抑制素A对牦牛颗粒细胞FSH、LH及其受体表达的影响  

Effects of Inhibin A on Expression of FSH,LH and Their Receptors in Yak Granulosa Cells

作　　者：王立斌[1] 王军乾 赵凌 黄振华[1] 王萌[1] 王靖雷 余四九[1,2] 潘阳阳 WANG Libin;WANG Junqian;ZHAO Ling;HUANG Zhenhua;WANG Meng;WANG Jinglei;YU Sijiu;PAN Yangyang(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou,Gansu 730070;Gansu Province Livestock Embryo Engineering Research Center,Lanzhou,Gansu 730070)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]甘肃省牛羊胚胎工程技术研究中心,甘肃兰州730070

出　　处：《核农学报》2023年第1期51-59,共9页Journal of Nuclear Agricultural Sciences

基　　金：甘肃省农牧厅农业生物技术研究项目(GNSW-2015-19);甘肃省教育厅产业支撑引导项目(2019C-03);甘肃农业大学盛彤笙基金(GSAU-STS-2018-23)

摘　　要：为了探明抑制素A(INHA)对牦牛卵泡颗粒细胞中促卵泡素(FSH)和促黄体素(LH)与其受体结合发挥的作用,通过体外培养牦牛卵泡颗粒细胞,采用免疫荧光技术(IF)检测促卵泡素受体(FSHR)和促黄体素受体(LHR)在颗粒细胞中的分布情况,用不同浓度外源性INHA(0、1.25、2.5、5、10、20 ng·mL^(-1))作用12 h后,利用实时荧光定量PCR(qRT-PCR)技术检测颗粒细胞中FSHβ、LHβ、FSHR、LHR基因的表达情况,采用酶联免疫分析试验(ELISA)检测细胞内外FSH、LH的含量。结果显示,FSHR和LHR在颗粒细胞质与细胞核中均有表达。FSHβ和LHβ基因在颗粒细胞中的表达随INHA浓度的增大呈先降后升的趋势;INHA浓度接近5 ng·mL^(-1)时,FSHβ表达最低;INHA浓度接近10 ng·mL^(-1)时,LHβ表达最低;FSHR和LHR基因的表达与INHA浓度呈负相关。INHA浓度接近5 ng·mL^(-1)时,颗粒细胞内外FSH含量最低;INHA浓度接近10 ng·mL^(-1)时,颗粒细胞内外LH含量最低;其他INHA浓度下,当颗粒细胞内容物中FSH和LH含量降低时,细胞上清中含量升高。综上所述,INHA对FSHR和LHR基因在颗粒细胞中的表达存在明显的抑制作用,并且影响FSHβ和LHβ基因在颗粒细胞中的表达。研究表明INHA可以抑制牦牛卵泡颗粒细胞中FSH、LH与其受体的结合;同时,INHA对颗粒细胞内外FSH和LH的含量具有重要调节作用。本研究为进一步探索抑制素对雌性牦牛生殖调控的影响提供了理论依据。To investigate the effect of inhibin A(INHA)on FSH and LH binding receptors in yak follicular granulosa cells.The distribution of Follicle-Stimulating Hormone Receptor(FSHR)and Luteinizing Hormone Receptor(LHR)in yak granulosa cells cultured in vitro are detected by Immunofluorescence(IF).After treatment with exogenous INHA at different concentrations(0,1.25,2.5,5,10,20 ng·mL^(-1))for 12 h,the expressions of FSHβ,LHβ,FSHR and LHR in granulosa cells are detected by Real-time fluorescence quantitative PCR(qRT-PCR),and the contents of FSH and LH in and outside the cells are detected by Enzyme Linked Immunoassay(ELISA).These results show that both FSHR and LHR are expressed in the cytoplasm and nucleus of granulosa cells.The expressions of FSHβand LHβgenes in granulosa cells are decreased at the first instance and then increased with increasing INHA concentration.The expression of FSHβis the lowest when INHA concentration is close to 5 ng·mL^(-1),and the expression of LHβis lowest when INHA concentration is close to 10 ng·mL^(-1).Expressions of FSHR and LHR genes are negatively correlated with INHA concentration.When INHA concentration is close to 5 ng·mL^(-1),the content of FSH in and out of granulosa cells is the lowest,and when INHA concentration is close to 10 ng·mL^(-1),the content of LH in and out of granulosa cells is also the lowest.Under other INHA concentrations,the content of FSH and LH in granulosa cells is decreased,but that in the cell supernatant is increased.It is concluded that INHA inhibit the expression of FSHR and LHR genes in granulosa cells,suggesting that INHA inhibit the binding of FSH and LH to their receptors in yak follicular granulosa cells.Meanwhile,INHA plays an important role in regulating the content of FSH and LH in and outside granulosa cells.This study provides a theoretical basis to further explore the effects of inhibin on reproductive regulation in female yaks.

关 键 词：颗粒细胞 抑制素A(INHA) 促卵泡素(FSH) 促黄体素(LH) 受体 

分 类 号：S823.85[农业科学—畜牧学]