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

doi:10.16431/j.cnki.1671-7236.2016.03.008

›› 2016, Vol. 43 ›› Issue (3): 615-621.doi: 10.16431/j.cnki.1671-7236.2016.03.008

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

任书强¹, 王嘉玮¹, 陈洪艳¹, 徐明强¹, 姜昊¹, 高妍¹, 周建忠², 曾凡明², 李成晟², 张嘉保¹, 陈承祯¹

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

收稿日期:2015-11-24 出版日期:2016-03-20 发布日期:2016-03-30
通讯作者: 张嘉保, 陈承祯 E-mail:jluzhangjiabao@163.com;1651001215@qq.com
作者简介:任书强(1991-),男,内蒙古牙克石人,硕士生,研究方向:动物营养与饲料科学,E-mail:waller_ren@163.com
基金资助:
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

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

REN Shu-qiang¹, WANG Jia-wei¹, CHEN Hong-yan¹, XU Ming-qiang¹, JIANG Hao¹, GAO Yan¹, ZHOU Jian-zhong², ZENG Fan-ming², LI Cheng-sheng², ZHANG Jia-bao¹, CHEN Cheng-zhen¹

1. College of Animal Science, Jilin University, Changchun 130062, China;
2. Nilka County Animal Husbandry and Veterinary Workstations, Nilka 835700, China

Received:2015-11-24 Online:2016-03-20 Published:2016-03-30

摘要/Abstract

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、500 μmol/L)处理细胞24 h。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Cx43 mRNA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100 μmol/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36 h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。

关键词: 维生素E; 颗粒细胞; Cx43; 增殖; 凋亡

Abstract: 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 500 μmol/L) for 24 h.After cultured,apoptotic cells were detected by FCM,mRNA expression levels of BCL2/BAX、P53 and Cx43 genes were determined by Real-time PCR and cell proliferation was detected by CCK8.The results showed that compared to control group,100 μmol/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,Cx43 genes (P<0.05).Vitamin E could significantly improve granulosa cells proliferation when granulosa cells were treated for 24 and 36 h (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.

Key words: vitamin E; granulosa cell; Connexin 43; proliferation; apoptosis

中图分类号:

Q566

任书强, 王嘉玮, 陈洪艳, 徐明强, 姜昊, 高妍, 周建忠, 曾凡明, 李成晟, 张嘉保, 陈承祯. 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[J]. , 2016, 43(3): 615-621.

REN Shu-qiang, WANG Jia-wei, CHEN Hong-yan, XU Ming-qiang, JIANG Hao, GAO Yan, ZHOU Jian-zhong, ZENG Fan-ming, LI Cheng-sheng, ZHANG Jia-bao, CHEN Cheng-zhen. Effect of Vitamin E on Bovine Granulosa Cells Apoptosis and Proliferation through Cx43[J]. , 2016, 43(3): 615-621.

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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

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