摘要
肿瘤转移抑制因子(Kisspeptin)在颗粒细胞上有表达,但其对颗粒细胞凋亡的作用尚不清楚,本试验旨在利用流式细胞术和荧光定量PCR的技术研究Kisspeptin对牛卵巢颗粒细胞凋亡的影响。向DMEM-F12培养液中分别添加0、100 nmol/L的Kp-10培养牛颗粒细胞,24 h后收集细胞,用流式细胞技术检测细胞凋亡和细胞周期,用荧光定量PCR技术检测Caspase-3、Bcl-2、Fas和Fasl的m RNA表达。结果表明:Kisspeptin可显著促进牛颗粒细胞凋亡,且伴随着Caspase-3、Fas和Fasl m RNA表达上调及Bcl-2 m RNA表达下调(P<0.01);流式细胞分析表明,Kisspeptin可使牛颗粒细胞在G1期增加、S期下降(P<0.05)。研究结果说明,Kisspeptin可促进牛颗粒细胞凋亡,这一作用可能通过上调促凋亡基因Caspase-3、Fas、Fasl和下调抗凋亡基因Bcl-2的m RNA表达实现。
Previous studies showed that Kisspeptin expresses in granulosa cells,however,its effect on apoptosis of bovine granulosa cells is still unknown.In the present study,flow cytometry analysis and q Real Time quantitative-PCR were used to study effect of Kisspeptin on apoptosis of bovine granulosa cells.Bovine granulosa cells were cultured with DMEM-F 12 solution added with 0 or 100 nmol/L of Kp-10 for 24 h,then flow cytometry was used to study apoptosis and cell cycleof the bovine granulosa cells,and Real 2,Fas and Fasl.The results showed that Time quantitative-PCR was used to Kisspeptin could promote apoptosis study mRNA expression of Caspase-3,Bcl-of bovine granulosa cells,accompanied with the decreased mRNA levels of Bcl-2 and the increased mRNA levels of Caspase-3,Fas and Fasl(P〈0.01).Additionally,results of flow cytometry analysis showed that Kisspeptin could significantly increase accumulation of cells in the G1 phase,decrease accumulation of cells in the S phase,and promote apoptosis of bovine granulosa cells(P〈0.05).Thus,our findings demonstrated for the first time that Kisspeptin promoted apoptosis of bovine ovarian granulosa cells,which may be achieved through regulation of Caspase-3,Fas,Fasl and Bcl-2 mRNA expression.
作者
王军
刘红羽
徐高青
吕文发
WANG Jun;LIU Hong-yu;XU Gao-qing;LV Wen-fa(Jilin Province Engineering Laboratory for Ruminant Reproductive Biotechnology and Healthy Production,College of Animal Science and Technology,Jilin Agricultural University,Jilin Changchun 130118,China)
出处
《中国畜牧杂志》
CAS
北大核心
2017年第9期57-61,共5页
Chinese Journal of Animal Science
基金
中国博士后基金(2015M581408)
吉林省科技厅项目(20150519018JH、20160209001NY)
