[1]王换换,申正杰,肖航,等.热应激对肝脏中Keap1-Nrf2信号通路及下游基因表达的影响[J].南京农业大学学报,2017,40(1):151-156.[doi:10.7685/jnau.201602023]
 WANG Huanhuan,SHEN Zhengjie,XIAO Hang,et al.Effects of heat stress on Keap1-Nrf2-ARE signal pathway of liver in dairy cows[J].Journal of Nanjing Agricultural University,2017,40(1):151-156.[doi:10.7685/jnau.201602023]
点击复制

热应激对肝脏中Keap1-Nrf2信号通路及下游基因表达的影响()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
40卷
期数:
2017年1期
页码:
151-156
栏目:
出版日期:
2017-01-08

文章信息/Info

Title:
Effects of heat stress on Keap1-Nrf2-ARE signal pathway of liver in dairy cows
作者:
王换换 申正杰 肖航 刘小倩 艾阳 张源淑
南京农业大学农业部动物生理生化重点开放实验室, 江苏 南京 210095
Author(s):
WANG Huanhuan SHEN Zhengjie XIAO Hang LIU Xiaoqian AI Yang ZHANG Yuanshu
Key Laboratory of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
奶牛热应激肝脏Keap1-Nrf2-ARE信号通路基因表达
Keywords:
dairy cowheat stressliverKeap1-Nrf2-ARE signal pathwaygenes expression
分类号:
S811.3
DOI:
10.7685/jnau.201602023
摘要:
[目的] 本试验旨在探讨热应激对泌乳期奶牛肝脏及抗氧化应激信号通路Keap1-Nrf2-ARE基因表达的影响。[方法] 6头泌乳期荷斯坦奶牛饲养于南京地区夏季炎热季节,试验从6至8月持续5周,在第1周(平均气温26℃)和最后1周(平均气温32℃)采集颈静脉血液和肝脏组织样品。检测血液中相关生化指标和皮质醇及肝脏组织中MDA等的含量,并检测奶牛肝脏HSP70、Keap1-Nrf2通路及其下游HO1、GCLM、NQO1、GCLC等基因的表达。[结果] 与常温时相比,热应激奶牛血液中皮质醇含量和AKP活性极显著升高(P<0.01);奶牛肝脏组织中MDA含量显著升高(P<0.05),CAT、SOD和GSH-Px活性有所升高;肝脏组织中HSP70基因表达极显著升高(P<0.01),Keap1Nrf2基因表达显著升高(P<0.05),其下游4个基因中,HO1NQO1表达显著提高(P<0.05),GCLMGCLC表达有升高趋势(P>0.05)。[结论] 高温致泌乳奶牛肝脏处于应激状态,肝脏中Keap1-Nrf2-ARE信号通路介导的Ⅱ相解毒酶[谷氨酰半胱氨酸合成酶催化亚单位(GCLC)、醌氧化还原酶1(NQO1)、谷氨酸-半胱氨酸连接酶修饰亚基(GCLM)]和抗氧化基因的转录通路被激活,提示Nrf2通路在抗热应激过程中可能发挥着重要作用。
Abstract:
[Objectives] The aim of this experiment was to investigate the liver function injury and the genes expression of Keap1-Nrf2-ARE signaling pathways during thermal stress of lactation cows. [Methods] The study lasted for 5 weeks(from June to August). Six lactation cows were selected. Blood samples were collected from jugular vein on both the first and last week to analyze the related blood composition and cortisol content,and malondialdehyde(MDA) content in the liver. The liver tissue samples were also collected by living donor. The gene expression of Keap1-Nrf2 pathway and its downstream HO1,GCLM,NQO1 and GCLC protein were tested. [Results] During heat stress,the cortisol and AKP increased significantly(P<0.01),the MDA content increased significantly(P<0.05),the activities of CAT,SOD and GSH-Px was increased in cows liver tissue. In liver,the gene expression of HSP70 increased significantly(P<0.01). The gene expression of Keap1 and Nrf2 increased significantly(P<0.05). And among the four downstream genes,HO1 and NQO1 increased significantly(P<0.05);GCLM and GCLC increased but with no significance. [Conclusions] Hyperthermia caused hepatic stress in dairy cows. The Keap1-Nrf2-ARE signal pathway mediated phase Ⅱ detoxification enzyme(GCLC,NQO1 and GCLM) and some antioxidant genes were motivated.

