分子营养 Molecular Nutrition

热处理对下丘脑食欲调控和腺苷酸活化蛋白激酶信号通路相关基因表达的影响

  • 张辉 ,
  • 李先磊 ,
  • 周华金 ,
  • 王继光 ,
  • 郝洋洋 ,
  • 宋志刚 ,
  • 苏鹏程
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  • 山东农业大学动物科技学院, 泰安 271018
张辉(1993-),男,山东章丘人,硕士研究生,研究方向为动物营养与饲料科学。E-mail:1033263349@qq.com

收稿日期: 2019-04-27

  网络出版日期: 2019-11-19

基金资助

山东省2018年度西部经济隆起带和省扶贫开发重点区域引进急需紧缺人才支持项目;山东省级家禽产业创新团队项目(STAIT-011-08)

Effects of Heat Treatment on Expression of Appetite Regulation- and Adenosine Monophosphate Activated Protein Kinase Signaling Pathway-Related Genes in Hypothalamus

  • ZHANG Hui ,
  • LI Xianlei ,
  • ZHOU Huajin ,
  • WANG Jiguang ,
  • HAO Yangyang ,
  • SONG Zhigang ,
  • SU Pengcheng
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  • College of Animal Science and Technology, Shandong Agricultural University, Tai'an 271018, China

Received date: 2019-04-27

  Online published: 2019-11-19

摘要

本试验旨在探讨热处理对下丘脑食欲调控和腺苷酸活化蛋白激酶(AMPK)信号通路相关基因表达的影响。试验采用6孔板进行肉鸡下丘脑神经元细胞体外培养,建立下丘脑神经元体外培养模型。将体外培养的神经元细胞分为对照组和热处理组(每组6个重复,每个重复对应1个孔),于培养箱(37℃、5% CO2)中培养。培养至第5天时进行热处理,热处理组培养温度为43℃,对照组培养温度为37℃,热处理1.5 h后收集神经元细胞,利用实时荧光定量PCR(RT-qPCR)技术检测下丘脑神经元细胞中食欲调控和AMPK信号通路相关基因的mRNA相对表达量。结果显示:与对照组相比,1)热处理显著上调下丘脑促食欲基因刺鼠色蛋白相关蛋白(AgRP)和厌食欲基因阿黑皮素原(POMC)mRNA相对表达量(P<0.05);2)热处理显著下调下丘脑AMPK信号通路相关基因AMPKα2和脂肪酸合成酶(FAS)mRNA相对表达量(P<0.05);3)热处理显著上调下丘脑AMPK信号通路上游因子丝氨酸/苏氨酸蛋白激酶1(AKT1)mRNA相对表达量(P<0.05)。综上可知,热处理能够显著上调下丘脑促食欲基因AgRP和厌食欲基因POMC的表达,并能够显著下调下丘脑AMPK信号通路中AMPKα2和FAS基因的表达,显著上调AKT1基因的表达。

本文引用格式

张辉 , 李先磊 , 周华金 , 王继光 , 郝洋洋 , 宋志刚 , 苏鹏程 . 热处理对下丘脑食欲调控和腺苷酸活化蛋白激酶信号通路相关基因表达的影响[J]. 动物营养学报, 2019 , 31(11) : 5221 -5228 . DOI: 10.3969/j.issn.1006-267x.2019.11.038

Abstract

This experiment was conducted to investigate the effects of heat treatment on the expression of appetite regulation- and adenosine monophosphate activated protein kinase (AMPK) signaling pathway-related genes in hypothalamus. Hypothalamus neuron cells of broilers were cultured in vitro with six porous plates, and the hypothalamus neuron cell culture model was established. Neuronal cells which cultured in vitro were divided into control group and heat treatment group, and cultured in incubator (37℃, 5%CO2). Each group had six replicates, and each replicate corresponded to a hole. Heat treatment was carried out on the 5th day of culture, the neuronal cells in the control group were cultured at 37℃, and the neuronal cells in the heat treatment group were cultured at 43℃. After 1.5 h of heat treatment, neuronal cells were collected and the mRNA relative expression levels of regulation- and AMPK signaling pathway-related genes in hypothalamus neuronal cells were detected by real-time quantitative PCR (RT-qPCR) technique. The results showed as follows:compared with the control group, 1) heat treatment significantly up-related the mRNA relative expression levels of hypothalamus appetite-stimulating gene agouti-related protein (AgRP) and appetite-suppressing gene proopiomelanocortin (POMC) (P<0.05); 2) heat treatment significantly down-related the mRNA relative expression levels of hypothalamus AMPK signaling pathway-related genes AMPKα2 and fatty acid synthase (FAS) (P<0.05); 3) heat treatment significantly up-related the mRNA relative expression levels of hypothalamus AMPK signaling pathway upstream factor serine/threonine protein kinase 1 (AKT1) (P<0.05). It is concluded that heat treatment can significantly up-regulate the expression of appetite-stimulating gene AgRP and appetite-suppressing gene POMC in hypothalamus, and it also can significantly down-regulate the expression of AMPKα2 and FAS genes and up-regulate the expression of AKT1 gene in AMPK signaling pathway in hypothalamus.

