姜黄素抑制PEDV感染Vero细胞的作用研究

Inhibitory effect of Curcumin on PEDV-infected Vero cells

为探究姜黄素是否具有抑制猪流行性腹泻病毒(PEDV)感染的作用,本研究以Vero细胞为实验对象,分别设置阴性(空白)对照组、PEDV感染组、姜黄素处理组、姜黄素预处理后感染PEDV组,在PEDV感染后观察细胞病变,利用RT-qPCR、Western-blot、IFA技术和病毒滴度测定法检测PEDV在Vero细胞中的增殖情况。结果显示,与阴性对照组相比,PEDV感染Vero细胞后有明显细胞病变,即细胞拉丝、细胞核固缩并空泡化。与PEDV感染组相比,姜黄素预处理后显著抑制了病毒在细胞内的增殖(P<0.05),降低了病毒感染后的滴度(P<0.05),显著上调了Vero细胞抗病毒细胞因子IFN-α、IFN-β的分泌水平及MX1、ISG15、ZAP的转录水平(P<0.05)。与阴性(空白)对照组相比,PEDV感染后抗病毒细胞因子IFN-α、IFN-β的分泌水平明显上调(P<0.05),但MX1、ISG15、ZAP的转录水平无显著变化(P>0.05);姜黄素处理后除IFN-α分泌水平无明显变化外,IFN-β分泌水平及MX1、ISG15、ZAP转录水平均显著上调(P<0.05)。结果表明,姜黄素能够增强Vero细胞抗病毒细胞因子的抗病毒效果,抑制PEDV在Vero细胞中的增殖,这首次证实了姜黄素具有抑制PEDV感染Vero细胞的抗病毒作用,为防控PEDV有效抗病毒药物的研发提供了参考。

To investigate whether Curcumin has the effect of inhibiting porcine epidemic diarrhea virus(PEDV)infection,Vero cells were used in this study,and negative(blank)control group,PEDV-infected group,Curcumin-treated group,and Curcumin-pretreated group were set up for PEDV infection.The cytopathic lesions were observed after PEDV infection,and the proliferation of PEDV in Vero cells was detected using RT-qPCR,Western-blot,IFA technique and virus titer assay.The results showed that,compared with the negative control group,PEDV infected Vero cells had significant cytopathic effects,namely,cell drawing,karyopycnosis and vacuolation.Compared with the PEDV-infected group,infection with PEDV after Curcumin pretreatment,significantly inhibited the intracellular proliferation of the virus(P<0.05)and decreased the titer of the virus after infection(P<0.05),significantly upregulated the secretion levels of antiviral cytokines IFN-αand IFN-βand the transcription levels of MX1,ISG15 and ZAP in Vero cells(P<0.05).Compared with the negative(blank)control group,the secretion levels of antiviral cytokines IFN-αand IFN-βwere significantly upregulated after PEDV infection(P<0.05),but the transcription levels of MX1,ISG15,and ZAP were not significantly changed(P>0.05),except for the secretion level of IFN-α,which was not significantly changed after Curcumin treatment,the secretion levels of IFN-βand the transcription levels of MX1,ISG15 and ZAP transcript levels were significantly upregulated after Curcumin treatment(P<0.05).The study showed that Curcumin could enhance the antiviral effect of antiviral cytokines in Vero cells and inhibit the proliferation of PEDV in Vero cells,which for the first time confirmed the antiviral effect of Curcumin in inhibiting PEDV infection in Vero cells and provided a reference for the development of effective antiviral drugs for the prevention and control of PEDV.