Effect of PKC signalling pathway and aldose reductase on expression of fibronectin induced by transforming growth factor-β1 in human mesangial cells.
- Author:
Ping HUANG
1
;
Yue-juan ZHANG
;
Yuan HUANG
;
Jing-jing ZHAO
;
Tao JIANG
;
Nong ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Aldehyde Reductase; antagonists & inhibitors; biosynthesis; genetics; Benzothiazoles; pharmacology; Carbazoles; pharmacology; Cells, Cultured; Fibronectins; metabolism; Gene Knockdown Techniques; Humans; Indoles; Maleimides; Mesangial Cells; cytology; metabolism; Phthalazines; pharmacology; Protein Kinase C; antagonists & inhibitors; metabolism; RNA, Messenger; metabolism; RNA, Small Interfering; genetics; Signal Transduction; Transfection; Transforming Growth Factor beta1; pharmacology; Up-Regulation
- From: Chinese Journal of Pathology 2010;39(6):405-409
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of PKC signalling pathway and aldose reductase (AR) on the expression of fibronectin (FN) induced by transforming growth factor-β1 (TGF-β1).
METHODSHuman mesangial cells (HMCs) were cultured and transfected with pcDNA3-AR, and subject to AR gene silencing with small interfering RNA (siRNA) and then the cell was treated with recombinant human TGF-β1. The AR mRNA expression in the HMCs was examined using real time RT-PCR and protein expression of AR and FN was detected by Western blotting.
RESULTSThe cultured HMC treated with TGF-β1 showed increased expression of AR and FN, the normal HMC showed not reduced expression of FN after incubation with single inhibitors of AR.Pre-incubation of cells with inhibitors of AR and PKC, then the different groups of cells were treated with TGF-β1, and the induction effect on FN expression was suppressed (34%) in HMC. HMCs transfected with AR showed a strong protein expression of FN, which was increased by 3.6-fold after treatment with TGF-β1 (P < 0.05), and the induction effect on FN expression was suppressed by GÖ6983 (42%) in HMCs (P < 0.05). The HMC with AR gene knock-down by siRNA showed a decreased expression of AR and 90% decrease of FN protein in HMCs (P < 0.01), and TGF-β1-induced up-regulation of FN was significantly suppressed by siRNA (12%) in HMCs (P < 0.01).
CONCLUSIONSAR is capable of regulating FN expression only in the presence of TGF-β1, and this reaction is possibly accomplished through the activation of PKC signalling pathway.