呼吸道合胞病毒诱导的NF-κB信号通路激活及PPARγ激动剂的抑制作用  

Study on activation of respiratory syncytial virus induced by NF-κB signaling pathway in human lung epithelial cells and the inhibitory effect of PPARγ agonists

作　　者：董琳[1] 张同强[1] 王志远[1] 万春杰[1] 徐月波[1] 刘琳[1] 陈小芳[1] 赵伟 

机构地区：[1]温州医科大学附属育英儿童医院呼吸科,浙江温州325027 [2]美国弗吉尼亚联邦大学医学院变态反应与免疫科,美国里士满23298

出　　处：《温州医学院学报》2013年第8期496-500,共5页Journal of Wenzhou Medical College

基　　金：浙江省自然科学基金资助项目(Y2090932);温州市科技计划对外合作交流基金资助项目(H20080054)

摘　　要：目的:研究呼吸道合胞病毒(RSV)感染后A549细胞NF-κB信号通路中NF-κB复合物抑制因子(IκB)α、IκB激酶(IKK)β、p65、p50 mRNA和蛋白表达的变化及PPARγ激动剂15脱氧前列腺素J2(15dPGJ2)和罗格列酮的干预作用。方法:建立RSV感染的细胞模型,将传代培养的细胞随机分成6组:15dPGJ2干预组(A组)、罗格列酮干预组(B组)、RSV感染模型组(C组)、PDTC组(D组)、GW9662组(E组)及对照组(F组)。各组在培养12 h、24 h和48 h后收获细胞,分别应用Western blot法和实时荧光定量PCR技术法检测IκBα、IKKβ、p65、p50蛋白和mRNA表达。结果:与F组相比,C组在12 h、24 h和48 h IKKβ、p65和p50蛋白及mRNA表达均明显升高,IκBα蛋白及mRNA表达明显降低(均P<0.01),其中IKKβ、p65和p50 mRNA在24 h达高峰,IκBαmRNA在24 h最低,与12 h、48 h相比差异有统计学意义(P<0.05或P<0.01),而IKKβ、p65、p50蛋白在48 h达高峰,IκBα蛋白在48 h最低,与12 h、24 h相比差异有统计学意义(P<0.05或P<0.01)。与C组相比,A组、B组和D组IKKβ、p65和p50蛋白及mRNA表达均明显降低,而IκBα蛋白及mRNA表达均明显升高(P<0.05或P<0.01),其中A组、B组IKKβ、p65和p50蛋白及mRNA在12 h最低,IκBα蛋白及mRNA在12 h最高,与24 h及48 h相比差异有统计学意义(P<0.05或P<0.01)。结论:RSV感染可导致NF-κB信号通路激活;15d-PGJ2和罗格列酮通过抑制NF-κB信号通路激活发挥抗炎作用,其作用在干预后12 h最强。Objective： To observe changes of the expression of Iκ B kinase β （IKKβ）,IκBα and NF-κB p65, NF- K B p50 which included in nuclear factor- n B （NF- K B） signaling pathway both at the levels of protein and mRNA in human lung epithelial cells （A549） infected with respiratory syncytial virus （RSV）, the changes differ- ence of intervention by 15-deoxy-deltal 2, 14 prostaglandin J2 （15d-PGJ2） and Rosiglitazone were observed, the inhibition effect to the activation of NF- K B signaling pathway in PPAR~ agonists was explored. Methods： RSV inoculation after A549 cells subculture＆ then A549 cells were randomly divided into six groups： group A （L5d- PGJ2＋RSV group）, group B （Rosiglitazone＋RSV group）, group C （RSV group）, group D （PDTC＋RSV group）, group E （GW9662＋Rosiglitazone＋RSV group） and group F （cell control group）. Cells were harvested after incubated for 12 h,24 h and 48 h and the expression of IκBα, IKBcc, P65 and P50 at protein and mRNA levels in the groups were determined by Western blotting and real-time RT-PCR. Results： The expression of IκBα, P65 and P50 at protein and mRNA levels in Group C were significantly higher than those in group F, and with peak at 48 h and 24 h, respectively （all P〈0.01）. Concomitantly, a decreased cellular amount of IKB~ protein and mRNA was observed in group C when compared with the group F at each time, and with bottom at 48 h and 24 h, respectively （P〈0.05 or P〈0.01）. The expression of IKKB, P65 and P50 at protein and mRNA levels in group A, B and D were significantly lower than group C （P〈0.05 or P〈0.01）. While the production of IκBα at protein and mRNA levels was significantly increased when compared with group C （P〈0.05 or P〈0.01）. Compared with 24 h and 48 h, the strongest effect to inhibit the expression ofIKK ~3, P65 and PS0 protein and mRNA was at 12 h （P 〈0.05 or P〈0.01） and the expression of I K B a at protein and mRNA levels was the highest at 12 h （P〈0.05

关 键 词：呼吸道合胞病毒 核转录因子-ΚB PPARΓ激动剂 

分 类 号：R725.6[医药卫生—儿科]