PRV感染PK-15细胞后相关炎性通路及炎症因子表达变化  

Changes of inflammatory pathways and expression of inflammatory factors in PK-15 cells infected with PRV

作　　者：张王芝 舒相华 张莹 杨广佳 全伟[2] 储宗秀 宋春莲 Zhang Wangzhi;Shu Xianghua;Zhang Ying;Yang Guangjia;Quan Wei;Chu Zongxiu;Song Chunlian(College of Animal Medicine,Yunnan Agricultural University,Kunming Yunnan 650201;Yunnan Rural Science and Technology Service Center,Kunming Yunnan 650021;Kunming Jinhangchen Biotechnology Co.,Kunming Yunnan 650501)

机构地区：[1]云南农业大学动物医学院,云南昆明650201 [2]云南省农村科技服务中心,云南昆明650021 [3]昆明金航辰生物科技有限公司,云南昆明650501

出　　处：《现代畜牧兽医》2024年第2期9-14,共6页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：云南省农业重大科技专项“威信县生猪健康养殖和粪污利用关键技术创新与示范(202202AE090027)”。

摘　　要：试验旨在了解猪伪狂犬病毒(PRV)感染宿主后炎症动态变化,以猪肾细胞为宿主,选用炎性通路中Toll样受体4(TLR4)、骨髓分化一级反应蛋白88(MyD88)、核因子kB(NF-kB)和炎症因子干扰素-α(IFN-α)、干扰素-β(IFN-β)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)为研究指标。通过PRV感染PK-15细胞和BHK-21细胞,对比分析两者细胞感染量(TCID_(50))、细胞病变(CPE)和病毒载量;选取PRV较敏感细胞,设计炎性通路因子特异性引物,用实时荧光定量PCR检测不同时间点TLR4、MyD88和NF-kB表达量,ELISA测定IFN-α、IFN-β、TNF-α、IL-1β和IL-6炎症因子含量。结果显示,PK-15细胞、BHK-21细胞的TCID_(50)分别为10^(-8.98)/0.1 mL和10^(-8.58)/0.1 mL;在感染PRV第18 h均见典型CPE。PK-15细胞病毒载量较高,且18 h内PK-15的CPE程度与病毒载量呈正相关。与未感染的PK-15细胞相比,PRV感染PK-15细胞后TLR4、MyD88和NF-kB表达量均在感染后0~6 h时升高,感染后第12 h时极显著降低(P<0.01);除IFN-β外的炎症因子含量总体呈先下降后上升趋势。研究表明,PRV感染宿主过程中可引起TLR4-MyD88-NF-kB通路表达量升高,促进炎症因子变化抵抗病毒感染,TLR4-MyD88-NF-kB炎症通路在抗PRV感染过程中发挥重要作用。The purpose of the study was to investigate the dynamic changes of inflammation in PRV infected host.TLR4,MyD88,NF-kB in the inflammatory pathway and inflammatory factors IFN-α,IFN-β,TNF-α,IL-1β,and IL-6 were selected as the research indicators.PK-15 cells and BHK-21 cells were infected by PRV,and their cell infection dose(TCID_(50)),cytopathic effect(CPE)and viral load were compared and analyzed.The expression of TLR4,MyD88 and NF-kB in PRV sensitive cells was detected by real-time quantitative PCR,and the contents of IFN-α,IFN-β,TNF-α,IL-1β,and IL-6 were detected by ELISA.The results showed that the TCID_(50) of PK-15 cells and BHK-21 cells were 10^(-8.98)/0.1 mL and 10^(-8.58)/0.1 mL,respectively.Typical CPE was found at 18 h after PRV infection.The viral load of PK-15 cells was high,and the CPE degree of PK-15 cells was positively correlated with the viral load within 18 h.Compared with uninfected PK-15 cells,the expressions of TLR4,MyD88,and NF-kB in PK-15 cells infected with PRV increased from 0 to 6 h after infection,but decreased significantly at 12 h after infection(P<0.01).The concentrations of inflammatory factors except IFN β decreased first and then increased.Studies have shown that PRV infection can increase the expression of TLR4-MyD88-NF-kB pathway and promote the change of inflammatory factors to resist virus infection.TLR4-MyD88-NF-kB inflammatory pathway plays an important role in the process of anti-PRV infection.

关 键 词：PRV PK-15细胞 炎症通路 炎症因子 RT-QPCR 

分 类 号：S855.3[农业科学—临床兽医学]