参考文献/References:

[1] Bohmanova J,Misztal I,Cole J B. Temperature-humidity indices as indicators of milk production losses due to heat stress[J]. Journal of Dairy Science,2007,90(4):1947-1956.
[2] Bernabucci U,Biffani S,Buggiotti L,et al. The effects of heat stress in Italian Holstein dairy cattle[J]. Journal of Dairy Science,2014,97(1):471-486.
[3] 吴文旋,段永邦.补饲精料对贵州黑山羊血浆抗氧化应激能力和生化指标的影响[J]. 家畜生态学报,2013,34(1):60-65. Wu W X,Duan Y B. Effects of supplemental concentrate on plasma anti-oxidative stress capative and boichemical metabolites for Guizhou black goats[J]. Acta Ecologiae Animalis Domastic,2013,34(1):60-65(in Chinese with English abstract).
[4] 付戴波,宋小珍,瞿明仁,等. 热应激对肉牛血清内分泌激素含量、抗氧化酶活性及生理生化指标的影响[J]. 动物营养学报,2012,24(12):62-65. Fu D B,Song X Z,Qu M R,et al. Effect of heat stress on hormone content,antioxidant enzyme activity and physiological and serum biochemical indices of beef cattle[J]. Chinese Journal of Animal Nutrition,2012,24(12):62-65(in Chinese with English abstract).
[5] 蔡维霞,张军,胡大海. 氧化和化学应激的防御性转导通路——Nrf2/ARE[J]. 中国生物化学与分子生物学报,2009,25(4):297-303. Cai W X,Zhang J,Hu D H. Defensive pathway of Nrf2/ARE involved in oxidative and chemical stress[J]. Chinese Journal of Biochemistry and Molecular Biology,2009,25(4):297-303(in Chinese with English abstract).
[6] 童海达,王佳茗,宋英. Keap1-Nrf2-ARE在机体氧化应激损伤中的防御作用[J]. 癌变·畸变·突变,2013,25(1):71-75. Dong H D,Wang J M,Song Y. The defensive role of Keap1-Nrf2-ARE in oxidative stress injury[J]. Carcinogenesis,Distortion and Mutagenesis,2013,25(1):71-75(in Chinese).
[7] 王菲,黄毅,李延森,等. Keap1/Nrf2/ARE通路相关基因在热诱导的氧化应激小鼠肝脏中的表达[J]. 中国生物化学与分子生物学报,2014,30(3):284-290. Wang F,Huang Y,Li Y S,et al. Expression of Keap1/Nrf2/ARE pathway-related genes in protective response to heat-induced oxidative stress in the liver of mice[J]. Chinese Journal of Biochemistry and Molecular Biology,2014,30(3):284-290(in Chinese with English abstract).
[8] 何钦. 热应激对不同泌乳阶段奶牛生产性能及其营养代谢的影响[D]. 重庆:西南大学,2012. He Q.Effects of heat stress on performance and nutrient metabolism of dairy cows in different lactation stages[D]. Chongqing:Southwest University,2012(in Chinese with English abstract).
[9] Velez J C,Donkin S S. Bovine somatotropin increases hepatic phosphoenolpyruvate carboxykinase mRNA in lactating dairy cows[J]. Journal of Dairy Science,2014,87:1325-1335.
[10] Wise M E,Armstrong D V,Huber J,et al. Hormonal alteration in the lactation dairy cow in response to thermal stress[J]. Journal of Dairy Science,1998,71:2480-2484.
[11] 李建国,桑润滋,张正珊,等. 热应激对奶牛生理常值、血液生化指标、繁殖及泌乳性能的影响[J]. 河北农业大学学报,1998,21(4):69-70. Li J G,Sang R Z,Zhang Z S,et al. Influence of heat stress on hysiological response,blood biochemical parameters,reproduction and milk performance of holstein cow[J]. Journal of Agricultural University of Hebei,1998,21(4):69-70(in Chinese with English abstract).
