鸽蛋低密度脂蛋白对冷休克和冻融猪精子HSP70及凋亡相关基因表达的影响  

Effect of LDL from Pigeon Egg Yolk on the Expression of HSP70 and Apoptosis-related Gene in Cold Shock and Frozen-thawed Boar Spermatozoa

作　　者：刘杨[1] 黄和璐[1] 董海涛[1] 金一[1] 

机构地区：[1]延边大学农学院,延吉133002

出　　处：《畜牧兽医学报》2015年第6期940-948,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31260529);吉林省科技厅重点科技攻关项目(20150204079NY)

摘　　要：旨在探究鸽蛋低密度脂蛋白(LDL)对冷休克和冻融猪精子HSP70及凋亡相关基因表达的影响。试验分为5组,分别是鲜精组、3%LDL(-80和-196℃)处理组、9%LDL(-80和-196℃)处理组。采用CASA系统分析不同处理组添加不同浓度鸽蛋LDL对冷休克及冻融前后猪精子生物学性状的影响;通过SDS-PAGE、Western blotting、PCR扩增检测分析HSP70蛋白及凋亡基因表达量的变化。结果表明:猪精子经过冻融处理后,热休克蛋白HSP70的表达量高于鲜精组,同时添加9%鸽蛋LDL并置于-196℃保存的冻融精子的热休克蛋白HSP70的表达量与鲜精组无显著差异(P>0.05);在抗凋亡基因Bcl-x1、Bcl-2l试验中,鲜精组的基因表达量均高于其他处理组,添加9%LDL(-196℃)处理组的基因表达量与鲜精组无显著差异(P>0.05);在促凋亡基因Bak试验中,鲜精组的基因表达量最低,且添加9%LDL(-196℃)处理组的Bak基因表达量与鲜精组无显著差异(P>0.05)。添加鸽蛋LDL可在一定程度上降低猪精子在冻融过程中造成的损伤,维持猪精子正常生物学性状,提高猪精子存活率。The purpose of this study is to investigate the effects of LDL from pigeon egg yolk on expression of HSP70 and apoptosis related genes in cold-shocked and frozen-thawed boar spermatozoa.This study divided into 5groups:fresh sperm group,3% LDL(-80and-196 ℃)treatment groups,9%LDL(-80and-196 ℃)treatment groups.The addition of LDL with different concentrations has effect on biological features of the spermatozoa before and after cold-shock and frozen-thawed treatments,which was analysed by CASA,and changes in expression of HSP70 and apoptosis-related genes were analysed and checked by SDS-PAGE,Western blotting and PCR.The results showed that:HSP70expression was higher than that of fresh sperm after freezing,there was no significant difference between HSP70 expression of frozen-thawed stored at-196 ℃ with addition of 9% pigeon egg and fresh spermatozoa in treatment group(P>0.05);in the experiment of anti-apoptosis gene Bcl-x1,Bcl-21,gene expression in fresh sperm treatment group was higher than other treatment groups(P>0.05);in the expression of pro-apoptotic Bak gene,the expression of fresh sperm was the lowest,there was no significant difference betweenBak gene of the 9% LDL(-196 ℃)treatment group and fresh spermatozoa in treatment group(P>0.05).Addition of LDL from pigeon egg yolk could reduce the damage caused by freezing and thawing,maintain biological features and improve the survival of the pig spermatozoa.

关 键 词：猪精子 冷休克 冻融 LDL HSP70 凋亡相关基因 

分 类 号：S828[农业科学—畜牧学]