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

Abstract

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 alpha1 collagen type IV mRNA, and a 29% reduction of fibronectin 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.

Adenovirus-Mediated Decorin Gene Transfection Has Therapeutic Effects in a Streptozocin-Induced Diabetic Rat Model

“…Our data support this possibility, as we observed that TGF-β 1 protein levels and the ratio of pSmad2,3 to Smad2,3 (index of TGF-β 1 activity) were decreased in the Ad-decorin group, as compared to diabetic-controls. The other possibility is decorin can inhibit the expression of TGF-β 1 mRNA, which has been confirmed in other studies [23,24]. Our results are also consistent with these studies.…”

Reduced dermatopontin expression is a molecular link between uterine leiomyomas and keloids

“…Furthermore, decorin is involved in normal tissue development and differentiation (Riquelme et al, 2001). Decorin activity also can regulate proteins associated with extracellular-matrix formation (Dudas et al, 2001;Costacurta et al, 2002). Dermatopontin competes with decorin for TGFB binding and also interacts with collagen directly (Kuroda et al, 1999;Okamoto et al, 1999).…”

A Novel Modulatory Mechanism of Transforming Growth Factor-β Signaling through Decorin and LRP-1

“…It has been shown that as a result of this interaction, decorin can inhibit as well as activate TGF-␤-dependent signaling depending on the cell type (12,37). It inhibits TGF-␤-dependent signaling in mesangial cells (38), glioma cells (39), and differentiated myotubes (30) and stimulates this signaling in osteoblasts (16) and non-differentiated myoblasts (18). Several proteoglycans have a well established role as co-receptors (19,40).…”