热应激造成奶牛乳腺上皮细胞损伤并影响乳合成相关载体的基因表达

doi:doi: 10.11843/j.issn.0366-6964.2016.08.023

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (8): 1704-1713.doi: doi: 10.11843/j.issn.0366-6964.2016.08.023

热应激造成奶牛乳腺上皮细胞损伤并影响乳合成相关载体的基因表达

权素玉^1，2，张源淑^1* ，卜登攀^2，3，4*

（1.南京农业大学农业部动物生理生化重点开放实验室，南京 210095； 2.中国农业科学院北京畜牧兽医研究所，动物营养学国家重点实验室，北京 100193； 3.CAAS-ICRAF农用林业与可持续畜牧业联合实验室，北京 100193； 4.东北农业大学食品安全与营养协同创新中心，哈尔滨 150030）

收稿日期:2015-09-23 出版日期:2016-08-23 发布日期:2016-09-08
通讯作者: 张源淑，教授，E-mail:zhangyuanshu@sina.com；卜登攀，研究员，E-mail:budengpan@126.com
作者简介:权素玉(1989-)，女，河北平山人，硕士生，主要从事反刍动物营养研究， E-mail:1203457176@qq.com
基金资助:
国家自然科学基金（31372341）；“十二五”科技支撑计划（2012BAD12B02-5）；中国农业科学院科技创新工程（ASTIP-IAS07）

Heat Stress-induced Cell Injury and Effects on the Gene Expression of Milk Synthesis-related Transporters in Dairy Cow

QUAN Su-yu^1，2，ZHANG Yuan-shu^1*，BU Deng-pan ^2，3，4*

(1.The Animal Physiology and Biochemistry Laboratory of the Ministry of Agriculture，Nanjing Agriculture
University，Nanjing 210095，China；2.State Key Laboratory of Animal Nutrition，Institute of Animal
Science，Chinese Academy of Agricultural Sciences，Beijing 100193，China；3.CAAS-ICRAF
Joint Laboratory on Agroforestry and Sustainable Animal Husbandry，Beijing 100193，China；4. Synergetic
Innovation Center of Food Safety and Nutrition，Northeast Agricultural University，Harbin 150030，China)

Received:2015-09-23 Online:2016-08-23 Published:2016-09-08

摘要/Abstract

摘要：

本试验以奶牛乳腺上皮细胞为模型，旨在寻找热应激下调奶牛泌乳功能的相关因素。42 ℃高温处理奶牛乳腺上皮细胞0、0.5、1、1.5、2、4、8 及12 h，检测其对细胞活率，线粒体膜电位，热休克分子及蛋白基因、氧化应激相关基因以及氨基酸和葡萄糖转运载体基因表达的影响。结果显示，不同时间热处理显著降低乳腺上皮细胞活率(P<0.05)，损伤细胞线粒体膜电位。HSP27基因表达显著下降(P<0.05)，HSP70基因表达显著上调(P<0.05)， HSP90基因表达在前4 h显著上调(P<0.05)，后恢复至正常水平，HSF-1基因表达在某些时间点显著上调(P<0.05)。氧化应激相关基因Keach样环氧氯丙烷相关蛋白-1 (Keap1)表达无显著变化 (P>0.05)，核因子E2相关因子2 (Nrf2) 表达显著上调(P<0.05)，NAD(P)H脱氢酶醌-1 (NQO1) 表达显著上调(P<0.05)。跨膜氨基酸转运载体SLC7A7基因表达在1 h和4 h显著上调(P<0.05)。葡萄糖转运载体GLUT1及GLUT8基因表达显著下调(P<0.05)。葡萄糖转运载体GLUT12在热处理0～2 h表达显著上调，4～12 h显著下调(P<0.05)。热应激能够显著降低奶牛乳腺上皮细胞活率，引起氧化应激，改变热休克蛋白和分子及跨膜转运载体的基因表达。

关键词: , 热应激, 牛乳腺上皮细胞, 细胞活力, 线粒体膜电位, 氧化应激, 热休克蛋白, 跨膜转运载体

Abstract:

Bovine mammary epithelial cell was chosen as the model to find the factors impairing lactation performance of dairy cow during heat stress.Cell vitality，mitochondrial membrane potential and the expression of genes related to heat shock molecule and proteins，oxidative stress，amino acids and glucose transporters were detected in the bovine mammary epithelial cells treated for 0，0.5，1，1.5，2，4，8 and 12 h at 42 ℃，respectively.The result showed that different heat treatment time significantly reduced mammary epithelial cell vitality (P<0.05) and damaged mitochondrial membrane potential.The expression of HSP27 decreased significantly (P<0.05)，while HSP70 increased significantly (P<0.05).The expression of HSP90 increased significantly in 0-4 h (P<0.05)，and returned to normal level after then.The expression of HSF-1 increased significantly in some time points (P<0.05).The gene expression of Keach like ECH-associated protein-1 (Keap1) showed no significant difference(P>0.05)，while nuclear factor-E2-related factor 2 (Nrf2) and NAD (P) H dehydrogenase quinone-1 (NQO1) increased significantly (P<0.05).The gene expression of transmembrane amino acid transporter SLC7A7 increased significantly in 1 h and 4 h(P<0.05).The gene expression of glucose transporters GLUT1 and GLUT8 decreased significantly (P<0.05).The expression of glucose transporter GLUT12 increased significantly during 0-2 h，and decreased significantly during 4-12 h (P<0.05).In conclusion，heat stress could significantly reduce the cell vitality，cause oxidative stress，change the gene expression of heat shock proteins and molecule and membrane transport carriers.

Key words: heat stress, bovine mammary epithelial cell, cell vitality, mitochondrial membrane potential, oxidative stress；heat shock protein, membrane transport carriers

中图分类号:

S852.23

权素玉，张源淑，卜登攀. 热应激造成奶牛乳腺上皮细胞损伤并影响乳合成相关载体的基因表达[J]. 畜牧兽医学报, 2016, 47(8): 1704-1713.

QUAN Su-yu，ZHANG Yuan-shu，BU Deng-pan . Heat Stress-induced Cell Injury and Effects on the Gene Expression of Milk Synthesis-related Transporters in Dairy Cow[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(8): 1704-1713.

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热应激造成奶牛乳腺上皮细胞损伤并影响乳合成相关载体的基因表达

Heat Stress-induced Cell Injury and Effects on the Gene Expression of Milk Synthesis-related Transporters in Dairy Cow

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