吉林大学学报(医学版) ›› 2026, Vol. 52 ›› Issue (4): 1043-1050.doi: 10.13481/j.1671-587X.20260416

• 基础研究 • 上一篇    

白藜芦醇对女性卵巢颗粒细胞功能的调控作用和调控过程中SIRT1的作用机制

柯慧(),颜礼征,张宇,毛记龙,姚宇飞   

  1. 海南医科大学第一附属医院,生殖健康及相关疾病研究与转化教育部重点实验室,海南省人类生殖与遗传重点实验室,海南省地方病(地中海贫血)临床医学研究中心,生殖医学中心,海南 海口 570102
  • 收稿日期:2024-08-24 接受日期:2024-10-04 出版日期:2026-07-28 发布日期:2026-07-27
  • 通讯作者: 柯慧 E-mail:kehui_kh@163.com
  • 作者简介:柯 慧(1987-),女,海南省海口市人,副主任医师,医学博士,主要从事高龄女性卵巢功能和卵巢低反应方面的研究。
  • 基金资助:
    海南省科技厅自然科学基金项目(821RC702);海南省卫健委省级临床医学中心项目(2021);海南省科技厅院士创新平台科研项目(2022)

Regulation effect of resveratrol on ovarian granulosa cell function in females and mechanism of SIRT1 in regulation process

Hui KE(),Lizheng YAN,Yu ZHANG,Jilong MAO,Yufei YAO   

  1. Key Laboratory of Reproductive Health Diseases Research and Translation of Ministry of Education,Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research,Hainan Provincial Clinical Research Center for Thalassemia,Department of Reproductive Medicine,The First Affiliated Hospital,Hainan Medical University,Haikou 570102,China
  • Received:2024-08-24 Accepted:2024-10-04 Online:2026-07-28 Published:2026-07-27
  • Contact: Hui KE E-mail:kehui_kh@163.com

摘要:

目的 探讨沉默信息调节因子1(SIRT1)对高龄女性卵巢颗粒细胞的调控作用,初步阐明SIRT1激活剂白藜芦醇(RESV)改善高龄女性卵巢颗粒细胞功能的机制。 方法 纳入60例接受体外受精-胚胎移植(IVF-ET)的患者,包括30例高龄患者(高龄组,年龄≥40岁)和30例非高龄患者(非高龄组,年龄<35岁)。采用密度梯度离心法收集高龄组和非高龄组女性患者卵泡液中的颗粒细胞样本各30例,每例高龄女性卵巢颗粒细胞分为对照组、RESV组和EX527(SIRT1抑制剂)组,对照组颗粒细胞不使用药物处理,RESV组颗粒细胞置于含100 μmol·L-1 RESV培养液中培养48 h,EX527组颗粒细胞置于含EX527的培养液中培养48 h,胰酶消化后收集颗粒细胞。采用实时荧光定量PCR(RT-qPCR)法检测各组细胞中SIRT1、类固醇激素合成急性调节蛋白(stAR)、细胞色素P450家庭19(CYP19)和细胞色素P450家族17(CYP17)mRNA表达水平,Western blotting 法检测各组细胞中含半胱氨酸的天冬氨酸蛋白酶3(Caspase-3)和B细胞淋巴瘤2(Bcl-2)蛋白表达水平,化学发光法检测人绒毛膜促性腺激素(HCG)注射日各组细胞培养液中雌二醇水平,脱氧核糖核苷酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测各组细胞凋亡率。 结果 RT-qPCR法,与非高龄组比较,高龄组患者颗粒细胞中SIRT1 mRNA和蛋白表达水平均降低(P<0.05)。与对照组比较,RESV组患者颗粒细胞中SIRT1 mRNA和蛋白表达水平均升高(P<0.05),EX527组患者颗粒细胞中SIRT1 mRNA和蛋白表达水平均降低(P<0.05)。与对照组比较,RESV组患者颗粒细胞中StARCYP19CYP17 mRNA及蛋白表达水平升高(P<0.05),EX527组患者颗粒细胞中StARCYP19CYP17 mRNA及蛋白表达水平降低(P<0.05)。化学发光法,与对照组比较,RESV组患者颗粒细胞培养液中雌二醇水平升高(P<0.05),EX527组患者颗粒细胞培养液中雌二醇水平降低(P<0.05)。TUNEL法和Western blotting法,与对照组比较,RESV组患者颗粒细胞凋亡率降低(P<0.05),颗粒细胞中Bcl-2蛋白表达水平升高(P<0.05),Caspase-3蛋白表达水平降低(P<0.05);EX527组颗粒细胞凋亡率升高(P<0.05),颗粒细胞中Bcl-2蛋白表达水平降低(P<0.05),Caspase-3蛋白表达水平升高(P<0.05)。 结论 高龄女性卵巢颗粒细胞中SIRT1表达降低。RESV通过促进高龄女性颗粒细胞中SIRT1表达,从而抑制颗粒细胞凋亡,增加颗粒细胞中类固醇激素合成酶表达,促进颗粒细胞合成雌二醇。

