Ye Hong, Zheng Yanfang, Ma Wanli, Ke Dan, Jin Xianrong, Liu Shengyuan, Wang Dixun
Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Pulmonary Laboratory of Ministry of Health of China, Wuhan 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2005;25(4):382-4, 407. doi: 10.1007/BF02828202.
Primary cell culture, techniques of gene transfection, gelatin zymography, and Western blot were used to investigate the effect of hypoxia on the secretion of MMP-2 and MMP-9 in pulmonary artery endothelial cells (PAEC) and smooth muscle cells (PASMC), and the role of HIF-1. Our results showed that (1) after exposure to hypoxia for 24 h, the protein content and activity of MMP-2 in the PAEC medium as well as these of MMP-2 and MMP-9 in PASMC medium (P < 0. 01) decreased significantly in contrast to those in normoxic group (P < 0.05); (2) after transfection of wild type EPO3'-enhancer, a HIF-1 decoy, the content and activity of MMP-2 and MMP-9 in hypoxic mediums became higher than those in normoxic group (P < 0.01), while transfection of mutant EPO3'-enhancer didn't affect the hypoxia-induced down-regulation. It is concluded that hypoxia could inhibit the secretion and activity of MMP-2 and MMP-9 in PAEC and PASMC, which could be mitigated by the transfection of EPO3'-enhancer and that HIF-1 pathway might contribute to hypoxia-induced down-regulation of MMP-2 and MMP-9.
采用原代细胞培养、基因转染技术、明胶酶谱法和蛋白质印迹法，研究缺氧对肺动脉内皮细胞（PAEC）和平滑肌细胞（PASMC）中基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）分泌的影响以及缺氧诱导因子-1（HIF-1）的作用。我们的结果表明：（1）缺氧暴露24小时后，与常氧组相比，PAEC培养基中MMP-2的蛋白含量和活性以及PASMC培养基中MMP-2和MMP-9的蛋白含量和活性均显著降低（P<0.01）；（2）转染野生型促红细胞生成素3'-增强子（一种HIF-1诱饵）后，缺氧培养基中MMP-2和MMP-9的含量和活性高于常氧组（P<0.01），而转染突变型促红细胞生成素3'-增强子对缺氧诱导的下调无影响。结论：缺氧可抑制PAEC和PASMC中MMP-2和MMP-9的分泌及活性，促红细胞生成素3'-增强子转染可减轻这种抑制作用，且HIF-1通路可能参与缺氧诱导的MMP-2和MMP-9下调。
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