为研究猪流行性腹泻病毒(PEDV)感染细胞后,细胞糖酵解的变化及其对PEDV复制的影响,本试验通过实时荧光定量PCR和Western blot检测PEDV感染猪小肠上皮细胞IPEC-J2 后36h糖酵解相关酶己糖激酶 2(HK2)的表达水平,然后通过添加糖酵解抑制剂2-脱氧-D-葡萄糖(2-DG)来进一步验证细胞糖酵解对PEDV复制的影响,最后检测了正常浓度葡萄糖(5.5 mmol/L)和高浓度葡萄糖(11.1 mmol/L 和22.2 mmol/L)培养条件下PEDV复制情况来探究葡萄糖浓度对PEDV复制的影响.结果表明,PEDV感染显著增加糖酵解限速酶HK2 的表达,PEDV感染可增强IPEC-J2 细胞的糖酵解,细胞糖酵解水平增强有利于PEDV的复制.在一定范围内提高葡萄糖浓度可促进PEDV复制.研究结果为初步阐明PEDV感染机制以及冠状病毒病的综合防控提供了理论依据.
This study was to investigate the changes in glucose metabolism in IPEC-J2 cells infected with porcine epidemic diarrhea virus(PEDV)and the effect of cell glycolysis on PEDV replication.The level of glycolysis related enzyme Hexokinase 2(HK2)in IPEC-J2 cells at 36 h post PEDV infection was detected by quantitative PCR and Western blot.Then,the effect of cell glycolysis on PEDV replication was further identified by adding glycolysis inhibitor 2-Deoxy-D-glucose(2-DG).Finally,the replication of PEDV under the conditions of nor-mal concentration of glucose(5.5 mmol/L)and high concentration of glucose(11.1 mmol/L and 22.2 mmol/L)conditions was detected to investigate the effect of glucose concentration on PEDV replication.The results showed that PEDV infection significantly increased the expres-sion of the glycolysis rate limiting enzyme HK2,which means that PEDV infection could enhance the glycolysis in IPEC-J2 cells.The en-hanced level of cell glycolysis was conducive to the replication of PEDV.Increasing glucose concentration within a certain range could pro-mote the replication of PEDV.This research provided a theoretical basis for preliminary elucidation of the infection mechanism of PEDV and comprehensive prevention and control of coronaviruses.