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作 者:王小康[1,2] 蔡亚非[1,2] 张凯[1,2] 范睿[1,2] 李建国[1,2] 吴国良[1,2] 王根林[3]
机构地区:[1]安徽师范大学安徽省重点资源保护与利用重点实验室,安徽芜湖241000 [2]安徽师范大学生物环境与生态安全安徽省高校重点实验室,安徽芜湖241000 [3]南京农业大学动物科技学院,江苏南京210095
出 处:《激光生物学报》2008年第5期569-576,共8页Acta Laser Biology Sinica
基 金:国家自然科学基金项目(30471243);安徽省自然科学研究项目(050410201);安徽师范大学博士科研启动计划项目(160-750517);安徽省教育厅自然基金项目(2006KJ183B);安徽省高校优秀中青年骨干教师基金项目(161-0605049)
摘 要:一个月大雄性小鼠24只,随机分为6组,用30μmol/kgCdCl2作用小鼠睾丸不同的时间(3h、6h、12h、18h、24h)后,利用DNA电泳、免疫组化和半定量RT-PCR技术,分析生殖细胞凋亡过程中三种关键物质Caspase-3、Bcl-2、Bax蛋白和mRNA的表达量变化。结果显示:1)DNA各组(除对照组外)均出现不同程度断裂。2)Caspase-3蛋白表达量一直上升,与对照组相比差异极显著;Bax蛋白在12h前一直上升,与对照组相比差异显著,12h后又开始下降,且与对照组相比无显著差异;Bcl-2蛋白在下降,与对照组相比差异显著。3)RT-PCR结果显示Caspase-3基因表达量减少;Bax基因表达量逐渐上升;Bcl-2基因表达量波动很大。综上所述,Caspase-3、Bcl-2和Bax三个基因可能参与了镉应激状态下小鼠睾丸组织细胞的凋亡过程。Juvenile ( 1 month old) mice were randomly divided into 6 groups. DNA electrophoresis, immunohistochemistry and semi-quantity RT-PCR methods were used to investigate the proteins and mRNA expression of Caspase-3, Bax and Bcl-2 which were involved in the apoptosis of mouse testis. The results were shown as follows : 1 ) DNA departed at all the groups except the control. 2 ) The expression of Caspase-3 protein was up-regulated and the difference with control was significant. The expression of Bax protein was up-regulated in the 3 h, 6 h and 12 h groups and down-regulated in 18 h, 24 h groups. The expression of Bcl-2 protein was down-regulated at all the groups. 3 )The result of RT-PCR showed that, Caspase-3 gene was slightly down-regulated, Bax gene was obviously up-regulated, but the Bcl-2 expression was greatly fluctuant. It was shown that Caspase-3, Bax and Bcl-2 gene maybe involved in the mouse testis apoptosis which induced by cadmium.
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