Changes of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expressions on cultured rat mesangial cells transfected with Smad 7 vector.
- Author:
Hong YU
1
;
Xiao-gang WANG
;
Yi WANG
;
Qi CHEN
;
Xiu-rong ZHANG
;
Mu-yi GUO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Blotting, Western; Cells, Cultured; DNA-Binding Proteins; genetics; metabolism; Gene Expression; Genetic Vectors; genetics; Glomerular Mesangium; cytology; metabolism; Matrix Metalloproteinase 2; genetics; metabolism; RNA, Messenger; genetics; metabolism; Rats; Reverse Transcriptase Polymerase Chain Reaction; Smad7 Protein; Tissue Inhibitor of Metalloproteinase-2; genetics; metabolism; Trans-Activators; genetics; metabolism
- From: Chinese Journal of Pathology 2003;32(6):544-547
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSLipofectin 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.
RESULTSTwo 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.
CONCLUSIONSIt 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.
