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过表达miR-21的骨髓间充质干细胞对化疗性卵巢早衰的修复作用
作者:汪庆如 陈小莹 李欣然 王清 付霞霏 
单位:510282 广州 南方医科大学珠江医院妇产科(汪庆如、陈小莹、李欣然、王清、付霞霏) 
关键词:miR-21 骨髓间充质干细胞 卵巢早衰 卵巢颗粒细胞 
分类号:R711.75
出版年,卷(期):页码:2019,44(3):203-209
摘要:

[摘要]  目的  探讨过表达miR-21的骨髓间充质干细胞(BMSCs)对化疗性卵巢早衰大鼠模型的修复作用。方法  构建miR-21慢病毒表达载体(LV-miR-21),转染BMSCs构建miR-21-BMSCs。实验分为5组:正常组、模型组、miR-21 组、BMSCs组、miR-21-BMSCs组,每组20只大鼠。大鼠腹腔内注射环磷酰胺(CTX)构建化疗性卵巢早衰大鼠模型。正常组及模型组大鼠双侧卵巢注射生理盐水,其余3组分别注射LV-miR-21BMSCsmiR-21-BMSCs。检测各组大鼠动情周期、雌二醇(E2)及卵泡刺激素(FSH)水平、卵巢重量及各级卵泡计数、卵巢颗粒细胞凋亡率。结果  正常组大鼠动情周期不变,其余4组注射CTX后均出现动情周期紊乱,其中miR-21-BMSCs组恢复动情周期的大鼠数量明显多于其余3组。在注射CTX304560dmiR-21-BMSCs组卵巢重量及各级卵泡计数明显高于模型组、miR-21BMSCs(P<0.05);各组大鼠基础E2FSH水平差异无统计学意义,正常组大鼠E2FSH在实验期间无明显改变,其余4组大鼠E2 浓度下降、FSH浓度升高,miR-21-BMSCs组激素水平与miR-21组及BMSCs组相比差异均有统计学意义[E2F30d=43.10F45d=43.57F60d=44.98P<0.05FSHF30d=14.71F45d=13.16F60d=15.20P<0.05)];与模型组、miR-21组、BMSCs组相比,miR-21-BMSCs组颗粒细胞凋亡率明显下降(F15d=27.20F30d=23.60F45d=21.80F60d=21.20P<0.05)结论  miR-21 能增强BMSCs对化疗性卵巢早衰的治疗作用,显著减轻卵巢结构及功能损伤。

[Abstract]  Objective  To explore the repair effects of bone marrow mesenchymal stem cells (BMSCs) over-expressing miR-21 on chemotherapeutic premature ovarian failure (POF) in rats. Methods  The expression vector of lentivirus-mediated miR-21 (LV-miR-21) was constructed, and BMSCs were transfected with LV-miR-21 to construct miR-21-BMSCs. A hundred rats were divided into 5 groups (20 each): normal group, model group, miR-21 group, BMSCs group and miR-21-BMSCs group. The chemotherapeutic POF model was established by intraperitoneal injection of cyclophosphamide (CTX) in rats of the latter 4 groups. Then LV-miR-21, BMSCs and miR-21-BMSCs were injected respectively into the bilateral ovaries of the rats in the latter 3 groups, and physiological saline was injected into the rats of normal group and model group. At 15, 30, 45 and 60 days after injection, the estrus cycle, estradiol (E2) and follicle stimulating hormone (FSH) level, ovarian weight, follicle count at all levels, and apoptotic rate of granulosa cells in each group were detected. Results  After injection of CTX, the estrus cycle of rats remained regular in normal group, and fell into disorder in the other four groups, of which the number of rats with recovered estrus cycle in the miR-21-BMSCs group was more than the other three groups; The ovarian weight and follicle count at all levels were significantly higher in miR-21- BMSCs group than in model group, miR-21 group and BMSCs group 30, 45 and 60 days after CTX injection (P<0.05); The basal levels of E2 and FSH showed no significant difference in each group during the experiment, while the E2 concentration decreased and FSH concentration increased in all the groups except the normal group. The hormone levels of E2 and FSH in miR-21-BMSCs group (F30d=43.10 and F30d=14.71) were significantly different when compared to those in miR-21 group and BMSCs group (F45d=43.57 and F45d=13.16, P<0.05; and F60d=44.98 and F60d=15.20, P<0.05); The apoptotic rate of ovarian granular cells in miR-21-BMSCs group was significantly lower (F60d=21.20) than that in model group, miR-21 group and BMSCs group (F15d=27.20, F30d=23.60 and F45d=21.80, P<0.05). Conclusion  miR-21 may enhance the therapeutic effect of BMMSCs on chemotherapeutic premature ovarian failure, and significantly improve the ovarian structure and function.

基金项目:
国家自然科学基金青年基金(81300462)
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