矽肺模型大鼠肺间质成纤维细胞MMP_药学论文.docVIP

矽肺模型大鼠肺间质成纤维细胞MMP_药学论文 矽肺模型大鼠肺间质成纤维细胞MMP_药学论文 作者：王海涛，刘昊，颜京霞，田艳霞，江小华，高俊玲 【关键词】 肺间质纤维化 Expression of matrix metalloproteinase2 and 9 in pulmonary fibroblasts of silicotic fibrosis rat model 　　【Abstract】 AIM: To study the expression of MMP2 and MMP9 in fibroblasts of rat silicotic fibrosis model and its significance. METHODS: Fifty pathogenfree Wistar rats were randomly divided into experimental group and control group. Experimental group was instillated with SiO2, while control group with normal saline. Pulmonary interstitial fibroblasts were isolated on day 1, 3, 7, 14 and 28 after SiO2 exposure. The protein and mRNA expression of MMP2 was evaluated by immunocytochemistry and RTPCR. RESULTS: ①The MMP2 protein expression in fibroblasts increased on day 1 after SiO2 exposure and reached 3.887 times（t=23.667，0.001） that of control group on day 7. The expression of MMP9 protein also increased on day 1 after SiO2 exposure and reached 3.103 times that of control group（t=11.399, 0.001）on day 7. ② The mRNA expression of MMP2 and MMP9 was enhanced in fibroblasts at 24 h after SiO2 instillation. CONCLUSION: The expression of MMP2 and MMP9 in fibroblasts is upregulated in silicotic fibrosis model. 　　【Keywords】 matrix metalloproteinase; fibroblast cells; pulmonary fibrosis; silicotic fibrosis 　　【摘要】 目的:探讨肺间质成纤维细胞MMP2和MMP9在SiO2致肺纤维化中的表达及意义. 方法: 清洁级Wistar大鼠50只，随机分为对照组和实验组，实验组气管内注射SiO2悬液，对照组代以等剂量生理盐水. 注射后1，3，7，14和28 d分离和提取肺间质成纤维细胞，应用免疫细胞化学和逆转录多聚酶链式反应（RTPCR）方法，观察其MMP2，MMP9蛋白及其mRNA的动态变化. 结果:①SiO2作用后1 d成纤维细胞MMP2表达增高，7 d达高峰，为对照组的3.887倍（t=23.667，0.001）；MMP9在SiO2作用后1 d表达增高，7 d达对照组的3.103倍（t=11.399，0.001）；②MMP2 mRNA和MMP9 mRNA的转录从SiO2作用后1 d就开始升高. 结论:受SiO2刺激后，肺间质成纤维细胞上调MMP2和MMP9的表达，损伤肺泡基底膜，启动肺纤维化的发生. 　　【关键词】 基质金属蛋白酶；肺间质成纤维细胞；肺间质纤维化；矽肺 　 　　0引言 　　基质金属蛋白酶及其组织抑制物（matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase, MMP/TIMP）是肺纤维化效应细胞分泌的生物活性分子，在细胞外基质(extracellular matrix, ECM)的合成和降解中具有重要意义. 在MMP家族中，MMP2和MMP9的特异性底物主要是Ⅳ型胶原，而后者是肺泡壁基底膜的重要成分. 肺泡上皮细胞和基底膜的损伤是肺间质纤维化过程的关键事件［1］. 我们在S