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【目的】探讨丹参酮ⅡA(TanshinoneⅡA)对人胎脐静脉上皮细胞(HUVEC)缝隙连接细胞通讯(GJIC)的影响及其机制。【方法】采用四甲基偶氮唑盐(MTT)法检测丹参酮ⅡA对HUVEC细胞生长的影响,荧光显微镜观察及流式细胞仪荧光示踪法分析GJIC功能变化,并与阳性对照药全反式维甲酸(ATRA)比较,采用荧光实时定量PCR(q RT-PCR)法分析Cx37、Cx40 m RNA的表达。【结果】丹参酮ⅡA作用HUVEC 24、48 h的半数致死量(IC50)分别为15、8.5μmol/L,32、64μmol/L丹参酮ⅡA作用HUVEC 48 h有明显细胞毒性,抑制率大于70%;选用0、4、8、16、32μmol/L丹参酮ⅡA作用HUVEC48 h,流式细胞仪检测结果显示:各组接受绿色荧光Calcein-AM的细胞数量与总细胞数比值明显升高,其中8、16、32μmol/L组可显著提高GJIC功能(P<0.01)。q RT-PCR分析结果显示:与空白对照组比较,8、16、32μmol/L丹参酮ⅡA组可显著提高Cx37、Cx40 m RNA表达(P<0.05或P<0.01),并有一定的浓度依赖关系。【结论】丹参酮ⅡA能够提高Cx37、Cx40 m RNA表达,这可能是丹参酮ⅡA增强上皮细胞GJIC功能的机制之一。
【Objective】 To investigate the effect and mechanism of TanshinoneⅡA on GJIC in human umbilical vein endothelial cells (HUVECs). 【Method】 The effects of tanshinone Ⅱ A on the growth of HUVECs were detected by MTT assay. The changes of GJIC function were observed by fluorescence microscopy and flow cytometry. Retinoic acid (ATRA), the expression of Cx37 and Cx40 m RNA was analyzed by real-time quantitative PCR (q RT-PCR). 【Results】 The results showed that the IC50 of HUVEC at 24h and 48h were 15 and 8.5μmol / L, respectively. The cytotoxicity of tanshinone ⅡA treated HUVEC at 48 h and 48 h was observed. The inhibition rate was more than 70% The effect of 0, 4, 8, 16, 32μmol / L tanshinone ⅡA on HUVEC 48h, flow cytometry results showed that: the number of cells receiving green fluorescent Calcein-AM cells and total cell number ratio was significantly increased, 32μmol / L group could significantly improve the function of GJIC (P <0.01). q RT-PCR analysis showed that 8,16,32μmol / L tanshinone ⅡA group could significantly increase the expression of Cx37 and Cx40 m RNA (P <0.05 or P <0.01), as well as in a concentration-dependent manner . 【Conclusion】 Tanshinone Ⅱ A can enhance the expression of Cx37 and Cx40 m RNA, which may be one of the mechanisms by which Tanshinone ⅡA enhances the function of GJIC in epithelial cells.