人脐带间充质干细胞联合高压氧对模拟高原环境下小鼠卵巢功能损伤的修复作用研究  

Study on the repair of ovarian tissue injury in mice in simulated plateau environment by human umbilical cord mesenchymal stem cell transplantation combined with high-pressure medical treatment

作　　者：杜静 常瑞亚 张宗翠 王玲 Du Jing;Chang Ruiya;Zhang Zongcui;Wang Ling(Reproductive Center of the 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army,Lanzhou 730050,China;First Clinical Medical College of Gansu University of Chinese Medicine,Lanzhou 730000,China;Reproductive Center of the 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army,Gansu Key Laboratory of Stem Cell and Gene Medicine,Lanzhou 730050,China)

机构地区：[1]中国人民解放军联勤保障部队第九四〇医院生殖医学中心,兰州730050 [2]甘肃省中医药大学第一临床医学院,兰州730000 [3]中国人民解放军联勤保障部队第九四〇医院生殖医学中心,甘肃省干细胞与基因药物重点实验室,兰州730050

出　　处：《中华生殖与避孕杂志》2025年第2期183-195,共13页Chinese Journal of Reproduction and Contraception

基　　金：军队后勤科研项目(2023HQZZ-04)。

摘　　要：目的探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)联合高压氧(hyperbaric oxygen,HBO)对模拟高原环境下小鼠卵巢功能损伤的修复作用及作用机制。方法按照随机数字表法将64只6周龄C57BL/6雌鼠随机分为模型组、hUCMSCs组、hUCMSCs+HBO组和对照组,每组16只。其中模型组、hUCMSCs组、hUCMSCs+HBO组小鼠放置在低压氧舱模拟海拔6500 m高原环境中持续饲养15 d,构建卵巢功能损伤模型。从造模成功后第1天开始给予对照组与模型组0.2 mL生理盐水尾静脉注射,hUCMSCs组、hUCMSCs+HBO组给予0.2 mL/只hUCMSCs(1×10^(6)个细胞)尾静脉注射,1次/周,共3次,持续干预15 d,hUCMSCs+HBO组每天予以HBO。记录小鼠一般情况、体质量、动情周期变化、性激素水平、双侧卵巢湿重、卵巢指数、各级卵泡计数情况、卵巢组织病理变化及超微结构改变,Western blotting检测卵巢组织中转化生长因子β1(transforming growth factor beta 1,TGF-β1)、Smad家族成员3(Smad family member 3,Smad3)蛋白的表达,以及各组小鼠产仔数、子代体质量,从而评估各组小鼠生育能力。结果①与hUCMSCs组比较,hUCMSCs+HBO组小鼠动情周期、体质量、双侧卵巢湿重、卵巢指数差异均无统计学意义(均P>0.05);血清抗苗勒管激素(anti-Müllerian hormone,AMH)、雌二醇、孕酮、卵泡刺激素(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)水平及各级卵泡计数差异均无统计学意义(均P>0.05),但hUCMSCs+HBO组卵巢组织颗粒细胞的核膜完整、个别线粒体轻度肿胀、粗面内质网无扩张、高尔基体无肿胀等,TGF-β1、Smad3蛋白表达水平均升高(P=0.010,P<0.001),子代情况及窝仔数差异均无统计学意义(均P>0.05)。②与对照组比较,hUCMSCs+HBO组小鼠体质量、双侧卵巢湿重、卵巢指数、血清AMH、雌二醇、孕酮水平略低,血清FSH、LH水平稍偏高,但差异均无统计学意义(均P>0.05),各级卵泡ObjectiveTo investigate the reparative effects and underlying mechanisms of human umbilical cord mesenchymal stem cells(hUCMSCs)in conjunction with hyperbaric oxygen(HBO)on ovarian dysfunction in mice exposed to simulated high-altitude conditions.MethodsA total of 64 six-week-old female C57BL/6 mice were randomly allocated according to random number table,assigned to control group,model group,hUCMSCs group and hUCMSCs+HBO group,with 16 mice in each group.Mice in model group,hUCMSCs group and hUCMSCs+HBO group were placed in a hypobaric oxygen chamber to simulate a high-altitude environment at an elevation of 6500 m for 15 d,thereby establishing a model of ovarian dysfunction.Beginning on the first day following model establishment,the control and model groups received intravenous injections of 0.2 mL saline via the tail vein.In contrast,the hUCMSCs group and the hUCMSCs+HBO group received 0.2 mL hUCMSCs(1×10^(6)cells)per mouse through tail vein injection,administered once a week for three weeks,with continuous intervention lasting 15 d.Furthermore,the hUCMSCs+HBO group were subjected to daily hyperbaric oxygen treatment.The recorded variables included general health status,body weight,estrous cycle changes,serum hormone levels,bilateral ovarian wet weight,ovarian index,follicular development assessment,pathological alterations of ovarian tissue,ultrastructural changes,and the expression levels of transforming growth factor beta 1(TGF-β1)and Smad family member 3(Smad3)detected by Western blotting in ovarian tissue.Additionally,litter size and offspring body weight were measured across all groups to evaluate the reproductive capacity of the mice.Results1)Compared with the hUCMSCs group,the hUCMSCs+HBO group exhibited no statistically significant differences in estrous cycle,body weight,bilateral ovarian wet weight,or ovarian index(all P>0.05).Furthermore,serum levels of anti-Müllerian hormone(AMH),estradiol,progesterone,follicle-stimulating hormone(FSH),and luteinizing hormone(LH),as well as counts of various st

关 键 词：高原 卵巢损伤 高压氧治疗 人脐带间充质干细胞 联合治疗 生育力保护 

分 类 号：R711.75[医药卫生—妇产科学]