[Effects of hepatocyte growth factor on TGF-beta1 triggered tubular epithelial-myofibroblast transdifferentiation by CTGF in vitro].
OBJECTIVE
To observe the effects of hepatocyte growth factor (HGF) on TGF-beta1 triggered tubular epithelial-myofibroblast transdifferentiation (TEMT) and on the expression of connective tissue growth factor (CTGF).


METHODS
The morphology of transdifferentiate tubular cells was observed using phase-contrast microscopy and scanning electron microscopy. alpha-SMA was assessed by immunohistochemistry and semiquantified by mean intergrated opitical density (IOD). The level of fibronectin (FN) in the culture supernatant was measured by ELISA. CTGF mRNA expression was examined by RT-PCR.


RESULTS
The TGF-beta1-induced TEMT characterized by expression of alpha-SMA was shown by immunohistochemistry. TGF-beta1 was also shown to stimulate the secretion of FN in cultured supernatant and the CTGF mRNA expression of NRK52E cells. There was no statistically significant difference between HGF-treated groups and control group in the result of alpha-SMA immunostaining and the level of FN, except that CTGF mRNA expression was slightly increased in the HGF-treated groups. The addition of HGF inhibited the TGF-beta1-induced TEMT, the secretion of FN, and the CTGF expression of NRK52E cells, there was a significant correlation between the expression of CTGF and the expression of alpha-SMA.


CONCLUSIONS
HGF could block TEMT and FN secretion triggered by TGF-beta1, which implies that HGF could participate in renal interstitial fibrosis as a negative regulator. The negative regulation of transdifferentiation of HGF may be partially achieved by attenuation of CTGF expression.
Zhong
Hui
H
Department of Nephrology, West China Hospital, Sichuan University, Chengdu 610041, China.
Fan
Jun-Ming
JM
Li
Zi
Z
Liu
Fang
F
Yang
Li-Chuan
LC
Ji
Ling
L
Sha
Zhao-Hui
ZH
Ma
Xing-Yi
XY
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor; D004847:Epithelial Cells / Q000166:cytology; D005353:Fibronectins / Q000096:biosynthesis; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000235:genetics; D007687:Kidney Tubules, Proximal / Q000166:cytology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
C526316:CCN2 protein, rat; D005353:Fibronectins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; C508962:Tgfb1 protein, rat; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor; D017228:Hepatocyte Growth Factor