Mechanism of Ginsenoside Rg1 Antagonizing the Secretion Disorder in Leydig Cells Induced by D-galactose
XIANG Yezhou1,2, WANG Ziling1, XIAO Hanxianzhi1, HOU Jiying1, XIANG Yue1, JIANG Rong1, WANG Lu1, HUANG Guoning2, WANG Yaping1*
This article aims to explore the mechanism of ginsenoside Rg1 antagonizing D-gal (D-galactose) on androgen secretion disorders in mouse Leydig cells. A mouse aging model was constructed by D-gal, with Rg1 injected in vivo to interfere with the aging process for observing the pathological changes of testicular tissues. According to the aging model of testicular stromal cells (TM3 cell line) induced by D-gal in vitro, Rg1 was added into the culture system to antagonize the aging effect of D-gal. The aging of mice testicular cells and TM3 cells cultured in vitro was observed by SA-β-Gal staining. The levels of testosterone secreted by TM3 cells and oxidative stress injury factors were detected by ELISA. Fluorescent probe DCFH-DA was used to detect the level of the ROS (reactive oxygen species). The expression of the key enzymes of testosterone synthesis and Nrf2/ARE pathway related proteins was detected by Western blot. mRNA expression of the key enzymes of testosterone synthesis and inflammatory factors was evaluated by qRT-PCR. The results showed that Rg1 injection could interfer with the aging process caused by D-gal, the senescent cells in the interstitium of the testes were significantly reduced. The addition of Rg1 in vitro could antagonize the aging effect of D-gal, and the secretion of testosterone in TM3 cells was not significantly reduced. The expression of IL-1, IL-6, IL-8 and other inflammatory factors was inhibited; the activity of GSH-Px and CAT in TM3 cells were increased, while the capability of cells producing MDA (malondialdehyde) and ROS was inhibited. The expression of genes and proteins of the key enzymes in testosterone synthesis, such as StAR, 3β-HSD and P450scc, was up-regulated. The antioxidant proteins such as Nrf2, HO-1 were up-regulated, while Keap1 was down-regulated. The results suggested that Rg1 might antagonize the oxidative stress injury to Leydig cells induced by D-gal through activating the Nrf2/ARE antioxidant signaling pathway, thus regulating the secretory function of the testis.
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