：国家自然科学基金(81960886);甘肃省高等学校创新基金(2020A-081);甘肃省重点研发计划项目(20YF3FA041)。

语种：中文

中文关键词：菖蒲郁金汤;抽动秽语综合征;突触体;SNARE蛋白复合体

外文关键词：Changpu Yujin Decoction;Tourette syndrome;synaptosome;SNARE protein complex

摘要：目的研究菖蒲郁金汤对抽动秽语综合征(Tourette syndrome,TS)模型大鼠突触体中SNARE蛋白复合体的调控作用,揭示菖蒲郁金汤治疗TS的药效机制。方法选择3周龄SD大鼠240只,依据随机数字表将SD大鼠分为空白组(60只)和造模组(180只),采用3,3'-亚氨基二丙腈(IDPN)腹腔注射法制备TS动物模型。造模成功后,再次随机将造模组大鼠分为模型组、硫必利组及菖蒲郁金汤组,每组60只。从造模完成后次日(即d0)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,每日1次,连续给药4周。各组大鼠分别于第0、7、14、21、28天(即d0,d7,d14,d21,d28),随机抽取12只大鼠,麻醉后取纹状体,采用Percoll密度梯度离心法制备纹状体中的突触体,并在透射电镜下进行形态学鉴定。ELISA法检测纹状体多巴胺(Dopamine,DA)含量,qPCR和Western blot法检测突触体中SNAP-25、Syntaxin-1a和VAMP-2 mRNA及蛋白的表达,免疫组织化学法检测纹状体上述指标的表达。结果与空白组相比,模型组各观察点纹状体中DA含量降低(P<0.01),突触体/纹状体中SNAP-25、Syntaxin-1a、VAMP-2 mRNA及蛋白的表达量显著降低(P<0.05,P<0.01);与模型组相比较,硫必利组及菖蒲郁金汤组在干预期间的各观察点(d7,d14,d21,d28)纹状体中DA含量以及突触体/纹状体内SNAP-25、Syntaxin-1a、VAMP-2 mRNA及蛋白的表达量呈逐渐升高趋势。在治疗结束后(d28),菖蒲郁金汤组突触体中SNAP-25、Syntaxin-1a、VAMP-2 mRNA,Syntaxin-1a蛋白及纹状体中SNAP-25、VAMP-2蛋白的表达高于硫必利组(P<0.05,P<0.01)。结论菖蒲郁金汤通过调控SNAP-25、Syntaxin-1a、VAMP-2的表达促进DA的释放,进而发挥其抗抽动的作用。
OBJECTIVE To study the regulatory effect of Changpu Yujin Decoction on the SNARE protein complex in the synaptosomes of Tourette syndrome(TS)model rats,and to reveal the pharmacodynamic mechanism of Changpu Yujin Decoction in the treatment of TS.METHODS 240 SPF SD male rats(3 weeks)were randomly divided into blank group(n=60)and model group(n=180).Model group received intraperitoneal injection of 3,3'-iminodipropionitrile to establish TS model.After the TS model was successfully established,the model rats were randomly divided into model group,tiapride group,and Changpu Yujin Decoction group,each with 60 rats.The intragastric administration was started from the next day after the TS model was made(d0).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 d0,d7,d14,d21 and d28,12 rats were randomly selected from each group.After anesthesia,the striatum was taken out,and the synaptosomes in the striatum were prepared by Percoll density gradient centrifugation.Morphological identification was performed under transmission electron microscope;The level of dopamine(DA)in the striatum was examined by ELISA;Expression of SNAP-25,Syntaxin-1a and VAMP-2 in the synaptosomes/striatumwere measured by qPCR,Western blot analysis and immunohistochemical staining.RESULTS Compared with the blank group,the DA content in the striatum in the model group was reduced(P<0.01),and the expressions of SNAP-25,Syntaxin-1a,VAMP-2 mRNA and protein in the synaptosome/striatum significantly reduced(P<0.05,P<0.01);Compared with the model group,the DA content in striatum and the mRNA and protein expression levels of SNAP-25,Syntaxin-1a,VAMP-2 in the tiapride group and Changpu Yujin group showed a gradual increase trend during the intervention period(d7,d14,d21,d28);At d28(after the end of treatment),the expressions of SNAP-25,Syntaxin-1a,VAMP-2 mRNA,Syntaxin-1a protein in synaptosome and SNAP-25 and VAMP-2 protein in the striatum were higher than those of tiapride group(P<0.05,P<0.01).CONCLUSION Changpu Yujin Decoction promotes the release of DA by regulating the expression of SNAP-25,Syntaxin-1a,and VAMP-2,thereby exerting its anti-tic effect.