中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (36): 9580-9588.doi: 10.12307/2026.913

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

班氏促卵助孕汤改善卵巢玻璃化冷冻氧化应激损伤

刘云佳1，岳晓蕾1，伍黎明1，周黄磊1，兰  想1，崔佳瑜2，白  琳1，廖彦博1，梁  静2，黎明星1，何国珍1，杨素芳2   

  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:2025-11-06 修回日期:2026-03-23 出版日期:2026-12-28 发布日期:2026-05-25
  • 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:刘云佳，1997年生，河南省平舆县人，汉族，广西中医药大学在读硕士，主要从事中西医防治不孕不育的基础研究。 共同第一作者：岳晓蕾，1998年生，河南省洛阳市人，汉族，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(2023BS048)，项目负责人：梁静；广西自然科学基金项目(2024JJA141299)，项目负责人：何国珍；广西中医药大学自然科学研究重点项目(2024ZD001)，项目负责人：何国珍；南宁市良庆区科技计划项目(202408)，项目负责人：杨素芳

Ban’s Culuan Zhuyun Decoction alleviates oxidative stress injury in vitrified ovarian tissue

Liu Yunjia1, Yue Xiaolei1, Wu Liming1, Zhou Huanglei1, Lan Xiang1, Cui Jiayu2, Bai Lin1, Liao Yanbo1, Liang Jing2, Li Mingxing1, He Guozhen1, Yang Sufang2#br#

#br#
  

  1. 1Basic Medical College, Guangxi University of Chinese Medicine, Nanning 530200, Guangxi Zhuang Autonomous Region, China; 2Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine, Nanning 530000, Guangxi Zhuang Autonomous Region, China
  • Received:2025-11-06 Revised:2026-03-23 Online:2026-12-28 Published:2026-05-25
  • 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:Liu Yunjia, MS candidate, 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 Liu Yunjia and Yue Xiaolei contributed equally to this work.
  • Supported by:
    National Natural Science Foundation of China, No. 81960794 (to HGZ); Introduced Doctoral Research Start-up Fund Project of Guangxi University of Chinese Medicine, No. 2023BS048 (to LJ); Guangxi Natural Science Foundation, No. 2024JJA141299 (to HGZ); Key Project of Natural Science Research of Guangxi University of Chinese Medicine, No. 2024ZD001 (o HGZ); Nanning Liangqing District Science and Technology Plan Project, No. 202408 (to YSF)

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摘要：



文题释义：
铁死亡：一种依赖铁离子的、以脂质过氧化为特征的调节性细胞死亡方式，其核心机制是铁依赖的脂质过氧化物积累超过细胞抗氧化能力，导致细胞膜损伤和细胞死亡。铁死亡相关标志物包括亚铁离子、丙二醛、谷胱甘肽、谷胱甘肽过氧化酶4等。肿瘤抑制蛋白p53/溶质载体家族7成员11/谷胱甘肽过氧化酶4信号通路是铁死亡的关键信号通路之一。
卵巢组织玻璃化冷冻：一种通过超快速降温使卵巢组织呈玻璃态固化的低温保存技术，其核心目标是保存女性生育力，尤其适用于肿瘤患者、青春期前女性、延迟生育女性。冷冻损伤机制包括冰晶形成导致的机械损伤、高浓度保护剂的化学毒性、氧化应激、细胞凋亡、铁死亡等。目前玻璃化冷冻的改良策略主要集中于冷冻保护剂配方的改良，而对机体进行提前药物干预和卵巢组织体外培养的改良策略则较少被关注。

背景：玻璃化冷冻导致卵巢组织发生冷冻损伤，而班氏促卵助孕汤对玻璃化冷冻卵巢组织的保护作用及分子机制亟待完善。
目的：探讨班氏促卵助孕汤对小鼠卵巢组织玻璃化冷冻的影响和分子机制。
方法：①玻璃化冷冻前实验：取60只6-8周龄雌性昆明小鼠，随机分为对照Ⅰ组、模型Ⅰ组和班氏促卵助孕汤低、中、高剂量组，连续给药21 d后取出卵巢组织，对照Ⅰ组为新鲜卵巢，其余各组卵巢玻璃化冷冻7 d。②玻璃化冷冻后实验：60只6-8周龄雌性昆明小鼠取出卵巢组织，对照Ⅱ组为新鲜卵巢，其余为玻璃化冷冻7 d后卵巢，随机分为模型Ⅱ组、含药血清低、中、高剂量组，分别于体积分数15%空白大鼠血清、不同剂量班氏促卵助孕汤含药血清中37 ℃体外培养1 h。苏木精-伊红染色观察卵巢形态，TUNEL法检测卵巢细胞凋亡，比色法检测亚铁离子、谷胱甘肽、丙二醛、谷胱甘肽过氧化酶4含量，RT-qPCR和Western blot检测溶质载体家族7成员11、肿瘤抑制蛋白p53、谷胱甘肽过氧化酶4的mRNA及蛋白表达水平。
结果与结论：①与对照Ⅰ、Ⅱ组相比，模型Ⅰ、Ⅱ组卵巢颗粒细胞排列紊乱且部分缺失，TUNEL阳性细胞率、亚铁离子含量、丙二醛含量、肿瘤抑制蛋白p53 mRNA及蛋白表达水平显著升高，谷胱甘肽含量、谷胱甘肽过氧化酶4含量与溶质载体家族7成员11、谷胱甘肽过氧化酶4 mRNA及蛋白表达水平显著降低(P < 0.05)。②与模型Ⅰ、Ⅱ组相比，班氏促卵助孕汤及其含药血清能减少卵巢组织闭锁卵泡数量，显著降低TUNEL阳性细胞率、亚铁离子含量、丙二醛含量、肿瘤抑制蛋白p53 mRNA及蛋白表达水平，显著提高谷胱甘肽含量、谷胱甘肽过氧化酶4含量与溶质载体家族7成员11、谷胱甘肽过氧化酶4 mRNA及蛋白表达水平(P < 0.05)。因此，班氏促卵助孕汤及其含药血清可改善玻璃化冷冻小鼠卵巢组织及卵泡的形态结构，抑制卵巢组织细胞凋亡及铁死亡水平，从而改善卵巢组织冷冻损伤，其机制可能与抑制肿瘤抑制蛋白p53/溶质载体家族7成员11/谷胱甘肽过氧化酶4信号通路的异常激活有关。  
https://orcid.org/0009-0002-8603-6890(刘云佳)；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)



