乌萸汤对高龄小鼠卵母细胞的能量代谢相关影响  

Effect of Wuyu decoction on energy metabolism of oocytes in aged mice

作　　者：郑凌琦 黄文玲[2] 刘艳霞[2] 徐彩[2] ZHENG Lingqi;HUANG Wenling;LIU Yanxia;XU Cai(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,100029 [2]北京中医药大学东方医院妇科

出　　处：《环球中医药》2021年第6期1020-1027,共8页Global Traditional Chinese Medicine

基　　金：国家自然科学基金青年项目(81704112)。

摘　　要：目的通过卵母细胞体外成熟(in vitro maturation,IVM)技术,探讨乌萸汤对体外培养的高龄小鼠卵母细胞能量代谢相关影响。方法6周龄C57BL/6J雌性小鼠促排后取卵母细胞培养于M2培养基为青年组,24周龄C57BL/6J雌性小鼠卵母细胞为实验组,将实验组卵母细胞分别培养于基础培养基、乌萸汤含药血清培养基、山茱萸环烯醚萜苷类提取物培养基中,并称其为高龄组、乌萸汤组、山茱萸提取物组。各组培养24小时后分别检测各组卵母细胞的第一极体排出率、ATP含量、线粒体数量及膜电位变化、超氧化物歧化酶(superoxide dismutase,SOD)活性、活性氧(reactive oxygen species,ROS)含量以及腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPKα)、磷酸化腺苷酸活化蛋白激酶(p-AMPKα)、线粒体增生有关的过氧化物酶体增殖物激活受体γ辅助活化因子1α(peroxisome proliferators-activated receptor gamma coactivator-1α,PCG-1α)蛋白的表达。结果(1)与青年组相比,高龄组第一极体排出率、ATP含量、线粒体数量及膜电位变化、SOD酶活性均有所降低,ROS含量升高,AMPKα、p-AMPKα、PGC-1α蛋白的表达降低(P<0.05)。(2)与高龄组相比,乌萸汤组第一极体排出率、ATP含量、线粒体数量、SOD酶活性均有所升高,AMPKα、PGC-1α蛋白表达量有所增多(P<0.05),但线粒体膜电位变化、ROS及p-AMPKα含量均无明显差异(P>0.05)。(3)与高龄组相比,山茱萸提取物组第一极体排出率、ATP含量、线粒体数量、SOD酶活性、p-AMPKα蛋白含量有明显升高(P<0.05),线粒体膜电位变化、ROS、AMPKα、PGC-1α蛋白表达量无明显差异(P>0.05)。结论乌萸汤能够通过影响高龄小鼠卵母细胞的线粒体功能,从而改善其能量代谢和氧化应激状态,促进卵母细胞培养。Objective To investigate the effect of Wuyu decoction on energy metabolism of oocytes from aged mice in vitro maturation(IVM).Methods The oocytes of 6-week-old C57BL/6J female mice were cultured in M2 culture medium(young group),and the oocytes of 24-week-old C57BL/6J female mice were cultured in basic culture medium(aged group),Wuyu decoction medicated serum culture medium(Wuyu decoction group)and cornel iridoid glycoside extract culture medium(Wuyu decoction extract group)respectively.After 24h,the oocytes of each group were collected for the detection of the first polar body extrusion rate,ATP content,mitochondrial number and membrane potential changes,SOD activity,ROS content and AMPKα,p-AMPKα,PGC-1αprotein expression.Results(1)Compared to the young group,the first polar body expulsion rate,ATP content,mitochondrial number,membrane potential changes and SOD activity in the elderly group were decreased,ROS content was increased,and the expression of AMPKα,p-AMPKαand PGC-1αprotein was significantly decreased(P<0.05).(2)Compared to the aged group,the first polar body expulsion rate,ATP content,mitochondrial number,SOD activity and protein expression of AMPKαand PGC-1αin the Wuyu decoction group were increased(P<0.05).However,there were no significant differences in the changes of mitochondrial membrane potential,ROS and p-AMPKαcontent(P>0.05).(3)Compared to that old group,the first polar body excretion rate,the content of ATP,the number of mitochondria,the activity of SOD and the content of p-AMPKαprotein increased significantly(P<0.05),but the changes of mitochondrial membrane potential,ROS,AMPKα,PGC-1αhad no significant difference(P>0.05).Conclusion Wuyu decoction can improve the energy metabolism and oxidative stress of oocytes and promote oocyte maturation by affecting the mitochondrial function of oocytes in elderly mice.

关 键 词：乌萸汤 卵母细胞 线粒体能量代谢 氧化应激能力 未成熟卵母细胞体外成熟 

分 类 号：R285.5[医药卫生—中药学]