摘要
为了揭示caspase-3等细胞凋亡相关基因及DNA甲基化水平在小鼠卵泡闭锁中的作用,小鼠经注射10 IU hCG后,在0、12、24、48、72和96 h解剖取出卵巢,用HE染色、DNA电泳、免疫组化、RT-PCR技术、甲基化酶切等试验方法,研究在卵泡发育及闭锁过程中caspase-3等凋亡相关基因的表达及DNA甲基化水平的变化。结果表明:bax、caspase-3的表达量在48 h以前持续减少,在48 h达到最低值,在72 h显著增多(P<0.05),且在卵腔液中大量表达;甲基化水平在48 h以前持续减少,在48 h达到最低值,在72 h显著增多(P<0.05);卵泡凋亡程度在48 h之前也持续减少,在72 h显著增多(P<0.05)。bcl-2的表达量在48 h之前持续增加,在72 h显著减少(P<0.05)。提示:卵泡的发育和闭锁可能受体细胞Caspase信号通路调控,并通过卵腔液发挥作用决定其存亡;另外,甲基化水平的改变可能与卵泡的闭锁直接相关。
To investigate the functions of caspase-3 gene,apoptosis-related genes and DNA methylation levels during the process of follicular atresia,follicles were collected at 0,12,24,48,72 and 96 h after mice were injected with 10 IU hCG.HE staining,DNA electrophoresis,immunohistochemistry,RT-PCR and methylation digestion methods were used to explore the follicular development.We investigated DNA methylation levels and the expression of apoptosis-related genes during follicular development and atresia.The results were shown as follows：The expression of bax,caspase-3 continued to decrease until 48 h,and then up-regulated significantly at 72 h（P0.05）,and the most caspase-3 positive reactions appeared in the antral liquid.At the same time,The expression of bcl-2 continued to increase until 48 h,and then significantly down-regulated at 72 h（P0.05）.The degree of apoptosis continued to decrease post 48 h,and then up-regulated significantly at 72 h（P0.05）.The methylation levels continued to decrease until at 48 h,and then increased significantly at 72 h（P0.05）.Taken together,the somatic Caspase signaling pathway may regulate the development of follicles through antral liquid,and the changes of methylation may induce follicular atresia.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2010年第3期88-93,共6页
Journal of Nanjing Agricultural University
基金
国家“十一五”科技支撑计划项目(2006BAD04A01,2006BAD04A12)
安徽省教育厅重点项目(KJ2009A039)
芜湖市科技计划重点项目(2008620)
作者简介
刘楠乔，硕士研究生。
通讯作者：蔡亚非，教授，研究方向为动物分子遗传学，E-mail：caiyafeinau@163．com。
