Chen Pingsheng, Zhai Weirong, Zhang Yue'e, Zhou Xiaomei, Zhang Jinsheng, Ling Yuqin, Gu Yinghong
Department of Pathology, Medical College, Fudan University, Shanghai, 200032, China.
Zhonghua Bing Li Xue Za Zhi. 2002 Aug;31(4):337-41.
To study the effects of hypoxia and hyperoxia on the expression and activity regulation of matrix metalloproteinase-2 (MMP-2) of the hepatic stellate cell (HSC).
The expression of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC under hypoxic or hyperoxic conditions were detected with immunocytochemistry (LSAB method), the contents of MMP-2, TIMP-2 in culture supernatant with ELISA and the activity of MMP-2 in supernatant with zymography.
(1) In the situation of hypoxia for 12 h, the expression of MMP-2 increased (hypoxia group positive indexes: 5.7 +/- 2.0; control: 3.2 +/- 1.0; P < 0.01), while TIMP-2 decreased (hypoxia group positive indexes: 2.5 +/- 0.7; control: 3.6 +/- 1.0; P < 0.05) in HSC, and the activity of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922; control: 17.277 +/- 7.424; P < 0.01). At the different time courses of hypoxia, the change of expression and activity of MMP-2 was most notable at 6 h. (2) In the situation of hyperoxia for 12 h, the protein contents of MMP-2, TIMP-2 in supernatant were both higher than those of the control, especially the TIMP-2 (hyperoxia group A(450): 0.050 +/- 0.014; control: 0.022 +/- 0.010; P < 0.01), and so was the activity of MMP-2 (hyperoxia group total A: 5.252 +/- 0.771; control: 4.304 +/- 1.083; P < 0.05). The expression of MT1-MMP was also increased.
The HSC is sensitive to the oxygen. Hypoxia accelerates the expression of MMP-2 and the effect is more marked at the early stage. Hyperoxia increases the activity of MMP-2.
研究缺氧和高氧对肝星状细胞（HSC）基质金属蛋白酶-2（MMP-2）表达及活性调节的影响。
采用免疫细胞化学法（LSAB法）检测缺氧或高氧条件下培养的大鼠HSC中MMP-2、基质金属蛋白酶组织抑制因子-2（TIMP-2）和膜型基质金属蛋白酶-1（MT1-MMP）的表达，用ELISA法检测培养上清液中MMP-2、TIMP-2的含量，用明胶酶谱法检测上清液中MMP-2的活性。
（1）缺氧12 h时，HSC中MMP-2表达增加（缺氧组阳性指数：5.7±2.0；对照组：3.2±1.0；P<0.01），而TIMP-2表达减少（缺氧组阳性指数：2.5±0.7；对照组：3.6±1.0；P<0.05），上清液中MMP-2活性明显下降（缺氧组：7.334±1.922；对照组：17.277±7.424；P<0.01）。在缺氧的不同时间点，MMP-2表达和活性变化在6 h时最为显著。（2）高氧12 h时，上清液中MMP-2、TIMP-2的蛋白含量均高于对照组，尤其是TIMP-2（高氧组A(450)：0.050±0.014；对照组：0.022±0.010；P<0.01），MMP-2活性也升高（高氧组总A：5.252±0.771；对照组：4.304±1.083；P<0.05）。MT1-MMP的表达也增加。
HSC对氧敏感。缺氧促进MMP-2表达，且早期作用更明显。高氧增加MMP-2活性。
