中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (14): 2958-2968.doi: 10.12307/2025.602

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

班氏促卵助孕汤改善多囊卵巢综合征小鼠卵母细胞的质量

黎明星1，岳晓蕾1，陈秀榕2，李康梅3，刘云佳1，伍黎明1，黄愉淋1，吴媛媛4，白  琳1，潘巧俐1，何国珍1，杨素芳5   

  1. 1广西中医药大学基础医学院，广西壮族自治区南宁市  530200；2福建中医药大学附属人民医院，福建省福州市  350004；3广东医科大学附属医院，广东省湛江市  524013；4广西中医药大学瑞康医院，广西壮族自治区南宁市  530011；5广西中医药大学附属国际壮医医院，广西壮族自治区南宁市  530000
  • 收稿日期:2024-04-08 接受日期:2024-06-11 出版日期:2025-05-18 发布日期:2024-09-28
  • 通讯作者: 何国珍，博士，教授，硕士生导师，广西中医药大学基础医学院，广西壮族自治区南宁市 530200 共同通讯作者：杨素芳，博士，教授，博士生导师，广西中医药大学附属国际壮医医院，广西壮族自治区南宁市 530000
  • 作者简介:黎明星，1986年生，2019年广西大学毕业，博士，讲师，主要从事中西医防治不孕不育的基础和临床研究。 共同第一作者：岳晓蕾，1998年生，2024年广西中医药大学毕业，硕士，主要从事中西医防治不孕不育的基础研究。
  • 基金资助:
    国家自然科学基金项目(81960794)，项目负责人：何国珍；广西中医药大学引进博士科研启动基金项目(2020BS032)，项目负责人：杨素芳；广西高校中青年教师科研基础能力提升项目(2022KY0290)，项目负责人：黄愉淋；南宁市良庆区科技计划项目(202408)，项目负责人：杨素芳；广西中医药大学大学生科研训练课题(2023DXS04)，项目负责人：潘巧俐

Ban’s Culuan Zhuyun Decoction improves oocyte quality in polycystic ovary syndrome mice

Li Mingxing1, Yue Xiaolei1, Chen Xiurong2, Li Kangmei3, Liu Yunjia1, Wu Liming1, Huang Yulin1, Wu Yuanyuan4, Bai Lin1, Pan Qiaoli1, He Guozhen1, Yang Sufang5   

  1. 1Basic Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 2Affiliated People’s Hospital of Fujian University of Traditional Chinese Medicine, Fuzhou 350004, Fujian Province, China; 3Affiliated Hospital of Guangdong Medical University, Zhanjiang 524013, Guangdong Province, China; 4Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530011, Guangxi Zhuang Autonomous Region, China; 5Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China 
  • Received:2024-04-08 Accepted:2024-06-11 Online:2025-05-18 Published:2024-09-28
  • Contact: He Guozhen, PhD, Professor, Master’s supervisor, Basic Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China Co-corresponding author: Yang Sufang, PhD, Professor, Doctoral supervisor, Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China
  • About author:Li Mingxing, PhD, Lecturer, Basic Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China Yue Xiaolei, MS, Basic Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China Li Mingxing and Yue Xiaolei contributed equally to this work.
  • Supported by:
    the National Natural Science Foundation of China, No. 81960794 (to HGZ); PhD Introduction Research Startup Foundation of Guangxi University of Chinese Medicine, No. 2020BS032 (to YSF); Young and Middle-aged Teachers Fundamental Ability Enhancement Project in Guangxi Universities, No. 2022KY0290 (to HYL); Nanning Liangqing District Science and Technology Program Project, No. 202408 (to YSF); Undergraduate Research Training Project of Guangxi University of Chinese Medicine, No. 2023DXS04 (to PQL)

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摘要：


文题释义：
卵母细胞分泌因子：指卵母细胞选择性或特异性产生的细胞因子。卵泡发育异常与下丘脑-垂体-卵巢轴及卵泡微环境的调节失常密切相关。在卵泡微环境信息交流系统中，卵母细胞通过自分泌和旁分泌的方式产生卵母细胞分泌因子，对卵泡发育有重要影响。生长分化因子9、骨形态发生蛋白15是经典的卵母细胞成熟相关因子。
卵母细胞体外成熟：指从卵巢中获取未成熟的卵母细胞，经过适宜的体外培养条件，卵母细胞发育成熟并具有受精和胚胎发育能力。卵母细胞成熟是一个高度调节和极度复杂的过程，包括细胞核成熟和细胞质成熟。当卵母细胞发生氧化应激、凋亡、ATP不足等病理状态时，可能会引发卵母细胞发育停滞，出现纺锤体组装异常和染色体分布、数量异常。

