慢病毒介导的miR-21对化疗所致大鼠卵巢早衰的治疗作用

发布时间：2018-04-24 07:43

本文选题：microRNA- + 卵巢早衰　；参考：《解放军医学杂志》2017年09期

【摘要】：目的探讨miR-21在化疗所致的卵巢早衰大鼠模型中的治疗潜能。方法采用分子生物学方法成功构建慢病毒介导的miR-21(LV-miR-21)载体。100只成年雌性Wistar大鼠随机分为:空白对照组、模型组、空载组以及miR-21组,每组25只。后3组采用环磷酰胺(CTX)腹腔注射构建化疗所致卵巢早衰大鼠模型,空载组及miR-21组于建模后分别在双侧卵巢注射空载体及LV-miR-21,空白对照组则注射生理盐水。于注射后第1、15、30、45、60天分批处死大鼠,留取血及卵巢标本。通过阴道脱落细胞涂片检测大鼠动情周期。采用化学发光法、放射免疫法检测雌二醇(E2)及卵泡刺激素(FSG)水平;测定卵巢重量、进行卵泡计数;采用TUNEL法测定卵巢颗粒细胞凋亡率。结果注射完成后15~30d、30~45d及45~60d,miR-21组分别有8、5、3只大鼠恢复规律的动情周期;注射完成后第15、30、45、60天,miR-21组的E2浓度高于模型组、空载组,而FSH浓度低于模型组及空载组(P=0.000);注射完成后第30、45、60天,miR-21组卵巢重量大于模型组及空载组(P=0.000);注射完成后第45、60天,miR-21组各级卵泡数目均高于模型组和空载组,但仍低于空白对照组(P=0.000);注射完成后第15、30、45、60天,miR-21组卵巢颗粒细胞凋亡率低于模型组和空载组(P=0.000)。结论上调miR-21能够部分修复CTX损伤的卵巢结构及功能。
[Abstract]:Objective to investigate the therapeutic potential of miR-21 in the model of premature ovarian failure induced by chemotherapy. Methods A total of 100 adult female Wistar rats were randomly divided into three groups: blank control group, model group, no-load group and miR-21 group, with 25 rats in each group. The model of premature ovarian failure induced by chemotherapy was established by intraperitoneal injection of Cyclophosphamide (CTX) in the latter three groups. The empty vector and LV-miR-21 were injected into both ovaries in the no-load group and miR-21 group, respectively, and normal saline was injected into the blank control group. The rats were killed in batches on day 1, 15, 30, 45 and 60 after injection, and blood and ovary samples were collected. The estrous cycle of rats was detected by smear of vaginal exfoliated cells. The levels of estradiol E _ 2 (E _ 2) and follicle stimulating hormone (FSG) were measured by chemiluminescence and radioimmunoassay. The ovarian weight and follicle count were measured. The apoptosis rate of ovarian granulosa cells was measured by TUNEL method. Results there were three regular estrus recovery cycles in the miR-21 group at 15: 30 and 60 days after injection, and the E _ 2 concentration in the miR-21 group was higher than that in the model group and the no-load group. However, the concentration of FSH was lower than that of the model group and the no-load group, and the ovarian weight of the control group was higher than that of the model group and the no-load group on the 60th day after the injection, and the number of follicles of all levels of the group was higher than that of the model group and the no-load group on the 450-60th day after the injection, and the ovarian weight of the control group was higher than that of the model group and the no-load group on the 60th day after the injection. However, the apoptosis rate of ovarian granulosa cells in the control group was still lower than that in the model group and the no-load group, and the apoptosis rate of ovarian granulosa cells in the control group was lower than that in the model group and the no-load group on the 60th day after injection. Conclusion upregulation of miR-21 can partially repair the ovarian structure and function of CTX.
【作者单位】：南方医科大学珠江医院妇产科;
【基金】：国家自然科学基金青年基金(81300462) 广东省科技计划项目(2013B021800145)~~
【分类号】：R711.75

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