特异性的VP1-siRNA对CVB3复制抑制作用的研究

目的 研究特异性的CVB3-VP1 siRNA对CVB3体外复制的抑制作用的量效关系与时效关系。方法 CVB3感染HeLa细胞，利用脂质体介导将CVB3-VP1siRNA转染HeLa细胞，用RT-PCR法检测CVB3-VP1 RNA水平，免疫荧光检测CVB3-VP1蛋白的表达水平。结果 60pmol／LCVB3-VP1 siRNA是抑制CVB3 RNA及VP1表达的最佳剂量；在转染后的48h，siRNA对CVB3-VP1的抑制作用达到最佳状态。结论 特异性的CVB3-VP1 siRNA对CVB3-VP1蛋白表达水平及CVB3-RNA复制水平呈明显的剂量依赖关系和时效关系，转染后48h呈现出完全抑制作用。

Inhibitory effect of specific siRNA on CVB3-VP1 expression in HeLa cells

Objective To investigate dose-dependence regulatory inhibition and time-effect relationship between CVB3-VP1 siRNA and the expression of CVB3-VP1 in HeLa cells. Methods Four differents dose siRNA were transfected into HeLa cells by liposome (IipofectAMINETM2000). 24, 48, 72, 96 h later, the CVB3-VP1 protein expression were detected by immunofluorescence, and CVB3 mRNA level was tested by semi-quantitative RT-PCR. Results Observations have showed that 60 pmol/L CVB3-VP1 siRNA was the optimal dose, and the inhibitory effect on CVB3-VP1 expression was the optimum after transfection 48 h. Conclusion Specific siRNA have the dose-dependence and time-effect relationship to the expression level of CVB3-VP1. These results suggested a feasibility of using RNAi to elucidate pathogenesis of CVB3 myocarditis and that RNAi might represent a new approach for the treatment of CVB3 myocarditis.