单月桂酸甘油酯对PEDV感染3D4/21巨噬细胞基因表达的影响

Effect of Glycerol Monolaurate on Gene Expression in PEDV-Infected 3D4/21 Macrophages

试验旨在研究单月桂酸甘油酯(ML)对猪流行性腹泻病毒(PEDV)感染3D4/21巨噬细胞基因表达的影响。通过ML对3D4/21细胞活力试验和PEDV在3D4/21细胞中的生长曲线确定最适浓度和最佳时间后,将3D4/21细胞随机分为3组(control组、PEDV组、PEDV+ML组),各组分别以全时添加10µmol/L ML和PEDV感染细胞后添加10µmol/L ML两种方式进行处理,并检测相关基因相对表达量。结果表明:添加10µmol/L的ML对3D4/21细胞生长具有显著的促进作用(P<0.05),病毒感染3D4/21细胞的48 h内PEDV-S、M、N基因相对表达量呈现先增加再降低的趋势,12 h时处于最高;与PEDV组相比,PEDV+ML组全时添加ML处理12 h使PEDV-S、M、N、IL-8、IFN-β、IFITM3基因相对表达量显著下调(P<0.05),处理24 h使PEDV-S、M、N、IL-8、IFN-β、MX1、ISG15、IFIT1和IFITM1基因相对表达量显著下调(P<0.05),KCNJ13基因相对表达量显著上调(P<0.05)。与PEDV组相比,PEDV+ML组在感染后添加ML处理12 h使MMP13基因相对表达量显著下调(P<0.05),PEDV-M基因相对表达量显著上调(P<0.05),处理24 h使IL-6、IL-8、IFN-β、MX1、ISG15、IFIT1、IFITM1、IFITM3基因相对表达量显著下调(P<0.05),PEDV-S、M、N、KCNJ13基因相对表达量显著上调(P<0.05)。综上所述,PEDV感染前ML预处理具有一定的抗病毒效果;PEDV感染使细胞处于免疫应激状态,ML有一定的缓解作用。

The experiment aims to investigate the effect of monoglyceride monolaurate(ML)on gene expression of 3D4/21 macrophages infected with porcine epidemic diarrhea virus(PEDV).After determining the optimal concentration and optimal time by ML on 3D4/21 cell viability experimental and PEDV growth curve in 3D4/21 cells,3D4/21 cells were randomly divided into three groups(control group,PEDV group,PEDV+ML group),and each group was treated with both full-time addition of 10μmol/L ML and 10μmol/L ML after PEDV-infected cells,respectively.And the expression of related genes were detected.The results showed that the addition of 10μmol/L ML significantly promoted the growth of 3D4/21 cells(P<0.05),and the relative expression of PEDV-S,M and N genes showed a tendency of increasing and then decreasing during 48 h of virus infection of 3D4/21 cells,and reached the highest at 12 h.Compared with the PEDV group,the relative expressions of PEDV-S,M,N,IL-8,IFN-β,and IFITM3 genes were significantly down-regulated in the full-time addition of ML treatment for 12 h in the PEDV+ML group(P<0.05);the relative expressions of PEDV-S,M,N,IL-8,IFN-β,MX1,ISG15,IFIT1,IFITM1 genes were significantly down-regulated(P<0.05)and the relative expression of KCNJ13 gene was significantly up regulated after treatment for 24 h(P<0.05).Compared with the PEDV group,the relative expression of MMP13 gene was significantly down-regulated(P<0.05),and the relative expression of PEDV-M gene was significantly up-regulated in the PEDV+ML group by adding ML for 12 h after infection(P<0.05);the relative expressions of IL-6,IL-8,IFNβ,MX1,ISG15,IFIT1,IFITM1 gene were significantly down-regulated(P<0.05)and the relative expressions of PEDV-S,M,N,KCNJ13 genes were significantly up-regulated after treatment for 24 h(P<0.05).In summary,ML pretreatment before PEDV infection has a certain antiviral effect;PEDV infection puts cells in an immune stress state,and ML has a certain alleviating effect.