表皮生长因子对牦牛卵丘细胞低氧诱导因子-1α表达的影响及与凋亡的关联性分析

doi:10.11843/j.issn.0366-6964.2016.06.010

摘要/Abstract

摘要：

旨在研究EGF是否通过调控HIF-1α抑制牦牛卵丘细胞凋亡。本研究在牦牛卵丘细胞体外培养时加入不同浓度的EGF，运用qRT-PCR和免疫荧光技术检测HIF-1α、Bcl-2和Bax的表达，用一步法TUNEL检测不同处理组卵丘细胞的凋亡情况。结果表明：(1)牦牛卵丘细胞体外培养液中添加不同浓度的EGF后，卵丘细胞HIF-1α和Bax mRNA的相对表达量降低，Bcl-2 mRNA的相对表达量增加，且具有浓度依赖性；当EGF浓度为50 ng•mL-1时，HIF-1α和Bax mRNA的相对表达量最低，Bcl-2 mRNA的相对表达量最高。(2)向其体外培养液中添加不同浓度的EGF后，卵丘细胞HIF-1α和Bax蛋白的相对表达量降低，Bcl-2蛋白的相对表达量增加，且具有浓度依赖性；当EGF浓度为50 ng•mL-1时，HIF-1α和Bax蛋白的相对表达量最低，Bcl-2蛋白的相对表达量最高。(3)TUNEL凋亡检测表明，对照组中卵丘细胞凋亡率最高，当EGF浓度为25 ng•mL-1时，细胞凋亡率显著降低(P＜0.05)；当EGF浓度为50 ng•mL-1时，细胞凋亡率最低(P＜0.05)，但随着EGF浓度的增加，卵丘细胞的凋亡率又升高。本研究结果表明，EGF可通过调控HIF-1α抑制卵丘细胞凋亡，其作用可能与线粒体介导的Bax和Bcl-2凋亡途径相关。

关键词: 表皮生长因子（EGF）, 低氧诱导因子-1&alpha, （HIF-1&alpha, ）, 凋亡, 卵丘细胞, 牦牛

Abstract:

The present study was carried out to verify whether EGF inhibited the apoptosis of cumulus cells of yak by regulating the expression of HIF-1α.The cumulus cells of yak were cultured in vitro with the supplement of different concentration of EGF.The expression levels of HIF-1α，Bax and Bcl-2 in different groups were detected by qRT-PCR and immunofluorescence.TUNEL was used to evaluate apoptosis rate.Results showed that：(1) After the addition of different concentration of EGF to the cumulus cells，there would be found a declined mRNA relative expression of HIF-1α and Bax and an increased mRNA relative expression of Bcl-2，with a characteristic manner of concentration-dependent.When the concentration of EGF was as high as 50 ng•mL-1，the mRNA relative expressions of HIF-1α and Bax were the lowest，while that of Bcl-2 was the highest.(2) The protein relative expressions of HIF-1α and Bax were decreased and that of Bcl-2 was increased，with a characteristic manner of concentration-dependent.When the concentration of EGF was as high as 50 ng•mL-1，the protein relative expressions of HIF-1α and Bax were the lowest，while that of Bcl-2 was the highest.(3) Apoptosis detection shown that the apoptosis rate from control group was the highest.It was extremely declined when the concentration of EGF was 25 ng•mL-1 (P＜0.05)，and down to its lowest level when EGF was 50 ng•mL-1 (P＜0.05)，while rose again when EGF was continues to increase.It is concluded from this study that EGF can inhibit the apoptosis of cumulus cells by regulating HIF-1α and may be related to mitochondrial Bax and Bcl-2 apoptotic pathway.

Key words: epidermal growth factor (EGF), hypoxia inducible factor-1&alpha, (HIF-1α), apoptosis, cumulus cells, yak

中图分类号:

S823.8+5.2

张译夫，潘阳阳，温泽星，余四九. 表皮生长因子对牦牛卵丘细胞低氧诱导因子-1α表达的影响及与凋亡的关联性分析[J]. 畜牧兽医学报, 2016, 47(6): 1154-1161.

ZHANG Yi-fu，PAN Yang-yang，WEN Ze-xing，YU Si-jiu. The Effect of Epidermal Growth Factor on the Expression of Hypoxia Inducible Factor-1α in Cumulus Cells of Yak (Bos grunniens) and Its Correlation Analysis with Apoptosis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(6): 1154-1161.

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