摘要
目的探讨补肾调经方改善体外培养小鼠卵母细胞核质成熟同步提高卵母细胞质量的作用机制。方法日龄10天的雌性ICR小鼠200只,随机分为补肾组(60只)、血清对照组(60只)和空白对照组(80只)。分别处死小鼠分离窦前卵泡,加入补肾调经方大鼠含药血清、正常大鼠血清和不加大鼠血清进行细胞培养,结束培养后分别收集MⅡ期卵母细胞计算卵母细胞第一极体排出率、活细胞荧光探针观察线粒体分布、实时荧光定量PCR检测卵母细胞线粒体DNA拷贝数(mtDNA)、解偶联蛋白(UCP2)和过氧化物酶体增殖物激活受体γ共激活因子-1α(PGC-1α)mRNA表达;收集卵丘颗粒细胞实时荧光定量PCR检测促凋亡基因Bax、抗凋亡基因Bcl-2以及缝隙连接蛋白Cx43、Cx37 mRNA表达。结果与空白对照组和血清对照组比较,补肾组卵母细胞第一极体排出率升高(P<0.05),线粒体均匀分布于细胞质中,mtDNA、PGC-1α、UCP2 mRNA表达升高(P<0.05),与空白对照组比较,血清对照组Cx37 mRNA表达升高(P<0.05),补肾组Cx43、Bcl-2、Bcl-2/Bax mRNA表达升高(P<0.05);与血清对照组比较,补肾组Cx37 mRNA表达降低(P<0.05),Cx43、Bcl-2、Bcl-2/Bax mRNA表达升高(P<0.05)。结论补肾调经方可促进体外培养小鼠卵母细胞核质成熟同步,提高卵母细胞质量,其机制可能与减少颗粒细胞凋亡、改善线粒体功能有关。
Objective To explore the mechanism of Bushen Tiaojing Decotion to improve the quality of mouse oocytes by improving the synchronizing of nucleocytoplasmic maturation in vitro.Methods Two hundred of female ICR mice,aged 10 days,were randomly divided into Shen-tonifying group(BS group,60 mice),serum control group(60 mice)and blank control group(80 mice).The mice were sacrificed to separate preantral follicles,and the cells were cultured with Bushen Tiaojing Decotion rat serum,normal rat serum or without rat serum.After culturing,MⅡoocytes were collected to calculate the first polar body excretion rate.Fluorescent probes for living-cell was used to observe the mitochondrial distribution.The mRNA expressions of mtDNA,uncoupling protein 2(UCP2)and peroxisome proliferator ctivated receptorγcoactivator-1α(PGC-1α)in oocytes were measured by real-time PCR assay;following collection of the cumulus granulosa cells,the mRNA expressions of pro-apoptotic gene Bax,anti-apoptotic gene Bcl-2,and connexin connexin43(Cx43),Cx37 were also assessed by real-time PCR.Results Compared with the blank control group and the serum control group,the first polar body excretion rate of oocytes in the BS group increased(P<0.05),the mitochondria were evenly distributed in the cytoplasm,and the mRNA expressions of mtDNA,PGC-1α,UCP2 increased(P<0.05).Compared with the blank control group,the expressions of Cx37 mRNA reduced in the serum control group(P<0.05),and the mRNA expression of Cx43,Bcl-2,and Bcl-2/Bax ratio in the cumulus granulosa cells of the BS group increased(P<0.05).Compared with the serum control group,the mRNA expression of Cx37 decreased(P<0.05);and the mRNA expression of Cx43,Bcl-2,and Bcl-2/Bax ratio in the BS group increased(P<0.05).Conclusion Bushen Tiaojing Decotion could promote oocyte quality of mouse oocytes by improving the synchronizing of nucleocytoplasmic maturation in vitro,which may be associated with the decrease of apoptosis in the granulosa cell,and improvement of mitochondrial function.
作者
吕金梦
耿静然
于志芳
杜惠兰
LYU Jin-meng;GENG Jing-ran;YU Zhi-fang;DU Hui-lan(Hebei Key Laboratory of Integrative Medicine on Liver-Kidney Patterns,Institute of Integrative Medicine,College of Integrative Medicine,Hebei University of Chinese Medicine,Shjiazhuang 050200)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2022年第9期1101-1106,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金资助项目(No.81774359)。
