畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (6): 940-948.doi: 10.11843/j.issn.0366-6964.2015.06.009

• 遗传繁育 • 上一篇    下一篇

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

刘杨,黄和璐,董海涛,金一*   

  1. (延边大学农学院,延吉 133002)
  • 收稿日期:2015-01-15 出版日期:2015-06-23 发布日期:2015-06-23
  • 通讯作者: 金一,教授,E-mail:yijin@ybu.edu.cn
  • 作者简介:刘杨(1990-),女,青海人,硕士生,主要从事动物繁殖与生物技术研究,E-mail:599295340@qq.com
  • 基金资助:

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

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

LIU Yang,HUANG He-lu,DONG Hai-tao,JIN Yi*   

  1. (Agricultural College of Yanbian University,Yanji 133002,China)
  • Received:2015-01-15 Online:2015-06-23 Published:2015-06-23

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。

Abstract:

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 5 groups:fresh sperm group,3% LDL(-80 and -196 ℃) treatment groups,9%LDL(-80 and -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:HSP70 expression 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 between Bak 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.

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