北京中医药大学学报2024，Vol.47Issue(1)：62-69,8.DOI:10.3969/j.issn.1006-2157.2024.01.010
基于卵母细胞线粒体探究补肾养精汤改善高龄不孕症的作用机制
Explore mechanism of Bushen Yangjing Soup in improving elderly infertility based on oocyte mitochondrion
摘要
Abstract
Objective We aimed to explore the mechanism by which Bushen Yangjing Soup improves elderly infertility based on oocyte mitochondrion.Methods C57BL/6J mice were used,including 15 young females(6-8 weeks old),45 elderly females(10 months old),and ten young males(6-8 weeks old).The female 6-8-week-old young mice were assigned to the young group,and the female 10-month-old elderly mice were randomly divided into the model group,the dehydroepiandrosterone(DHEA)group,and the Bushen Yangjing Soup group.Mice in the DHEA and Bushen Yangjing Soup groups were given DHEA[11.26 mg/(kg·d)]and Bushen Yangjing Soup[11.41 g/(kg·d)].Mice in the other groups were given normal saline.After 45 days,oocytes were collected,the number of oocytes and the proportion of mature oocytes were recorded,and in vitro fertilization(IVF)was carried out,followed by calculation of the successful fertilization rate.Real-time PCR was used to determine the copy number of mitochondrial DNA in oocytes,mito-tracker green was used to observe the distribution of mitochondria,and the mitochondrial membrane potential(MMP)of oocytes was determined using the JC-1 kit.The mRNA and protein levels of mitofusin 2(Mfn2)in oocytes were determined by real-time PCR and Western blotting,respectively.Results Compared with the young group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly reduced(P＜0.01),the copy number of mitochondrial DNA in oocytes was decreased,the rate of abnormal mitochondrial distribution was increased,the MMP was decreased(P＜0.01),and the mRNA and protein levels of Mfn2 were decreased(P＜0.01)in the elderly model mice.Compared with the elderly model group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were significantly increased(P＜0.05),the copy number of mitochondrial DNA in oocytes was increased(P＜0.05),the rate of abnormal mitochondrial distribution was decreased,the MMP was increased(P＜0.05),and the mRNA and protein levels of Mfn2 were increased(P＜0.01)in the Bushen Yangjing Soup group.Compared with the DHEA group,the number of oocytes,the proportion of mature oocytes,and the IVF rate were increased,the copy number of mitochondrial DNA in oocytes was increased,the rate of abnormal mitochondrial distribution was decreased,the MMP was increased,the mRNA and protein levels of Mfn2 were increased in the Bushen Yangjing Soup group,and there was a significant difference in the number of oocytes and the expression of Mfn2 protein(P＜0.05).Conclusion Bushen Yangjing Soup may improve the quantity,distribution,and function of oocyte mitochondria by increasing the mRNA and protein levels of Mfn2,thereby improving oocyte quality.This is one of the possible mechanisms for clinical application of Bushen Yangjing Soup to improve ovarian function and treat elderly infertility.关键词
高龄不孕症/卵母细胞/线粒体/线粒体融合蛋白2/补肾养精汤/小鼠Key words
elderly infertility/oocyte/mitochondrion/mitofusin 2/Bushen Yangjing Soup/mice分类
医药卫生引用本文复制引用
王昭,于昕,武丽丽,刘鹏飞,于潇,刘金星..基于卵母细胞线粒体探究补肾养精汤改善高龄不孕症的作用机制[J].北京中医药大学学报,2024,47(1):62-69,8.基金项目
国家自然科学基金项目(No.82104917) （No.82104917）
山东省自然科学基金项目(No.ZR2019PH053) （No.ZR2019PH053）
山东省中医药科技项目(No.2020Q030,Q-2023009) （No.2020Q030,Q-2023009）
山东省中医药重点学科(鲁卫中医药科教字[2022]4号)National Natural Science Foundation of China(No.82104917) （鲁卫中医药科教字[2022]4号）
Shandong Natural Science Foundation(No.ZR2019PH053) （No.ZR2019PH053）
Shandong TCM Science and Technology Project(Nos.2020Q030 and Q-2023009) （Nos.2020Q030 and Q-2023009）