背景：班氏促卵助孕汤改善多囊卵巢综合征卵母细胞质量的分子机制亟待完善补充。
目的：探讨班氏促卵助孕汤对多囊卵巢综合征小鼠卵母细胞质量的影响和分子机制。
方法：21 d龄雌性昆明小鼠颈部皮下注射硫酸脱氢表雄酮构建多囊卵巢综合征模型，连续给药21 d，记录动情周期及妊娠情况，ELISA检测血清性激素水平，Annexin V染色检测卵母细胞凋亡率，DCFH-DA荧光探针检测卵母细胞内活性氧水平，免疫荧光法观察卵母细胞纺锤体及染色体情况，网络药理学及分子对接验证班氏促卵助孕汤核心有效成分与卵母细胞成熟相关因子(生长分化因子9和骨形态发生蛋白15)结合活性，实时荧光定量PCR和Western blot检测卵母细胞中生长分化因子9和骨形态发生蛋白15的mRNA及蛋白表达水平。
结果与结论：①班氏促卵助孕汤中的成分(槲皮素、山奈酚、β-谷甾醇)与生长分化因子9、骨形态发生蛋白15具有良好的结合活性；②班氏促卵助孕汤能恢复小鼠动情期，改善性激素紊乱和妊娠情况，降低细胞凋亡率、活性氧水平、纺锤体组装异常率、染色体丢失率(P < 0.01，P < 0.05)，促进生长分化因子9、骨形态发生蛋白15 mRNA和蛋白表达(P < 0.01，P < 0.05)。结果表明，班氏促卵助孕汤可能通过调控生长分化因子9和骨形态发生蛋白15的基因表达，改善多囊卵巢综合征小鼠卵母细胞质量，提高生育力。
https://orcid.org/0000-0002-9833-2673(黎明星)；https://orcid.org/0000-0002-7450-6413(岳晓蕾)；
https://orcid.org/0009-0000-1902-2566(何国珍)；https://orcid.org/0000-0003-2926-2314(杨素芳)

关键词: 班氏促卵助孕汤, 多囊卵巢综合征, 小鼠卵母细胞, 体外成熟, 生长分化因子9, 骨形态发生蛋白15, 妊娠, 不孕

Abstract: BACKGROUND: It is urgent to improve the study on the molecular mechanism of Ban’s Culuan Zhuyun Decoction improving oocyte quality in polycystic ovary syndrome.
OBJECTIVE: To observe the effects of Ban’s Culuan Zhuyun Decoction on oocyte quality in a mouse model of polycystic ovary syndrome and to explore the underlying mechanisms of its intervention in polycystic ovary syndrome.
METHODS: Subcutaneous injection of dehydroepiandrosterone sulfate was used to establish the polycystic ovary syndrome model in 21-day-old female Kunming mice, and the treatment was conducted for 21 consecutive days. The estrous cycle and pregnancy was recorded. ELISA was used to detect serum sex hormone levels. The rate of apoptosis in oocytes was detected using Annexin V staining. The level of reactive oxygen species in oocytes was detected using dichlorodihydrofluorescein diacetate. The condition of spindle bodies and chromosomes in oocytes were detected using the immunofluorescence method. Network pharmacology and molecular docking were used to verify the binding properties of Ban’s Culuan Zhuyun Decoction core active components and oocyte maturation-related factors (growth differentiation factor 9 and bone morphogenetic protein 15). Real-time fluorescence quantitative PCR and western blot were used to detect the mRNA and protein expression levels of growth differentiation factor 9 and bone morphogenetic protein 15 in oocytes, respectively.
RESULTS AND CONCLUSION: (1) Ban’s Culuan Zhuyun Decoction core active components (quercetin, kaempferol, and β-sitosterol) showed good binding activities with growth differentiation factor 9 and bone morphogenetic protein 15. (2) Ban’s Culuan Zhuyun Decoction ameliorated the estrous cycle, regulated serum hormone, increased the pregnancy, decreased the rate of apoptosis, declined the level of reactive oxygen species, diminished the rate of abnormal spindle assembly and chromosome loss (P < 0.01, P < 0.05); and promoted the mRNA and protein expression of growth differentiation factor 9 and bone morphogenetic protein 15 (P < 0.05). Therefore, Ban’s Culuan Zhuyun Decoction may improve the oocyte quality and increase the fertility of polycystic ovary syndrome mice by regulating the gene expression of growth differentiation factor 9 and bone morphogenetic protein 15.

Key words: Ban’s Culuan Zhuyun Decoction, polycystic ovary syndrome, mouse oocyte, in vitro maturation, growth differentiation factor 9, bone morphogenetic protein 15, pregnancy, infertility

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