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【目的】研究丹参素钠(salvianic acid A sodium,SAAS)对小鼠黑色素瘤细胞株B16细胞间缝隙连接(GJIC)的影响及其机制。【方法】采用四甲基偶氮唑盐(MTT)法检测SAAS对B16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结合分析缝隙连接(GJ)功能变化,并与阳性对照药全反式维甲酸(ATRA)比较,采用western blot法分析缝隙连接蛋白Cx32和Cx43的表达。【结果】以0～8μmol/L SAAS处理B16细胞48 h不影响其生存;用0、1、2、4、8μmol/L SAAS处理细胞48 h后,荧光显微镜下观察,SAAS能明显提高B16细胞Calcein传递,流式细胞术分析对照组和试验组的绿色荧光细胞(G4)与双阴性受体细胞(G3)比值(G4/G3)分别是0.10±0.01、0.16±0.02、0.20±0.01、0.21±0.02和0.25±0.02,各试验组G4/G3值显著高于对照组(P<0.01);western blot分析结果显示:用0、1、2、4、8μmol/L SAAS处理细胞48 h能显著提高Cx32、Cx43的表达。【结论】体外较低浓度SAAS能够促进B16细胞细胞间缝隙连接功能,但在一定药物浓度作用后,不能再提高其功能。SAAS促进GJ机制可能是通过上调Cx32、Cx43蛋白表达途径。
【Objective】 To investigate the effect of salvianic acid sodium (SAAS) on the gap junction (GJIC) of mouse melanoma cell line B16 and its mechanism. 【Methods】 The effect of SAAS on the growth of B16 cells was detected by MTT assay. Fluorescent microscopy and flow cytometry (FCM) were used to analyze the functional changes of gap junction (GJ) and were compared with the positive control Drug ATRA (ATRA) comparison, Western blot analysis of connexin Cx32 and Cx43 expression. 【Results】 B16 cells treated with 0-8 μmol / L SAAS for 48 h did not affect their survival. After treatment with 0, 1, 2, 4 and 8 μmol / L SAAS for 48 h, SAAS significantly increased B16 cells Calcein transmission, flow cytometry analysis of the control group and the experimental group of green fluorescent cells (G4) and double negative recipient cells (G3) ratio (G4 / G3) were 0.10 ± 0.01,0.16 ± 0.02,0.20 ± 0.01,0.21 ± 0.02 and 0.25 ± 0.02, respectively. The G4 / G3 values in each experimental group were significantly higher than those in the control group (P <0.01). Western blot analysis showed that the cells treated with 0, 1, 2, 4 and 8μmol / Improve Cx32, Cx43 expression. 【Conclusion】 Lower concentrations of SAAS in vitro can promote the cell gap junctional function of B16 cells. However, after a certain drug concentration, SAAS could not enhance its function. SAJ promote GJ mechanism may be through up-regulation of Cx32, Cx43 protein expression pathway.