南方医科大学学报 ›› 2012, Vol. 32 ›› Issue (10): 1423-.

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星形胶质细胞中缝隙连接蛋白connexin 43的表达及其功能调控

董淑英,童旭辉,蒋国君,谷昱琛,焦浩,李俊   

  • 出版日期:2012-10-20 发布日期:2012-10-20

Expression of connexin43and functional modulation of gap junction in neonatal rat astrocytesin vitro

  • Online:2012-10-20 Published:2012-10-20

摘要: 目的研究星形胶质细胞中缝隙连接蛋白connexin43(Cx43)的表达及由其形成的缝隙连接通讯功能的药物调控。方法
实验分为正常对照组、全反式维甲酸组(10µmol/L全反式维甲酸作用24h)及油酸酰胺组(25µmol/L油酸酰胺作用2h)。
western blotting法检测各组星形胶质细胞中Cx43总蛋白的表达;细胞免疫荧光法检测各组星形胶质细胞Cx43胞膜蛋白的表
达;荧光示踪法检测各组星形胶质细胞的缝隙连接通讯功能。结果与正常对照组相比,全反式维甲酸能增强星形胶质细胞中
Cx43总蛋白的表达(P<0.01),油酸酰胺则能减弱其表达(P<0.01);全反式维甲酸能增强Cx43胞膜蛋白表达,油酸酰胺能减弱其
表达;全反式维甲酸能增强星形胶质细胞缝隙连接通讯功能(P<0.01),而油酸酰胺则可显著降低该功能(P<0.01)。结论药物
全反式维甲酸及油酸酰胺可以对星形胶质细胞缝隙连接通讯功能进行调控,其机制可能与其影响星形胶质细胞Cx43总蛋白和
胞膜蛋白的表达有关。

Abstract: Abstract: ObjectiveTo determine the expression of connexin43(Cx43) protein and explore the functional modulation of gap
junction intercellular communication in astrocytes. MethodsCultured neonatal SD rat astrocytes were divided into normal
control group, all-trans retinoic acid (ATRA) group (treated with10μmol/LATRAfor24h) and oleamide group (treated with
25μmol/Loleamidefor2h). Western blotting and immunofluorescence assay were used to detect total cellular Cx43protein
expression and Cx43expression on the surface of the astrocytes, respectively. Parachute assay was used to evaluate the
functional changes of gap junction intercellular communication of the astrocytes. ResultsCompared with the normal control
cells, ATRA treatment resulted in a significantly increased expression of total Cx43protein in the astrocytes (P<0.01), and
oleamide significantly suppressed its expression (P<0.01). Similarly, ATRA obviously enhanced while oleamide suppressed
Cx43protein expression on the surface of the astrocytes. The gap junction intercellular communication of the astrocytes was
enhanced by ATRA (P<0.01) and inhibited by oleamide (P<0.01). ConclusionATRA and oleamide can modulate gap junction
intercellular communication of the astrocytes possibly by regulating the expression of Cx43protein.