Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China; Laboratory of Animal Fat Deposition & Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China.
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China; Laboratory of Animal Fat Deposition & Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, China.
Theriogenology. 2023 Dec;212:19-29. doi: 10.1016/j.theriogenology.2023.08.025. Epub 2023 Aug 29.
During growth, proliferation, differentiation, atresia, ovulation, and luteinization, the morphology and function of granulosa cells (GCs) change. Estrogen and progesterone are steroid hormones secreted by GCs that regulate the ovulation cycle of sows and help maintain pregnancy. miR-10a-5p is highly expressed in GCs and can inhibit GC proliferation. However, the role of miR-10a-5p in the steroid hormone synthesis of porcine GCs is unclear. In this study, miR-10a-5p agomir or antagomir was transfected into GCs. Overexpression of miR-10a-5p in GCs inhibited steroid hormone secretion and significantly downregulated steroid hormone synthesis via 3β-hydroxy steroid dehydrogenase and cytochrome P450 family 19 subfamily A member 1. Interference with miR-10a-5p had the opposite effect. Bodipy and Oil Red O staining showed that overexpression of miR-10a-5p significantly reduced the formation of lipid droplets. Overexpression significantly inhibited the content of total cholesterol esters in GCs. The mRNA and protein levels of 3-hydroxy-3-methylglutaryl-CoA reductase and scavenger receptor class B member 1 decreased significantly, and the opposite effects were seen by interference with miR-10a-5p. Bioinformatic analysis of potential targets identified cAMP-responsive element binding protein 1 as a potential target and dual-luciferase reporter system analysis confirmed that miR-10a-5p directly targets the 3' untranslated region. These findings suggest that miR-10a-5p inhibits the expression of 3β-hydroxy steroid dehydrogenase and cytochrome P450 family 19 subfamily A member 1 to inhibit the synthesis of steroid hormones in GCs. In addition, miR-10a-5p inhibits the cholesterol metabolism pathway of GCs to modulate steroid hormone synthesis.
在生长、增殖、分化、闭锁、排卵和黄体化过程中，颗粒细胞 (GCs) 的形态和功能发生变化。雌激素和孕激素是 GCs 分泌的甾体激素，调节母猪的排卵周期，有助于维持妊娠。miR-10a-5p 在 GCs 中高度表达，可抑制 GC 增殖。然而，miR-10a-5p 在猪 GCs 甾体激素合成中的作用尚不清楚。在本研究中，将 miR-10a-5p agomir 或 antagomir 转染到 GCs 中。GC 中 miR-10a-5p 的过表达抑制了甾体激素的分泌，并通过 3β-羟类固醇脱氢酶和细胞色素 P450 家族 19 亚家族 A 成员 1 显著下调了甾体激素的合成。干扰 miR-10a-5p 则产生相反的效果。Bodipy 和油红 O 染色表明，miR-10a-5p 的过表达显著减少了脂滴的形成。过表达显著抑制了 GCs 中总胆固醇酯的含量。3-羟-3-甲基戊二酰辅酶 A 还原酶和清道夫受体 B 类成员 1 的 mRNA 和蛋白水平显著降低，干扰 miR-10a-5p 则产生相反的效果。对潜在靶标的生物信息学分析鉴定 cAMP 反应元件结合蛋白 1 为潜在靶标，双荧光素酶报告系统分析证实 miR-10a-5p 可直接靶向 3'非翻译区。这些发现表明，miR-10a-5p 通过抑制 3β-羟类固醇脱氢酶和细胞色素 P450 家族 19 亚家族 A 成员 1 的表达抑制 GCs 中甾体激素的合成。此外，miR-10a-5p 抑制 GCs 的胆固醇代谢途径，调节甾体激素的合成。
