热应激对肉仔鸡肝AMPKα1及脂肪代谢相关分子基因表达的影响

doi:10.11843/j.issn.0366-6964.2018.01.012

摘要/Abstract

摘要：

旨在探讨热应激对肉仔鸡肝脂肪代谢的影响及其可能的内在分子机制。试验选取体重相近的健康28日龄AA肉仔鸡96只，随机分成4组，每组4个重复，每个重复6只鸡，分别为24 h热应激组、24 h对照组和72 h热应激组、72 h对照组。热应激组设定温度为（33±1）℃，对照组设定温度为（23±1）℃。热应激24和72 h后，2个时间点各组分别取8只鸡采集血浆并屠宰收集肝样品。统计生产性能，使用分光光度计测定血浆中葡萄糖（GLU）、尿酸（UA）、总胆固醇（TCHO）、总胆汁酸（TBA）、甘油三脂（TG）含量，利用RT-PCR检测肝中SREBP-1c、FATP-1、LXR、APOB、CYP7A1、FXR、AMPKα1的基因表达量。试验结果表明：1）24和72 h热应激均显著降低肉仔鸡平均体增重与采食量（P<0.05）。2）24 h热应激显著提高肉仔鸡血浆中葡萄糖和总胆汁酸含量（P<0.05），72 h热应激显著提高肉仔鸡血浆中葡萄糖、甘油三酯和总胆固醇含量（P<0.05）。3）脂肪调控因子FATP-1、SREBP-1c、APOB和LXR基因表达检测发现，24 h热应激处理下没有出现显著差异，72 h热应激处理时则均显著升高（P<0.05）；胆汁酸合成相关基因FXR和CYP7A1在24 h热应激处理下没有出现显著差异，在72 h热应激处理下则有升高的趋势（0.05 < P < 0.1）；24 h热应激处理对肝AMPKα1基因表达无显著影响，72 h热应激处理则显著提高AMPKα1在肝的基因表达量（P<0.05）。由此可知，热应激显著降低肉仔鸡生产性能；短期热应激对肉仔鸡肝脂肪代谢影响有限，而长期热应激则显著影响肉仔鸡肝脂肪代谢；热应激时，肝AMPKα1与脂肪代谢相关分子基因表达的变化趋势相似，提示AMPK信号通路可能参与热应激影响肉仔鸡脂肪代谢的调控过程。

Abstract:

The study was conducted to investigate the effect of heat stress on lipid metabolism and the underlying molecular mechanisms in liver of broiler chickens. A total of 96 AA broilers at the age of 28 with similar body weight were randomly allocated into 4 groups with 4 replicates of 6 chicks. The birds in two experimental groups were administered with 24 or 72 h heat stress with the temperature of (33±1)℃. The birds in two control groups were raised under normal temperature of (23±1)℃. After 24 or 72 hours high temperature exposure, 8 chicks from each group were sacrificed, respectively. The samples of liver and plasma were collected and stored in -80℃ for further determination. The growth performance was calculated. The levels of plasma glucose (GLU), uric acid (UA), total cholesterol (TCHO), total bile acid (TBA), triglyceride (TG) in plasma were measured by spectrophotometer. The relative mRNA levels of SREBP-1c, FATP-1, LXR, APOB, CYP7A1, FXR and AMPKα1 in liver were detected by RT-PCR method. The results showed that:1) Heat stress significantly reduced body weight gain and feed intake of broiler chickens (P<0.05). 2) The content of glucose and total bile acid in plasma were significantly increased after 24 hours high stress (P<0.05), and the content of glucose, triglyceride and total cholesterol in plasma were significantly increased after 72 hours heat stress (P<0.05). 3) The mRNA levels of FATP-1, SREBP-1c, APOB, LXR genes were significantly increased after 72 hours heat stress treatment(P<0.05). Heat exposure of 72 hours tended to increase genes expression of liver FXR and CYP7A1 (0.05<P<0.1). The mRNA level of AMPKα1 gene was significantly increased after 72 hours heat stress(P<0.05).In conclusion, heat stress significantly reduced the growth performance of broiler chickens. Short-term heat stress showed little or no effect on liver lipid metabolism, however long-term heat exposure significantly changed liver lipid metabolism at the molecular levels. The similar alteration profiles of the genes expression of AMPKα1 and genes related to lipid metabolism indicated that AMPK signal pathway might play important roles in the liver lipid metabolism of broiler chickens under heat stress.

