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。
Objective To explore the therapeutic potential of miR-21 in rat model of chemotherapy-induced premature ovarian failure （POF）. Methods Lentivirus-mediated miR-21 （LV-miR-21） was constructed successfully in vitro with molecular biology methods. Rats were divided into 4 groups named control group, model group, blank vector group and miR-21 group. Rat models of chemotherapy-induced POF were established in the latter 3 groups by intraperitoneal injection of cytoxan （CTX）. Bilateral ovaries of rats in miR-21 group were iniected with LV-miR-2 lp of rats in blank vector group were iniected with lentivirus vector, and rats in model group received no treatment. At 1, 15~ 30, 45 and 60 days after the last injection, blood sample was collected, the rats were then sacrificed and the ovaries were removed. The estrous cycle was observed by vaginal smears. The E2 level was detected by chemiluminescent immunoassay, and the follicle stimulating hormone （FSH） level was detected by homologous double antibody radiation immunoassay. Ovary weights were measured, and the follicle count was conducted through observing paraffin section under microscope. The apoptosis of ovarian granulosa cells was analyzed by TUNEL assay. Results During 15-30 days, 30-45 days and 45-60 days after the last injection, regular estrous cycle was recovered respectively in 8, 5 and 3 rats in miR-21 group. At the 15th, 30th, 45th and 60th day after the last injection, the E2 level was higher in miR-21 group than in model group and blank vector group, but the FSH level showed the opposite trend （P=0.000）. At the 45th and 60th day after the last injection, the follicle numbers at all stages increased markedly in miR-21 group than in model group and blank vector group （P=0.000）. At the 30th, 45th and 60th day after the last iniection, the ovary weights were higher in miR-21 group than in model group and blank vector group. At the 1Sth, 30th, 45th and 60th day after the last injection, the apoptosis rate of ovarian granulosa cell were signif
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2017年第9期759-763,共5页
Medical Journal of Chinese People's Liberation Army
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)~~
