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为了研究慢病毒介导的shRNA(Short hairpin RNA,shRNA)在柯萨奇B组3型病毒(Coxsackievirus B3,CVB3)导致的心肌炎小鼠模型中的抗病毒作用,合成针对CVB3基因组3753～3771区域的慢病毒Lenti-sh3753,感染HeLa细胞后感染CVB3病毒,通过荧光显微镜观测shRNA的表达和病毒致细胞病变效应,并测定培养上清中的病毒滴度,将慢病毒Lenti-sh3753感染BALB/c小鼠后感染CVB3病毒,观察小鼠的存活率,心脏组织中的病毒滴度和病理变化。结果发现Lenti-sh3753能在HeLa细胞中表达shRNA,并能有效抑制细胞中病毒RNA的复制。在小鼠模型上,Lenti-sh3753能提高小鼠的存活率,降低心脏中的病毒含量,从而减轻病理反应。这些结果提示,Lenti-sh3753在细胞和动物模型中能针对性地降解CVB3病毒RNA,明显降低病毒滴度,有效控制病毒感染。
To investigate the antiviral effect of lentivirus-mediated short hairpin RNA (shRNA) in a mouse model of myocarditis induced by Coxsackievirus B3 (CVB3), a recombinant plasmid targeting the region 3753-3771 of the CVB3 genome Of lentivirus Lenti-sh3753, infected with CVB3 virus after infection of HeLa cells. The expression of shRNA and the cytopathic effect of virus were observed by fluorescence microscopy. The virus titer in the culture supernatant was determined. Lenti-sh3753 lentivirus infected BALB / c After the mice were infected with CVB3 virus, the survival rate of mice was observed, and the virus titer and pathological changes in heart tissues were observed. The results showed that Lenti-sh3753 can express shRNA in HeLa cells, and can effectively inhibit the replication of viral RNA in cells. In the mouse model, Lenti-sh3753 can improve the survival rate of mice, reduce the virus content in the heart, thereby reducing the pathological response. These results suggest that Lenti-sh3753 can specifically degrade CVB3 virus RNA in cell and animal models, significantly reducing virus titer and effectively controlling viral infection.