Sun Jiandong, Xu Weiwei, Zheng Shuyuan, Lv Chengyu, Lin Jianmin, Chen Siqi, Qiu Yonghong, Jiang Xia, Draz Eman, Wang Shie
Key Laboratory of Stem Cell Engineering and Regenerative Medicine of Fujian Province University, Fujian Medical University, Fuzhou 350122, PR China.
Key Laboratory of Stem Cell Engineering and Regenerative Medicine of Fujian Province University, Fujian Medical University, Fuzhou 350122, PR China; Undergraduate Class, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350122, PR China.
Toxicol Appl Pharmacol. 2022 Apr 15;441:115969. doi: 10.1016/j.taap.2022.115969. Epub 2022 Mar 5.
Icariin (ICA), extracted from Epimedium, is a flavonoid used in traditional Chinese medicine. Di(2-ethylhexyl) phthalate (DEHP) is a phthalate used in commercial products as a plasticizer that can influence the human endocrine and reproduction system. We previously found that ICA reversed DEHP-induced damage through the prevention of reactive oxygen species accumulation and promotion of testosterone secretion. Here we investigated the mechanisms of ICA in promoting testosterone secretion from murine Leydig cells. We used ICA, DEHP, the Akt agonist SC-79, the Akt inhibitor MK2206, and the Creb inhibitor KG501 to determine the effect of these treatments on the expression levels of the steroidogenic enzymes, Cyp11a1 and Hsd3b, which play critical roles in androgen production, in Leydig cells. Bioinformatic analysis was used to search for ICA-targeted proteins and their associated pathways. We found that icariin interacted with estrogen receptor on the cell membrane, leading to increased phosphorylation levels of Akt and Creb proteins and enhanced transcription of genes encoding steroidogenic enzymes and testosterone synthesis. We further investigated ICA activity in vivo using male mice pretreated with 100 mg/kg ICA and then treated with 750 mg/kg DEHP. ICA pretreatment reversed the reduced protein expression levels of Cyp11a1 and Hsd3b induced by DEHP in Leydig cells in vivo. Furthermore, while the phosphorylation levels of Akt and Creb were decreased in testes of mice exposed to DEHP alone, these effects were reversed by ICA pretreatment. These findings indicate that ICA promotes testosterone synthesis via the Esr1/Src/Akt/Creb/Sf-1 signaling pathway.
淫羊藿苷（ICA）是从淫羊藿中提取的一种黄酮类化合物，用于传统中药。邻苯二甲酸二（2-乙基己基）酯（DEHP）是一种用于商业产品的增塑剂邻苯二甲酸盐，可影响人体内分泌和生殖系统。我们之前发现，ICA通过防止活性氧积累和促进睾酮分泌来逆转DEHP诱导的损伤。在此，我们研究了ICA促进小鼠睾丸间质细胞分泌睾酮的机制。我们使用ICA、DEHP、Akt激动剂SC-79、Akt抑制剂MK2206和Creb抑制剂KG501来确定这些处理对睾丸间质细胞中类固醇生成酶Cyp11a1和Hsd3b表达水平的影响，这两种酶在雄激素生成中起关键作用。利用生物信息学分析来寻找ICA靶向的蛋白质及其相关途径。我们发现淫羊藿苷与细胞膜上的雌激素受体相互作用，导致Akt和Creb蛋白的磷酸化水平升高，并增强了编码类固醇生成酶和睾酮合成的基因的转录。我们进一步使用经100mg/kg ICA预处理然后用750mg/kg DEHP处理的雄性小鼠在体内研究了ICA的活性。ICA预处理逆转了DEHP在体内诱导的睾丸间质细胞中Cyp11a1和Hsd3b蛋白表达水平的降低。此外，虽然单独暴露于DEHP的小鼠睾丸中Akt和Creb的磷酸化水平降低，但ICA预处理可逆转这些作用。这些发现表明ICA通过Esr1/Src/Akt/Creb/Sf-1信号通路促进睾酮合成。
