温胆汤含药血清对星形胶质细胞Cx43和GJ的影响  

Effect of Wendan decoction medicated serum on Cx43 and GJ in astrocytes

作　　者：朱金华[1] 刘丹丹[1] 魏妍妍[1] 熊怀亮 戎文娟[1] 万红娇[1] 

机构地区：[1]江西中医药大学,南昌330004

出　　处：《中药药理与临床》2015年第5期5-8,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目;基金号:81160423

摘　　要：目的:探讨温胆汤含药血清对星形胶质细胞缝隙连接(GJ)和连结蛋白43(Cx43)表达的影响。方法:40只SD大鼠随机分为5组:正常组、氯氮平20mg/kg组、温胆汤40g/kg组、温胆汤30g/kg组、温胆汤20g/kg组。氯氮平组ig氯氮平20 mg/kg,温胆汤40g/kg、30g/kg、20g/kg组分别ig温胆汤生药40、30、20 g/kg,正常组用等量生理盐水ig,1次/d,8天后处死大鼠,取血,离心取血清,灭活,过滤除菌备用。将星形胶质细胞分为6组:对照组(普通培养基培养)、正常鼠血清组、氯氮平20mg/kg血清组、温胆汤40g/kg血清组、温胆汤30g/kg血清组、温胆汤20g/kg血清组,按照相应的分组更换含药血清进行培养24h、36h、48h后,采用划痕染料标记示踪技术(SLDT)在荧光显微镜下,通过荧光黄扩散距离测定星形胶质细胞间GJ水平;Western blotting法检测星形胶质细胞中Cx43磷酸化和非磷酸化的表达。结果:温胆汤40g/kg、30g/kg、20g/kg血清组作用于星形胶质细胞24h、36h、48h后,能使荧光扩散面积及强度不同程度的显著增加;并显著提高Cx43磷酸化和非磷酸化的表达。结论:温胆汤能够上调星形胶质细胞Cx43磷酸化和非磷酸化的表达,增强GJ水平,从而改善细胞间隙连接通讯(GJIC)功能。Objective： To studyeffect of Wendan decoction medicated serum on connexin 43（ Cx43） and gap junction（ GJ） in astrocytes. Method：40 SD rats were randomly divided into five groups： normal group,clozapine 20 mg / kg group,Wendan decoction 40 g / kg group,Wendan decoction 30 g / kg group and Wendan decoction 20 g / kg group,eight rats in each group. Normal group rats were given normal saline every day,clozapine 20 mg / kg group given original drug clozapine 20 mg / kg,Wendan decoction 40 g / kg,30 g / kg,20 g / kg groups were respectively given Wendan decoction（ 40,20,20 g / kg） every day. After 8 days,the rats were sacrificed to preparation containing serum. Astrocytes were divided into six groups： control group,normal rat serum group,clozapine 20 mg / kg medicated serum group,Wendan decoction 40 g / kg medicated serum,Wendan decoction 30 g / kg medicated serum and Wendan decoction 20 g / kg medicated serum. According to the cultivating group,to respectively replace the corresponding medicated serum of 24 h,36 hand 48 h. Under the fluorescence microscope,the GJ of astrocytes was detected through fluorescent yellow diffusion distance by scrape-loading and dye transfer（ SLDT）. The expression of Cx43 phosphorylated and non-phosphorylated in astrocytes was determined by Western blotting method. Result： Wendan decoction 40 g / kg、30g / kg、20g / kg medicated serumon astrocyte for 24 h,36 h and 48 h,it could significantly increase diffusion area andfluorescence intensity（ P〈0. 05）,which could also increase the expression of phosphorylation and non-phosphorylated Cx43（ P〈0. 05） in astrocytes. Conclusions： Wendan decoction medicated serum could obviously increasethe expression of phosphorylation and non-phosphorylated Cx43,enhance GJ,which thus play a regulatory role in the function of GJIC.

关 键 词：温胆汤含药血清 星形胶质细胞 连结蛋白43 缝隙连接 

分 类 号：R285[医药卫生—中药学]