2003PubMedRequires access

[Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expressions on cultured rat mesangial cells transfected with Smad 7 vector].

Hong Yu, Xiaogang Wang, Yi Wang, Qi Chen, Xiurong Zhang, Muyi Guo

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Abstract

OBJECTIVE: To investigate matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions in the cultured rat mesangial cells (MsC) transfected with Smad 7 vector and to elucidate the mechanism of Smad 7 in blocking tissue fibrosis. METHODS: Lipofectin method was used to transfect Smad 7 vector into MsC. Western blot and RT-PCR analyses were then used to detect Smad 7 protein and mRNA expression levels. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and zymography assay. RESULTS: Two MsC clones (S-22, S-26) with Smad 7 overexpression were successfully established. The two clones showed an increased expression of MMP-2 protein and enhanced enzyme activity. The expressions of TIMP-2 protein and mRNA however were suppressed. CONCLUSIONS: It is possible that Smad 7 can alleviate the development of tissue fibrosis by upregulating the expression of MMP-2 and downregulating the expression of TIMP-2 in mesangial cells.

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OBJECTIVE: To investigate matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions in the cultured rat mesangial cells (MsC) transfected with Smad 7 vector and to elucidate the mechanism of Smad 7 in blocking tissue fibrosis. METHODS: Lipofectin method was used to transfect Smad 7 vector into MsC. Western blot and RT-PCR analyses were then used to detect Smad 7 protein and mRNA expression levels. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and zymography assay. RESULTS: Two MsC clones (S-22, S-26) with Smad 7 overexpression were successfully established. The two clones showed an increased expression of MMP-2 protein and enhanced enzyme activity. The expressions of TIMP-2 protein and mRNA however were suppressed. CONCLUSIONS: It is possible that Smad 7 can alleviate the development of tissue fibrosis by upregulating the expression of MMP-2 and downregulating the expression of TIMP-2 in mesangial cells.

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Available abstract

OBJECTIVE: To investigate matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions in the cultured rat mesangial cells (MsC) transfected with Smad 7 vector and to elucidate the mechanism of Smad 7 in blocking tissue fibrosis. METHODS: Lipofectin method was used to transfect Smad 7 vector into MsC. Western blot and RT-PCR analyses were then used to detect Smad 7 protein and mRNA expression levels. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and zymography assay. RESULTS: Two MsC clones (S-22, S-26) with Smad 7 overexpression were successfully established. The two clones showed an increased expression of MMP-2 protein and enhanced enzyme activity. The expressions of TIMP-2 protein and mRNA however were suppressed. CONCLUSIONS: It is possible that Smad 7 can alleviate the development of tissue fibrosis by upregulating the expression of MMP-2 and downregulating the expression of TIMP-2 in mesangial cells.

Key concepts: SMAD, Western blot, Transfection, Matrix metalloproteinase, Molecular biology, Tissue inhibitor of metalloproteinase, Chemistry, Messenger RNA

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[Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expressions on cultured rat mesangial cells transfected with Smad 7 vector]. — Research Paper | ScholarLens