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目的:通过在小鼠病毒性心肌炎动物模型研究短双链小干扰RNA(small interfering RNA,siRNA)对病毒感染和复制的抑制作用,研究RNAi在治疗病毒性疾病的可行性。方法:利用质粒载体将siRNA转染至HeLa细胞和Balb-c小鼠后感染病毒,荧光显微镜观察GFP表达量观察细胞内质粒转染效率和持续时间,通过病毒致细胞病变作用(CPE)保护实验病毒空斑形成实验检测病毒受抑制程度,动物模型中观察动物死亡率和易感组织病理变化评价siRNA的保护作用。结果:在HeLa细胞中针对CVB3 2B区的siRNA能显著抑制柯萨奇病毒B3的感染和复制,抑制率可达90%。动物模型中siRNA质粒可改善动物存活率(30%),并降低易感脏器中病毒含量,减轻病理反应。结论:针对CVB3基因组2B区的siRNA在病毒性心肌炎动物模型中具有保护作用。
OBJECTIVE: To study the feasibility of RNAi in the treatment of viral diseases by studying the inhibitory effect of small interfering RNA (siRNA) on viral infection and replication in an animal model of viral myocarditis in mice. Methods: The siRNA was transfected into HeLa cells and Balb-c mice using plasmid vector. The expression of GFP was observed under a fluorescence microscope to observe the transfection efficiency and the duration of the plasmids. The virus cytopathic effect (CPE) protection experiment Virus plaque formation assay was used to detect the degree of virus inhibition. Animal models were observed for animal mortality and susceptibility histopathological changes to evaluate the protective effect of siRNA. Results: siRNA targeting CVB3 2B in HeLa cells significantly inhibited infection and replication of coxsackievirus B3 with an inhibitory rate of up to 90%. SiRNA plasmid in animal model can improve the animal survival rate (30%), and reduce the virus content in susceptible organs, reduce the pathological response. CONCLUSION: siRNA targeting region 2B of the CVB3 genome has a protective effect in animal models of viral myocarditis.