卵巢组织冻存移植后卵泡丢失的机制及N-乙酰半胱氨酸的保护作用研究

doi:10.3969/j.issn.1006-7795.2026.04.003

首都医科大学学报 ›› 2026, Vol. 47 ›› Issue (4): 650-657.doi: 10.3969/j.issn.1006-7795.2026.04.003

• 更年期妇科内分泌与生育力保护 • 上一篇下一篇

卵巢组织冻存移植后卵泡丢失的机制及N-乙酰半胱氨酸的保护作用研究

李扬璐¹, 阮祥燕^1*, 程姣姣¹, 杜娟¹, 谷牧青¹, 金凤羽¹, 李妍秋¹, 王泽铖¹, 张明珍¹, Alfred O. Mueck^1,2

首都医科大学附属北京妇产医院/北京妇幼保健院内分泌科，北京 100026；2.德国图宾根大学妇产医院妇女健康部与妇女健康研究中心，图宾根 D-72076

收稿日期:2026-04-29 修回日期:2026-06-06 出版日期:2026-08-21 发布日期:2026-07-26
通讯作者: 阮祥燕 E-mail:ruanxiangyan@ccmu.edu.cn
基金资助:
国家自然科学基金项目（82101822）,首都医科大学附属北京妇产医院中青年学科骨干培养专项（FCYY202015）,北京市属医院科研培育计划项目（PX2021053）,首都医科大学校培育基金（PYZ20058）,北京市自然科学基金项目（7202047）。

Mechanisms of follicle loss after ovarian tissue cryopreservation and transplantation and the protective effect of N-acetylcysteine

Li Yanglu¹, Ruan Xiangyan^1*, Cheng Jiaojiao¹， Du Juan¹， Gu Muqing¹，Jin Fengyu¹，Li Yanqiu¹，Wang Zecheng¹，Zhang Mingzhen¹，Alfred O. Mueck^1,2

1.Department of Gynecological Endocrinology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University. Beijing Maternal and Child Health Care Hospital, Beijing 100026, China ；2.Department of Women's Health, Research Centre for Women's Health and University Women's Hospital of Tuebingen, University Hospitals of Tuebingen, Tuebingen D-72076, Germany

Received:2026-04-29 Revised:2026-06-06 Online:2026-08-21 Published:2026-07-26
Supported by:
This study was supported by National Natural Science Foundation of China(82101822), Beijing Obstetrics and Gynecology Hospital, Capital Medical University (FCYY202015), Beijing Municipal Administration of Hospitals Incubating Program (PX2021053), Capital Medical University (PYZ20058), Natural Science Foundation of Beijing (7202047).

摘要/Abstract

摘要： 目的本研究旨在探讨卵巢组织冻存移植后卵泡丢失的机制，并评估N-乙酰半胱氨酸（N-acetylcysteine，NAC）对原始卵泡的保护作用。方法将冻融人卵巢组织移植于鸡胚绒毛尿囊膜（chorioallantoic membrane，CAM）上，构建短期移植模型。实验分为4组：新鲜卵巢组织组（fresh group）、标准化冻存移植组（control group）、磷脂酰肌醇3-激酶抑制剂（phosphatidylinositol 3-kinase inhibitor, PI3Ki）组（PI3Ki group）和NAC处理组（NAC group）。采用苏木精-伊红染色计数卵泡，流式细胞术检测线粒体膜电位（ΔΨm, JC-1绿/红比值），Western blotting检测PI3K-Akt-Foxo3a信号通路关键蛋白［磷酸化蛋白激酶(phosphorylated protein kinase B, p-Akt)、磷酸化叉头框蛋白(phospholated forhead box o3a，p-Foxo3a)］的表达。结果与fresh group相比，各移植组卵泡数量均显著减少（P<0.008 3），线粒体去极化程度显著升高（P<0.001）。与PI3Ki group相比，control group中p-Akt和p-Foxo3a表达浓度升高，提示PI3K-Akt-Foxo3a通路被激活；PI3Ki group中该通路被显著抑制；NAC group中p-Akt和p-Foxo3a表达量较control group显著降低（P<0.008 3），且NAC组中线粒体去极化程度在各移植组中最低（P<0.008 3）。结论 NAC可通过下调PI3K-Akt-Foxo3a信号通路，抑制移植后原始卵泡的过度激活，减轻线粒体损伤，从而在卵巢组织冻存移植中发挥保护作用。

关键词: 卵巢组织冻存与移植, 卵泡丢失, N-乙酰半胱氨酸, PI3K-Akt-Foxo3a信号通路, 鸡胚绒毛尿囊膜模型, 卵泡活化

Abstract: Objective To investigate the mechanisms underlying follicle loss after ovarian tissue transplantation and evaluate the protective effects of N-acetylcysteine (NAC) on primordial follicles.Methods Frozen-thawed human ovarian tissue was transplanted onto the chorioallantoic membrane (CAM) of chick embryos. Four groups were established: fresh ovarian tissue (fresh group), standardized cryopreservation and transplantation (control group), PI3K inhibitor treatment (PI3Ki group), and NAC treatment (NAC group). Follicle counts were assessed by hematoxylin-eosin staining (HE) staining, mitochondrial membrane potential (ΔΨm) was measured by JC-1 staining using flow cytometry, and the expression of phosphorylated protein kinase B (p-Akt) and phospholated forhead box o3a (p-Foxo3a) in the PI3K-Akt-Foxo3a pathway was detected by Western blotting.Results Compared with the fresh group, all transplantation groups showed significantly reduced follicle counts (P<0.008 3) and increased mitochondrial depolarization (P<0.001). Compared with PI3Ki group, the control group exhibited activation of the PI3K-Akt-Foxo3a pathway, as indicated by elevated p-Akt and p-Foxo3a levels. This activation was suppressed in the PI3Ki group. In the NAC group, the expression of p-Akt and p-Foxo3a was significantly lower than in the control group (P<0.008 3), and the mitochondrial depolarization was the lowest among all transplanted groups(P<0.008 3).Conclusion NAC protects primordial follicles during ovarian tissue transplantation by downregulating the PI3K-Akt-Foxo3a signaling pathway, thereby inhibiting excessive follicle activation and reducing mitochondrial damage.

Key words: ovarian tissue cryopreservation and transplantation, follicle loss, N-acetylcysteine, PI3K-Akt-Foxo3a signaling pathway, chick embryo chorioallantoic membrane model, follicle activation

中图分类号:

李扬璐, 阮祥燕, 程姣姣, 杜娟, 谷牧青, 金凤羽, 李妍秋, 王泽铖, 张明珍, Alfred O. Mueck. 卵巢组织冻存移植后卵泡丢失的机制及N-乙酰半胱氨酸的保护作用研究[J]. 首都医科大学学报, 2026, 47(4): 650-657.

Li Yanglu, Ruan Xiangyan, Cheng Jiaojiao, Du Juan, Gu Muqing, Jin Fengyu, Li Yanqiu, Wang Zecheng, Zhang Mingzhen, Alfred O. Mueck, . Mechanisms of follicle loss after ovarian tissue cryopreservation and transplantation and the protective effect of N-acetylcysteine[J]. Journal of Capital Medical University, 2026, 47(4): 650-657.

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卵巢组织冻存移植后卵泡丢失的机制及N-乙酰半胱氨酸的保护作用研究

Mechanisms of follicle loss after ovarian tissue cryopreservation and transplantation and the protective effect of N-acetylcysteine

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