罗格列酮拮抗博莱霉素致大鼠肺纤维化的实验研究

摘要

Objective To explore the role of rosiglitazone in inhibiting pulmonary fibrosis and its mechanism. Methods 24 rats were divided into physiological saline control group, Rosiglitazone control group,bleomycin group and rosiglitazone interference group, bleomycin was adopted to induce pulmonary fibrosis in rats. Hematoxylin and eosin (HK) staining and Masson staining were used to perform pathological examine. Kit was used to measure hydroxyproline content of pulmonary tissue. Western blot and realtime fluorescent quantitative polymerase chain reaction ( RT-PCR) were employed to probe expression of peroxisome proliferator-activated receptor 7 (PPARy) ,matrix metalloproteinase 9 (MMP-9) and transforming growth factor β1 (TGF-β1 ). Enzyme-linked immunosorbent assay (ELISA) was applied to detect the concentration of TGF-β1 in bronchoalveolar lavage fluid (BALF). Results After intratracheal infusion of bleomycin, Ashcroft score,collagen deposition and hydroxyproline content of pulmonary tissue,level of TGF-β1 in BALF,and expression of TGF-β1 ,PPARγ,MMP-9 in pulmonary tissue were increased compared to those in physiological saline group. However,the indicators above were decreased after rosiglitazone interference,except PPARy expression showing further increased. Levels of PPARy protein expression had negative correlation to levels of TGF-β1 and MMP-9 Mrna expression. Conclusion Rosiglitazone shows antagonism against bleomycin-induced pulmonary fibrosis which mechanism probably related to up-regulation of PPARy protein expression and inhibition of TGF-β1 and MMP-9 Mrna transcription.%目的 探讨罗格列酮抑制肺纤维化的作用及其机制.方法 将24只大鼠分为生理盐水对照组、罗格列酮对照组、博莱霉素组、罗格列酮干预组,予博莱霉素诱导大鼠肺纤维化.HE染色和Masson染色行病理学检查;试剂盒检测肺组织中羟脯氨酸含量;应用 Westen blot及荧光定量实时逆转录聚合酶链反应(RT-PCR)检测过氧化物酶体增殖物激活受体γ(PPARγ)、基质金属蛋白酶9(MMP-9)和转化生长因子β1(TGF-β1)的表达;采用酶联免疫吸附法(ELISA)检测支气管肺泡灌洗液(BALF)中TGF-β1浓度.结果 博莱霉素气管内注入后,Ashcroft评分,肺组织胶原沉积、羟脯氨酸含量,BALF中TGF-β1水平及肺组织中TGF-β1、PPARγ、MMP-9表达较生理盐水对照组增加;给予罗格列酮干预后,除PPARγ表达进一步增加外,上述指标均下降.博莱霉素组PPARγ蛋白表达水平与TGF-β1 mRNA、MMP-9 mRNA表达水平分别呈负相关.结论 罗格列酮对博莱霉素诱导的肺纤维化进程有拮抗作用,其具体机制可能与上调PPARγ蛋白表达,抑制TGFβ1、MMP-9 mRNA转录有关.