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目的 探讨人脐带间充质干细胞外泌体(HUMSCs-EXOs)对早发性卵巢功能不全(POI)模型小鼠的治疗效果及治疗时机。方法 采用腹腔注射环磷酰胺(CTX,120 mg/kg)建立POI小鼠模型。选取动情周期正常的C57B6雌性小鼠60只随机分为模型组、治疗组(注射HUMSCs-EXOs 2μg/g, 1次/d)[包括预防组(造模前)、早期治疗组(造模后第1～7天)、中期治疗组(造模后第8～14天)、晚期治疗组(造模后第15～21天)]和对照组，每组10只。造模成功后第28天将小鼠麻醉，眼球取血后处死小鼠取双侧卵巢。ELISA法检测小鼠血清生殖激素水平，微板法检测血清抗氧化指标水平，HE染色观察小鼠卵巢结构形态变化，免疫组织化学法检测SIRT1蛋白表达情况。每组随机选取4只小鼠与雄性小鼠以2∶1比例进行合笼饲养21 d,记录生育子代数量。结果 与对照组相比，POI组小鼠建模成功。与POI组相比，治疗组的小鼠一般情况好转，体重增加较多，抗苗勒管激素(AMH)、血清雌二醇(E2)、LH水平上升，FSH水平下降，抗氧化应激因子谷胱甘肽过氧化物酶(GSHPx)、超氧化物歧化酶(SOD)上升，卵巢指数升高，卵巢正常卵泡数量增多，闭锁卵泡数量减少，SIRT1蛋白在卵巢的表达增多，差异均有统计学意义(P<0.05)。4个治疗组间比较显示，早、中期治疗组在一般情况、体重增加、性激素水平、氧化应激水平、卵巢指数、卵巢正常卵泡和SIRT1蛋白表达方面均显著优于治疗晚期组和预防组(P<0.05)。结论 HUMSCs-EXOs能够改善POI小鼠的卵巢功能，可能与氧化应激有关，且早期进行HUMSCs-EXOs治疗的效果更好。
Abstract：Objective:To investigate the therapeutic effect and timing of human umbilical cord mesenchymal stem cell exosomes(HUMSCs-EXOs) on model mice with premature ovarian insufficiency(POI).Methods:A mouse model of POI was established by intraperitoneal injection of cyclophosphamide(CTX,120 mg/kg). Sixty female C57B6 mice with normal estrous cycle were randomly divided into model group, treatment group(injection of 2 μg/g HUMSCs-EXOs once a day) [including prevention group(before modeling),early treatment group(day 1 to 7 after modeling),middle treatment group(day 8 to 14 after modeling),late treatment group(day 15 to 21 after modeling)] and control group, with 10 mice in each group. On the 28th day after the successful modeling, the mice were anesthetized, blood was taken from the eyeballs, and the mice were killed to take bilateral ovaries. Serum reproductive hormone levels were detected by enzyme-linked immunosorbent assay(ELISA),serum antioxidant index levels were detected by microplate method, ovarian structural and morphological changes were observed by hematoxylin and eosin(HE) staining, and SIRT1 protein expression level was detected by immunohistochemistry. Four mice in each group were randomly selected and kept in a cage for 21 days with male mice at a ratio of 2∶1,and the number of offspring was recorded.Results:The POI group mice showed successful modeling compared to the control group. Compared with the POI group, the mice in the treatment group were generally better, with more weight gain, increased hormone levels of anti-Müllerian hormone(AMH),serum estradiol(E2),and LH,decreased hormone levels of FSH,increased antioxidant stress factors in terms of glutathione peroxidase(GSHPx) and superoxide dismutase(SOD),increased ovarian index, increased number of normal ovarian follicles, decreased number of atretic follicles, and increased expression level of SIRT1 protein in the ovary, all showing statistical significance(P<0.05). The comparison between the four treatment groups showed that the mice in the early and middle treatment groups were significantly better than those of the late treatment group and the prevention group in terms of general condition, weight gain, sex hormone level, oxidative stress level, ovarian index, the amount of normal ovarian follicles and SIRT1 protein expression level(P<0.05).Conclusions:HUMSCs-EXOs can improve the ovarian function of POI mice, which may be related to oxidative stress, and early treatment with HUMSCs-EXOs has a better effect.
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基本信息:
中图分类号:R711.75;R-332
引用信息:
[1]刘利贞,刘萍,周超,等.人脐带间充质干细胞外泌体对POI小鼠模型的治疗效果及治疗时间窗的探讨[J].生殖医学杂志,2025,34(06):817-825.
基金信息:
海南省自然科学基金高层次人才项目(821RC732); 儋州市科技和工业信息发展局项目(儋科工信{2024}79号); 山东省医药卫生科技项目(202405010551); 山东省中医药科技项目(2020M085); 滨州医学院“临床+X”项目(BY2021LCX26)
2024-12-22
2024
2025-02-28
2025-05-11
2025
1
2025-06-15
2025-06-15