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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-VP1 siRNA转染HeLa细胞,用RT-PCR法检测CVB3-VP1 RNA水平,免疫荧光检测CVB3-VPl蛋白的表达水平。结果60 pmol／L CVB3-VP1 siRNA是抑制CVB3 RNA及VP1表达的最佳剂量;在转染后的48 h,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48 h呈现出完全抑制作用。
Objective To investigate the dose-response and time-dependence of specific CVB3-VP1 siRNA on the in vitro replication of CVB3. Methods CVB3 was transfected into HeLa cells. CVB3-VP1 siRNA was transfected into HeLa cells by lipofectamine. CVB3-VP1 RNA was detected by RT-PCR. The expression of CVB3-VP1 protein was detected by immunofluorescence. Results 60 pmol / L CVB3-VP1 siRNA could inhibit the expression of CVB3 RNA and VP1 at optimal concentration. At 48 h after transfection, the inhibition of CVB3-VP1 was best. Conclusion The specific CVB3-VP1 siRNA has a significant dose-dependent and time-dependent relationship with the expression level of CVB3-VP1 and the level of CVB3-RNA replication. The complete inhibitory effect of CVB3-VP1 siRNA was observed 48 h after transfection.