中图分类号:

S831.4

周华金, 胡希怡, 杨家昶, 丁祥文, 王玉, 宋志刚. 热应激对肉仔鸡肝AMPKα1及脂肪代谢相关分子基因表达的影响[J]. 畜牧兽医学报, 2018, 49(1): 102-110.

ZHOU Hua-jin, HU Xi-yi, YANG Jia-chang, DING Xiang-wen, WANG Yu, SONG Zhi-gang. Effects of Heat Stress on Gene Expression of AMPKα1 and Lipid Metabolism Related Molecules in the Liver of Broiler Chickens[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 102-110.

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参考文献

[1] SONG Z G, LIU L, SHEIKHAHMADI A, et al. Effect of heat exposure on gene expression of feed intake regulatory peptides in laying hens[J]. J Biomed Biotechnol, 2012, 2012:484869.
[2] NICOL C. Behaviour as an indicator of animal welfare[M]//Webster J. Management and Welfare of Farm Animals:The UFAW Farm Handbook. Chichester, UK:Blackwell Wiley, 2011:31-67.
[3] SUN X L, ZHANG H C, SHEIKHAHMADI A, et al. Effects of heat stress on the gene expression of nutrient transporters in the jejunum of broiler chickens (Gallus gallus domesticus)[J]. Int J Biometeorol, 2015, 59(2):127-135.
[4] GERAERT P A, PADILHA J C, GUILLAUMIN S. Metabolic and endocrine changes induced by chronic heat exposure in broiler chickens:Biological and endocrinological variables[J]. Brit J Nutr, 1996, 75(2):205-216.
[5] 刘梅. 急性热应激对肉仔鸡生长性能及脂肪代谢的影响[J]. 动物营养学报, 2011, 23(5):862-868.
LIU M. Effects of acute heat stress on growth performance and lipid metabolism of broilers[J]. Chinese Journal of Animal Nutrition, 2011, 23(5):862-868. (in Chinese)
[6] JASTREBSKI S F, LAMONT S J, SCHMIDT C J. Chicken hepatic response to chronic heat stress using integrated transcriptome and metabolome analysis[J]. PLoS One, 2017, 12(7):e0181900.
[7] 王会敏, 王正平, 董旻岳. 胆汁酸代谢与调控研究进展[J]. 国际消化病杂志, 2010, 30(2):79-82.
WANG H M, WANG Z P, DONG M Y. Research progress about metabolism and regulation of bile acids[J]. International Journal of Digestive Diseases, 2010, 30(2):79-82. (in Chinese)
[8] ROMANSKI K W. The role and mechanism of action of bile acids within the digestive system-bile acids in the liver and bile[J]. Adv Clin Exp Med, 2007, 16(6):793-799.
[9] LI T G, CHIANG J Y L. Bile acid signaling in metabolic disease and drug therapy[J]. Pharmacol Rev, 2014, 66(4):948-983.
[10] PRAWITT J, CRON S, STAELS B. Bile acid metabolism and the pathogenesis of type 2 diabetes[J]. Curr Diab Rep, 2011, 11:160.
[11] THOMAS C, PELLICCIARI R, PRUZANSKI M, et al. Targeting bile-acid signalling for metabolic diseases[J]. Nat Rev Drug Discovery, 2008, 7:678-693.
[12] KHESHT F A, HASSANABADI A. Effects of sterol regulatory element-binding protein (SREBP) in chickens[J]. Lipids Health Dis, 2012, 11:20.
[13] 朱玉萍,周平,李蛟龙,等.低蛋白氨基酸平衡日粮添加半胱胺对生长猪肉质和相关基因表达的影响[J].畜牧兽医学报,2017,48(4):660-668.
ZHU Y P, ZHOU P, LI J L, et al. Effects of low protein level diets supplemented with essential amino acids and Cysteamine on meat quality and related genes expression of growing pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(4):660-668.(in Chinese)
[14] 齐仁立, 黄金秀, 杨飞云, 等. 脂肪酸转运蛋白家族及其介导的脂肪酸跨膜转运[J]. 动物营养学报, 2013, 25(5):905-911.
QI R L, HUANG J X, YANG F Y, et al. Fatty acid transport protein family and their mediation in transmembrane transportation of fatty acids[J]. Chinese Journal of Animal Nutrition, 2013, 25(5):905-911. (in Chinese)
[15] ZHANG S, SHI H, LI H. Cloning and tissue expression characterization of the chicken APOB gene[J]. Anim Biotechnol, 2007, 18(4):243-250.
