RNA干扰在柯萨奇病毒致心肌炎小鼠模型中的抗病毒研究

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。

Inhibition of Virus Infection in Coxsackievirus-induced Myocarditis Model by RNA Interference

To evaluate the effects of lentivirus-delivered short hairpin RNA(shRNA) on CVB3 infection in an animal model by RNA interference technique,we constructed a recombinant lentivirus expressing shRNA-3753 against the viral genome region 3753-3771,then transduced Lenti-sh3753 into mice infected with CVB3.We evaluated the antiviral ability of lenti-sh3753 by cytopathic effect(CPE),viral plaque assay and histological analysis of mice hearts.The results showed that Lenti-sh3753 exhibited a significant protective effect on cell viability and reduction of viral titers in supernatant of cell culture by specific inhibition on viral replication.Lenti-sh3753 also prolonged the mice survival and limited the viral production in mice hearts.These data proposed that Lenti-sh3753 can effectively inhibit CVB3 infection in a coxsackievirus-induced myocarditis model,suggesting its potential role in prevention and therapy of viral diseases.