Effect of TIMP-1 on expression of FN and type Ⅳ collagen in rat glomerular mesangial cells
- VernacularTitle:金属蛋白酶组织抑制剂1基因转染对系膜细胞表达纤连蛋白和Ⅳ型胶原的影响
- Author:
Hongli LIN
;
Xiangmei CHEN
;
Zhiheng YU
;
Al ET
;
- Publication Type:Journal Article
- Keywords:
Tissue inhibitor of metalloproteinase-1;
Fibronectin;
Collagen Ⅳ type;
Genetransfection
- From:
Chinese Journal of Nephrology
1997;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
ObjectⅣe To investigate the effect of sense and antisense TIMP-1 on the expression of FN and type Ⅳ collagen of rat glomerular mesangial cells in vitro. Methods Two recombinant retroviral vectors, pLXIN-TIMP-1 (PLT)encoding sense TIMP-1 and pLXIN-ATIMP-1 (PLA)encoding antisense TIMP-1 were constructed by using DNA recombining techniques. These two vectors were then introduced into the PA317 packaging cell line with lipofectin DOTAP. The high-titer retroviral supematants containing sense or antisense TIMP-1 were used to infect rat glomerular mesangial cells. PCR and Northern blot were used to detect the integration and expression of human TIMP-1. Northern blot and ELISA were employed to investigate the expression of endogenous TIMP-1, FN, and Ⅳ collagen. Results Both sense and antisense TIMP-1 were successfully integrated into rat mesangial cells and expressed the sense and antisense ITMP-1 RNA. Overexpression of TIMP-1 induced by sense TIMP-1 caused upregulation of FN and Ⅳ type collagen in protein level, in contrast, supprssion of TIMP-1 expression by antisense TIMP-1 caused downregulation of FN and Ⅳ type collagen protein; but neither sense nor antisense TIMP-1 infection had effects on the RNA level of FN and Ⅳ type collagen. Conclusion TIMP-1 suppresses the degradation of ECM in rat glomerular mesangial cells and antisense TIMP-1 may be available for renal fibrosis in the future.
