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目的探讨HBX特异性RNA小干扰抑制乙型肝炎病毒复制的作用。方法用脂质体转染法将干扰质粒pSilencer3.1-shHBX和通用阴性对照质粒分别转染HepG2.2.15细胞,并设立未转染的空白对照组。分别于转染后24、487、2 h收集细胞培养上清液和细胞总蛋白,Western blot检测HBx蛋白表达情况,ELISA检测3组细胞上清中HBsAg和HBeAg表达情况。结果 Western blot检测显示,干扰质粒转染组HBx蛋白表达量逐渐降低,各时间点的表达量与空白对照组和通用阴性对照组比较差异均有统计学意义(P<0.01);ELISA检测显示,各时间点干扰质粒转染组细胞培养液上清中HBsAg和HBeAg的表达均下调,表达量与空白对照组和通用阴性对照组比较差异均具有统计学意义(P<0.01)。结论 HBX特异性小干扰RNA可抑制HBV复制,为siRNA作为抗乙肝病毒感染制剂提供了实验依据。
Objective To investigate the effect of HBX-specific RNA interference on hepatitis B virus replication. Methods Interfering plasmids pSilencer3.1-shHBX and universal negative control plasmids were transfected into HepG2.2.15 cells respectively by lipofection method, and untransfected blank control group was established. The cell culture supernatant and total cellular protein were collected 24,487 and 2 h after transfection respectively. The expression of HBx protein was detected by Western blot. The expression of HBsAg and HBeAg in the supernatant of the three groups was detected by ELISA. Results The results of Western blot showed that the expression of HBx protein in the plasmid transfected group decreased gradually, and the expression of HBx protein at each time point was significantly different from that in the blank control group and the common negative control group (P <0.01) The expression of HBsAg and HBeAg in the cell culture supernatant of the plasmid transfection group were all down-regulated at each time point, and the difference was statistically significant compared with the blank control group and the common negative control group (P <0.01). Conclusion HBX-specific small interfering RNA can inhibit HBV replication and provide an experimental basis for siRNA as an anti-HBV infection preparation.