热应激通过诱导奶牛乳腺细胞凋亡减少乳蛋白

高胜涛; 郭江; 权素玉; 南雪梅; 卜登攀

doi:10.3969/j.issn.1006-267x.2016.05.040

动物营养学报 >

2016 , Vol. 28 >Issue 5: 1615 - 1625

DOI: https://doi.org/10.3969/j.issn.1006-267x.2016.05.040

研究简报 Short Communications

热应激通过诱导奶牛乳腺细胞凋亡减少乳蛋白

高胜涛 ,
郭江 ,
权素玉 ,
南雪梅 ,
卜登攀

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1. 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;
2. 中国农业科学院-世界农用林业中心, 农用林业与可持续畜牧业联合实验室, 北京 100081;
3. 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: 2015-12-15

网络出版日期: 2016-05-24

基金资助

国家自然科学基金(31372341);十二五国家科技支撑计划(2012BAD12B02-05);中国农业科学院科技创新工程(ASTIP-IAS07)

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Heat-Stress Decreases Milk Protein through Induction of Mammary Cells Apoptosis of Cows

GAO Shengtao ,
GUO Jiang ,
QUAN Suyu ,
NAN Xuemei ,
BU Dengpan

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1. State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. CAAS-ICRAF Joint Laboratory on Agroforestry and Sustainable Animal Husbandry, Beijing 100081, China;
3. Synergetic Innovation Center of Food Safety and Nutrition, Harbin 150030, China

Received date: 2015-12-15

Online published: 2016-05-24

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摘要

本试验旨在研究热应激时乳蛋白含量和产量降低与泌乳相关激素、乳蛋白合成相关信号通路及乳腺细胞凋亡的关系,并揭示热应激引起乳蛋白含量和产量下降的原因。选取4头泌乳日龄、体重、产奶量相近的经产健康荷斯坦奶牛作为试验动物。采取2×2交叉试验设计,试验共2期,每期18 d(其中预试期和试验期各9 d),2期之间间隔30 d。预试期为热中性环境,自由采食。试验期奶牛随机分为2组(n=2),分别为热应激组和配对限饲组。结果表明:1)热应激显著降低了乳蛋白的含量和产量(P<0.05)。2)热应激对酪氨酸激酶(JAK)-信号转导及转录激活因子(STAT)信号通路及κ-酪蛋白(CSN3)基因表达量没有显著影响(P>0.05)。3)热应激显著增加了哺乳动物雷帕霉素靶蛋白(mTOR)信号通路中的mTOR的基因表达量(P<0.05),且有增加核糖体S6蛋白激酶(S6K1)基因表达量的趋势(0.05≤P<0.10)。4)热应激增加了乳腺细胞中与细胞凋亡相关的半胱氨酸蛋白酶3(CASP3)、环氧合酶-2(COX2)基因的表达量(P<0.05),且有增加B淋巴细胞瘤-2相关X蛋白(BAX)基因表达量的趋势(0.05≤P<0.10)。综合可知,热应激可能并不是通过调控单个乳腺细胞合成乳蛋白的能力来影响乳蛋白的含量和产量,而是通过诱导乳腺细胞的凋亡,减少可用于乳蛋白合成的乳腺细胞的数量来影响的。

关键词： 热应激; 乳蛋白; JAK-STAT; mTOR; 细胞凋亡

本文引用格式

高胜涛 , 郭江 , 权素玉 , 南雪梅 , 卜登攀 . 热应激通过诱导奶牛乳腺细胞凋亡减少乳蛋白[J]. 动物营养学报, 2016 , 28(5) : 1615 -1625 . DOI: 10.3969/j.issn.1006-267x.2016.05.040

Abstract

This experiment was conducted to investigate the relationship among the decrease of milk protein content and yield under heat stress, hormones related to lactation, pathways involved in milk protein synthesis, and apoptosis of mammary cells, and to find reasons for milk protein decline caused by heat stress. Four healthy multiparous lactating Holstein cows with similar days in milk, body weight and milk yield were used as experiment animals. Crossover design was applied, there were two experimental periods, and each period lasted for 18 days (9 d of pretrial period, and 9 d of trail period) with 30 d between the two periods. Cows in pretrial period were exposed to thermal neutral conditions and allowed to eat ad libitum. Cows in trial period were randomly divided into two groups (n=2), which were heat stress group and pair-restricted feeding group. The results showed as follows:1) heat stress significantly decreased milk protein yield and content (P<0.05). 2) Heat stress had no significant influence on Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway and κ-casein (CSN3) gene expression (P>0.05). 3) In mTOR pathway, heat stress significantly increased the expression of mammalian target of rapamycin (mTOR) gene (P<0.05), and had the tendency to increase the expression of ribosomal protein S6 kinase (S6K1) gene (0.05≤P<0.10). 4) Heat stress significantly increased the expressions of Caspase-3(CASP3) and cyclooxygenase-2(COX2) genes involved in apoptosis (P<0.05), and had the tendency to increase the expression of B-cell lymphoma-2-associated X protein (BAX) (0.05≤P<0.10). It is concluded that heat stress may not be able to decline milk protein content and yield through regulating the capacity of individual mammary cells, but thorough induction of apoptosis of the mammary cells and decrease of the quantity of mammary cells used for milk protein synthesis.