中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 班氏促卵助孕汤, 卵巢组织玻璃化冷冻, 冷冻损伤, 铁死亡, 生育力保存, 肿瘤抑制蛋白p53, 溶质载体家族7成员11 (SLC7A11), 谷胱甘肽过氧化酶4, 信号通路

Abstract: BACKGROUND: Vitrification causes cryoinjury to ovarian tissue. The protective effect and molecular mechanism of Ban’s Culuan Zhuyun Decoction on vitrified ovarian tissue urgently need to be further elucidated.  
OBJECTIVE: To investigate the effect and molecular mechanism of Ban’s Culuan Zhuyun Decoction on vitrified mouse ovarian tissue.  
METHODS: (1) Pre-vitrification experiment: Sixty female Kunming mice, 6-8 weeks old, were randomly divided into control group I, model group I, and low, medium-, and high-dose Ban’s Culuan Zhuyun Decoction groups. After continuous administration for 21 days, ovarian tissues were collected. Ovaries in the control group I were fresh, while those in the other groups were vitrified for 7 days. (2) Post-vitrification experiment: Ovarian tissues were collected from another 60 female Kunming mice, 6-8 weeks old. Ovaries in the control group II were fresh. The remaining ovaries, after vitrification for 7 days, were randomly divided into model group II and low-, medium-, and high-dose Ban’s Culuan Zhuyun Decoction-containing serum groups, and then cultured in vitro at 37 °C for 1 hour in medium containing 15% blank rat serum or different doses of Ban’s Culuan Zhuyun Decoction-containing serum. Ovarian morphology was observed by hematoxylin-eosin staining. Ovarian cell apoptosis was detected by TUNEL assay. The levels of ferrous ion glutathione, malondialdehyde, and glutathione peroxidase 4 were measured by colorimetric methods. The mRNA and protein expression levels of solute carrier family 7 member 11, tumor suppressor protein p53, and glutathione peroxidase 4 were detected by RT-qPCR and western blot, respectively.  
RESULTS AND CONCLUSION: (1) Compared with control groups I and II, model groups I and II showed disorganized and partially absent ovarian granulosa cells, a significant increase in TUNEL-positive cell rate, ferrous ion content, malondialdehyde content, and tumor suppressor protein p53 mRNA and protein expression levels, as well as a significant reduction in glutathione content, glutathione peroxidase 4 content, and solute carrier family 7 member 11 and glutathione peroxidase 4 mRNA and protein expression levels (P < 0.05). (2) Compared with model groups I and II, Ban’s Culuan Zhuyun Decoction and its drug-containing serum reduced the number of atretic follicles in ovarian tissue, significantly decreased the TUNEL-positive cell rate, ferrous ion content, malondialdehyde content, and tumor suppressor protein p53 mRNA and protein expression levels, and significantly increased glutathione content, glutathione peroxidase 4 content, and solute carrier family 7 member 11 and glutathione peroxidase 4 mRNA and protein expression levels (P < 0.05). Therefore, Ban’s Culuan Zhuyun Decoction and its drug-containing serum can improve the morphological structure of ovarian tissue and follicles in vitrified mice, inhibit ovarian cell apoptosis and ferroptosis, thereby alleviating cryoinjury in ovarian tissue. Its mechanism may be related to inhibiting the abnormal activation of the tumor suppressor protein p53/solute carrier family 7 member 11/glutathione peroxidase 4 signaling pathway.  

Key words: Ban’s Culuan Zhuyun Decoction, ovarian tissue vitrification, cryoinjury, ferroptosis, fertility preservation, tumor suppressor protein p53, solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4, signaling pathway

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