[12] 穆玉云.乳牛热应激指标的检测[J]. 安徽农业大学学报,1993,20(1):66-67. Mu Y Y. Detection of cows heat stress index[J]. Journal of Anhui Agricultural University,1993,20(1):66-67(in Chinese with English abstract).
[13] 林谦,邱磊,云龙,等. 核因子E2相关因子2调控机体抗氧化途径特性及其与畜禽的健康和肉品质的关系[J]. 动物营养学报,2014,26(6):1421-1429. Lin Q,Qiu L,Yun L,et al. Nuclear factor erythroid-2-related factor 2 mediated antioxidant pathway character and its relation on health and meat quality of livestock and poultry.[J]. Chinese Journal of Animal Nutrition,2014,26(6):1421-1429(in Chinese with English abstract).
[14] Lin H,Decuypere E,Buyse J. Acute heat stress induces oxidative stress in broiler chickens[J]. Comparative Biochemistry and Physiology Part A:Molecular and Integrative Physiology,2006,144(1):11-17.
[15] Yang L,Tan G Y,Fu Y Q,et al. Effects of acute heat stress and subsequent stress removal on function of hepatic mitochondrial respiration,ROS production and lipid peroxidation in broiler chickens[J]. Comparative Biochemistry and Physiology Part C:Toxicology and Pharmacology,2010,151(2):204-208.
[16] 陈强.奶牛HSP70基因5’-侧翼区PCR-SSCP分析及其与生产性能的关系[D]. 南京:南京农业大学,2006. Chen Q.PCR-SSCR analysis in 5’-flanking region of heat shock protein 70 gene and their association with production traits in dairy cows[J]. Nanjing:Nanjing Agricultural University,2006(in Chinese with English abstract).
[17] 彭立颖,张思学,王洪伟,等. 不同程度奶牛乳腺炎乳中碱性磷酸酶和酸性磷酸酶的变化[J]. 中国兽医杂志,2011,47(4):40-41. Peng L Y,Zhang S X,Wang H W,et al. The change of milk alkaline and acid phosphatase in different types of mastitis in dairy cattle[J]. Chinese Journal of Veterinary Medicine,2011,47(4):40-41(in Chinese).
[18] Ardeshirpour L,Dumitru C,Dann P,et al. OPG treatment prevents bone loss during lactation but does not affect milk production or maternal calcium metabolism[J]. Endocrinology,2015,11:12-32.
[19] 薛瑞益. 围产期奶牛血中某些生化指标与脂类代谢病关系的研究[D]. 呼和浩特:内蒙古农业大学,2006. Xue R Y.Studies on the relations between lipid metabolism and some biochemical indexes in blood of cow during perinatal period[J]. Hohhot:Inner Mongolia Agricultural University,2006(in Chinese with English abstract).
[20] Thimmulappa R K,Mai K H,Srisuma S,et al. Idetification of Nrf2-regulated genes induced by the chemopreventive agent sulforaphane by oligonucleotide microarray[J]. Cancer Res,2002,62:5196-5203.
[21] 李锦春,徐同锏,钟刚,等. γ-氨基丁酸对热应激奶牛泌乳性能和血液抗氧化酶、激素及离子的影响[J]. 畜牧与兽医,2016,48(5):12-18. Li J C,Xu T J,Zhong G,et al. Effects of coaled γ-aminobutyric acid on laction performance blood antioxidant enzymes hormones and ions in dairy cows[J]. Animal Husbandry and Veterinary Medicine,2016,48(5):12-18(in Chinese with English abstract).
[22] 坚哲. Nrf2-ARE信号通路在白癜风氧化应激发病中的作用和机制研究[D]. 西安:第四军医大学,2013. Jian Z. The role of Nrf2-ARE pathway in the oxidative stress pathogenesis of vitiligo and its mechanism[J]. Xi’an:The Fourth Military Medical University,2013(in Chinese with English abstract).

