26种中药抗PEDV感染IPEC-J2细胞的筛选效果分析  

Analysis of Screening Effect of 26 Kinds of Traditional Chinese Medicine against PEDV-infected IPEC-J2 Cells

作　　者：任涛 李俊贤 郭杨 文安林 张依裕[1] 欧德渊[1,2] REN Tao;LI Jun-xian;GUO Yang;Wen An-lin;ZHANG Yi-yu;OU De-yuan(College of Animal Science,Guizhou University,Guiyang,Guizhou,550025,China;Guizhou Provincial Key Laboratory of Animal Disease and Veterinary Public Health,Guiyang,Guizhou,550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病与兽医公共卫生重点实验室,贵州贵阳550025

出　　处：《动物医学进展》2025年第6期47-56,共10页Progress In Veterinary Medicine

基　　金：贵州省生猪产业高质量发展项目(黔财农[2022]183号,黔财农[2023]78号)。

摘　　要：由于猪流行性腹泻病毒(PEDV)具有较高的变异性并引起仔猪的高死亡率,且疫苗免疫对变异毒株的保护率低,西药的治疗成本高,因此,目前寻找抗PEDV中药已成为当今研究关键。通过TCID 50法检测病毒滴度,结果显示PEDV感染猪肠上皮细胞(IPEC-J2)随时间的增加病毒滴度逐渐升高;HE观察发现PEDV感染IPEC-J2细胞后发生皱缩,细胞间隙变大,拉丝及变圆的现象;CCK8法检测中药液在IPEC-J2细胞的最大安全浓度,以及通过计算病毒抑制率得出,在阻断PEDV作用上穿心莲水煎液(CXL)最佳,病毒抑制率高达57.8%;炒白术水煎液(CBZ)对病毒直接灭活效果最佳,病毒抑制率高达51.9%;党参水煎液(DS)的抑制效果最佳,病毒抑制率高达60.7%;设计不同剂量DS组(DS 1.25、2.5、5 mg/mL),进行RT-PCR检测DS对PEDV感染IPEC-J2细胞中PEDV N、IL-1β、IL-2、IL-6、TNF-αmRNA表达水平,发现DS对PEDV N基因呈浓度依赖性抑制,DS能极显著降低PEDV感染IPEC-J2细胞中PEDV N mRNA表达水平(P<0.01);与低、中剂量DS(DS 1.25、2.5 mg/mL)比,高剂量DS更能降低PEDV感染IPEC-J2细胞中的IL-1β、IL-2、IL-6、TNF-αmRNA表达水平;ELISA检测IL-1β、IL-2、IL-6、TNF-α蛋白含量,与低、中剂量DS比,高剂量DS更能降低PEDV感染IPEC-J2细胞后IL-1β、IL-2、IL-6、TNF-α蛋白含量。结果表明,DS对病毒抑制作用最佳,其中高剂量DS对PEDV复制有着显著的抑制作用,能缓解PEDV对IPEC-J2细胞的炎症反应。As the porcine epidemic diarrhea virus(PEDV)features high variability and causes a high mortality rate among piglets,the protection rate of vaccine immunity against variant strains is low,and the treatment cost of Western medicine is high.Currently,seeking Chinese herbal medicines against PEDV has become a crucial research point nowadays.Through the TCID 50 method to observe the viral titer,it was concluded that as the infection time of PEDV on intestinal porcine epithelial cell line-J2(IPEC-J2)increased,the viral titer gradually rose.HE observation revealed that after PEDV infected IPEC-J2 cells,shrinkage occurred,the intercellular space enlarged,and the phenomena of drawing silk and rounding up were observed.The CCK8 method was employed to detect the maximum safe concentration of the Chinese medicinal liquid in IPEC-J2 cells.Through calculating the virus inhibition rate,it was found that the water decoction of Andrographis paniculata(CXL)was the best in blocking the effect of PEDV,with a virus inhibition rate as high as 57.8%.The water decoction of roasted Atractylodes Rhizome(CBZ)had the best effect on directly inactivating the virus,with a virus inhibition rate as high as 51.9%.The water decoction of Codonopsis pilosula(DS)had the best inhibitory effect,with a virus inhibition rate as high as 60.7%.DS groups with different doses(DS 1.25,2.5 and 5 mg/mL)were designed to detect the mRNA expression levels of PEDV N,IL-1β,IL-2,IL-6 and TNF-αin PEDV-infected IPEC-J2 cells by RT-PCR.It was found that DS inhibited PEDV N gene in a concentration-dependent manner,and DS significantly decreased PEDV N mRNA expression level in PEDV-infected IPEC-J2 cells(P<0.01).Compared with low and medium doses of DS(DS 1.25,2.5 mg/mL),high dose of DS could significantly reduce the mRNA expression levels of IL-1β,IL-2,IL-6 and TNF-αin PEDV-infected IPEC-J2 cells.The contents of IL-1β,IL-2,IL-6 and TNF-αproteins were detected by ELISA.Compared with low and medium doses of DS,it was concluded that high-dose DS could better reduce

关 键 词：中药 猪流行性腹泻病毒 猪肠上皮细胞 病毒抑制率 抗病毒作用 炎症 

分 类 号：S852.659.6[农业科学—基础兽医学] S853.74[农业科学—兽医学]