基于microRNA探讨补肾养精颗粒对高龄小鼠卵母细胞质量的影响  

Effect of granule for tonifying kidney and replenishing essence on oocyte quality of aged mice based on MicroRNA

作　　者：王昭[1] 付凌宇 武丽丽[1] 刘鹏飞 于潇 刘金星[2] WANG Zhao;FU Lingyu;WU Lili;LIU Pengfei;YU Xiao;LIU Jinxing(Shandong College of Traditional Chinese Medicine,Yantai 264199,Shandong,China;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,Shandong,China)

机构地区：[1]山东中医药高等专科学校,山东烟台264199 [2]山东中医药大学附属医院,山东济南250014

出　　处：《现代中西医结合杂志》2023年第14期1919-1924,1939,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：山东省自然科学基金项目(ZR2019PH053);山东省中医药科技项目(2020Q030,Q-2023009);国家自然科学基金项目(82104917);山东省中医药重点学科(鲁卫中医药科教字〔2022〕4号)。

摘　　要：目的基于颗粒细胞miR-23a和miR-27a探讨补肾养精颗粒对高龄小鼠卵母细胞质量的影响。方法采用C57BL/6J小鼠,雌性6~8周龄10只、10月龄30只,雄性6~8周龄10只。雌性6~8周龄小鼠作为青年组,将10月龄小鼠随机分为高龄组、补肾养精低剂量组、补肾养精高剂量组各10只。补肾养精低、高剂量组分别给予补肾养精颗粒5.5 g/kg、11 g/kg灌胃,青年组和高龄组给予同等体积生理盐水灌胃,均连续干预45 d。收集卵母细胞,记录获卵数、卵母细胞成熟率;进行体外受精,培养至4.5 d,记录体外受精率和囊胚率;用JC-1试剂盒检测卵母细胞线粒体膜电位,用活性氧(ROS)试剂盒检测卵母细胞线粒体ROS含量;RT-PCR法检测颗粒细胞miR-23a和miR-27a mRNA表达量。结果与青年组比较,高龄组小鼠获卵数、卵母细胞成熟率、体外受精率和囊胚率、卵母细胞线粒体膜电位均明显降低(P均<0.05),卵母细胞线粒体ROS含量和颗粒细胞miR-23a、miR-27a mRNA表达量均明显增高(P均<0.05)。与高龄组比较,补肾养精高剂量组小鼠获卵数、卵母细胞成熟率、体外受精率、卵母细胞线粒体膜电位均明显增高(P均<0.05),补肾养精高剂量组卵母细胞线粒体ROS含量和补肾养精低、高剂量组颗粒细胞miR-23a、miR-27a mRNA表达量均明显降低(P均<0.05)。结论补肾养精颗粒可能通过下调miR-23a和miR-27a表达,减少颗粒细胞凋亡,促进颗粒细胞增殖,从而提高卵母细胞质量。Objective It is to explore the effect of granule for tonifying kidney and replenishing essence on oocyte quality of aged mice based on miR-23a and miR-27a in granulosa cells.Methods C57BL/6J mice were used,including 10 females aged 6-8 weeks,30 females aged 10 months,and 10 males aged 6-8 weeks.The female mice aged 6-8 weeks were as young group,and the ones aged 10 months were randomly divided into aged group,low dose and high dose groups of granule for tonifying kidney and replenishing essence.The low dose and high dose groups of granule for tonifying kidney and replenishing essence were given granule for tonifying kidney and replenishing essence 5.5 g/kg and 11 g/kg by gavage,respectively,the young group and the aged group were given the same volume of normal saline by gavage,all the groups were continuously treated for 45 days.The oocytes were collected,the number of oocytes and the rate of mature oocytes were recorded;in vitro fertilization(IVF)was carried out,the culture lasted for 4.5 days,then the IVF rate and blastocyst rate were recorded;the oocyte mitochondrial membrane potential(MMP)of oocyte was detected by JC-1 kit,and the content of reactive oxygen species(ROS)in oocyte mitochondria was detected by ROS kit;the expressions of mRNA of miR-23a and miR-27a in granulosa cells were detected by Real-time PCR.Results Compared with the young group,the number of oocytes,the rate of mature oocytes and the rate of IVF,and the oocyte MMP of mice in the aged group were significantly decreased(all P<0.05),the mitochondrial ROS content in oocytes and the expressions of miR-23a and miR-27a mRNA in granulosa cells were significantly increased(all P<0.05).Compared with the aged group,the number of oocytes,the rate of mature oocytes and the rate of IVF,and the oocyte MMP of the mice in the high dose group of granule for tonifying kidney and replenishing essence were significantly increased(all P<0.05),the mitochondrial ROS content in oocytes of mice in the high dose group of granule for tonifying kidney and replenishin

关 键 词：高龄小鼠 卵母细胞 颗粒细胞 miR-23a miR-27a 补肾养精颗粒 

分 类 号：R-332[医药卫生]