参考文献

[1] GAO Q,HORVATH T L.Cross-talk between estrogen and leptin signaling in the hypothalamus[J].American Journal of Physiology-Endocrinology and Metabolism,2008,294(5):E817-E826.
[2] 徐彦博.芍药苷对离体大鼠下丘脑腹内侧核神经元电生理活动的影响[D].硕士学位论文.西安:第四军医大学,2008.
[3] 唐焰.下丘脑神经元在体活动参与摄食调控[D].博士学位论文.上海:华东师范大学,2016.
[4] ELMQUIST J K,MARCUS J N.Rethinking the central causes of diabetes[J].Nature Medicine,2003,9(6):645-647.  
[5] SUZUKI K,JAYASENA C N,BLOOM S R.Obesity and appetite control[J].Experimental Diabetes Research,2012,2012:824305.
[6] DENBOW D M,MEADE S,ROBERTSON A,et al.Leptin-induced decrease in food intake in chickens[J].Physiology & Behavior,2000,69(3):359-362.  
[7] GERETS H H J,PEETERS K,ARCKENS L,et al.Sequence and distribution of pro-opiomelanocortin in the pituitary and the brain of the chicken (Gallus gallus)[J].Journal of Comparative Neurology,2000,417(2):250-262.  
[8] KRISTENSEN P,JUDGE M E,THIM L,et al.Hypothalamic CART is a new anorectic peptide regulated by leptin[J].Nature,1998,393(6680):72-76.  
[9] HARDIE D G,SCOTT J W,PAN D A,et al.Management of cellular energy by the AMP-activated protein kinase system[J].FEBS Letters,2003,546(1):113-120.  
[10] DZAMKO N,VAN DENDEREN B J W,HEVENER A L,et al.AMPK β1 deletion reduces appetite,preventing obesity and hepatic insulin resistance[J].Journal of Biological Chemistry,2010,285(1):115-122.  
[11] XIAO B,SANDERS M J,UNDERWOOD E,et al.Structure of mammalian AMPK and its regulation by ADP[J].Nature,2011,472(7342):230-233.  
[12] MINOKOSHI Y,ALQUIER T,FURUKAWA N,et al.AMP-kinase regulates food intake by responding to hormonal and nutrient signals in the hypothalamus[J].Nature,2004,428(6982):569-574.  
[13] AHIMA R S,OSEI S Y.Molecular regulation of eating behavior:new insights and prospects for therapeutic strategies[J].Trends in Molecular Medicine,2001,7(5):205-213.  
[14] CEJAS P J,MARTINEZ M,KARMALLY S,et al.Lumbar transplant of neurons genetically modified to secrete brain-derived neurotrophic factor attenuates allodynia and hyperalgesia after sciatic nerve constriction[J].Pain,2000,86(1/2):195-210.
[15] BURDYGA G,DE LARTIGUE G,RAYBOULD H E,et al.Cholecystokinin regulates expression of Y2 receptors in vagal afferent neurons serving the stomach[J].Journal of Neuroscience,2008,28(45):11583-11592.  
[16] 魏凤,郭广君,吕素芳,等.猪原代神经细胞的体外培养与鉴定[J].中国兽医学报,2016,36(8):1367-1370.
[17] MAKIMURA H,MIZUNO T M,ROBERTS J,et al.Adrenalectomy reverses obese phenotype and restores hypothalamic melanocortin tone in leptin-deficient ob/ob mice[J].Diabetes,2000,49(11):1917-1923.  
[18] TSIGOS C,CHROUSOS G P.Hypothalamic-pituitary-adrenal axis,neuroendocrine factors and stress[J].Journal of Psychosomatic Research,2002,53(4):865-871.  
[19] JEONG J H,LEE D K,LIU S M,et al.Activation of temperature-sensitive TRPV1-like receptors in ARC POMC neurons reduces food intake[J].PLoS Biology,2018,16(4):e2004399.
[20] 周华金,孙晓蕾,钟光,等.大麻素受体1拮抗剂利莫那班对肉仔鸡下丘脑食欲调控因子和腺苷酸激活蛋白激酶基因表达的影响[J].动物营养学报,2019,31(1):258-265.
[21] LEVINE A S,MORLEY J E.Stress-induced eating in rats[J].American Journal of Physiology-Regulatory,Integrative and Comparative Physiology,1981,241(1):R72-R76.
[22] KOLA B,HUBINA E,TUCCI S A,et al.Cannabinoids and ghrelin have both central and peripheral metabolic and cardiac effects via AMP-activated protein kinase[J].Journal of Biological Chemistry,2005,280(26):25196-25201.  
[23] 余春莲.AMPK信号通路负向调节热应激诱导仔猪睾丸支持细胞乳酸分泌[D].硕士学位论文.重庆:西南大学,2017.
[24] LEE-YOUNG R S,BONNER J S,MAYES W H,et al.AMP-activated protein kinase (AMPK)α2 plays a role in determining the cellular fate of glucose in insulin-resistant mouse skeletal muscle[J].Diabetologia,2013,56(3):608-617.  
[25] 王启军.高温环境对不同生长阶段北京油鸡脂肪沉积及脂质代谢的影响[D].硕士学位论文.杨凌:西北农林科技大学,2006.
[26] BAZIZ H A,GERAERT P A,PADILHA J C F,et al.Chronic heat exposure enhances fat deposition and modifies muscle and fat partition in broiler carcasses[J].Poultry Science,1996,75(4):505-513.  
[27] 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.  
[28] 秦玉辉.蛋鸡体内AMPK酶活分布及应激对AMPK酶活影响研究[D].硕士学位论文.雅安:四川农业大学,2003.
[29] YANG C S,LAM C K L,CHARI M,et al.Hypothalamic AMP-activated protein kinase regulates glucose production[J].Diabetes,2010,59(10):2435-2443.  
[30] CHEN L,JIAO Z H,ZHENG L S,et al.Structural insight into the autoinhibition mechanism of AMP-activated protein kinase[J].Nature,2009,459(7250):1146-1149.  
[31] YAMAUCHI T,KAMON J,MINOKOSHI Y,et al.Adiponectin stimulates glucose utilization and fatty-acid oxidation by activating AMP-activated protein kinase[J].Nature Medicine,2002,8(11):1288-1295.  
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