相似文献/References:

[1]孙建全,赵红波,刘春林,等.改变日粮蛋白质和赖氨酸含量对奶牛生产性能和氮利用及代谢激素的影响[J].南京农业大学学报,2011,34(2):124.[doi:10.7685/j.issn.1000-2030.2011.02.022]
 SUN Jian-quan,ZHAO Hong-bo,LIU Chun-lin,et al.Effects of changing diet protein and lysine content on milk performance,nitrogen utilization and blood metabolic hormones for dairy cows[J].Journal of Nanjing Agricultural University,2011,34(1):124.[doi:10.7685/j.issn.1000-2030.2011.02.022]
[2]丁月云,张莉莉,陈莎莎,等.组织块种植法体外培养奶牛乳腺上皮细胞[J].南京农业大学学报,2008,31(4):91.[doi:10.7685/j.issn.1000-2030.2008.04.017]
 DING Yue-yun,ZHANG Li-li,CHEN Sha-sha,et al.Explants culture of cow mammary epithelial cells in vitro[J].Journal of Nanjing Agricultural University,2008,31(1):91.[doi:10.7685/j.issn.1000-2030.2008.04.017]
[3]杨震,黄克和,候加法,等.分娩前后酮病奶牛脂代谢指标的分析[J].南京农业大学学报,1993,16(00):134.[doi:10.7685/j.issn.1000-2030.1993.00.031]
 Yang Zheng Huang Kehe Hou Jiafa Mao Xinzhi Yue Zehua.STUDIES ON LIPID MOBILIZATION IN DAIRY COWS WITH KETOSIS BEFORE AND AFTER PARTURITION[J].Journal of Nanjing Agricultural University,1993,16(1):134.[doi:10.7685/j.issn.1000-2030.1993.00.031]
[4]佘鹏湘,陈振旅,王小龙.南京地区奶牛酮病病因和诊断的研究[J].南京农业大学学报,1990,13(02):106.[doi:10.7685/j.issn.1000-2030.1990.02.020]
 She Pengxiang Chen Zhenlu Wang Xiaolong.STUDY ON ETIOLOGY AND DIAGNOSIS OF KETOSIS IN DAIRY COW IN NANJING AREA[J].Journal of Nanjing Agricultural University,1990,13(1):106.[doi:10.7685/j.issn.1000-2030.1990.02.020]
[5]郭爱珍,陈万芳.不孕母牛的染色体畸变分析[J].南京农业大学学报,1989,12(03):66.[doi:10.7685/j.issn.1000-2030.1989.03.014]
 Guo Aizheng,Chen Wanfang.CHROMOSOME ABNORMALITIES IN INFERTILE COWS[J].Journal of Nanjing Agricultural University,1989,12(1):66.[doi:10.7685/j.issn.1000-2030.1989.03.014]
[6]杨利国,刘荣军,王元兴.不孕奶牛用复方缩宫素乳剂处理后外周血中雌二醇和孕酮水平的变化[J].南京农业大学学报,1998,21(2):73.[doi:10.7685/j.issn.1000-2030.1998.02.014]
 Yang Liguo,Liu Rongjun,Wang Yuanxing.Estradiol and progesterone levels in sera of infertile dairy cows after treatment with Oxytocin Compound Emulsion[J].Journal of Nanjing Agricultural University,1998,21(1):73.[doi:10.7685/j.issn.1000-2030.1998.02.014]
[7]吴金节,章孝荣,刘亚,等.奶牛子宫内膜炎防治前后血液流变学参数的变化[J].南京农业大学学报,2000,23(2):65.[doi:10.7685/j.issn.1000-2030.2000.02.016]
