为了探讨冷应激对雏鸡肝脏中热休克蛋白(HSP)及炎性因子的影响,试验将120只1日龄伊莎公鸡饲喂15 d后随机分成12组进行急慢性冷应激处理。急性冷应激处理分为1个对照组和5个试验组,慢性冷应激处理分为3个对照组和3个试验组。急性冷应激条件下分别饲养0,1,3,6,12,24 h,慢性冷应激条件下分别饲养5,10,20 d,应激处理结束后迅速剖杀伊莎公鸡,取肝脏组织,采用总RNA的提取及反转录、实时荧光定量PCR反应和Western-blot技术检测HSP60、HSP70、HSP90、葡萄糖调节蛋白78(GRP78)及炎性因子PGE合成酶(PGE synthase)、环氧化酶-2(Cox-2)、核因子κB(NF-κB)、肿瘤坏死因子α(TNFα)、诱导型一氧化氮合酶(iNOS)、血红素氧合酶-1(HO-1)mRNA及蛋白的表达情况。结果表明:RNA的纯度和完整性符合要求。急慢性冷应激均可导致鸡肝脏HSP60、HSP70、HSP90、GRP78 mRNA表达明显上调,HSP70、GRP78蛋白表达与相应mRNA表达趋势一致;同时急慢性冷应激均可导致鸡肝脏炎性相关因子mRNA及蛋白表达明显上调。说明急慢性冷应激可导致肝脏炎性因子及HSP的表达增加。
To investigate the effects of cold stress on heat shock proteins and inflammatory factors in chick liver,one hundred and twenty one-day-old ISA cocks were fed to 15 days and randomly divided into twelve groups for treatment with acute and chronic cold stress.Acute cold stress was divided into one control group and five experimental groups.Chronic cold stress was divided into 3 control groups and 3 experimental groups.Acute cold stress conditions were reared for 0,1,3,6,12,24 h.And chronic cold stress conditions were fed for 5,10,20 d respectively.After that,the chicks were sacrificed immediately for extracting liver tissue.Heat shock protein 60(HSP60),heat shock protein 70(HSP70),heat shock were detected by total RNA extraction and reverse transcription,real-time fluorescence quantitative polymerase chain reaction(qPCR)and Western-blot.Protein 90(HSP90),glucose-regulated protein 78(GRP78)and inflammatory factor PGE synthase,cyclooxygenase-2(Cox-2),nuclear factor kappa B(NF-κB),tumor necrosis factor alpha Expression of(TNFα),inducible nitric oxide synthase(iNOS),and heme oxygenase-1(HO-1).The results show that the purity and integrity of RNA meet the requirements.Acute and chronic cold stress can significantly up-regulate the expression of HSP60,HSP70,HSP90 and GRP78 mRNA in chicken liver.The expression of HSP70 and GRP78 protein is consistent with the expression of relative mRNA.At the same time,both acute and chronic cold stress can cause Chicken liver inflammatory related factor mRNA and protein expression were significantly up-regulated.It indicated that acute and chronic cold stress can lead to increased expression of inflammatory factors and heat shock proteins in the liver.
Heilongjiang Animal Science And veterinary Medicine
acute and chronic cold stress
heat shock proteins