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Animal Reproduction and Physiology
Asian-Australasian Journal of Animal Sciences 2009;22(5): 618-625.
DOI: https://doi.org/10.5713/ajas.2009.80695    Published online April 30, 2009.
Effects of Daidzein on Testosterone Synthesis and Secretion in Cultured Mouse Leydig Cells
Liuping Zhang, Sheng Cui*
Correspondence:  Sheng Cui,
Abstract
The objective of this work was to study the direct effects of daidzein on steroidogenesis in cultured mouse Leydig cells. Adult mouse Leydig cells were purified by Percoll gradient centrifugation, and the cell purity was determined using a 3-hydroxysteroid dehydrogenase (3-HSD) staining method. The purified Leydig cells were exposed to different concentrations (10-7 M to 10-4 M) of daidzein for 24 h under basal and human chorionic gonadotropin (hCG)-stimulated conditions. The cell viability and testosterone production were determined, and the related mechanisms of daidzein action were also evaluated using the estrogen receptor antagonist ICI 182,780 and measuring the mRNA levels of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), and 3-HSD-1 involved in testosterone biosynthesis. The results revealed that daidzein did not influence cell viability. Daidzein increased both basal and hCG-stimulated testosterone production in a dose-dependent manner, and this effect was statistically significant at concentrations of 10-5 M and 10-4 M daidzein (p<0.05). ICI 182,780 had no influence on daidzein action. RT-PCR results revealed that 10-5 M and 10-4 M daidzein did not exert any obvious influence on the mRNA level of P450scc in Leydig cells. However, in the presence of hCG, these concentrations of daidzein significantly increased the StAR and 3-HSD-1 mRNA levels (p<0.05), but in the absence of hCG, only 10-5 M and 10-4 M daidzein up-regulated the StAR and 3-HSD-1 mRNA expression (p<0.05), respectively. These results suggest that daidzein has direct effect on Leydig cells. Daidzein-induced increase of testosterone production is probably not mediated by the estrogen receptor but correlates with the increased mRNA levels of StAR and 3-HSD-1.
Keywords: Daidzein; Testosterone; StAR; P450scc; 3b-HSD-1; Mouse Leydig Cells


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