缺氧条件下巨噬细胞移动抑制因子对肉鸡心肌脂肪酸代谢的影响

doi:10.11843/j.issn.0366-6964.2018.01.022

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (1): 195-202.doi: 10.11843/j.issn.0366-6964.2018.01.022

缺氧条件下巨噬细胞移动抑制因子对肉鸡心肌脂肪酸代谢的影响

任昊¹, 李丽芳², 王彦美³, 黄楠¹, 裴芳樱¹, 李嘉威¹, 孙耀贵¹, 段智变¹, 李宏全¹, 王文魁^1*

1. 山西农业大学动物科技学院, 太谷 030801;
2. 山西农业大学信息学院环境科学与食品工程系, 太谷 030801;
3. 滨州学院生物工程学院, 滨州 256600

收稿日期:2017-07-28 出版日期:2018-01-23 发布日期:2018-01-23
通讯作者: 王文魁,教授,博士生导师,E-mail:wenkui2009@yeah.net
作者简介:任昊(1992-),男,山西临汾人,硕士生,主要从事炎症因子及其作用机制研究,E-mail:renbuhaof@163.com
基金资助:
山西省科技厅自然基金（2014011028-2）

The Effects of Macrophage Migration Inhibitory on Metabolism of Free-Fatty-Acid in Broiler Chicken Cardiocytes in Hypoxia

REN Hao¹, LI Li-fang², WANG Yan-mei³, HUANG Nan¹, PEI Fang-ying¹, LI Jia-wei¹, SUN Yao-gui¹, DUAN Zhi-bian¹, LI Hong-quan¹, WANG Wen-kui^1*

1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
2. Department of Environmental Science and Food Engineering, College of Information, Shanxi Agricultural University, Taigu 030801, China;
3. School of Biological Engineering, Binzhou University, Binzhou 256600, China

Received:2017-07-28 Online:2018-01-23 Published:2018-01-23

摘要/Abstract

摘要：

旨在探讨缺氧条件下巨噬细胞移动抑制因子（MIF）对肉鸡心肌脂肪酸代谢的影响。取9~12日龄鸡胚，采用胰蛋白酶和Ⅱ型胶原酶联合消化法分离培养心肌细胞，镜下观察细胞形态及活力，并对其进行免疫细胞化学鉴定。以低氧诱导因子1α（HIF-1α）为缺氧指标，化学缺氧模拟剂氯化钴（CoCl₂）处理鸡胚心肌细胞，构建细胞缺氧模型。为了确认MIF与磷酸化腺苷酸活化蛋白激酶（p-AMPK）的关系，在缺氧模型上用MIF抑制剂（ISO-1）阻断MIF，观测p-AMPK是否随MIF阻断而变化。利用Western bolt检测脂肪酸代谢关键酶脂肪酸转运酶（FAT/CD₃₆）、磷酸化乙酰辅酶A羧化酶（p-ACC）及肉毒碱棕榈酰转移酶（CPT-1A）是否在缺氧状态下发生变化。结果：经差速培养的鸡心肌细胞纯度达90%以上；镜下观察，细胞呈梭形和不规则形，并可自发节律性搏动，频率80~130次·min^-1。Western bolt检测显示，经600 μmol·L^-1 CoCl₂处理心肌细胞24 h，HIF-1α表达较正常对照组极显著升高（P<0.01）；100 μmol·L^-1 ISO-1阻断MIF后，MIF及p-AMPK表达量均明显降低（P<0.01）。与正常组相比，缺氧组心肌MIF、p-AMPK、FAT/CD₃₆、p-ACC和CPT-1A表达均极显著升高（P<0.01）；ISO阻断组心肌各目的蛋白质的表达较缺氧组均极显著下降（P<0.01）。缺氧条件下，心肌自分泌MIF增多，通过激活AMPK促进脂肪酸相继进入细胞质和线粒体，加强氧化供能，为缺氧提供代偿性保护。

Abstract:

The aim of this study was to investigate the effects of macrophage migration inhibitory (MIF) on metabolism of Free-Fatty-Acid in broiler chicken cardiocytes in hypoxia. Firstly, chicken embryonic cardiomyocytes were isolated by trypsin and type Ⅱ collagenase, and cultured in DMEM, then identified by α-actinin immunocytochemistry. Secondly, hypoxia model of chicken cardiocyte induced by cobalt chloride was established, which was identified by the hypoxia sign protein, HIF-1α. Thirdly, the MIF blocker ISO-1 was used to test the relationship between MIF and p-AMPK. The expression levels of FAT/CD₃₆, p-ACC and CPT-1A in cardiomyocytes were determined by western blot during hypoxia and normoxia. Results were as follows:The chicken cardiocytes, above 90% purity, showed spherical and polygon shape in vivo, and they beat spontaneously about 80-130 times per minute. HIF-1α expression reached the highest at 24 h with the 600 μmol·L^-1 CoCl₂. The p-AMPK levels decreased significantly (P<0.01) along with the MIF which was blocked by 100 μmol·L^-1 ISO-1. The levels of MIF, p-AMPK, FAT/CD₃₆, p-ACC and CPT-1A were all increased significantly (P<0.01) in the hypoxia group compared with the normal one, while the levels of the target proteins were all decreased significantly (P<0.01) in the inhibitor group compared with the hypoxia one. MIF, a myocardial autocrine factor, plays an important protective role in the fatty acid adapted metabolism of chicken cardiomyocytes during hypoxia by activating the AMPK signal pathway.

中图分类号:

S852.2

任昊, 李丽芳, 王彦美, 黄楠, 裴芳樱, 李嘉威, 孙耀贵, 段智变, 李宏全, 王文魁. 缺氧条件下巨噬细胞移动抑制因子对肉鸡心肌脂肪酸代谢的影响[J]. 畜牧兽医学报, 2018, 49(1): 195-202.

REN Hao, LI Li-fang, WANG Yan-mei, HUANG Nan, PEI Fang-ying, LI Jia-wei, SUN Yao-gui, DUAN Zhi-bian, LI Hong-quan, WANG Wen-kui. The Effects of Macrophage Migration Inhibitory on Metabolism of Free-Fatty-Acid in Broiler Chicken Cardiocytes in Hypoxia[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(1): 195-202.

