Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expressions on cultured rat mesangial cells transfected with Smad 7 vector
Yu, H.; Wang, X-gang.; Wang, Y.; Chen, Q.; Zhang, X-rong.; Guo, M-yi.
Zhonghua Bing Li Xue Za Zhi 32(6): 544-547
ISSN/ISBN: 0529-5807 PMID: 14761601 Accession: 048490660
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. 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. 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. 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.