Huang Fen, Hua Xiuguo, Yang Shixing, Yuan Congli, Zhang Wen
Zoonosis Laboratory, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, PR China.
Antiviral Res. 2009 Sep;83(3):274-81. doi: 10.1016/j.antiviral.2009.06.008. Epub 2009 Jul 1.
RNA interference (RNAi) is a natural mechanism for suppressing or silencing expression of aberrant or foreign genes. It is a powerful antiviral strategy that has been widely employed to protect hosts from viral infection. Hepatitis E (HE) is an acute fulminant hepatitis in adults that has particularly high mortality in pregnant women. At this point in time, there is no vaccine or antiviral treatment that is effective against the infectious agent, HEV. The nonstructural polyprotein region possesses an RNA-dependent RNA polymerase (RdRp) that is responsible for the replication of the viral RNA genome. RdRp is therefore regarded as one of the most attractive candidates for RNA interference (RNAi). In the present study, the high efficiency and specificity of siRNA were evaluated by Real-Time quantitative PCR and Western blot assays. Protective effects against HEV infection were achieved in A549 cells and in piglets. In piglets treated with a shRNA-RdRp-1 expression plasmid prior to HEV inoculation, HEV antigens were significantly reduced in the liver, spleen, and kidneys, and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL) were clearly decreased. These results suggested that RNAi is a potentially effective antiviral strategy against HEV replication and infection.
RNA干扰（RNAi）是一种抑制或沉默异常或外源基因表达的天然机制。它是一种强大的抗病毒策略，已被广泛用于保护宿主免受病毒感染。戊型肝炎（HE）是一种成人急性暴发性肝炎，在孕妇中死亡率特别高。目前，尚无针对戊型肝炎病毒（HEV）这种感染因子的有效疫苗或抗病毒治疗方法。非结构多聚蛋白区域拥有一种依赖RNA的RNA聚合酶（RdRp），它负责病毒RNA基因组的复制。因此，RdRp被认为是RNA干扰（RNAi）最具吸引力的候选靶点之一。在本研究中，通过实时定量PCR和蛋白质印迹分析评估了小干扰RNA（siRNA）的高效性和特异性。在A549细胞和仔猪中实现了对HEV感染的保护作用。在用shRNA-RdRp-1表达质粒预处理的仔猪中，接种HEV后，肝脏、脾脏和肾脏中的HEV抗原显著减少，丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和总胆红素（TBIL）的活性明显降低。这些结果表明，RNAi是一种针对HEV复制和感染的潜在有效抗病毒策略。
