CAJ | 학술논문

目的探讨 A549细胞呼吸道合胞病毒(RSV)感染后Toll样受体3(TLR3)及肿瘤坏死因子-α(TNF-α) mRNA和蛋白表达及PPARγ激动剂15-脱氧前列腺素J2(15d-PGJ2)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:15d-PGJ2干预组(A)、罗格列酮干预组(B)、RSV组(C)、PDTC组(D)、GW9662组(E)及对照组(F).各组在培养12、24和48 h后收获上清液和细胞,实时荧光RT-PCR检测TLR3和TNF-α mRNA表达,Western Blot检测TLR3蛋白表达,ELISA法检测上清中TNF-α蛋白水平.结果与F组相比,各时间点C组TLR3和TNF-α mRNA及蛋白表达均明显升高(均P＜0.01);与C组相比,A组、B组和D组TLR3和TNF-α mRNA及蛋白表达均明显降低(均P＜0.01).15d-PGJ2和罗格列酮对TLR3和TNF-α mRNA和蛋白表达的抑制作用在12 h最明显;同一时间点A组和B组TLR3和TNF-α mRNA和蛋白的表达量均呈剂量依赖性下降.结论 RSV感染后TLR3和TNF-α mRNA及蛋白表达升高;罗格列酮和15d-PGJ2能以剂量依赖性方式抑制TLR3和TNF-α mRNA和蛋白表达.
목적 탐토 A549세포호흡도합포병독(RSV)감염후Toll양수체3(TLR3)급종류배사인자-α(TNF-α) mRNA화단백표체급PPARγ격동제15-탈양전렬선소J2(15d-PGJ2)화라격렬동적간예작용.방법 건립RSV감염적세포모형,장전대배양적세포수궤분성6조:15d-PGJ2간예조(A)、라격렬동간예조(B)、RSV조(C)、PDTC조(D)、GW9662조(E)급대조조(F).각조재배양12、24화48 h후수획상청액화세포,실시형광RT-PCR검측TLR3화TNF-α mRNA표체,Western Blot검측TLR3단백표체,ELISA법검측상청중TNF-α단백수평.결과 여F조상비,각시간점C조TLR3화TNF-α mRNA급단백표체균명현승고(균P＜0.01);여C조상비,A조、B조화D조TLR3화TNF-α mRNA급단백표체균명현강저(균P＜0.01).15d-PGJ2화라격렬동대TLR3화TNF-α mRNA화단백표체적억제작용재12 h최명현;동일시간점A조화B조TLR3화TNF-α mRNA화단백적표체량균정제량의뢰성하강.결론 RSV감염후TLR3화TNF-α mRNA급단백표체승고;라격렬동화15d-PGJ2능이제량의뢰성방식억제TLR3화TNF-α mRNA화단백표체.
Objective To study the change of toll-like receptor 3 (TLR3) and tumor necrosis factoralpha (TNF-α) both at the levels of protein and mRNA in human lung epithelial cells (A549) infected with respiratory syncytial virus (RSV) and the effects of peroxisome proliferator-activated receptor γ (PPARγ) agonists Rosiglitazone and 15-deoxy-delta12,14 prostaglandin J2 (15d-PGJ2).Methods RSV inoculation after A549 cells subcultured,then A549 cells were randomly divided into six groups:15d-PGJ2 + RSV group (group A),Rosiglitazone + RSV group (group B),RSV group (group C),PDTC + RSV group (group D),GW9662 + Rosiglitazone + RSV group (group E) and cell control group (group F).Cells were harvested after incubated for 12 h,24 h and 48 h respectively.The expression of TLR3 and TNF-α at mRNA was determined by real-time RT-PCR,and the protein level of TLR3 and TNF-α was determined by Western Blot and ELISA respectively.Results The expression of TLR3 and TNF-α at mRNA and protein at virous time in group C were significantly higher than those of group F (all P ＜ 0.01).While,the expression of TLR3 and TNF-α mRNA and protein in group A,B and D were significantly lower than than those of group C (all P ＜ 0.01).15d-PGJ2 and Rosiglitazone had the strongest effect to inhibit the expression of TLR3 and TNF-αmRNA and protein at 12 h and the expression of TLR3 and TNF-α mRNA and protein decreased as the increasing dose of 15d-PGJ2 and Rosiglitazone.Conclusion RSV infection induces increased expression of TLR3 and TNF-α mRNA and protein,which can be inhibited by Rosiglitazone and 15d-PGJ2 in a dose-dependent manner.