原代黄素化卵泡颗粒细胞氧化应激模型的建立与评价  

Establishment and evaluation of oxidative stress model of follicular granulosa cells

作　　者：邢鹏[1] 袁博[2] 王娜[1] 李澎涛[1] 孟娜娜 Xing Peng;Yuan Bo;Wang Na;Li Pengtao;Meng Nana(Departmen of Reproductive Medicine,Baoding Maternal and Child Health Hospital,Baoding Hebei 071000,P.R.China;Department of pediatrics,Affiliated Hospital of Hebei University,Baoding Hebei 071000,P.R.China)

机构地区：[1]保定市妇幼保健院生殖医学科,河北保定071000 [2]河北大学附属医院儿科,河北保定071000

出　　处：《中国计划生育和妇产科》2022年第2期69-72,共4页Chinese Journal of Family Planning & Gynecotokology

基　　金：保定市科技计划项目(项目编号:2041ZF137)。

摘　　要：目的建立过氧化氢(H_(2)O_(2))诱导的原代黄素化卵泡颗粒细胞氧化应激模型。方法收集2019年10月至12月于保定市妇幼保健院生殖医学科行体外受精患者取卵后废弃的卵泡液,经密度梯度离心收集卵泡颗粒细胞,进行培养。不同浓度H_(2)O_(2)处理颗粒细胞12 h,采用最大半数抑制浓度(half maximal inhibitory concentration,IC50)建立颗粒细胞氧化应激损伤模型。以未加H_(2)O_(2)的颗粒细胞为对照组,通过测定模型组丙二醛(malondialdehyde,MDA)含量、超氧化物歧化酶(superoxide dismutase,SOD)水平、雌二醇(estradiol,E_(2))分泌量和细胞周期,评价模型组是否建模成功。结果使用IC50浓度H_(2)O_(2)=65μmol/L,建立颗粒细胞氧化损伤模型。与对照组比较,模型组的MDA含量升高、SOD水平下降、E_(2)分泌量下降、细胞周期表现为sub G1峰增高、G1/G0期和G2/M期的细胞数量减少(P<0.05)。结论65μmol/L H_(2)O_(2)处理颗粒细胞12 h,建立了稳定的颗粒细胞氧化应激损伤模型,为进一步开展颗粒细胞抗氧化研究提供了细胞模型。Objective To establish a model of oxidative stress in follicular granulosa cells induced by hydrogen peroxide(H_(2)O_(2)).Methods The discarded follicular fluid of patients undergoing in vitro fertilization in Department of Reproductive Medicine of Baoding Maternal and Child Health Hospital from October 2019 to December 2019 was collected and cultured by density gradient centrifugation.Granulosa cells were treated with different concentrations of(H_(2)O_(2))for 12 hours,and the oxidative stress injury model of granulosa cells was established by the maximum inhibitory concentration(IC50).The granulosa cells without(H_(2)O_(2))were used as the control group.The content of malondialdehyde(MDA),the level of superoxide dismutase(SOD),the secretion of estradiol(E_(2))and the cell cycle in the model group were measured to evaluate whether the model group was successful or not.Results The oxidative damage model of granulosa cells was established by using IC50 concentration(H_(2)O_(2)=65μmol/L).Compared with the control group,the content of MDA increased,the level of SOD decreased,the secretion of E_(2) decreased,the G1 peak of sub increased and the number of cells in G1/G0 and G2/M phase decreased in the model group(P<0.05).Conclusion Granulosa cells were treated with 65μmol/L(H_(2)O_(2))for 12 hours,and a stable model of oxidative stress injury of granulosa cells was established,which provided a cell model for further study of antioxidation of granulosa cells.

关 键 词：颗粒细胞 过氧化氢 氧化应激 细胞模型 

分 类 号：R71[医药卫生—妇产科学]