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Effects of hypoxia,hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 in hepatic stellate cells

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World Journal of Gastroenterology 2001年第5期7卷 647-651页

作者： Ping-Sheng Chen~(1,2) Wei-Rong Zhai~1 Xiao-Mei Zhou~3 Jin-Sheng Zhang~1 Yue-E Zhang~1 Yu-Qin Ling~1 Ying-Hong Gu~1 1 Department of Pathology,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China2 Ping-Sheng Chen now works in the Department of Pathology,School of Basic Medical Sciences the (former Nanjing Railway Medical College),Southeast University,Nanjing 210009,China3 Institute of Cancer Research,Shanghai 200032,China Department of Pathology School of Basic Medical Sciences Fudan University 138 Yixueyuan Road Shanghai 200032 China.

AIM: To study the effects of hypoxia, hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 (MMP-2) in hepatic stellate cells (HSC). METHODS: The expressions of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC were detected by immunocytochemistry (ICC) and in situ hybridization (ISH). The contents of MMP-2 and TIMP-2 in culture supernatant were detected with ELISA and the activity of MMP-2 in supernatant was revealed by zymography. RESULTS: In the situation of hypoxia for 12h, the expression of MMP-2 protein was enhanced (hypoxia group positive indexes: 5.7 +/- 2.0, n=10; control: 3.2 +/- 1.0, n = 7; P<0.05), while TIMP-2 protein was decreased in HSC (hypoxia group positive indexes: 2.5 +/- 0.7, n = 10; control: 3.6 +/- 1.0, n = 7; P < 0.05), and the activity (total A) of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922, n = 9; control: 17.277 +/- 7.424, n = 11; P < 0.01). Compared the varied duration of hypoxia, the changes of expressions including mRNA and protein level as well as activity of MMP-2 were most notable in 6h group. The highest value(A(hypoxia)-A(control)) of the protein and the most intense signal of mRNA were in the period of hypoxia for 6h, along with the lowest activity of MMP-2. In the situation of hyperoxia for 12h, the contents (A(450)) of MMP-2 and TIMP-2 in supernatant were both higher than those in the control, especially the TIMP-2 (hyperoxia group: 0.0499 +/- 0.0144, n = 16; control: 0.0219 +/- 0.0098, n = 14; P < 0.01), and so was the activity of MMP-2 (hyperoxia group: 5.252 +/- 0.771, n = 14; control: 4.304 +/- 1.083, n = 12; P < 0.05), and the expression of MT1-MMP was increased. CONCLUSION: HSC is sensitive to the oxygen, hypoxia enhances the expression of MMP-2 and the effect is more marked at the early stage; hyperoxia mainly raises the activity of MMP-2.

关键词： Animals Cell Division Cell Hypoxia Cells, Cultured Gelatinase A Gene Expression Regulation, Enzymologic Hepatocytes Hyperoxia metalloendopeptidases RNA, Messenger Rats Rats, Sprague-Dawley Research Support, Non-U.S. Gov't Tissue Inhibitor of Metalloproteinase-2

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