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AIM To study the effects of hypoxia,hyperoxia on theregulation of expression and activity of matrixmetalloproteinase-2 (MMP-2) in hepatic stellate cells(HSC).METHODS The expressions of MMP-2,tissue inhibitor ofmatrix metalloproteinass-2 (TIMP-2) and membrane typematrix matalloproteinass-1 (MT1-MMP) in cultured rat HSCwere detected by immunocytochemistry (ICC) and in situhybridization (ISH).The contents of MMP-2 and TIMP-2 inculture supernatant were detected with ELISA and theactivity of MMP-2 in supernatant was revealed byzymography.RESULTS In the situation of hypoxia for 12h,theexpression of MMP-2 protein was enhanced (hypoxiagroup positive indexes:5.7±2.0,n=10;control:3.2±1.0,n=7;P<0.05),while TIMP-2 protein was decreasedin HSC (hypoxla 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 suparnatant declined obviously(hypoxla group:7.334±1.922,n=9;control:17.277±7.424,n=11;P<0.01).Compared the varied duration ofhypoxia,the changes of expressions Including mRNA andprotein level as well as activity of MMP-2 were mostnotable in 6 h group.The highest value (Ahypoxla~-Acontrol) ofthe protein and the most intense signal of mRNA were inthe period of hypoxia for 6 h,along with the lowestactivity of MMP-2.In the situation of hyparoxia for 12 h,the contents (A_(450)) of MMP-2 and TIMP-2 in supernatantwere both higher than those in the control,especially theTIMP-2 (hyperoxla group:0.0499±0.0144,n=16;control:0.0219±0.0098,n=14;P<0.01),and so wasthe 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,hypoxiaenhances the expression of MMP-2 and the effect is moremarked at the early stage;hyperoxia mainly raises theactivity of MMP-2.
AIM To study the effects of hypoxia, hyperoxia on thereulation and expression of activity of matrix metalloproteinase-2 (MMP-2) in hepatic stellate cells (HSC). METHODS The expressions of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (MT1-MMP) in cultured rat HSC Were detected by immunocytochemistry (ICC) and in situ hybridization (ISH). The contents of MMP-2 and TIMP-2 inoculum supernatant were detected with ELISA and the activity of MMP- 2 in supernatant was revealed byzymography.RESULTS In the situation of hypoxia for 12h, theexpression of MMP-2 protein was enhanced (hypoxiagroup positive indexes: 5.7 ± 2.0, n = 10; control: 3.2 ± 1.0, n = 7; P <0.05 ), while the activity (total A) of MMP-2, while the activity of the TIMP-2 protein was decreased in HSC (hypoxa group positive indexes: 2.5 ± 0.7, n = 10; control: 3.6 ± 1.0, n = 7; in suparnatant declined obviously (hypoxla group: 7.334 ± 1.922, n = 9; control: 17.277 ± 7.424, n = 11; P <0.01) .Compared the varied duration of hypoxia, the chang es of expressions Including mRNA and protein levels as well as activity of MMP-2 were most notable in 6 h group. the highest value (Ahypoxla ~ -Acontrol) of the protein and the most intense signal of mRNA were inthe period of hypoxia for 6 h, along with the lowest activity of MMP-2. the situation of hyparoxia for 12 h, the contents (A_ (450)) of MMP-2 and TIMP-2 in supernatantwere higher than those in the control, especially theTIMP-2 : 0.0499 ± 0.0144, n = 16; control: 0.0219 ± 0.0098, n = 14; P <0.01) and so wasthe 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.