[16] PIEKARSKI A, DECUYPERE E, BUYSE J, et al. Chenodeoxycholic acid reduces feed intake and modulates the expression of hypothalamic neuropeptides and hepatic lipogenic genes in broiler chickens[J]. Gen Comp Endocrinol, 2016, 229:74-83.
[17] LÓPEZ M, LELLIOTT C J, VIDAL-PUIG A. Hypothalamic fatty acid metabolism:A housekeeping pathway that regulates food intake[J]. Bioessays, 2007, 29(3):248-261.
[18] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitativeI PCR and the 2^-△△CT method[J]. Methods, 2001, 25(4):402-408.
[19] LIN H, BUYSE J, DU R, et al. Response of rectal temperature of broiler chickens to thermal environment factors[J]. Archiv Fur Geflugelkunde, 2004, 68(3):126-131.
[20] LIN H, DECUYPERE E, BUSE J. Oxidative stress induced by corticosterone administration in broiler chickens (Gallus gallus domesticus):2.
Short-term effect[J]. Comp Biochem Physiol Biochem Mol Biol, 2004, 139(4):745-751.
[21] LIN H, ZHANG H F, JIAO H C, et al. Thermoregulation responses of broiler chickens to humidity at different ambient temperatures. I. One week of age[J]. Poultry Sci, 2005, 84(8):1166-1172.
[22] QUINTEIRO-FILHO W M, RIBEIRO A, FERRAZDE-PAULA V, et al. Heat stress impairs performance parameters, induces intestinal injury, and decreases macrophage activity in broiler chickens[J]. Poultry Sci, 2010, 89(9):1905-1914.
[23] BRANU E J, SWEAZEA K L. Glucose regulation in birds[J]. Comp Biochem Physiol Biochem Mol Biol, 2008, 151(1):1-9.
[24] VIRDEN W S, KIDD M T. Physiological stress in broilers:ramifications on nutrient digestibility and responses[J]. J Appl Poult Res, 2009, 18(2):338-347.
[25] LIN H, DECUYPERE E, BUYSE J. Oxidative stress induced by corticosterone administration in broiler chickens (Gallus gallus domesticus):1.
Chronic exposure[J]. Comp Biochem Physiol Biochem Mol Biol, 2004, 139(4):737-744.
[26] GREENE E, KHALDI S, ISHOLA P, et al. Heat and oxidative stress alter the expression of orexin and its related receptors in avian liver cells[J]. Comp Biochem Physiol A Mol Integr Physiol, 2015, 191:18-24.
[27] EVANS A J. In vitro lipogenesis in the liver and adipose tissues of the female Aylesbury duck at different ages[J]. Br Poult Sci, 1972, 13(6):595-602.
[28] HIRSCH D, STAHL A, LODISH H F. A family of fatty acid transporters conserved from mycobacterium to man[J]. Proc Natl Acad Sci U S A, 1998, 95(15):8625-8629.
[29] DAVIS R A, BOOGAERTS J R, BORCHARDT R A, et al. Intrahepatic assembly of very low density lipoproteins. Varied synthetic response of individual apolipoproteins to fasting[J]. J Biol Chem, 1985, 260(26):14137-14144.
[30] HATCH G M, SMITH A J, XU F Y, et al. FATP1 channels exogenous FA into 1,2,3-triacyl-sn-glycerol and down-regulates sphingomyelin and cholesterol metabolism in growing 293 cells[J]. J Lipid Res, 2002, 43(9):1380-1389.
[31] LOBO S, WICZER B M, SMITH A J, et al. Fatty acid metabolism in adipocytes:functional analysis of fatty acid transport proteins 1 and 4[J]. J Lipid Res, 2007, 48(3):609-620.
[32] COMMERFORD S R, PENG L, DUBÉ J J, et al. In vivo regulation of SREBP-1c in skeletal muscle:effects of nutritional status, glucose, insulin, and leptin[J]. Am J Physiol Regul Integr Compol Physiol, 2004, 287(1):R218-R227.
