两株H5N1亚型禽流感病毒NS1基因诱导细胞凋亡的研究

NS1 genes of two H5N1 avian influenza virus isolates induce aoptosis

为了研究H5N1亚型禽流感病毒NS1基因诱导的细胞凋亡作用,我们将两株H5N1亚型禽流感病毒A/Duck/Guangdong/40/2000(H5N1)(简称DKGD/40/00)和A/Duck/Guangxi/35/2001(H5N1)(简称DKGX/35/01)的NS1基因克隆到真核表达载体pIRES2-EGFP,转染Hela细胞后,应用荧光显微镜检测转染表达效率,应用TUNEL和流式细胞仪观察NS1基因产生的细胞凋亡作用。结果两株病毒NS1蛋白表达均能诱导Hela细胞凋亡,凋亡率无明显差异。表明单一的NS1并不能完全阐明凋亡的产生与病毒毒力强弱和病毒扩散之间的关系。

The objective of this study was to elucidate the mechanism of H5NI avian influenza virus-induced apoptosis. The NS 1 genes from two H5N 1 viruses A/Duck/Guangdong/40/2000 （HSN 1） （DKGD/40/00） and A/DucldGuangxi/35/2001 （H5N 1） （DKGX/35/01） were separately cloned into the eukaryotic expression vector pIRES2-EGFP, and transfected into Hela cells. The transfection efficiency of NSI genes were tested by fluorescent microscope, and the NSl-induced apoptosis was observed by TUNEL and flow cytomentry. The results demonstrated that the expression of NSI genes of the AIV isolates is sufficient to induce apoptosis in Hela cell, though there is no significant difference as to apoptosis rate between the two NSI genes. Our results implied that NSI alone may not clarify the interplay between NSl-induced apoptosis and viral titers or viral spread.