Key words： heat stress; milk protein; JAK-STAT; mTOR; apoptosis

参考文献

[1] BERNABUCCI U,BASIRICÕ L,MORERA P,et al.Effect of summer season on milk protein fractions in Holstein cows[J].Journal of Dairy Science,2015,98(3):1815-1827.

[2] MCDOWELL R E,MOODY E G,VAN SOEST P J,et al.Effect of heat stress on energy and water utilization of lactating cows[J].Journal of Dairy Science,1969,52(2):188-194.

[3] 马露,卜登攀,高胜涛,等.热应激影响奶牛乳腺酪蛋白合成的机制[J].动物营养学报,2015,27(11):3319-3325.
[4] NEVILLE M C,MCFADDEN T B,FORSYTH I.Hormonal regulation of mammary differentiation and milk secretion[J].Journal of Mammary Gland Biology and Neoplasia,2002,7(1):49-66.

[5] BIONAZ M,LOOR J J.Gene networks driving bovine mammary protein synthesis during the lactation cycle[J].Bioinformatics and Biology Insights,2011,5:83-98.
[6] 童娜.热应激对草鱼肾细胞凋亡的影响[D].硕士学位论文.武汉:华中农业大学,2014.
[7] 周振峰,崔瑞莲,王加启,等.热应激对体外培养奶牛乳腺上皮细胞生长、凋亡及其热休克蛋白mRNA转录的影响[J].畜牧兽医学报,2010,41(5):600-607.
[8] 蔡亚非,李莲,刘庆华,等.热应激奶牛外周血淋巴细胞凋亡和BAX-α基因表达[J].南京农业大学学报,2005,28(1):66-70.
[9] 胡菡,王加启,李发弟,等.高温诱导体外培养奶牛乳腺上皮细胞的应激响应[J].农业生物技术学报,2011,19(2):287-293.
[10] BANDARANAYAKA D D,HOLMES C W.Changes in the composition of milk and rumen contents in cows exposed to a high ambient temperature with controlled feeding[J].Tropical Animal Health and Production,1976,8(1):38-46.

[11] RHOADS M L,RHOADS R P,VANBAALE M J,et al.Effects of heat stress and plane of nutrition on lactating Holstein cows:Ⅰ.Production,metabolism,and aspects of circulating somatotropin[J].Journal of Dairy Science,2009,92(5):1986-1997.

[12] COWLEY F C,BARBER D G,HOULIHAN A V,et al.Immediate and residual effects of heat stress and restricted intake on milk protein and casein composition and energy metabolism[J].Journal of Dairy Science,2015,98(4):2356-2368.

[13] ARMSTRONG D V.Heat stress interaction with shade and cooling[J].Journal of Dairy Science,1994,77(7):2044-2050.

[14] NRC.Nutrient requirements of dairy cattle[M].7th ed.Washington,D.C.:The National Academies Press,2001.
[15] 王加启.反刍动物营养学研究方法[M].北京:现代教育出版社,2011.
[16] VAN SOEST P J,ROBERTSON J B,LEWIS B A.Methods for dietary fiber,neutral detergent fiber,and nonstarch polysaccharides in relation to animal nutrition[J].Journal of Dairy Science,1991,74(10):3583-3597.

[17] DE MORAES C N,MAIA L,DE LIMA P F,et al.Temporal analysis of prostaglandin F2α receptor,caspase 3,and cyclooxygenase 2 messenger RNA expression and prostaglandin F2α receptor and cyclooxygenase 2 protein expression in endometrial tissue from multiparous Nelore (Bos taurus indicus) cows treated with cloprostenol sodium during puerperium[J].Theriogenology,2015,83(2):276-284.

[18] KAMEMORI Y,WAKAMIYA K,NISHIMURA R,et al.Expressions of apoptosis-regulating factors in bovine retained placenta[J].Placenta,2011,32(1):20-26.

