原花青素A1抗猪流行性腹泻病毒作用及其对细胞自噬和线粒体功能的影响OA

To investigate the effects of porcine epidemic diarrhea virus(PEDV)infection on cellular autophagy,mitochondrial dynamics,and mitophagy,and to evaluate the antiviral activity of procyanidin A1,this study examined the autophagy-related proteins microtubule-associated protein 1A/1B light chain 3-Ⅱ(LC3-Ⅱ)and ubiquitin-binding protein p62(p62),mitochondrial dynamics-related proteins mitofusin 1(MFN1)and dynamin-related protein 1(DRP1),and mitophagy-related proteins PTEN-induced putative kinase 1(PINK1)and Parkin RBR E3 ubiquitin protein ligase(Parkin)after PEDV infection and procyanidin A1 treatment using cell viability assays,Western blot(WB),indirect immunofluorescence assay(IFA),and mCherry-GFP-LC3 dual-fluorescence plasmid transfection.The results showed that procyanidin A1 treatment at concentrations below 50 μg/mL had no significant effect on the viability of Vero cells.Compared with the Mock control group,PEDV infection significantly increased LC3-Ⅱ protein expression(P<0.001),decreased p62 protein expression(P<0.05 or P<0.001),and promoted autophagic flux.PEDV infection also significantly increased DRP1 protein expression(P<0.01 or P<0.001),decreased MFN1 protein expression(P<0.05 or P<0.001),and upregulated the expression of PINK1(P<0.05)and Parkin(P<0.01 or P<0.001).After treatment with 30 μg/mL procyanidin A1,viral expression was significantly reduced compared with the Mock control group(P<0.001).Compared with the PEDV infection group,p62 protein expression was restored(P<0.01),LC3-Ⅱ protein expression was significantly downregulated(P<0.01),which was consistent with the results of the mCherry-GFP-LC3 dual-fluorescence plasmid transfection assay.In addition,DRP1 protein expression was significantly decreased(P<0.001),and Parkin protein expression was significantly reduced(P<0.01).These results indicate that PEDV infection affects autophagy-related proteins,mitochondrial dynamics-related proteins,and mitophagy-related proteins,whereas procyanidin A1 treatment reverses these alterations.This study provides new insights into potential therapeutic targets for PEDV infection and the molecular mechanisms underlying the antiviral activity of procyanidin.