维生素E通过间隙连接蛋白Cx43对牛颗粒细胞凋亡及增殖的影响  

Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43

作　　者：任书强 王嘉玮[1] 陈洪艳[1] 徐明强[1] 姜昊[1] 高妍[1] 周建忠 曾凡明 李成晟 张嘉保[1] 陈承祯[1] 

机构地区：[1]吉林大学动物科学学院,长春130062 [2]尼勒克县畜牧兽医工作站,尼勒克835700

出　　处：《中国畜牧兽医》2016年第3期615-621,共7页China Animal Husbandry & Veterinary Medicine

基　　金：现代农业产业技术体系建设专项资金资助(CARS-38);吉林省重点科技攻关项目(20140204077NY);吉林省肉牛产业体系

摘　　要：本研究旨在研究维生素E对间隙连接蛋白Cx43的影响和其对牛颗粒细胞增殖与凋亡的作用及其机制。将从牛卵巢中分离获得颗粒细胞进行体外培养,用不同浓度的维生素E(0、25、50、100、200、500μmol/L)处理细胞24h。采用流式细胞术检测颗粒细胞的凋亡率、实时荧光定量PCR法检测凋亡标志基因BCL2/BAX、P53及Cx43mRNA的表达变化、CCK8法检测颗粒细胞的增殖变化。结果表明:流式细胞术检测显示,与对照组相比,体外培养过程中添加100μmol/L维生素E可显著抑制颗粒细胞凋亡(P<0.05),实时荧光定量PCR检测显示维生素E能引起BCL2/BAX、P53及Cx43基因表达显著变化(P<0.05),CCK8检测显示维生素E处理细胞24和36h时可以显著促进颗粒细胞增殖(P<0.05)。此研究结果为深入解析维生素E通过影响颗粒细胞增殖、凋亡进而影响卵母细胞发育、成熟的机制及提高雌性动物繁殖能力提供了理论依据。The aim of this study was to determine the effect of vitamin E on Cx43,the mechanism and function of vitamin E on bovine granulosa cells apoptosis and proliferation.In this study,granulosa cells were isolated from bovine ovary and cultivated in vitro by adding different concentration of vitamin E（0,25,50,100,200 and 500μmol/L）for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100μmol/L vitamin E could significantly inhibit the apoptosis of granulosa cells（P〈0.05）.Real-time PCR detection results showed that vitamin E significantly changed the mRNA expression levels of BCL2/BAX,P53,Cx43genes（P〈0.05）.Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36h（P〈0.05）.The results provided a theoretical basis on further analysis for studing the influence mechanism of vitamin E on oocytes development and maturity,and improvement of female animal reproduction by influencing granulosa cells proliferation and apoptosis.

关 键 词：维生素E 颗粒细胞 CX43 增殖 凋亡 

分 类 号：Q566[生物学—生物化学]