受精过程中不同激活力强度对小鼠卵子线粒体三磷酸腺苷和活性氧生成的影响  

Effects of different activation intensities on mitochondrial adenosine triphosphate and reactive oxygen species production in mouse oocytes during fertilization

作　　者：孙婷也 周怡雯[1] 李文治 闫铮[1] 燕志光[1] 于韦娜 吕祁峰[1] Laboratory StudyEffects of different activation intensities on mitochondrial adenosine triphosphate and reactive oxygen species production in mouse oocytes during fertilizationSun Tingye;Zhou Yiwen;Li Wenzhi;Yan Zheng;Yan Zhiguang;Yu Weina;Lyu Qifeng(Department of Assisted Reproduction,Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院辅助生殖科,200011

出　　处：《中华生殖与避孕杂志》2019年第12期995-999,共5页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金(81571486)。

摘　　要：目的通过对三磷酸腺苷(ATP)含量及活性氧(ROS)水平的检测,观察在小鼠卵子受精过程中不同水平的激活强度对卵子线粒体代谢的影响。方法采用我们已经建立的弱激活力小鼠精子对卵子进行卵胞质内单精子显微注射(ICSI)后,施以不同浓度的离子霉素(0μmol/L、2.5μmol/L、10μmol/L)处理10 min进行辅助激活,并以正常小鼠精子ICSI作为对照。根据处理方式分为4组:正常精子组(normal,N组)、激活力减弱精子(weak sperm,W组)、激活力减弱精子施以适度辅助激活剂组(weak sperm+moderate AOA,M组)、激活力减弱精子施以过度辅助激活剂组(weak sperm+excessive AOA,E组)。在ICSI后1.5~2 h内对卵子内ATP含量和ROS的水平进行检测。对未进行ICSI卵子组(oocyte,O组)进行ATP检测。结果①ATP含量检测显示:随着激活力的上升,O组[(2.005±0.053)pmol]、W组[(2.485±0.064)pmol]、M组[(3.763±0.075)pmol]的卵子内平均ATP含量逐步提高,且差异有统计学意义(P<0.0001);其中M组与N组[(3.573±0.068)pmol]平均ATP含量接近(P>0.05);E组虽较O组[(3.204±0.070)pmol]高,但其卵子的平均ATP含量显著小于N组(P<0.001)。②ROS水平检测显示:随着激活力的增加,W组(0.420±0.028,n=9)、M组(1.097±0.037,n=9)、E组(1.361±0.033,n=10)表现出ROS水平的逐步升高(P<0.001),其中M组与N组ROS水平(1.000±0.026,n=10)也比较接近,差异有统计学意义(P=0.044)。结论受精过程中激活力水平将对卵子线粒体代谢产生明显影响,对弱激活力小鼠精子显微受精后,行适度激活时,ATP产量增加,ROS生成较少;而过高的激活强度反而会引起ATP产量的下降和ROS生成的增多。Objective To observe the effect of different levels of activation intensity on mitochondrial metabolism in mouse oocyte during fertilization by detecting adenosine triphosphate(ATP)content and reactive oxygen species(ROS)level.Methods After intracytoplasmic sperm injection(ICSI)with weak sperm,different concentrations of ionomycin(0μmol/L,2.5μmol/L,10μmol/L)were applied for 10 min to assist the oocyte activation,and ICSI with normal mouse sperm was used as the control.According to the treatment methods,they were divided into four groups:normal sperm group(normal,N group),weak sperm group(W group),weak sperm+moderate AOA group(M group),weak sperm+excessive assisted oocyte activation(AOA)group(E group).ATP and ROS levels in oocyte were measured within 1.5-2 h after ICSI.In addition,ATP level of oocyte groups(O group)without ICSI was detected.Results 1)With the increase of activation power,the average ATP content in oocyte of O group[(2.005±0.053)pmol],W group[(2.485±0.064)pmol],M group[(3.763±0.075)pmol]gradually increased,with a significant difference(P<0.0001);the average ATP content in M group was close to that in N group[(3.573±0.068)pmol](P>0.05).In addition,the average ATP content of eggs in E group[(3.204±0.070)pmol]was significantly lower than that in N group(P<0.001),although it was higher than that in O group.2)The level of ROS showed that with the increase of activation power,the levels of ROS in W group(0.420±0.028,n=9),M group(1.097±0.037,n=9),E group(1.361±0.033,n=10)increased gradually(P<0.001),and the level of ROS in M group only showed slight difference with that of N group(1.000±0.026,n=10)(P=0.044).Conclusion This study preliminarily found that the level of activation power during fertilization will have a significant impact on mitochondrial metabolism of oocyte.In mouse,when oocyte was moderately activated after ICSI with weak acitivtion ability sperm,ATP production will increase,while ROS production will be less;however,excessive activation intensity will lead to the decrease of ATP

关 键 词：受精 卵胞质内单精子注射 活性氧 三磷酸腺苷 辅助激活 

分 类 号：R321.2[医药卫生—人体解剖和组织胚胎学]