Effect of transforming growth factor beta1/Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in cultured rat mesangial cells.
- Author:
Chen YANG
1
;
Lu DAI
;
Xue-guang LIU
;
Qi CHEN
;
Xiu-rong ZHANG
;
Mu-yi GUO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Blotting, Western; Cells, Cultured; DNA-Binding Proteins; genetics; physiology; Fluorescent Antibody Technique; Gene Expression; Genetic Vectors; genetics; Glomerular Mesangium; cytology; drug effects; metabolism; Male; Matrix Metalloproteinase 2; genetics; metabolism; RNA, Messenger; genetics; metabolism; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Smad2 Protein; Smad3 Protein; Smad7 Protein; Tissue Inhibitor of Metalloproteinase-2; genetics; metabolism; Trans-Activators; genetics; physiology; Transfection; Transforming Growth Factor beta; pharmacology; Transforming Growth Factor beta1
- From: Chinese Journal of Pathology 2003;32(6):553-557
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of transforming growth factor (TGF) beta1/Smad signaling pathway on the expression and enzymatic activity of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in cultured rat mesangial cells (MsC).
METHODSLipofectin method was used to transfect Smad 2, Smad 3 and Smad 7 vectors into MsC; and immunofluorescence, RT-PCR and Western blot analysis were used to detect their transfection efficiency. The expression and enzymatic activity of MMP-2 and TIMP-2 were determined by Western blot, zymography or reverse zymography assay.
RESULTSMsC transfected with Smad 2 gene showed slightly increased expression and enzymatic activity of both MMP-2 and TIMP-2, which was more obvious upon stimulation by TGF-beta1. MsC transfected with Smad 3 gene showed a slight upregulation of TIMP-2 expression and its enzymatic activity, which was enhanced after TGF-beta1 stimulation. There was however no change in MMP-2 expression and its enzymatic activity. On the other hand, MsC transfected with Smad 7 gene showed a decrease in MMP-2 and TIMP-2 expression and enzymatic activity, which was especially obvious after stimulation by TGF-beta1.
CONCLUSIONSTGF-beta1/Smad signaling pathway may play an important role in the pathogenesis of glomerulosclerosis, probably via MMP-2 and TIMP-2 expression and the associated enzymatic activity.
