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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 investigate the effect of Wen Dan Tang containing serum on the expression of GJ and Cx43 in astrocytes. Methods: Forty SD rats were randomly divided into 5 groups: normal group, clozapine 20mg / kg group, Wendan decoction 40g / kg group, Wendan decoction group 30g / kg group and Wendan decoction group 20g / kg group. Clozapine group ig clozapine 20 mg / kg, Wendan Decoction 40g / kg, 30g / kg, 20g / kg group were ig Wendan Tang crude drug 40,30,20 g / kg, the normal group with the same amount of saline ig, 1 time / d, 8 days after the rats were sacrificed, take blood, centrifuged serum, inactivated, filter sterilized spare. Astrocytes were divided into 6 groups: control group (normal medium), normal rat serum group, clozapine 20mg / kg serum group, Wendan Decoction 40g / kg serum group and Wendan Decoction 30g / kg serum Group and Wendan Decoction 20g / kg serum group. The serum containing serum was changed according to the corresponding group and cultured for 24h, 36h and 48h. The fluorescence dye-labeled tracer technique (SLDT) GJ level in astrocytes; Western blotting was used to detect phosphorylation and non-phosphorylation of Cx43 in astrocytes. Results: After treated with 40g / kg, 30g / kg and 20g / kg serum of Wendan Decoction for 24h, 36h and 48h, the area and intensity of fluorescence diffusion could be significantly increased, and Cx43 phosphate And non-phosphorylated expression. Conclusion: Wendan Decoction can up-regulate the expression of Cx43 phosphorylation and non-phosphorylation in astrocytes, enhance the GJ level and improve the function of GJIC.