Role of connective tissue growth factor in renal tubular epithelial-myofibroblast transdifferentiation and extracellular matrix accumulation in vitro

Connective tissue growth factor (CTGF) has been reported to play an important role in mediating the profibrotic effects of transforming growth factor-beta (TGF-beta) in the pathogenesis of renal fibrosis. To further elucidate the role of CTGF in renal tubular trans differentiation and extracellular matrix (ECM) metabolism, we examined the time-course of CTGF, alpha-smooth muscle actin (alpha-SMA), fibronectin and plasminogen activator inhibitor-1(PAI-1) gene expression upon the stimulation of TGF-beta1 (5 mug/L) in cultured human proximal tubular epithelial cell line (HKC), and further investigated the effects of endogenous CTGF blockade. On reverse transcriptional-polymerase chain reaction (RT-PCR) analysis, TGF-beta1 upregulated CTGF gene expression, preceding that of alpha-SMA, fibronectin and PAI-1. The alpha-SMA, fibronectin and PAI-1 mRNA expression induced by TGF-betal were significantly inhibited by CTGF antisense oligodeoxynucleotide (ODN) transfection. With prolonged incubation time, CTGF antisense ODN also inhibited intracellular alpha-SMA and PAI-1 protein synthesis, lowered the level of fibronectin and PAI-1 protein secreted into the media, as confirmed by indirect immuno-fluorescence, flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot methods respectively. These results suggested that CTGF may play a crucial role in the renal tubular epithelial-transdifferentiation and the following deposition/degradation process of ECM during tubulointerstitial fibrosis. (C) 2004 Elsevier Inc. All rights reserved.