糖代谢在肠道病毒EV71复制中的作用研究  

The role of glucose metabolism in the replication of enterovirus EV71

作　　者：杨秀文 陆剑云[2] 卢紫欣 陈嘉仪 郭彦彤 梁羽芃 王佳颐 朱利 张宝 YANG Xiu-wen;LU Jian-yun;LU Zi-xin;CHEN Jia-yi;GUO Yan-tong;LIANG Yu-peng;WANG Jia-yi;ZHU Li;ZHANG Bao(Biosafety Level 3 Laboratory,School of Health,Southern Medical University,Guangzhou 510515,China;Guangzhou Center for Disease Control and Prevention)

机构地区：[1]南方医科大学公共卫生学院三级生物安全实验室,广东广州510515 [2]广东省广州市疾病预防控制中心

出　　处：《中国病原生物学杂志》2022年第2期143-148,共6页Journal of Pathogen Biology

基　　金：广东省自然科学基金项目(No.2018A030313767)。

摘　　要：目的研究细胞内糖代谢对EV71病毒复制的影响。方法用MOI=100的EV71感染HUVECs,观察HUVECs在4h、8h和12h时存活率和EV71复制水平变化,确定EV71复制峰值时间点。用不含糖的DMEM培养基培养感染EV71的HUVECs 4h、8h和12h,检测EV71的RNA和蛋白水平改变。在EV71复制峰值时间点使用糖代谢抑制剂KAN0438757干扰糖代谢途径,检测EV71复制的改变;采用靶向代谢组学技术检测EV71感染或KAN0438757处理HUVEC后糖代谢关键代谢产物丙酮酸的含量变化。结果用MOI=100的EV71感染HUVECs 8h和12h后,EV71mRNA分别为4h的123.8倍和298.9倍(均P<0.05);在感染后4h、8h、12h细胞存活率为对照组的81%、66%和48%(均P<0.05);感染后12h,细胞内的丙酮酸含量明显降低(P<0.05)。糖代谢抑制剂KAN0438757处理HUVECs后丙酮酸含量降低(P<0.05),糖代谢水平受到抑制,EV71复制受到抑制(P<0.05)。感染EV71的HUVECs使用不含糖的DMEM培养基培养4h、8h和12h,相同时间点的EV71复制水平低于含糖细胞培养组(P<0.05)。结论EV71病毒感染HUVECs后利用HUVECs内糖代谢提供的能量促进自身的复制,抑制HUVECs内糖代谢水平可抑制EV71的复制。Objective Effect of intracellular glucose metabolism on EV71virus replication.Methods HUVECs were infected with EV71with MOI=100.The changes of survival rate and EV71replication level of HUVECs at 4h,8h and 12hwere observed to determine the peak time point of EV71replication.HUVECs infected with EV71were cultured in sugar free DMEM medium for 4h,8hand 12h,and the changes of RNA and protein levels of EV71were detected.At the peak time point of EV71replication,the glucose metabolism inhibitor KAN0438757was used to interfere with the glucose metabolism pathway and detect the changes of EV71replication;The changes of pyruvate,a key metabolite of glucose metabolism,after EV71infection or KAN0438757treatment of HUVEC were detected by targeted metabolomics.Results After HUVECs were infected with EV71with MOI=100for 8hand 12h,EV71mRNA was 123.8times and 298.9times higher than that at 4h(all P<0.05);The cell survival rates at 4h,8hand 12hafter infection were 81%,66%and 48%of those in the control group(all P<0.05);The level of pyruvate in cells decreased significantly at 12hafter EV71infection(P<0.05).The level of pyruvate decreased(P<0.05),the level of glucose metabolism was inhibited and the replication of EV71was inhibited(P<0.05)after treatment of HUVECs with KAN0438757.HUVECs infected with EV71were cultured in sugar free DMEM medium for 4h,8hand 12h.The replication level of EV71at the same time point was lower than that in sugar containing cell culture group(P<0.05).Conclusion After EV71virus infects HUVECs,it uses the energy provided by glucose metabolism in HUVECs to promote its own replication.Inhibiting the level of glucose metabolism in HUVECs can inhibit EV71replication.

关 键 词：糖代谢 EV71 丙酮酸 

分 类 号：R373.2[医药卫生—病原生物学]