Liu Mengyu, Zhu Hanfei, Zhou Xiaowei, Duan Jingru, Shen Yuhong, Zhang Aijun
Reproductive Medical Center, Department of Obstetrics and Gynecology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 197 Ruijin 2nd Road, Shanghai, 200025, China.
State Key Laboratory of Female Fertility Promotion, Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China.
Stem Cell Res Ther. 2025 May 30;16(1):269. doi: 10.1186/s13287-025-04383-6.
Age-related diminished ovarian reserve (DOR) leads to declining fertility, miscarriage, and systemic health issues. Cell-free fat extract (CEFFE) has demonstrated therapeutic effects in various tissues. In our previous study, CEFFE successfully improved ovarian function in mice without adverse effects; however whether it can prevent DOR remains unclear. This study aimed to determine if early intervention with CEFFE can delay DOR and extend reproductive lifespan, exploring its potential as a novel clinical approach for women with DOR.
The mice in the Prophylactic group received tail vein injections of 200 μL of CEFFE per mouse every 3 days for three months; the Control group were administered an equivalent volume of saline injections. Post-treatment outcomes assessed included body weight, ovarian weight, follicle count, embryo quality, production rates, and levels of serum hormones. Safety was evaluated via organ weights and hematoxylin and eosin staining in parents and offspring. The impact of CEFFE on cell proliferation, hormone synthesis, oxidative stress, and senescence was assessed via Cell counting Kit-8 assays, enzyme-linked immunosorbent assays, and reverse transcription quantitative real-time PCR, 1,1',3,3'-tetraethyl-5,5',6,6'-tetrachloroimidacarbocyanine iodide and Senescence-associated beta-galactosidase staining.
Compared with the Control group, CEFFE treatment effectively preserved ovarian function in aging mice, improving ovarian weight, hormone levels, and follicular development, and reduced follicular atresia. CEFFE treatment enhanced embryo quality and development, induced a higher pregnancy rate, reduced the number of abnormal pregnancies, and increased the litter size and the number of live births. CEFFE demonstrated favorable safety in the Prophylactic group and their offspring, with no significant adverse effects on organ morphology or coefficients, except for increased liver weight in treated mice. Transcriptomic analysis suggested CEFFE influences cell proliferation, hormone response, and oxidative stress pathways in ovarian granulosa cells, which was verified in primary mouse granulosa cells. In vitro studies demonstrated that CEFFE pre-treatment alleviated the phenotype of Control mice by inhibiting oxidative stress, promoting proliferation, and enhancing hormone secretion.
Early prophylactic administration of CEFFE in adult mice can prevent DOR and extend their reproductive lifespan by inhibiting granulosa cell senescence.
年龄相关的卵巢储备功能减退（DOR）会导致生育能力下降、流产和全身健康问题。无细胞脂肪提取物（CEFFE）已在多种组织中显示出治疗效果。在我们之前的研究中，CEFFE成功改善了小鼠的卵巢功能且无不良反应；然而，它是否能预防DOR仍不清楚。本研究旨在确定早期使用CEFFE干预是否能延缓DOR并延长生殖寿命，探索其作为DOR女性新型临床治疗方法的潜力。
预防组小鼠每3天接受一次尾静脉注射，每只小鼠注射200μL CEFFE，共注射三个月；对照组注射等量的生理盐水。评估治疗后的结果包括体重、卵巢重量、卵泡计数、胚胎质量、产仔率和血清激素水平。通过对亲代和子代的器官重量以及苏木精和伊红染色评估安全性。通过细胞计数试剂盒-8检测、酶联免疫吸附测定、逆转录定量实时聚合酶链反应、1,1',3,3'-四乙基-5,5',6,6'-四氯咪唑羰花青碘化物和衰老相关β-半乳糖苷酶染色评估CEFFE对细胞增殖、激素合成、氧化应激和衰老的影响。
与对照组相比，CEFFE治疗有效保留了衰老小鼠的卵巢功能，改善了卵巢重量、激素水平和卵泡发育，并减少了卵泡闭锁。CEFFE治疗提高了胚胎质量和发育，诱导了更高的妊娠率，减少了异常妊娠的数量，并增加了窝产仔数和活产仔数。CEFFE在预防组及其子代中显示出良好的安全性，除了治疗小鼠的肝脏重量增加外，对器官形态或系数没有显著不良影响。转录组分析表明，CEFFE影响卵巢颗粒细胞中的细胞增殖、激素反应和氧化应激途径，这在原代小鼠颗粒细胞中得到了验证。体外研究表明，CEFFE预处理通过抑制氧化应激、促进增殖和增强激素分泌减轻了对照组小鼠的表型。
成年小鼠早期预防性给予CEFFE可通过抑制颗粒细胞衰老来预防DOR并延长其生殖寿命。
