急性冷应激对绵羊免疫功能和不同组织热休克蛋白70家族基因表达的影响  

Effects of Acute Cold Stress on Immune Function and Expression of Heat Shock Protein 70 Family Genes in Different Tissues in Sheep

作　　者：彭孝坤 张宇[1] 黄晓瑜 周广琛 邢晓南 张恩平[1] PENG Xiaokun;ZHANG Yu;HUANG Xiaoyu;ZHOU Guangchen;XING Xiaonan;ZHANG Enping(College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物科技学院

出　　处：《畜牧兽医学报》2019年第8期1625-1634,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划项目(2016YFD0500508);陕西省重点计划研发项目(2017TSCXL-NY-04-02);陕西省农业科技创新项目(NYKJ-2018-Yl16)

摘　　要：旨在研究急性冷应激对绵羊免疫功能和不同组织热休克蛋白70(HSP70)家族基因表达的影响。本研究选取8只健康、体况接近的(12±0.5)月龄小尾寒羊×湖羊杂交F1母羊,单笼饲养于保温舍内(风寒温度(-7.14±2.53)℃),适应7 d。第8天将母羊移置舍外(风寒温度(-27.40±3.12)℃)急性冷应激12 h,采集冷应激前后试验个体血样和组织样。利用ELISA法测定血清免疫指标(细胞因子和免疫球蛋白G含量),通过实时荧光定量PCR法测定不同组织(肝、肾、脾、心、背最长肌和十二指肠)HSP70家族基因(HSPA1A、HSPA6和HSPA8)和热休克转录因子1基因(HSF1)的表达量。结果显示:1)与冷应激前相比,冷应激后试验羊血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL^-1β)、白细胞介素-2(IL-2)和白细胞介素-6(IL-6)浓度显著升高(P<0.05);血清白细胞介素-4(IL-4)和免疫球蛋白G(IgG)浓度显著下降(P<0.05)。2)冷应激后,试验羊HSPA1A mRNA表达量在肝、肾、心和背最长肌中显著升高(P<0.05);HSPA6 mRNA表达量在心和背最长肌中显著升高(P<0.05);HSPA8 mRNA表达量在背最长肌中显著升高(P<0.05),在脾和十二指肠中显著下降(P<0.05);HSF1 mRNA表达量在肝、心和背最长肌中显著升高(P<0.05),在脾和十二指肠中显著下降(P<0.05)。在本试验条件下,急性冷应激能抑制绵羊的免疫功能;HSP70家族基因(HSPA1A、HSPA6和HSPA8)在各组织中的表达水平不同,其中HSPA1A对温度更敏感,宜作为绵羊冷应激的生物标记物;肝、心和背最长肌等组织中HSP70家族基因表达量升高可能与其保护组织细胞维持正常产热有关。This experiment was conducted to explore the effect of acute cold stress on immune function and expression of HSP70 family genes in different tissues in sheep.Eight(12±0.5)-month-old healthy female sheep(F1 of Small Tail Han sheep×Hu sheep)were selected and raised separately in cage in heat-preserved sheep house(wind chill temperature:(-7.14±2.53)℃).The adaptation period lasted for 7 d.At the 8 th day,female sheep were treated with acute cold stress for 12 h in outdoor(wind chill temperature:(-27.40±3.12)℃),and then blood and tissue samples were collected before and after cold stress.Serum immune indexes(cytokine and immunoglobulin G content),the expression of HSP70 family genes(HSPA1 A,HSPA6 and HSPA8)and heat shock transcription factor 1(HSF1)in different tissues(liver,kidney,spleen,heart,longissimus dorsi muscle and duodenum)were measured by ELISA and RT-PCR method,respectively.The results showed as the follows:1)Compared with before acute cold stress,the serum levels of TNF-α,IL^-1β,IL-2 and IL-6 increased significantly after acute cold stress(P<0.05),while the serum levels of IL-4 and IgG decreased significantly(P<0.05).2)The mRNA expression of HSPA1 A increased significantly in liver,kidney,heart and longissimus dorsi muscle after acute cold stress(P<0.05).HSPA6 expression level increased significantly in the longissimus dorsi muscle and heart(P<0.05).HSPA8 expression level increased significantly in longissimus dorsi muscle(P<0.05),and decreased significantly in spleen and duodenum(P<0.05).HSF1 expression level increased significantly in liver,heart and longissimus dorsi muscle(P<0.05),and decreased significantly in spleen and duodenum(P<0.05).In this experiment,acute cold stress could inhibit the immune function,and the expression levels of HSP70 family genes(HSPA1 A,HSPA6 and HSPA8)were different in different tissues.The HSPA1 A was more sensitive to temperature and could be suitable to as a biomarker for cold stress.The increased expression of HSP70 family genes in liver,heart and longissimus

关 键 词：急性冷应激 绵羊 风寒温度 免疫功能 热休克蛋白70家族基因 

分 类 号：S826.4[农业科学—畜牧学]