过表达 miRNA-320 对顺铂诱导卵巢早衰大鼠卵巢功能的影响

摘要:目的 探讨过表达 miRNA-320 对顺铂诱导卵巢早衰大鼠卵巢功能的影响。 方法 选择清洁级 SD 雌性大鼠32 只,随机分为 4 组:正常对照组、模型组、阴性对照组和 miRNA-320 过表达组,每组 8 只。 在实验过程中,每天观察各组大鼠饮食、活动和体毛等外在表现的差异。 采用 ELISA 检测各组大鼠血清中 E2、FSH、LH 的含量;采用 HE染色,光镜下观察各组大鼠卵巢组织形态学变化;采用 PCR 检测各组大鼠卵巢组织 miRNA-320、PGRMC1、BMP15的表达水平;采用 Western blot 检测各组大鼠卵巢组织中 PGRMC1、BMP15 的蛋白表达水平。 结果 与正常对照组相比,模型组大鼠活动迟缓、无精神、反应迟钝、食欲下降、体质量下降以及体毛暗淡无光泽;组织形态学分析表明,卵巢组织明显萎缩、皮质变薄、闭锁卵泡数目多以及间质纤维组织增多;血清 FSH、LH 显著升高,而血清 E2 及卵巢组织中 PGRMC1、BMP15 的蛋白和 mRNA 表达水平均降低,差异具有统计学意义( P< 0. 05) 。 与阴性对照组相比,miRNA-320 过表达组大鼠活动量增加、反应较为敏捷、进食增多、体毛逐渐有光泽;间质纤维化明显减轻、成熟卵泡数量明显增加、黄体数增多;血清 E2 的含量、卵巢组织中 PGRMC1、BMP15 的蛋白和 mRNA 的表达水平均升高,而FSH、LH 的含量降低( P<0. 05) 。 结论 顺铂可诱导卵巢早衰大鼠卵巢功能障碍,过表达 miRNA-320 可改善顺铂诱导的卵巢早衰大鼠的卵巢功能。

Effect of Overexpression of miRNA-320 on Ovarian Function in Cisplatin Induced Premature Ovarian Failure Rats

Abstract: Objective To investigate the effect of overexpression of miRNA-320 on ovarian function in cisplatin induced premature ovarian failure rats. Method Thirty two SD female rats were randomly divided into 4 groups: normal control group, model group, negative control group and miRNA-320 overexpression group. During the experiment, the differences of diet, activity and body hair were observed every day. The contents of E2, FSH and LH in serum of each group were detected by ELISA. HE staining was used to observe the histological changes of ovaries. The expression levels of miRNA-320, PGRMC1 and BMP15 in ovaries of rats in each group were detected by qRT-PCR. Western blot was used to detect the protein expression of PGRMC1 and BMP15 in ovarian tissue of each group. Result Compared with the normal control group, the rats in the model group were slow in activity and in response, loss of appetite and weight, dull and lusterless body hair; histomorphological analysis showed that the ovarian tissue was obviously atrophic, the cortex became thinner, the number of atretic follicles was more, the interstitial fibrous tissue was more; the serum FSH and LH were significantly increased, while the protein and mRNA levels of PGRMC1 and BMP15 were decreased in serum E2 and ovarian tissues ( P<0. 05) . Compared with the negative control group, the activity of overexpression group was increased, the response was more agile, the food intake was increased, the body hair was gradually glossy; interstitial fibrosis was significantly reduced, the number of mature follicles was significantly increased, the number of corpus luteum was increased; the content of serum E2, the protein and mRNA expression level of PGRMC1 and BMP-15 in ovarian tissue were increased, while the content of FSH and LH were decreased ( P<0. 05) . Conclusion Cisplatin induces ovarian dysfunction in rats with premature ovarian failure; overexpression miRNA-320 improves ovarian function in cisplatin-induced premature ovarian failure rats.