黄体生成素干预大鼠卵巢颗粒细胞的蛋白质组学分析  

Proteomic Analysis of the Effects of Luteinizing Hormone on Rat Primary Ovarian Granulosa Cells

作　　者：董玉婷 陈泰任 叶晓锋 黑常春[1] 蔡玉芳[1] 孔斌[1] 赵承军[1] 常青[1] DONG Yuting;CHEN Tairen;YE Xiaofeng;HEI Changchun;CAI Yufang;KONG Bin;ZHAO Chengjun;CHANG Qing(Department of Human Anatomy,Histology and Embryology,School of Basic Medical Sciences,Ningxia Medical University/Fertility Conservation Key Laboratory of the Ministry of Education,Yinchuan 750004,China;Cancer Hospital of General Hospital of Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院人体解剖与组织胚胎学系,生育力保持教育部重点实验室,银川750004 [2]宁夏医科大学总医院肿瘤医院,银川750004

出　　处：《宁夏医科大学学报》2021年第6期569-576,584,共9页Journal of Ningxia Medical University

基　　金：国家自然科学基金(81460230);宁夏自然科学基金(2018AAC0325B)。

摘　　要：目的分析黄体生成素(luteinizing hormone,LH)干预卵巢颗粒细胞后差异蛋白的表达,探讨LH对颗粒细胞作用中重要蛋白及其生物学功能。方法体外分离培养大鼠原代卵巢颗粒细胞,给予LH处理3 h为实验组,生理盐水处理作为对照组。通过液相色谱-质谱(liquid chromatography-mass spectrometry,LC-MS)联用法得到蛋白样品二级质谱,利用质谱数据库筛选显著差异表达蛋白,对差异表达蛋白进行生物信息学分析,并通过Western blot对部分差异蛋白进行验证。结果与对照组相比,LH干预组共有57个蛋白表达有显著差异,其中25个上调,32个下调;生物信息学分析显示多数差异蛋白具有结合活性和催化活性,主要富集于酶代谢过程中及参与调控细胞代谢过程;Western blot验证发现调控细胞增殖与极性的蛋白Pard3、Mark3表达上调,抗氧化应激蛋白Gstp1和Nqo1的表达量下调,与蛋白质组学结果一致。结论LH可引起卵巢颗粒细胞多种差异蛋白表达,影响颗粒细胞间的结合方式、激素合成能力、氧化应激及细胞周期和增殖等功能,且差异蛋白涉及多条相关细胞信号通路传导,提示LH从多方面影响卵巢颗粒细胞的功能。Objective To analyze the expression of differential proteins after LH intervention in ovarian granulosa cells,in order to find the important proteins and biological functions of LH action on granulocyte cells,and to provide a basis for further mechanism research.Methods Primary rat ovarian granulocytes were isolated and cultured in vitro and treated with LH for 3 h as the experimental group and saline as the control group.Protein samples were obtained by liquid chromatography-mass spectrometry(LC-MS),and the differentially expressed proteins were screened by mass spectrometry database.Bioinformatics analysis was performed on differentially expressed proteins.Western blot was used to verify some of the differential proteins.Results Compared with the control group,there were significant differences in the expression of 57 proteins in the LH intervention group,25 of which were up-regulated and 32 of which were down-regulated.Bioinformatics analysis showed that most of the differential proteins had binding and catalytic activities,which were mainly concentrated in enzyme metabolism and involved in the regulation of cell metabolism.Western blot results verified the differential proteins,and the expressions of Pard3 and Mark3,which regulate cell proliferation and polarity,and the expressions of anti-oxidative stress proteins Gstp1 and Nqo1 were down-regulated,which was consistent with the proteomics results.Conclusion LH can induce the expression of a variety of differential proteins in ovarian granulosa cells,affect the binding mode between granulosa cells,hormone synthesis ability,oxidative stress,cell cycle and proliferation and other functions.And the differential proteins are involved in multiple cell signaling pathways,suggesting that LH affects the function of ovarian granulosa cells in many ways.

关 键 词：黄体生成素 卵巢颗粒细胞 蛋白质组学 差异表达蛋白 

分 类 号：R711.75[医药卫生—妇产科学]