[33] CHU K, MIYAZAKI M, MAN W C, et al. Stearoyl-coenzyme A desaturase 1 deficiency protects against hypertriglyceridemia and increases plasma high-density lipoprotein cholesterol induced by liver X receptor activation[J]. Mol Cell Biol, 2006, 26(18):6786-6798.
[34] BORÉN J, WETTESTEN M, RUSTAEUS S, et al. The assembly and secretion of APOB-100-containing lipoproteins[J]. Biochem Soc Trans, 1993, 21(2):487-493.
[35] WANG S Y, PARK S, KODLI V K, et al. Identification of protein disulfide isomerase 1 as a key isomerase for disulfide bond formation in apolipoprotein B100[J]. Mol Biol Cell, 2014, 26(4):594-604.
[36] WÍJCICKA G, JAMROZ-WISNIEWSKA A, HOROSZEWICZ K, et al. Liver X receptors (LXRs). Part I:structure, function, regulation of activity, and role in lipid metabolism[J]. Postepy Hig Med Dosw, 2007, 61:736-759.
[37] SATO K, KAMADA T. Regulation of bile acid, cholesterol, and fatty acid synthesis in chicken primary hepatocytes by different concentrations of T0901317, an agonist of liver X receptors[J]. Comp Biochem Physiol A Mol Integr Physiol, 2011, 158(2):201-206.
[38] DEMEURE O, DUBY C, DESERT C, et al. Liver X receptor α regulates fatty acid synthase expression in chicken[J]. Poultry Sci, 2009, 88(12):2628-2635.
[39] PEET D J, TURLEY S D, MA W Z, et al. Cholesterol and bile acid metabolism are impaired in mice lacking the nuclear oxysterol receptor LXRα[J]. Cell, 1998, 93(5):693-704.
[40] RUDLIG M, ANGELIN B, STAHLE L, et al. Regulation of hepatic low-density lipoprotein receptor, 3-hydroxy-3-methylglutaryl coenzyme A reductase, and cholesterol 7α-hydroxylase mRNAs in human liver[J]. J Clin Endocrinol Metabol, 2002, 87(9):4307-4313.
[41] BJÖRKHEM I, ARAYA Z, RUDLING M, et al. Differences in the regulation of the classical and the alternative pathway for bile acid synthesis in human liver. No coordinate regulation of CYP7A1 and CYP27A1[J]. J Biol Chem, 2002, 277(30):26804-26807.
[42] CALKIN A C, TONTONOZ P. Transcriptional integration of metabolism by the nuclear sterol-activated receptors LXR and FXR[J]. Nat Rev Mol Cell Biol, 2012, 13(4):213-224.
[43] KIM I, AHN S -H, INAGAKI T, et al. Differential regulation of bile acid homeostasis by the farnesoid X receptor in liver and intestine[J]. J Lipid Res, 2007, 48(12):2664-2672.
[44] SINAL C J, TOHKIN M, MIYATA M, et al. Targeted disruption of the nuclear receptor FXR/BAR impairs bile acid and lipid homeostasis[J]. Cell, 2000, 102(6):731-744.
[45] ANDERSSON U, FILIPSSON K, ABBOTT C R, et al. AMP-activated protein kinase plays a role in the control of food intake[J]. J Biol Chem, 2004, 279(13):12005-12008.
[46] ALEXANDER A, WALKER C L. The role of LKB1 and AMPK in cellular responses to stress and damage[J]. FEBS Lett, 2011, 585(7):952-957.
[47] VIOLLET B, FORETZ M, GUIGAS B, et al. Activation of AMP-activated protein kinase in the liver:a new strategy for the management of metabolic hepatic disorders[J]. J Physiol, 2006, 574(1):41-53.
[48] VIOLLET B, GUIGAS B J, HÉBRARD S, et al. AMP-activated protein kinase in the regulation of hepatic energy metabolism:from physiology to therapeutic perspectives[J]. Acta Physiol, 2009, 196(1):81-98.
[49] ZHANG B B, ZHOU G C, LI C. AMPK:an emerging drug target for diabetes and the metabolic syndrome[J]. Cell Metab, 2009, 9(5):407-416.
[50] RUDERMAN N B, SAHA A K, KRAEGEN E W. Minireview:malonyl CoA, AMP-activated protein kinase, and adiposity[J]. Endocrinology, 2004, 144(12):5166-5171.