FSH对缺血缺氧诱导的小鼠卵巢颗粒细胞凋亡的保护作用  

Protective Effect of FSH on Apoptosis of Ovarian Granulosa Cells Induced by Ischemia and Hypoxia in Mice

作　　者：王菲[1] 田媛 郝耀 刘心蕊 张淑雅[1] 杨延周[1] 裴承斌 王燕蓉[1] 裴秀英[1] WANG Fei;TIAN Yuan;HAO Yao;LIU Xinrui;ZHANG Shuya;YANG Yanzhou;PEI Chengbin;WANG Yanrong;PEI Xiuying(School of the Basic Medical Sciences,Ningxia Medical University,Key Laboratory of Fertility Preservation and Maintenance,Ministry of Education Key Laboratory of Reproduction and Heredity of Ningxia Hui Autonomous Region,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,生育力保持教育部重点实验室,宁夏生殖与遗传重点实验室,银川750004

出　　处：《宁夏医科大学学报》2022年第6期590-596,共7页Journal of Ningxia Medical University

基　　金：宁夏回族自治区重点研发项目(2019BFG02007);国家自然科学基金项目(81760286,82160290);宁夏自然科学基金项目(2022AAC02031,2021AAC03133)。

摘　　要：目的探究促卵泡刺激素(FSH)对缺血缺氧损伤的小鼠原代卵巢颗粒细胞的保护作用。方法21日龄雌性ICR小鼠注射孕马血清(PMSG)42 h后,体外分离并培养小鼠原代卵巢颗粒细胞,分为对照组、单纯缺血缺氧组、缺血缺氧FSH干预组(0.1、0.2、0.4、0.6 IU·mL^(-1) FSH)。对单纯缺血缺氧组及缺血缺氧FSH干预组的细胞分别进行缺血缺氧处理及不同浓度FSH干预,通过TUNEL染色及流式细胞术检测细胞凋亡情况;通过Western blot检测抗凋亡蛋白Bcl-2、凋亡蛋白Bax的表达情况。结果TUNEL检测细胞结果显示:单纯缺血缺氧组TUNEL阳性细胞数最多、细胞凋亡率最高,FSH干预能够减少细胞凋亡,其中0.4、0.6 IU·mL^(-1) FSH干预组TUNEL阳性细胞较其他干预组减少,细胞凋亡率降低(P均<0.01);流式细胞术结果显示:除对照组外,其余各组凋亡率均有所增加(P均<0.01),在添加不同浓度FSH干预后,细胞凋亡被抑制,其中0.4 IU·mL^(-1) FSH干预组细胞凋亡率低于单纯缺血缺氧组和0.1 IU·mL^(-1) FSH干预组(P均<0.05);Western blot检测凋亡相关蛋白结果显示:与对照组相比,其余各组Bax蛋白表达均上调,Bcl-2蛋白表达量、Bcl-2/Bax比值均降低,0.4 IU·mL^(-1) FSH干预组的Bcl-2蛋白表达量、Bcl-2/Bax比值均高于单纯缺血缺氧组和其他干预组(P均<0.01)。结论添加FSH能抑制缺血缺氧诱导的小鼠颗粒细胞凋亡,降低细胞凋亡率,增强细胞的抗凋亡能力。Objective To investigate the protective role of recombinant human follicle-stimulating hormone(FSH)on ischemia and hypoxia induced injury in mouse primary ovarian granulosa cells.Methods The 21-day-old ICR female mice were injected with pregnant mare serum(PMSG).After 42 h,primary ovarian granulosa cells were isolated and cultured in vitro,and divided into control group,ischemic-hypoxic group and ischemic-hypoxic with FSH intervention group(0.1,0.2,0.4 and 0.6 IU·mL^(-1) FSH)group.Then the cells in the ischemic-hypoxic group and the ischemic-hypoxic with FSH intervention group were treated with ischemic-hypoxic treatment and different concentrations of FSH,respectively,and the cellular apoptosis was detected by TUNEL staining and flow cytometry;the expression of anti-apoptotic protein Bcl-2 and apoptotic protein Bax was detected by Western blot.Results The results of TUNEL detection showed that the number of TUNEL positive cells and the apoptotic rate were the highest in the ischemia-hypoxia group.FSH intervention could reduce the apoptotic rate of cells,and the TUNEL positive cells in 0.4 and 0.6 IU·mL^(-1) FSH intervention groups were less than those in other intervention groups,and the apoptotic rate was lower(P all<0.01);Flow cytometry results showed that apoptotic rate was increased in all groups except the control group(P all<0.01),and apoptosis was inhibited after the intervention of different concentrations of FSH.The apoptotic rate in the 0.4 IU·mL^(-1) FSH intervention group was lower than that in the ischemia-hypoxia group and the 0.1 IU·mL^(-1) FSH intervention group(P all<0.05).Western blot analysis of apoptotic-related proteins showed that compared with the control group,Bax protein expression was upregulated and Bcl-2 protein expression and Bcl-2/Bax ratio were decreased in the remaining groups,and Bcl-2 protein expression and Bcl-2/Bax ratio were higher in the 0.4 IU·mL^(-1) FSH intervention group than in the ischemia-hypoxia group and other intervention groups(P all<0.01).Conclusion FSH can

关 键 词：小鼠原代卵巢颗粒细胞 缺血缺氧 促卵泡刺激素 细胞凋亡 

分 类 号：R714.13[医药卫生—妇产科学]