The Changes of Matrix Metalloproteinase-2 and Tissue Inhibitor of Metalloproteinase-2 Expressions on Cultured Rat Mesangial Cells Transfected with Smad2 Vector

Purpose To elucidate the mechanism of the role of TGF-β in the development of glomeru-losclerosis by observing the matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) expressions on cultured rat mesangial cells (MsC) transfected with Smad2 vector. Methods Calcium phosphate method was used to transfect Smad2 vector into MsC, Westem blot analysis for detecting Smad2 protein. The expressions of MMP-2 and TIMP-2 were determined by Western blot, RT-PCR and Zy-mography assay respectively. Results Overexpresion of Smad2 on four positive MsC clones (T-12,T-31, T-35 and T-40) were successfully established. Two clones showed increased expression of MMP-2 protein and enhanced enzyme activity,associated with upregulation of TIMP-2 protein and its mRNA expression. Conclusions It is possible that TGF-β mediates the development of glomerular injury and glomerulosclerosis by upregulating Smad2 expression and increasing the expressions of MMP-2 and TIMP-2.