论文部分内容阅读
目的 研究基质金属蛋白酶 2 (MMP2 )在肺纤维化中的作用。方法 (1)用免疫组织化学方法 (ABC法 )观察博莱霉素所诱导的实验性大鼠纤维化肺组织内MMP2 表达的动态变化 ;(2 )用逆转录 聚合酶链反应 (RT PCR)观察博莱霉素所诱导的肺纤维化不同时期间质成纤维细胞MMP2 mRNA的动态变化。 (3)用Northern印迹杂交和免疫细胞化学方法观察转化生长因子 β1(TGFβ1)对体外培养的大鼠肺成纤维细胞MMP2 mRNA和蛋白表达的影响。结果 (1)诱导纤维化组 1～ 7d ,病灶内浸润的单核巨噬细胞及肺泡间质细胞数量增多 ,MMP2 免疫组织化学染色阳性。 14d以后 ,阳性单核巨噬细胞减少。用博莱霉素 2 8d时 ,阳性间质细胞亦减少。 (2 )博莱霉素作用后 1d ,肺成纤维细胞MMP2基因转录即明显增强 ,为对照组的 2 0 5倍 ,2 8d时下降。 (3)TGFβ1作用后 ,大鼠肺成纤维细胞MMP2mRNA及其蛋白表达增强。结论 肺组织内MMP2 的过度表达 ,可能是肺纤维化启动的重要原因。
Objective To investigate the role of matrix metalloproteinase 2 (MMP2) in pulmonary fibrosis. Methods (1) The dynamic changes of MMP2 expression in the experimental rat fibrosis lung tissue induced by bleomycin were observed by immunohistochemical method (ABC method); (2) By reverse transcription polymerase chain reaction (RT PCR) To observe the dynamic changes of MMP2 mRNA in mesenchymal fibroblasts induced by bleomycin at different stages of pulmonary fibrosis. (3) The effect of transforming growth factor-β1 (TGFβ1) on the expression of MMP2 mRNA and protein in rat lung fibroblasts was observed by Northern blotting and immunocytochemistry. Results (1) The numbers of infiltrating mononuclear macrophages and alveolar interstitial cells in the lesion increased from 1 to 7 days after induction of fibrosis, and MMP2 immunohistochemical staining was positive. After 14d, positive mononuclear macrophages decreased. With bleomycin 2 8d, positive stromal cells also decreased. (2) 1 d after bleomycin treatment, the transcription of MMP2 gene in lung fibroblasts was significantly enhanced, which was 25 times of the control group and decreased on the 28th day. (3) After treated with TGFβ1, the expression of MMP2mRNA and its protein in rat lung fibroblasts increased. Conclusion Overexpression of MMP2 in lung tissue may be an important reason for pulmonary fibrosis.