猪流行性腹泻病毒结构蛋白对细胞极性分子Par6表达的影响OA

Tight junctions(TJs)are the major junctional structures among intestinal epithelial cells,which can resist the invasion of external pathogens and selectively regulate the entry of small molecules and ions into the body.In intestinal epithelial cells,polarity complexes can interact with tight junctions to maintain cell polarity.To investigate the effects of porcine epidemic diarrhea virus(PEDV)structural proteins on the expression of the cell polarity molecule Par6,eukaryotic expression plasmids carrying PEDV N,S1,S2,M and E genes were constructed.Using Lipofectamine 3000 transfection reagent,time-course and dose-response groups were set up for transfection into porcine jejunum epithelial cell line(IPEC-J2).Specifically,in the time-course group,cells were transfected with a fixed dose of 5 μL and samples were collected at 24,36 and 48 hours post-transfection;in the dose-response group,cells were transfected with 1,3 and 5 μL of the reagent and samples were collected at 24 hours post-transfection.The expression levels of Par6 mRNA and protein in the cells were then detected.The results showed that transfection with eukaryotic expression plasmids carrying PEDV N,S1,S2,M and E proteins all up-regulated the expression of Par6 mRNA and protein in the cells,with the N protein exerting the most significant up-regulatory effect.This study provides a reference for revealing the pathogenic mechanism of PEDV.