【摘要】 旨在分析中草药淫羊藿苷(Icariin, ICA)及其代谢物对奶山羊睾丸间质细胞(Leydig cells, LCs)睾酮合成的影响。利用代谢组学分析ICA在奶山羊体内的代谢物，在此基础上，用ICA和代谢物分别对分离培养的奶山羊LCs进行处理。CCK-8检测细胞活率，ELISA检测睾酮含量的变化，并进一步用qPCR和Western blot分析睾酮合成关键酶的表达水平。结果表明：本研究中共检测到358个代谢物，ICA在奶山羊体内的主要代谢物是淫羊藿素，且主要在瘤胃液中代谢。CCK-8检测发现，ICA和淫羊藿素对奶山羊LCs的毒性较低，而淫羊藿Ⅱ和去甲淫羊藿素毒性较大。ELISA、qPCR和Western blot进一步分析发现，ICA和代谢物主要通过上调睾酮合成关键酶17β-HSD和Cyp17a1的表达水平提高雄激素睾酮的合成水平。综上所述，ICA在奶山羊瘤胃液中代谢为淫羊藿素，ICA和代谢物淫羊藿素能够通过上调睾酮合成酶的表达水平显著促进睾酮的合成。更多还原

【Abstract】 The aim of this study was to investigate the effect of Chinese herb icariin(ICA) and its metabolites on testosterone synthesis in Leydig cells(LCs) of dairy goats.We analyzed the main metabolites of ICA in the rumen fluid, serum, and small intestine, as well as the differential metabolites between ICA group and control group.The cultured primary LCs of dairy goats were treated with ICA and its metabolites, respectively.The cell viability of LCs was assessed by CCK-8,the content of testosterone in supernatant was determined by ELISA,and the expression levels of key enzymes involved in testosterone synthesis were further analyzed by qPCR and Western blot.The results showed that a total of 358 metabolites were detected in this study.The primary metabolite of ICA in dairy goats was icaritin, which was mainly metabolized in the rumen fluid microorganisms.The CCK-8 assay found that ICA and icaritin had low toxicity to LCs of dairy goats, while icarisid II and desmethyl icaritin had high toxicity.Further analysis by ELISA,qPCR and Western blot revealed that ICA and its metabolites significantly increased the testosterone synthesis by up-regulating the expression levels of 17β-HSD and Cyp17a1.In conclusion, ICA is metabolized into icaritin in the rumen fluid of dairy goats, and both ICA and its metabolite icaritin can promote testosterone synthesis by up-regulating the expression levels of testosterone synthesis enzymes.更多还原