 Wu Jinjie,Zhang Xiaorong,Liu Ya,et al.Changes of hemorrheology parameters of dairy cows with endometritis before and after prevention and treatment[J].Journal of Nanjing Agricultural University,2000,23(1):65.[doi:10.7685/j.issn.1000-2030.2000.02.016]
[8]韩博,梁俭,张一贤,等.日粮添加硒碘铜对宁夏地区奶牛生产和繁殖性能的影响[J].南京农业大学学报,2000,23(3):76.[doi:10.7685/j.issn.1000-2030.2000.03.020]
 Han Bo,Liang Jian,Zhang Yixian,et al.Effect of diets supplemented with selenium,iodine and copper on the production and reproduction of dairy cowsin Ningxia area[J].Journal of Nanjing Agricultural University,2000,23(1):76.[doi:10.7685/j.issn.1000-2030.2000.03.020]
[9]蔡亚非,李莲,刘庆华,等.热应激奶牛外周血淋巴细胞凋亡和bax2α基因表达[J].南京农业大学学报,2005,28(1):66.[doi:10.7685/j.issn.1000-2030.2005.01.015]
 CAI Ya-fei,LI Lian,LIU Qing-hua,et al.Analysis of apoptosis and expression of bax-α of lymphocytes in peripheral blood of dairy cows[J].Journal of Nanjing Agricultural University,2005,28(1):66.[doi:10.7685/j.issn.1000-2030.2005.01.015]
[10]廖旭东,崔群维,周璇,等.公牛HSP 基因多态性与其精液品质及后裔性能的相关分析[J].南京农业大学学报,2011,34(4):105.[doi:10.7685/j.issn.1000-2030.2011.04.019]
 LIAO Xu-dong,CUI Qun-wei,ZHOU Xuan,et al.The correlation analysis between bulls HSP gene polymorphism and semen quality and progeny performance[J].Journal of Nanjing Agricultural University,2011,34(1):105.[doi:10.7685/j.issn.1000-2030.2011.04.019]
[11]张丽,韩兆玉.甜菜碱对夏季热应激奶牛产奶性能和HSP70及其mRNA表达的影响[J].南京农业大学学报,2013,36(6):89.[doi:10.7685/j.issn.1000-2030.2013.06.014]
 ZHANG Li,HAN Zhaoyu.Effects of supplementing betaine on milk performance, HSP70 and HSP70 mRNA level of cows under heat stress[J].Journal of Nanjing Agricultural University,2013,36(1):89.[doi:10.7685/j.issn.1000-2030.2013.06.014]
[12]徐同锏,钟刚,王希春,等.脂肪包被γ-氨基丁酸对持续热应激奶牛HPA轴和部分免疫指标的影响[J].南京农业大学学报,2014,37(1):127.[doi:10.7685/j.issn.1000-2030.2014.01.021]
 XU Tongjian,ZHONG Gang,WANG Xichun,et al.Effects of lipid coated γ-aminobutyric acid on HPA axis and some immune indicators of cows in heat stress[J].Journal of Nanjing Agricultural University,2014,37(1):127.[doi:10.7685/j.issn.1000-2030.2014.01.021]

备注/Memo

备注/Memo:
收稿日期:2016-02-23。
基金项目:国家重大基础研究发展计划项目(2011CB100802)
作者简介:王换换,硕士研究生。
通信作者:张源淑,教授,博导,研究方向为营养生物化学,E-mail:zhangyuanshu@njau.edu.cn。
更新日期/Last Update: 1900-01-01