[Effects of ginsenoside Rb1 on TGF-beta1 induced p47phox expression and extracellular matrix accumulation in rat renal tubular epethelial cells].
OBJECTIVE
To investigate the effects of Ginsenoside Rb1 (G-Rb1) on the oxidative damage and extracellular matrix accumulation in rat renal tubular epethelial cells induced by transforming growth factor-beta1 (TGF-beta1).


METHODS
Cultured normal rat renal tubular epethelial cells (NRK-52E) were divided into control group, 10 ng/mL TGF-beta1-induced group, G-Rb1 treated groups in which rat renal tubular epethelial cells were treated with different concentration of G-Rb1 (10 ng/mL, 20 ng/mL, 40 ng/mL) after TGF-beta1 induction, G-Rb1 40 ng/mL group and 100 nmol/L DPI(diphenyleneiodonium, an inhibitor of NADPH oxidase) group. Intracellular reactive oxidative species (ROS) level was measured by flowcytometry. p47phox protein expression was assessed by immunohistochemistry and western blotting method. The expressions of collagen I (Col-I) and fibronectin(FN) gene were measured by real-time PCR analysis. The protein level of Col-I and FN were quantitatively detected by enzyme-linked immunosorbent assay.


RESULTS
TGF-beta1 at 10 ng/mL significantly increased the intercellular ROS production and p47phox expression (P < 0.05). The levels of Col-I and FN were also significantly up-regulated with the stimulation of 10 ng/mL TGF-beta1 (P < 0.05). Compared to TGF-beta1-induced group, G-Rb1 and DPI depressed TGF-beta1-induced ROS production and p47phox overexpression. Meanhile, G-Rb1 and DPI decreased the levels of Col-I and FN.


CONCLUSIONS
G-Rb1 could inhibit TGF-beta1 induced ROS production and decrease the levels of Col-I and FN in a dose-dependent manner. The mechanism might be partly related to the suppression of p47phox expression.
Xie
Xi-Sheng
XS
Department of Nephrology, West China Hospital, Sichuan University, Chengdu 610041, China.
Liu
Heng-Chuan
HC
Fan
Jun-Ming
JM
Li
Hui-Juan
HJ
D000818:Animals; D002460:Cell Line; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D036145:Ginsenosides / Q000494:pharmacology*; D007684:Kidney Tubules / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
D004365:Drugs, Chinese Herbal; D036145:Ginsenosides; D017382:Reactive Oxygen Species; D053773:Transforming Growth Factor beta1; D019255:NADPH Oxidases; C060505:neutrophil cytosolic factor 1; C035054:ginsenoside Rg1