小鼠卵泡非黄体化颗粒细胞体外培养体系构建
Modeling In Vitro Culture of Non-Luteinized Mouse Follicular Granulosa Cells
温静茹 1赵梦瑶 1赵玉芬 1齐盟 1郝绍瑜 1杜晨光 1李海军1
作者信息
- 1. 内蒙古农业大学兽医学院,呼和浩特 010018;动物胚胎与发育工程自治区高等学校重点实验室,呼和浩特 010018
- 折叠
摘要
对小鼠卵泡颗粒细胞黄体化状态进行研究,使用黄体生成素受体(LHCGR)抑制剂BAY899建立非黄体化颗粒细胞体外培养体系.ELISA检测孕酮(P4)变化水平,荧光定量PCR检测促卵泡素受体(FSHR)、类固醇急性调节蛋白(STAR)、胆固醇侧链切割酶(CYP11A1)及LHCGR基因表达水平,Western Blot检测STAR蛋白水平.在0～72h P4水平激增(P＜0.01);STAR、CYP11A1 mRNA 表达显著升高(P＜0.01),FSHR mRNA 表达显著降低(P＜0.01),LHCGR mRNA表达显著升高(P＜0.01);BAY899对细胞增殖活性无影响,且能够持续下调P4激素的分泌(P＜0.01),以及显著下调STAR基因和蛋白的表达(P＜0.01).小鼠颗粒细胞体外培养过程中会发生自发黄体化,而BAY899能够显著抑制这种黄体化的发生.
Abstract
To investigate the luteinization of mouse follicular granulosa cells,we established an in vitro culture system for non-luteinized granulosa cells using the luteinizing hormone receptor(LHCGR)inhibitor BAY899.Progesterone(P4)levels were measured using ELISA,while quantita-tive PCR was employed to determine the expression levels of follicle-stimulating hormone receptor(FSHR),steroidogenic acute regulatory protein(STAR),cholesterol side-chain cleavage enzyme(CYP11A1),and LHCGR genes.Western Blot was conducted to assess STAR protein levels.From 0 hours to 72 hours,there was a significant increase in P4 levels(P＜0.01);mRNA expression of STAR and CYP11A1 also significantly increased(P＜0.01).In contrast,FSHR mRNA expression significantly decreased(P＜0.01),and LHCGR mRNA expression significantly increased(P＜0.01).BAY899 did not affect cell proliferation but continuously suppressed P4 secretion(P＜0.01)and significantly downregulated both STAR gene and protein expression(P＜0.01).In conclusion,during the in vitro culture of mouse granulosa cells,spontaneous luteinization occurs.BAY899 sig-nificantly inhibits this luteinization process.
关键词
颗粒细胞/黄体化/孕酮/BAY899Key words
granulosa cells/luteinization/progesterone/BAY899引用本文复制引用
出版年
2024
NETL
NSTL
万方数据