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. 2001 Oct;7(5):647-51.
doi: 10.3748/wjg.v7.i5.647.

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

P S Chen  1 , W R Zhai, X M Zhou, J S Zhang, Y E Zhang, Y Q Ling, Y H Gu
Affiliations

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

P S Chen et al. World J Gastroenterol. 2001 Oct.

Abstract

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.

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Figures

Figure 1
Figure 1
Zymographic analyses. Lane1: hypoxia group; Lane 2: control; Lane 3-5: hyperoxia group; Lane 6-8: control.
Figure 2
Figure 2
MMP-2 protein in HSC presented intense positive in hypoxia group. LSAB (DAB), × 200
Figure 3
Figure 3
MMP-2 protein in HSC presented positive in hypoxia control. LSAB (DAB), × 200
Figure 4
Figure 4
TIMP-2 protein in HSC presented weak positive in hypoxia group. LSAB (DAB), × 200
Figure 5
Figure 5
TIMP-2 protein in HSC presented intense positive in hypoxia control. LSAB (DAB), × 200
Figure 6
Figure 6
MT1-MMP protein in HSC presented positive in hyperoxia group. LSAB (DAB), × 200
Figure 7
Figure 7
MT1-MMP protein in HSC presented weak positive in hyperoxia control. LSAB (DAB), × 200
Figure 8
Figure 8
Signal of MMP-2 mRNA in HSC was intense in hypoxia for 6 h. ISH (NBT/BCIP), × 400
Figure 9
Figure 9
Signal of MMP-2 mRNA in HSC was weak in hypoxia for 12 h. ISH (NBT/BCIP), × 400
Figure 10
Figure 10
Signal of MMP-2 mRNA in HSC was intense in hypoxia for 24 h. ISH (NBT/BCIP), × 400
Figure 11
Figure 11
Signal of MT1-MMP mRNA in HSC was intense in hyperoxia group. ISH (NBT/BCIP), × 100
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