RSV 感染诱导的 RANTES、FKN、IP-10表达及 PPARγ激动剂抑制作用的体外研究  

In vitro study of expressions of RANTES, FKN and IP-10 induced by RSV infection and the inhibito-ry effects of PPARγagonists

作　　者：刘琳[1] 董琳[1] 徐月波[1] 陈兆兴[1] 樊节敏[1] 陈小芳[1] 

机构地区：[1]温州医科大学附属育英儿童医院呼吸科,325027

出　　处：《中华微生物学和免疫学杂志》2013年第9期659-665,共7页Chinese Journal of Microbiology and Immunology

基　　金：浙江省自然科学基金（5（2090932）;温州市科技计划对外合作交流基金（H20080054）

摘　　要：目的研究A549细胞呼吸道合胞病毒（ RSV）感染12 h、24 h、48 h后调节激活正常T细胞表达和分泌细胞因子（RANTES）、分形趋化因子（FKN）、干扰素诱导蛋白-10（IP-10） mRNA和蛋白水平的变化及不同剂量过氧化物酶体增殖物激活受体γ（PPARγ）激动剂15-脱氧前列腺素J2（15d-PGJ2）和罗格列酮及其拮抗剂（ GW9662）的干预作用。方法建立RSV感染A549细胞的体外模型，将传代培养的细胞随机分成5组：A组（15d-PGJ2＋RSV组），B组（罗格列酮＋RSV组）、C组（DMSO＋RSV组）、D组（ GW9662＋罗格列酮＋RSV组）、E组（细胞对照组）。各组分别在培养12 h、24 h、48 h收获细胞及上清液待测。应用ELISA检测各组上清液RANTES、FKN、IP-10蛋白水平，实时荧光定量RT-PCR检测各组RANTES、FKN、IP-10 mRNA表达。结果与细胞对照组相比， RSV感染组RAN-TES、FKN、IP-10 mRNA和蛋白的表达在12 h、24 h和48 h均明显升高（P均＜0．05），其中RANTES、FKN、IP-10 mRNA的表达量在24 h达高峰，48 h有所下降，与12 h的表达量比较差异无统计学意义（P均＞0．05）；而3种趋化因子蛋白的表达量均在48 h达高峰，与12 h、24 h比较差异有统计学意义（P均＜0．05）。在同一作用时间点上，随着15d-PGJ2和罗格列酮药物浓度的增加，RANTES、FKN、IP-10 mRNA和蛋白的表达量均呈剂量依赖性下降，与 RSV 感染组相比差异有统计学意义（ P 均＜0．05），其中以20μmol/L 15d-PGJ2和30μmol/L罗格列酮干预后RANTES、FKN和IP-10 mRNA和蛋白的表达量最低。结论 RSV感染可导致RANTES、FKN和IP-10 mRNA及蛋白表达升高，其中mRNA水平在感染后24 h达到高峰，蛋白的表达自感染后48 h达到高峰；而PPARγ激动剂能以剂量依赖性方式抑制上述趋化因子mRNA和蛋白的表达，从而起到减轻炎症的作用。Objective To observe the expressions of RANTES , FKN and IP-10 at mRNA and pro-tein levels in human lung epithelial cells （A549） infected by respiratory syncytial virus （RSV）, and to eval-uate the changes of them interfered with 15-deoxy-delta12,14prostaglandin J2（15d-PGJ2）, rosiglitazone or 2-chloro-5-nitrobenzanilide （ GW9662 ） .Methods A549 cells were seeded in 6-well culture plates and cul-tured overnight in F12K culture solution.Then they were randomly divided into five groups , including group A （15d-PGJ2＋RSV group）, group B （rosiglitazone＋RSV group）, group C （DMSO＋RSV group）, group D （GW9662＋rosiglitazone＋RSV group） and group E （cell control group）.Cells and supernatants were harves-ted from each group at different time points （12 h, 24 h and 48 h） of culture.The expressions of RANTES , FKN and IP-10 at mRNA and protein levels were measured by ELISA and real-time quantitative RT-PCR analysis.Results The expressions of RANTES , FKN and IP-10 at mRNA and protein levels in group C were significantly higher than those in group E at time points of 12 h, 24 h and 48 h （all P0.05）.Moreo-ver, the expressions at protein level were peaked at 48h, and had significant difference with those expressed at 12 h and 24 h （all P〈0.05）.Compared with group C, the expressions of the three chemokines both at mRNA level and protein level were decreased in group A and B as the dose was increased （all P〈0.05）,＆nbsp;and the lowest levels were observed with the intervention of 20 μmol/L of 15d-PGJ2 in group A and 30μmol/L of rosiglitazone in group B .Conclusion The expressions of RANTES , FKN and IP-10 at mRNA and protein levels were increased with RSV infection , and the peaks of mRNA level and protein level were respectively achieved at 24 h and 48 h after infection.PPARγagonists played an anti-inflammatory role through inhibiting the expressions of the three chemokines both at mRNA level and protein level in a dose -de-pendent manner .

关 键 词：PPARΓ激动剂 呼吸道合胞病毒 趋化因子 A549细胞 

分 类 号：R725.6[医药卫生—儿科]