氧化应激对PRRSV体外感染PAMs后诱导TLR3/NF-kB信号通路相关蛋白的影响  

Effects of PRRSV infection in vitro on related proteins of TLR3/NF-kB signaling pathway in PAMs treated by oxidative stress

作　　者：闫晓霞 谢丽君[1] 刘欢欢[1] 臧雅婷[1] 高洪[1] 严玉霖[1] 

机构地区：[1]云南农业大学动物科技学院,云南昆职650201

出　　处：《中国兽医学报》2016年第11期1916-1922,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31160496,31360599);云南省高校动物性食品安全与人兽共患病科技创新团队项目(云教科[2011]14号);云南农业大学2015年研究生科技创新项目(2015ykc4)

摘　　要：为探讨氧化应激对猪繁殖与呼吸征病毒(PRRSV)体外感染猪肺泡巨噬细胞(PAMs)后诱导TLR3/NF-kB信号转导中相关蛋白的影响,从PRRSV阴性的健康仔猪肺脏中分离和培养PAMs,随后分为正常对照组、PRRSV各试验组、NAC+PRRSV组以及H_2O_2+PRRSV组,于感染后6、12、24、48、72h收集细胞,Western blot检测各组不同时间段PAMs中TLR3、TRIF、TRAF6以及NF-kB蛋白表达水平的动态变化。结果显示,在PRRSV各试验组,TLR3、TRIF、TRAF6及NF-κB蛋白水平表达与正常对照组相比均升高(P<0.05),而且其表达量与感染之间有时间相关性;在NAC+PRRSV组中,TLR3、TRIF、TRAF6及NF-κB的蛋白表达虽有升高,但表达量都比正常接毒组明显下降;在H_2O_2+PRRSV组中,TLR3、TRIF、TRAF6及NF-κB的蛋白表达比正常接毒组表达量明显升高(P<0.05)。结果表明,抗氧化剂和促氧化剂可引起PAMs氧化应激状态的改变,从而导致PRRSV感染PAMs后TLR3/NF-κB信号传导通路的相关蛋白水平发生了变化。To investigate the effects of PRRSV infection in vitro on related proteins of TLR3/NF-κB signaling pathway in PAMs treated by oxidative stress,the PAMs,which were isolated from a lung of PRRS negative healthy piglets and cultured in cell culture plates, were randomly divided into control group, PRRSV-infected group, NAC ＋ PRRSV group and H202 ＋ PRRSV. These PAMs were collected at 6,12,24,48,72 h post infection. The dynamic changes of TLR3,TRIF, TRAF6 and NF-κB protein at different time points were detected by Western blot. The results showed that the expression levels of TLR3,TRIF,TRAF6 and NF-κB proteins were significantly up-regulated by PRRSV infection at different time points （P〈0. 05） ,with a time-dependent maner; the expression levels of TLR3,TRIF, TRAF6 and NF-κB proteins were significantly up-regulated in NAC＋PRRSV group,but were significantly lower control group ;their expression levels in H2O2＋PRRSV group were increased significantly compared to the control group（P〈0.05）. It could concluded that the antioxidant and oxidant could cause the change of oxidative stress in PAMs,so that PRRSV infection results in related protein levels of TLR3/NF-κB signaling pathway.

关 键 词：氧化应激 PRRSV PAMs TLR3/NF-κB信号通路 相关蛋白 

分 类 号：S852.65[农业科学—基础兽医学]