[19] OKUDA K,SAKUMOTO R,OKAMOTO N,et al.Cellular localization of genes and proteins for tumor necrosis factor-α (TNF),TNF receptor types Ⅰ and Ⅱ in bovine endometrium[J].Molecular and Cellular Endocrinology,2010,330(1/2):41-48.
[20] ZIMBLEMAN R B,RHOADS R P,RHOADS,M L,et al.A re-evaluation of the impact of temperature humidity index (THI) and black globe humidity index (BGHI) on milk production in high producing dairy cows[C]//Proceedings of the 24th annual southwest nutrition conference.Arizona:Department of Animal Sciences,the University of Arizona,2009.
[21] 艾阳,曹洋,谢正露,等.热应激时奶牛血液中游离氨基酸流向与乳蛋白下降的关系研究[J].食品科学,2015,36(11):38-41.
[22] 程建波,王伟宇,郑楠,等.自然生产条件下热应激周期变化揭示泌乳中期奶牛出现"热应激乳蛋白降低征"[J].中国畜牧兽医,2014,41(10):73-84.
[23] WEST J W,MULLINIX B G,BERNARD J K.Effects of hot,humid weather on milk temperature,dry matter intake,and milk yield of lactating dairy cows[J].Journal of Dairy Science,2003,86(1):232-242.

[24] JOHNSON H D,VANJONACK W J.Effects of environmental and other stressors on blood hormone patterns in lactating animals[J].Journal of Dairy Science,1976,59(9):1603-1617.

[25] BARBER M C,TRAVERS M T,FINLEY E,et al.Growth-hormone-prolactin interactions in the regulation of mammary and adipose-tissue acetyl-CoA carboxylase activity and gene expression in lactating rats[J].Biochemical Journal,1992,285:469-475.
[26] BASSETT N S,CURRIE M J,BREIER B H,et al.The effects of ovine placental lactogen and bovine growth hormone on hepatic and mammary gene expression in lactating sheep[J].Growth Hormone & IGF Research,1998,8(6):439-446.

[27] 王皓宇,秦彤,郝海生,等.胰岛素对体外培养奶牛乳腺上皮细胞乳蛋白、乳脂肪合成相关基因mRNA表达的影响[J].畜牧兽医学报,2013,44(5):710-718.
[28] 宋代军,何钦,姚焰础.热应激对不同泌乳阶段奶牛生产性能和血清激素浓度的影响[J].动物营养学报,2013,25(10):2294-2302.
[29] HERBEIN J H,AIELLO R J,ECKLER L I,et al.Glucagon,insulin,growth hormone,and glucose concentrations in blood plasma of lactating dairy cows[J].Journal of Dairy Science,1985,68(2):320-325.

[30] ITOH F,OBARA Y,ROSE M T,et al.Insulin and glucagon secretion in lactating cows during heat exposure[J].Journal of Animal Science,1998,76(8):2182-2189.
[31] 朱国标,辛火,李继红,等.高热与皮质醇、泌乳素、LPO及SOD的关系[J].中国公共卫生学报,1994,13(4):218-219.
[32] MOHAMMED M E,JOHNSON H D.Effect of growth hormone on milk yields and related physiological functions of Holstein cows exposed to heat stress[J].Journal of Dairy Science,1985,68(5):1123-1133.

[33] MCGUIRE M A,BEEDE D K,COLLIER R J,et al.Effects of acute thermal stress and amount of feed intake on concentrations of somatotropin,insulin-like growth factor (IGF)-Ⅰ and IGF-Ⅱ,and thyroid hormones in plasma of lactating Holstein cows[J].Journal of Animal Science,1991,69(5):2050-2056.
[34] LIU X,ROBINSON G W,HENNIGHAUSEN L.Activation of STAT5A and STAT5B by tyrosine phosphorylation is tightly linked to mammary gland differentiation[J].Molecular Endocrinology,1996,10(12):1496-1506.
[35] HENNIGHAUSEN L,ROBINSON G W.Interpretation of cytokine signaling through the transcription factors STAT5A and STAT5B[J].Genes & Development,2008,22(6):711-721.

[36] SHEKAR P C,GOEL S,RANI S D S,et al.Kappa-casein-deficient mice fail to lactate[J].Proceedings of the National Academy of Sciences,2006,103(21):8000-8005.

[37] IADEVAIA V,HUO Y L,ZHANG Z,et al.Roles of the mammalian target of rapamycin,mTOR,in controlling ribosome biogenesis and protein synthesis[J].Biochemical Society Transactions,2012,40(1):168-172.

[38] KIMBALL S R,JEFFERSON L S.New functions for amino acids:effects on gene transcription and translation[J].The American Journal of Clinical Nutrition,2006,83(2):500S-507S.
[39] YOSHIHARA T,NAITO H,KAKIGI R,et al.Heat stress activates the Akt/mTOR signaling pathway in rat skeletal muscle[J].Acta Physiologica,2013,207(2):416-426.

[40] KAKIGI R,NAITO H,OGURA Y,et al.Heat stress enhances mTOR signaling after resistance exercise in human skeletal muscle[J].Journal of Physiological Sciences,2011,61(2):131-140.

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