姜黄素抑制轮状病毒感染猪肠上皮细胞的作用研究

Inhibitory Effect of Curcumin against Rotavirus Infection in Intestinal Porcine Epithelial Cells

本研究旨在探讨姜黄素对猪轮状病毒(PRV)感染猪肠上皮细胞(IPEC-J2细胞)的抗病毒作用。以IPEC-J2细胞为试验对象,分别设置阴性对照组、感染PRV(感染复数=0.1)组和感染PRV后姜黄素(20μmol/L)处理组。在感染PRV后观察细胞病变,利用流式细胞术检测细胞内活性氧(ROS)含量,并通过实时荧光定量PCR(qRT-PCR)技术和病毒滴度测定法检测PRV在IPEC-J2细胞内的复制与增殖。结果表明:与阴性对照组相比,1)PRV感染导致IPEC-J2细胞病变,显著降低细胞活力(P<0.05),极显著上调细胞内ROS含量(P<0.01);2)姜黄素可显著抑制PRV在细胞内的复制与增殖(P<0.05);3)在PRV吸附细胞阶段添加姜黄素显著抑制了病毒的复制与增殖(P<0.05);4)姜黄素在感染前与PRV直接孵育能显著降低感染后病毒的滴度(P<0.05);5)PRV感染显著提高了细胞内黑色素瘤分化相关基因5、干扰素诱导蛋白44样蛋白抗体和干扰素β的mRNA相对表达量(P<0.05),显著降低了细胞内Toll样受体适配器分子1、线粒体抗病毒信号蛋白和髓样分化因子88的mRNA相对表达量(P<0.05),而添加姜黄素能显著降低或提高这些受病毒影响的免疫相关因子的表达(P<0.05)。综上所述,本研究证实姜黄素可抑制PRV对IPEC-J2细胞的感染与复制,可为预防和治疗仔猪病毒性腹泻提供重要的参考。

This study was explored the antiviral effect of curcumin on porcine rotavirus(PRV)-infected intestinal porcine epithelial cells(IPEC-J2 cells). IPEC-J2 cells were used as the experimental object and were assigned to the negative control group,the PRV infection(multiplicity of infection = 0.1)group,and the PRV infection with curcumin treatment(20μmol/L)group,respectively. After PRV infection,the cytopathic changes were observed,and the content of reactive oxygen species in IPEC-J2 cells was detected by flow cytometry,and the replication and proliferation of PRV in IPEC-J2 cells were detected by real-time fluorescence quantification(qRT-PCR)and virus titer assay. The results showed as follows:compared with a negative control group,1)PRV infection caused lesions of IPEC-J2 cells,significantly reduced cell viability(P<0.05),and significantly upregulated the content of intracellular ROS(P<0.01);2)curcumin could inhibit the replication and proliferation of PRV in IPEC-J2 cells(P<0.05);3)adding curcumin at the stage of PRV adsorption cells significantly inhibited the replication and proliferation of the virus(P<0.05);4)direct incubation of curcumin with PRV before infection could significantly reduce the titer of the virus after infection(P<0.05);5)PRV infection significantly increased the mRNA relative expression levels of melanoma differentiation related gene 5,interferon-induced protein 44-like protein antibody and interferon-β (P<0.05),and significantly reduced the mRNA relative expression levels of Toll-like receptor adapter molecule 1,mitochondrial antiviral signaling protein and myeloid differentiation factor 88 in IPEC-J2 cells(P<0.05),however,adding curcumin could significantly reduce or increase the expression of these immune gene affected by the virus(P<0.05). In summary,this study confirmed for the first time that curcumin can inhibit PRV infection and replication in intestinal porcine epithelial cells,providing important approach for prevention and treatment of piglet viral diarrhea.[Chinese Journal of Animal Nutrition,2022,34(2):1285-1295]