脐带间充质干细胞移植对卵巢早衰模型小鼠卵巢功能的影响

Effects of transplantation of umbilical cord mesenchymal stem cells on ovarian function in mice with premature ovarian failure

目的探讨人脐带间充质干细胞(hUCMSCs)移植小鼠卵巢修复化学治疗药物诱导的卵巢早衰(POF)。方法选择雌性ICR小鼠40只,鼠龄8周,体质量25~30 g。将ICR小鼠采用SPSS生成随机数字分为空白对照组、POF组、h UCMSCs组。空白对照组不做任何处理,POF组与hUCMSCs组每天腹腔注射环磷酰胺(CTX)70 mg/mL,连续15 d,建立POF模型。建模成功后hUCMSCs组小鼠经尾静脉注射hUCMSCs进行治疗,POF组注射同等剂量的0.9%氯化钠溶液(生理盐水)。观察小鼠一般情况及体质量,并在移植后的7 d、14 d测定血清中雌激素(E2)、卵泡刺激素(FSH)、抗苗勒管激素(AMH)含量及观察卵巢组织形态改变。结果建模完成后,与空白对照组比较,POF组和hUCMSCs组血清E2、AMH均都降低,血清FSH升高,其差异均有统计学意义(P <0.05);hUCMSCs移植后7 d,与POF组比较[(94.39±14.99) mIU/mL、(60.92±14.60) pg/mL、(2 870.00±801.74) pg/mL],hUCMSCs组血清FSH[(80.15±13.36) mIU/mL]、血清E2[(72.38±8.12) pg/mL]差异均无统计学意义(P> 0.05),血清AMH[(3 514.50±833.93) pg/mL]差异也无统计学意义(P> 0.05)。hUCMSCs移植后14 d,与POF组比较[(2 224.50±117.08) pg/mL、(90.00±10.48) mIU/mL、(53.77±7.61) pg/mL],hUCMSCs组血清AMH[(2 934.16±210.62) pg/mL]升高,血清FSH[(77.52±6.477) mIU/mL]降低,差异均有统计学意义(P <0.05);而血清E2[(60.73±3.79) pg/mL]比较,差异无统计学意义(P> 0.05)。苏木精-伊红(HE)染色结果显示:hUCMSCs移植后,与POF组比较,hUCMSCs组原始卵泡、初始卵泡、次级卵泡均有所上升,闭锁卵泡减少,差异有统计学意义(P <0.05)。POF组小鼠子宫内膜发育不良,间质血管较少、腺体数量少及结构混乱,移植后血管数量明显改善。结论 hUCMSCs移植治疗POF可改善小鼠血清E2、AMH的水平,降低血清FSH的水平,促进卵泡的生长,修复卵巢内分泌功能。

Objective To investigate the effect of human umbilical cord mesenchymal stem cells(hUCMSCs) transplanted into mice ovaries for repairing chemotherapy induced premature ovarian failure(POF). Methods Forty female Institute of Cancer Research(ICR) mice, aged 8-week with body mass of 25-30 g, were selected. The ICR mice were randomly divided into blank control group, POF group and hUCMSCs group using SPSS. The blank control group without any treatment, POF group and h UCMSCs group were intraperitoneally injected cyclophosphamide(CTX) with 70 mg/mL per day for 15 days to establish POF model. After successful modeling, mice in hUCMSCs group were injected with hUCMSCs via tail vein, and POF group were injected with the same dose of 0.9 % sodium chloride solution(normal saline). The general condition and body mass of mice were observed. The serum level of estrogen(E2), follicle stimulating hormone(FSH), and anti mullerian hormone(AMH) were measured on the 7 th and 14 th days after transplantation, and morphological changes of ovarian tissue were observed. Results After modeling, compared with blank control group, serum E2 and AMH of POF group and hUCMSCs group decreased but serum FSH increased, and the differences were statistically significant(P < 0.05). Seven days after hUCMSCs transplantation, com-pared with POF group[(94.39 ± 14.99) mIU/mL,(60.92 ± 14.60) pg/mL,(2 870.00 ± 801.74) pg/mL], serum FSH[(80.15 ±13.36) mIU/mL], serum E2[(72.38 ± 8.12) pg/mL] and serum AMH[(3 514.50 ± 833.93) pg/mL] in hUCMSCs group had no statistical significant difference with their counterparts(P > 0.05). Fourteenth days after hUCMSCs transplantation, compared with POF group[(2 224.50 ± 117.08) pg/mL,(90.00 ± 10.48) mIU/mL,(53.77 ± 7.61) pg/mL], serum AMH[(2 934.16 ± 210.62) pg/mL]of h UCMSCs group increased, serum FSH[(77.52 ± 6.477) mIU/mL] decreased, and the differences were statistically significant(P < 0.05);while, serum E2 of hUCMSCs group[(60.73 ± 3.79) pg/mL] was no statistically different with its counterpart(P > 0.05).The results of hematoxylin-eosin(HE) staining showed that compared with POF group, after hUCMSCs transplantation, primordial follicles, primary follicles, and secondary follicles were increased and atretic follicles decreased, the differences were statistically significant(P < 0.05). In POF group, endometrium of mice was poorly developed with fewer interstitial blood vessels and glands, and the structural was chaotic, and the number of blood vessels was significantly increased after transplantation.Conclusion It is demonstrated that hUCMSCs transplantation for POF could improve the levels of serum E2 and AMH,reduced levels of serum FSH, which promote follicle growth and repair ovarian endocrine function.