脂肪间充质干细胞对小鼠化疗性卵巢功能不全的治疗作用及机制研究

Study on the therapeutic effect and mechanism of adipose mesenchymal stem cells on chemotherapy-induced premature ovarian insufficiency in mice

目的探讨脂肪间充质干细胞(adipose-derived mesenchymal stem cells,mADSCs)对化疗药物表柔比星诱导的化疗性卵巢功能不全(premature ovarian insufficiency,POI)的影响及其作用机制。方法将动情期同步的雌性昆明白小鼠分为空白对照组、模型组(POI组)和治疗组(m ADSC1、m ADSC2),观察小鼠的动情周期;卵泡刺激素(follicle-stimulation hormone,FSH)水平;计数各级卵泡数量;RT-qPCR及蛋白印迹法检测各组小鼠卵巢组织中相关基因及Bax、Casepase3、Casepase9蛋白的表达情况。结果尾静脉或卵巢原位注射mADSCs移植后,与POI组比较,治疗组小鼠生存质量提高,血清AMH、E_(2)值显著升高,FSH值显著降低(P均<0.05);与POI组比较,治疗组小鼠卵巢切片中初级、次级卵泡数量明显增多,闭锁卵泡数显著减少(P均<0.05);治疗组Gdf9、Figla、Foxl2、Inhibin的表达明显增加,Bax、Casepase3、Casepase9的表达明显减少;与POI组比较,治疗组Bax、Casepase3、Casepase9的表达显著下降,其中卵巢原位注射组优于尾静脉注射组(P均<0.05)。结论应用表柔比星成功诱导构建POI小鼠模型。移植m ADSCs能够有效地修复表柔比星化疗所致的卵巢功能损害,分子机制可能是促进颗粒细胞的增殖、分化,抑制其凋亡,促进卵泡发育。

Objective To explore the therapeutic effect of adipose mesenchymal stem cell transplantation on premature ovarian insufficiency(POI)induced by epirubicin chemotherapy.Methods Female Kunming mice with synchronous estrus were divided into blank control group,model group(POI group)and treatment group(mADSC1,m ADSC2).The estrous cycle of mice was observed,follicle-stimulation hormone(FSH)in serum was measured,the number of follicles at all levels was counted,and the expression of related genes and Bax,Casepase3 and Casepase9 proteins in ovarian tissue of mice in each group was detected.Results After in situ injection of mADSCs into caudal vein or ovary,the quality of life in the treatment group was higher than that in the POI group,the serum AMH and E_(2) levels in the treatment group were significantly higher than those in POI group,while the FSH values in the treatment group were significantly lower than those in the POI group(P<0.05).Compared with POI,the number of primary and secondary follicles in ovarian sections increased significantly in the treatment group,the expression of Gdf9,Figla,Foxl2 and Inhibin was significantly increased,while the expression of Bax,Casepase3 and Casepase9 was significantly decreased in the treatment group.Western blotting showed that the expression of Bax,Casepase3 and Casepase9 in the treatment group was significantly lower than that in the POI group,and the ovarian in situ injection group was better than the tail vein injection group.Conclusions The POI mouse model was successfully induced by epirubicin.Transplantation of mADSCs can effectively repair ovarian function damage caused by epirubicin chemotherapy.The molecular mechanism may be to promote the proliferation and differentiation of granulosa cells,inhibit their apoptosis,and promote follicular development.