甲型H1N1流感病毒致宿主细胞氧化与凋亡之间的关系研究

Study on the relationship between oxidative damage and apoptosis of MDCK cells infected by influenza A virus H1N1

目的:观察甲型H1N1流感病毒诱导的犬肺(MDCK)细胞氧化与凋亡之间的关系。方法:MDCK细胞培养后,甲型H1N1流感病毒感染0h、1h、3h、6h、12h、24h、48h,在各时间点进行检测:采用流式细胞仪Annexin V-FITC/PI双染料法检测细胞的凋亡率;丙二醛(MDA)法观察膜的氧化损伤程度。结果:(1)甲型H1N1流感病毒感染后0、1、3、6h组细胞凋亡率与正常对照组无显著性差异(P>0.05),感染后12、24、48h组的凋亡率显著高于正常对照组(P<0.01);(2)实验各组MDA含量随着感染时间的延长呈下降趋势,0、1、3、6、12h时MDA含量显著高于正常对照组(P<0.01),而24、48h时与正常对照组无显著性差异(P>0.05);(3)MDA含量与凋亡率呈高度的负相关,回归分析进一步证明二者间的相关关系。结论:甲型H1N1流感病毒感染对细胞造成的氧化损伤是极早期事件,可能是细胞凋亡的重要因素。

Objective：To investigate the relationship between apoptosis and oxidative damage of the Madin-Darby canine kidney （MDCK）cell infected by influenza A virus.Metbods：MDCK cells were cultured and infected with H1N1.The apoptosis and the oxidative damage of the host cells were detected by Annexin V-FITC/ PI double staining with flow cytometry and malondialdehyde （MDA） detection respectively after being infected for 0,1,3,6,12,24 and 48 hours.Results： （1）There were no significant differences between the experimental groups＇apoptosis rates and the control groups＇for the first 6 hours,but the 12,24 and 48h groups had significant high apoptosis rates than the corresponding control ones （P〈0.01）.（2）MDA test showed that its amount was the highest when the cells were infected,and was declining with the time going to the normal level at 24h after being infected. Statistically significant differences could be observed between the experimental groups and the corresponding control ones at 0,1,3,6 and 12h （P〈0.01）. （3 ）Extremely high negative correlation was found between the MDA amount and the apoptosis rate（spearman correlation coefficient.Conclusion：The oxidative damage caused by the H1N1 infection to the cells is the extremely early event,and may be one of the most important reasons leading to apoptosis.