Joint Laboratory of Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, Changchun 130118, China; Key Laboratory of Animal Production, Product Quality and Security, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
Joint Laboratory of Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, Changchun 130118, China; Key Laboratory of Animal Production, Product Quality and Security, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
Life Sci. 2020 Apr 1;246:117431. doi: 10.1016/j.lfs.2020.117431. Epub 2020 Feb 13.
Melatonin is an endogenous indoleamine hormone involved in various physiological processes. However, the mechanism of melatonin in mediating Leydig cells function has not been fully explained. In this study, we investigated the mechanism through which melatonin inhibits apoptosis in mouse Leydig cells by activating the retinoic acid-related orphan nuclear receptor (ROR) α/p53 signaling pathway. We confirmed the expression and localization of RORα in mouse Leydig cells using immunofluorescence. After treatment with 10 ng/mL melatonin for 36 h, RORα mRNA and protein levels were significantly increased (P < 0.01). TUNEL and flow cytometry showed that melatonin significantly decreased the TUNEL-positive cell ratio and apoptosis rate (P < 0.05). Moreover, melatonin decreased BAX expression and increased BCL-2 expression (P < 0.05). However, the RORα inhibitor SR1001 reversed the inhibitory effects of melatonin on apoptosis (P < 0.05). Additionally, analysis of p53 expression showed that melatonin inhibited p53 mRNA and protein expression (P < 0.05), whereas SR1001 reversed these effects. p53 reversed the anti-apoptotic process involving RORα-mediated melatonin in mouse Leydig cells. Collectively, our findings suggested that melatonin inhibited apoptosis via the RORα/p53 pathway.
褪黑素是一种内源性吲哚胺激素，参与多种生理过程。然而，褪黑素介导睾丸间质细胞功能的机制尚未完全阐明。在这项研究中，我们通过激活维甲酸相关孤儿核受体（ROR）α/p53 信号通路，研究了褪黑素抑制小鼠睾丸间质细胞凋亡的机制。我们使用免疫荧光法证实了 RORα在小鼠睾丸间质细胞中的表达和定位。用 10ng/mL 褪黑素处理 36 小时后，RORα mRNA 和蛋白水平显著增加（P<0.01）。TUNEL 和流式细胞术显示，褪黑素显著降低了 TUNEL 阳性细胞比例和凋亡率（P<0.05）。此外，褪黑素降低了 BAX 的表达，增加了 BCL-2 的表达（P<0.05）。然而，RORα抑制剂 SR1001 逆转了褪黑素对凋亡的抑制作用（P<0.05）。此外，对 p53 表达的分析表明，褪黑素抑制了 p53 mRNA 和蛋白的表达（P<0.05），而 SR1001 则逆转了这些作用。p53 逆转了涉及 RORα介导的褪黑素的抗凋亡过程。总之，我们的研究结果表明，褪黑素通过 RORα/p53 通路抑制细胞凋亡。
