苦参素对HeLa细胞自噬性死亡的影响

Impact of Sophora on the Autophagy of HeLa Cells

目的探讨苦参素对人宫颈癌HeLa细胞自噬的影响,初步探讨其作用机制。方法不同浓度(1.0、1.5、2.0 g·L^(-1))的苦参素作用于HSV-2感染的人宫颈癌HeLa细胞,RFP-GFP-LC3和HPV16 DNA共转染后12 h和36 h收集细胞,透射电子显微镜观察细胞自噬活性的改变,SDS-聚丙烯酰胺凝胶电泳以及Western Blot检测在HeLa细胞中HSV-2感染引起自噬相关蛋白LC3-Ⅱ的变化。结果病毒感染组自噬小体明显增多,多于对照组(P<0.05);病毒感染组自噬相关蛋白LC3-Ⅱ明显升高,高于对照组(P<0.05);与对照组比较,作用72 h后,1.0、1.5、2.0 g·L^(-1)苦参素组均能不同程度提高HeLa细胞的自噬活性,均可见到自噬小体的增加,其自噬相关蛋白LC3-Ⅱ蛋白表达量明显增加(P<0.05),HSV-2释放滴度明显降低(P<0.05)。结论苦参素能通过增强HeLa细胞自噬活性,减少HSV-2感染后的释放与产生。

Objective To explore the impact of sophora on human cervical cancer HeLa cell autophagy and to explore its action mechanism.Methods Different concentrations(1.0,1.5,2.0 g·L^(-1)) of sophora was used for human cervical cancer HeLa cells after HSV-2 infection,The cells were collected in a total of 12 hours and 36 hours after RFP-GFP-LC3 and HPV16 DNA transfection,the autophagy activity change was observed by using transmission electron microscopy,SDS polyacrylamide gel electrophoresis and Western Blot detection were used to detect the autophagy related protein LC3-Ⅱ changes in the HeLa cells after HSV-2 infection.Results The autophagosome was obviously increased in the virus infection group,compared with the control group(P<0.05);the autophagy related proteins LC3-Ⅱ content was increased significantly in the virus infection group,compared with the control group(P<0.05).After 48 hours and 72 hours,compared with the control group,1.0,1.5,2.0 g·L^(-1) of sophora could improve the autophagy activity of HeLa cells(P<0.05);the autophagosome was significant increased(P<0.05);the autophagy related protein LC3-Ⅱ was significantly increased(P<0.05),the HSV-2 release drop degree was decreased obviously(P<0.05).Conclusion Sophora can improve the autophagy activity of HeLa cells,reduce the release of HSV-2 after virus infection.