[Effect of PKC signalling pathway and aldose reductase on expression of fibronectin induced by transforming growth factor-β1 in human mesangial cells].
OBJECTIVE
To 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).


METHODS
Human 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.


RESULTS
The 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).


CONCLUSIONS
AR 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.
Department of Pathology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Ping
P
Huang
Yue-juan
YJ
Zhang
Yuan
Y
Huang
Jing-jing
JJ
Zhao
Tao
T
Jiang
Nong
N
Zhang
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D052160:Benzothiazoles / Q000494:pharmacology; D002227:Carbazoles / Q000494:pharmacology; D002478:Cells, Cultured; D005353:Fibronectins / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D007211:Indoles; D008301:Maleimides; D050527:Mesangial Cells / Q000166:cytology / Q000378:metabolism*; D010793:Phthalazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D015854:Up-Regulation
C465664:2-(1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl)-3-(1H-indol-3-yl)maleimide; D052160:Benzothiazoles; D002227:Carbazoles; D005353:Fibronectins; D007211:Indoles; D008301:Maleimides; D010793:Phthalazines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; C067172:zopolrestat; D000449:Aldehyde Reductase; D011493:Protein Kinase C