冷热应激对牛PBMC和MDBK细胞活力及HSP70表达的影响  

Effect of cold and heat stress on cell vitality and HSP70 expression in PBMC and MDBK of cattle

作　　者：康玲 胡丽蓉 李玮[2] 鄢新义 亓建刚 徐青[1] 王雅春[2] KANG Ling;HU Li-rong;LI Wei;YAN Xin-yi;QI Jian-gang;XU Qing;WANG Ya-chun(Institute of Biological Science and Bioengineering,Beijing Jiaotong University,Beijing 100044,China;College of Animal Science and Technology,China Agricultural University,Beijing 100193,China;College of Animal Science,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]北京交通大学生命科学与生物工程研究院,北京100044 [2]中国农业大学动物科技学院,北京100193 [3]新疆农业大学动物科学学院,新疆乌鲁木齐830052

出　　处：《中国兽医学报》2018年第7期1359-1365,共7页Chinese Journal of Veterinary Science

基　　金：现代农业产业技术体系建设专项资金资助项目(CARS-37);北京市科技计划资助项目(Z151100001015012)

摘　　要：以奶牛原代培养细胞PBMC（peripheral blood mononuclear cells）和商业化细胞系MDBK（madin-darby bovine kidney）为细胞模型，采用MTS、实时荧光定量PCR（quantitative real-time PCR）及Western blot等方法检测不同程度的冷热应激对牛PBMC和MDBK细胞活力及HSP70表达水平的影响，分析不同温度的冷热应激条件下细胞活力和HSP70表达变化的规律，以及不同类型细胞应答冷热应激的差异。结果显示，冷应激和热应激均会使细胞活力降低，应激强度越大，细胞活力越低；冷热应激会调控细胞中HSP70的表达水平，重度冷应激（10℃）下调HSP70的表达，而热应激则上调HSP70的表达，但若超过细胞耐受极限度HSP70表达开始下降；HSP70表达水平应对冷热应激的变化与细胞的类型和冷热应激强度有关。结果表明，冷热应激会降低细胞活力，调控细胞HSP70的表达水平，HSP70的表达变化能够反映细胞冷热应激的强度，可作为评价奶牛冷热应激反应的潜在分子标记在奶牛个体中进行深入研究。The present study was conducted to investigate the effect of different temperature stress on cell vitality and HSP70 expression in PBMC（peripheral blood mononuclear ceils） and MDBK （madin-darby bovine kidney） from cattle. Using MTS, quantitative real-time PCR, Western blot, and so on,analyzed the changes of cell viability and HSP70 expression in PBMC and MDBK treated with different temperature, and and the differences between the two types of cell in response to cold and heat. The result suggested that both cold and heat stress caused the decrease of cell viability. Also cold and heat stress changed the expression of HSP70 in two types of cell, reducting HSP70 under severe cold stress（10℃）, increasing HSP70 level to heat stress at first, then decreased when the heat stress exceeded the cell tolerance. The expression variation of HSP70 was associated with cell types and thermal stress conditions. The results revealed heat and cold stress changed the cell viability and expression of HSP70 in cattle cells,and different levels of HSP70 appeared in different thermal conditions,so further study should be carried out using HSP70 as the potential molecular marker to assess the heat and cold stress of cattle.

关 键 词：冷热应激 HSP70 细胞活力 PBMC细胞 MDBK细胞 基因表达 

分 类 号：S856.9[农业科学—临床兽医学]