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

doi:10.11843/j.issn.0366-6964.2015.06.009

摘要/Abstract

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。

关键词: 猪精子, 冷休克, 冻融, LDL, HSP70, 凋亡相关基因

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.

Key words: boar spermatozoa, cold shock, LDL, frozen-thawed, HSP70, apoptosis-related gene

中图分类号:

S813.3

刘杨，黄和璐，董海涛，金一. 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[J]. 畜牧兽医学报, 2015, 46(6): 940-948.

LIU Yang，HUANG He-lu，DONG Hai-tao，JIN Yi. Effect of LDL from Pigeon Egg Yolk on the Expression of HSP70 and Apoptosis-related Gene in Cold Shock and Frozen-thawed Boar Spermatozoa[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(6): 940-948.

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