热休克预处理间充质干细胞移植治疗化疗性卵巢早衰的效果

Experiment of transplanting the heat shock pretreated MSCs to premature ovary caused by chemotherapy

目的探讨热休克预处理间充质干细胞(MSCs)对环磷酰胺损伤大鼠卵巢的修复作用。方法分离、培养大鼠骨髓MSCs,移植前以42℃预处理1 h。实验分为4组:正常组、模型组、MSCs组、热休克组,每组25只大鼠。正常组不处理,其余3组建立化疗性卵巢早衰动物模型,MSCs组大鼠双侧卵巢内注射1×106个MSCs,热休克组大鼠双侧卵巢内注射1×106个热休克预处理的MSCs。阴道涂片监测大鼠的动情周期,以化学发光法检测雌二醇(E2),放射免疫法检测卵泡刺激素(FSH)。每组在建模后第1、15、30、45、60天采血及各组分别处死5只大鼠,观察卵巢重量、卵巢结构及各级卵泡数。结果热休克组大鼠比MSCs组更快恢复动情周期;热休克组、MSCs组的卵巢重量、卵泡总数及各级卵泡数与模型组相比差异有统计学意义(P<0.05),热休克组与MSCs组间相比亦差异有统计学意义(P<0.05);移植后第30、45、60天,MSCs组、热休克组E2浓度、FSH浓度与模型组相比,差异有统计学意义(P<0.05),热休克组与MSCs组之间比较差异亦有统计学意义(P<0.05)。结论热休克预处理MSCs对化疗性卵巢早衰具有更强的修复作用。

Objective To investigate the repair effect of mesenchymal stem cells（ MCSs） pre-treated by thermal stress on cyclophosphamide-induced ovarian damage. Methods Bone marrow-derived MSCs were isolated from Wistar rats and cultured in vitro and exposed to 42℃ for 1 hour before transplantation. The experiment consisted of 4 groups,control group,model group,MCSs group and heat shock group（ n = 25）. Chemotherapy-induced premature ovarian failure models were established in model group,MCSs group and heat shock group. MCSs were transplanted into both ovaries of rats in MCSs group,while MCSs pre-treated by thermal stress were transplanted into both ovaries of rats in heat shock group. Vaginal smear was used to determinate the estrous cycle. The E2 was assessed by chemiluminescence immunoassay. The FSH was detected by radioimmunoassay. The blood samples were collected. Each 5 rats in every group were sacrificed 1,15,30,45 and 60 days after animal model establishment. The ovaries were collected to recorded their weight,structures and follicles of different phases. Results Rats in heat shock group recycled its estrous cycle more quickly than MCSs group. There were significant differences in ovarian weight,structures and follicles of different phases among MCSs group,heat shock group and model group. There were significant differences in levels of E2 and FSH among MCSs group,heat shock group and model group 30,45 and 60 days after transplantation. Conclusion MCSs pre-treated by thermal stress have stronger repair effect on chemotherapy-induced premature ovarian in rats.