关键词: 沉默信息调节因子1, 高龄, 颗粒细胞, 细胞凋亡, 白藜芦醇

Abstract:

Objective To discuss the regulatory effect of silent information regulator 1 (SIRT1) on ovarian granulosa cells of advanced-age women, and to preliminarily clarify the mechanism by which SIRT1 activator resveratrol (RESV) improves the function of ovarian granulosa cells of advanced-age women. Methods Sixty patients underwent in vitro fertilization-embryo transfer (IVF-ET) were enrolled, including 30 advanced-age patients (advanced age group, age ≥40 years) and 30 non-advanced-age patients (non-advanced age group, age <35 years). Density gradient centrifugation was used to collect the granulosa cell samples from follicular fluid of female patients in advanced age group(n=30) and non-advanced age group (n=30). The granulosa cells from each advanced-age female patient were divided into control group, RESV group, and EX527 (SIRT1 inhibitor) group. The granulosa cells in control group were treated without any drug; the granulosa cells in RESV group were cultured in medium containing 100 μmol·L-1 RESV for 48 h; the granulosa cells in EX527 group were cultured in medium containing EX527 for 48 h. After trypsin digestion, the granulosa cells were collected. Real-time fluorescence quantitative PCR(RT-qPCR) method was used to detect the expression levels of SIRT1, steroidogenic acute regulatory protein (StAR), cytochrome P450 family 19 (CYP19), and cytochrome P450 family 17 (CYP17) mRNA in the cells in various groups; Western blotting method was used to detect the expression levels of cysteine-containing aspartate protease 3 (Caspase-3) and B-cell lymphoma 2 (Bcl-2) proteins in the cells in various groups; chemiluminescence method was used to detect the estradiol level in the cell culture medium of the cells in various groups on the day of human chorionic gonadotropin (HCG) injection; terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL) method was used to detect the apoptotic rate of the cells in various groups. Results The RT-qPCR results showed that compared with non-advanced age group, the expression levels of SIRT1 mRNA and protein in the granulosa cells of the patients in advanced age group were decreased (P<0.05). Compared with control group, the expression levels of SIRT1 mRNA and protein in granulosa cells of the patients in RESV group were increased (P<0.05), while those in EX527 group were decreased (P<0.05). Compared with control group, the expression levels of StARCYP19, and CYP17 mRNA and protein in granulosa cells of the patients in RESV group were increased (P<0.05), while those in EX527 group were decreased (P<0.05). The chemiluminescence method results showed that compared with control group, the estradiol level in granulosa cell culture medium of the patients in RESV group was increased (P<0.05), while that in EX527 group was decreased (P<0.05). The TUNEL method and Western blotting method results showed that compared with control group, the apoptotic rate of the granulosa cells of the patients in RESV group was decreased (P<0.05), the expression level of Bcl-2 protein in the granulosa cells was increased (P<0.05), and the expression level of Caspase-3 protein was decreased (P<0.05); compared with control group, the apoptotic rate of granulosa cells of the patients in EX527 group was increased (P<0.05), the expression level of Bcl-2 protein in the granulosa cells was decreased (P<0.05), and the expression level of Caspase-3 protein was increased (P<0.05). Conclusion The expression of SIRT1 in ovarian granulosa cells of advanced-age women is decreased. RESV inhibits granulosa cell apoptosis, increases the expression of steroidogenic enzymes in granulosa cells, and promotes estradiol synthesis in granulosa cells by promoting the expression of SIRT1 in granulosa cells of advanced-age women.

Key words: Silent information regulator 1, Advanced age, Granulosa cells, Apoptosis, Resveratrol

中图分类号: 

  • R711.75