骨髓间充质干细胞移植修复化疗所致卵巢损伤的实验研究

目的:探讨大鼠骨髓间充质干细胞(MSCs)移植对化疗所致卵巢损伤的影响。方法:采用腹腔注射环磷酰胺(CTX)建立化疗所致大鼠卵巢损伤模型,并随机分为模型组、实验对照组及移植组。同龄正常大鼠作为空白对照组。移植组在建模结束后每侧卵巢注射绿色荧光标记约1×106个MSCs。于移植后1、15、30、45天及60天,留取各组血标本,并于后4个时间点分批处死大鼠,留取卵巢标本。荧光显微镜下观察绿色荧光及其分布情况。阴道脱落细胞涂片观察大鼠动情周期的变化;以化学发光法检测雌二醇(E2)浓度,放射免疫法检测卵泡刺激素(FSH)浓度;计数各级卵泡数。结果:移植MSCs后15天,移植组卵巢组织内可见到明亮的绿色荧光,持续至移植后2月。绿色荧光分布在卵巢间质组织。移植组30%(6/20)的大鼠在移植后15~30天内恢复正常的动情周期,在移植后30~60天内又有4只大鼠恢复正常的动情周期。移植后15、30、45天和60天时,移植组E2浓度高于实验对照组相应时间点E2浓度,而FSH浓度则低于实验对照组相应时间点FSH浓度,差异有显著性。移植45、60天时,移植组各级卵泡数目均高于模型组和实验对照组,低于空白对照组。结论:骨髓间充质干细胞可以在环磷酰胺损伤的大鼠卵巢组织中存活、迁移,但其分化为卵泡组分的可能性小。骨髓间充质干细胞移植可修复环磷酰胺损伤的卵巢组织,部分改善卵巢内分泌功能。

In vivo effects of bone marrow derived mesenchymal stem cells transplantation on ovarian recovery from chemotherapy-induced damage

Objective: To investigate the influence of rat bone marrow derived mesenchymal stem cells transplantation on chemotherapy-induced ovarian damage in vivo.Methods:Cyclophosphamide(CTX) was injected i.p.to establish the chemotherapy-induced ovarian damage model.The rats then were randomly divided into 4 groups of 20 rats each,named chemotherapy-induced ovarian damage model group(model group),model group with ovary saline injection(experimental control group),and model group with MSC transplantation(transplantation group).20 normal rats served as controls(normal control group).Immediately after the last injection of CTX,labeled MSCs(2×106 cells in 20μl volume) were injected into the bilateral ovaries of the rats in transplantation group.Immediately after the last CTX injection and 15,30,45 and 60 days after transplantation,1 ml blood was collected from the rats for E2 and FSH examination.At 15,30,45 and 60 days after the transplantation,5 rats from each group were sacrificed to remove the bilateral ovaries.The sections of the ovaries were also examined to track the injected GFP-labeled MSCs.Ovarian follicle populations were determined.Results:Fluorescent cells were found in interstitial tissues of rat ovaries 15 days after labeled MSC transplantation,which was persisted for 2 months.In transplantation group,6/20(30%) of the rats recovered the regular estrous cycle during 15~30 days after MSCs transplantation,and the estrous cycles of 4 rats became normal after 30~60 days.At 15,30,45 and 60 days after transplantation,the E2 levels in the transplantation group were significantly higher than those of the experimental control group.The FSH concentrations in transplantation group at 15,30,45 and 60 days after transplantation were lower than those of the experimental control group.At 45 and 60 days after MSCs transplantation,the numbers of follicles in the transplantation group were higher than those of the model and experimental control group.Conclusion:The transplanted MSCs can survival,migrate in the rat ovaries damaged by CTX.However,it is not likely that these MSCs may differentiate into follicles including oocytes and granulosa cells.Direct injection of MSCs into ovary enhances ovarian endocrine function in rats with chemotherapy-induced ovarian damage.