Decorin transfection in human mesangial cells downregulates genes playing a role in the progression of fibrosis

被引:22
作者
Costacurta, A
Priante, G
D'Angelo, A
Chieco-Bianchi, L
Cantaro, S
机构
[1] Univ Padua, Policlin, Dept Med & Surg Sci, Renal Lab, I-35128 Padua, Italy
[2] Univ Padua, Dept Surg & Oncol Sci, I-35128 Padua, Italy
[3] Univ Hosp Padova, Clin Div Nephrol 1, Padua, Italy
关键词
human mesangial cell transfection; RT-PCR; chronic renal fibrosis; gene therapy; alpha 1 collagen type IV; fibronectin; TGF-beta; 1;
D O I
10.1002/jcla.10038
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
The proteoglycan decorin inhibits TGF-beta; therefore, it could antagonize progression of fibrotic diseases associated with activation of TGF-beta(1). The effect of decorin transfection in human mesangial cells (HMCs) on the expression of genes related to kidney fibrosis was investigated. HMCs, isolated from glomeruli of healthy portions of human kidneys removed due to carcinoma, were histochemically typed. Decorin cDNA cloned in a eukaryotic expression vector was transfected into HMCs. Gene expression of fibrogenetic cytokines and fibrotic proteins TGF-beta(1), PDGF-beta, alpha(1) collagen type IV, alpha(1) collagen type I, fibronectin, and tenascin was analyzed, by reverse transcription polymerase chain reaction (RT-PCR), 24 hr after transfection. Immunoblotting analysis of protein extracts using anti-decorin IgG, revealed a positive signal of about 52 MDa, corresponding to the molecular weight of decorin, in cultures transfected with the decorin gene. Decorin mRNA increased about 12 times in cultures transfected with the construct pCR3.1-Deco. Cells with increased decorin synthesis showed a 61% decrease of TGF-beta(1) mRNA, a 71% reduction of alpha(1) collagen type IV mRNA, and a 29% reduction of fibrenectin mRNA. This study is the first to investigate decorin transfection into human mesangial cells, and supports the use of the decorin gene to control the progression of glomerular and interstitial fibrosis in kidney diseases. (C) 2002 Wiley-Liss, Inc.
引用
收藏
页码:178 / 186
页数:9
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