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参考文献

[1] SHEN Y R, SHI S R, TONG H B, et al. Metabolomics analysis reveals that bile acids and phospholipids contribute to variable responses to low-temperature-induced ascites syndrome[J]. Mol Biosyst, 2014, 10(6):1557-1567.
[2] ZHANG J J, FENG X J, ZHAO L H, et al. Expression of hypoxia-inducible factor 1α mRNA in hearts and lungs of broiler chickens with ascites syndrome induced by excess salt in drinking water[J]. Poult Sci, 2013, 92(8):2044-2052.
[3] YANG F, CAO H B, XIAO Q Y, et al. Transcriptome analysis and gene identification in the pulmonary artery of broilers with ascites syndrome[J]. PLoS One, 2016, 11(6):e0156045.
[4] LI H Y, WANG Y M, CHEN L L, et al. The role of MIF, cyclinD1 and ERK in the development of pulmonary hypertension in broilers[J]. Avian Pathol, 2017, 46(2):202-208.
[5] TAN X, HU S H, WANG X L. Possible role of nitric oxide in the pathogenesis of pulmonary hypertension in broilers:a synopsis[J]. Avian Pathol, 2007, 36(4):261-267.
[6] LI K, ZHAO L H, GENG G R, et al. Increased calcium deposits and decreased Ca²⁺-ATPase in erythrocytes of ascitic broiler chickens[J]. Res Vet Sci, 2011, 90(3):468-473.
[7] WALLHAUS T R, TAYLOR M, DEGRADO T R, et al. Myocardial free fatty acid and glucose use after carvedilol treatment in patients with congestive heart failure[J]. Circulation, 2001, 103(20):2441-2446.
[8] RAMANNA H, ELVAN A, WITTKAMPF F H, et al. Increased dispersion and shortened refractoriness caused by verapamil in chronic atrial fibrillation[J]. J Am Coll Cardiol, 2001, 37(5):1403-1407.
[9] NEUBAUER S. The failing heart-an engine out of fuel[J]. N Engl J Med, 2007, 356(11):1140-1151.
[10] STANLEY W C, RECCHIA F A, LOPASCHUK G D. Myocardial substrate metabolism in the normal and failing heart[J]. Physiol Rev, 2005, 85(3):1093-1129.
[11] MA H, WANG J J, THOMAS D P, et al. Impaired macrophage migration inhibitory factor (MIF)-AMPK activation and ischemic recovery in the senescent heart[J]. Circulation, 2010, 122(3):282-292.
[12] MILLER E J, LI J, LENG L, et al. Macrophage migration inhibitory factor stimulates AMP-activated protein kinase in the ischaemic heart[J]. Nature, 2008, 451(7178):578-582.
[13] SIMPSON P, SAVION S. Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells. Cross-striations, ultrastructure, and chronotropic response to isoproterenol[J]. Circ Res, 1982, 50(1):101-116.
[14] WIDEMAN R F, RHOADS D D, ERF G F, et al. Pulmonary arterial hypertension (ascites syndrome) in broilers:a review[J]. Poult Sci, 2013, 92(1):64-83.
[15] OLKOWSKI A A. Pathophysiology of heart failure in broiler chickens:structural, biochemical, and molecular characteristics[J]. Poult Sci, 2007, 86(5):999-1005.
[16] HASSANZADEH M, BUYSE J, TOLOEI T, et al. Ascites syndrome in broiler chickens:a review on the aspect of endogenous and exogenous factors interactions[J]. J Poult Sci, 2014, 51(3):229-241.
[17] BAGHBANZADEH A, DECUYPERE E. Ascites syndrome in broilers:physiological and nutritional perspectives[J]. Avian Pathol, 2008, 37(2):117-126.
[18] MAXWELL P H, WIESENER M S, CHANG G W, et al. The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis[J]. Nature, 1999, 399(6733):271-275.
[19] JAAKKOLA P, MOLE D R, TIAN Y M, et al. Targeting of HIF-α to the von hippel-lindau ubiquitylation complex by O₂-regulated prolyl hydroxylation[J]. Science, 2001, 292(5516):468-472.
[20] KALLIO P J, WILSON W J, O'BRIEN S, et al. Regulation of the hypoxia-inducible transcription factor 1α by the ubiquitin-proteasome pathway[J]. J Biol Chem, 1999, 274(10):6519-6525.
[21] WELFORD S M, BEDOGNI B, GRADIN K, et al. HIF1α delays premature senescence through the activation of MIF[J]. Genes Dev, 2006, 20(24):3366-3371.
[22] RUSSELL Ⅲ R R, LI J, COVEN D L, et al. AMP-activated protein kinase mediates ischemic glucose uptake and prevents postischemic cardiac dysfunction, apoptosis, and injury[J]. J Clin Invest, 2004, 114(4):495-503.
[23] OAKHILL J S, STEEL R, CHEN Z P, et al. AMPK is a direct adenylate charge-regulated protein kinase[J]. Science, 2011, 332(6036):1433-1435.
[24] ADACHI Y, KANBAYASHI Y, HARATA I, et al. Petasin activates AMP-activated protein kinase and modulates glucose metabolism[J]. J Nat Prod, 2014, 77(6):1262-1269.
[25] WANG J Y, TONG C, YAN X Y, et al. Limiting cardiac ischemic injury by pharmacological augmentation of macrophage migration inhibitory factor-AMP-activated protein kinase signal transduction[J]. Circulation, 2013, 128(3):225-236.
[26] MU J, BROZINICK J T Jr, VALLADARES O, et al. A role for AMP-activated protein kinase in contraction-and hypoxia-regulated glucose transport in skeletal muscle[J]. Mol Cell, 2001, 7(5):1085-1094.
[27] 韩丽娟. 肉鸡腹水综合征(AS)中MIF与AMPK表达关系的研究[D]. 太谷:山西农业大学, 2015.
HAN L J. The relationship between MIF and AMPK expression in ascites syndrome (AS)[D]. Taigu:Shanxi Agricultural University, 2015. (in Chinese)
[28] PIRRO M, MAURIÈGE P, TCHERNOF A, et al. Plasma free fatty acid levels and the risk of ischemic heart disease in men:prospective results from the Québec cardiovascular study[J]. Atherosclerosis, 2002, 160(2):377-384.

缺氧条件下巨噬细胞移动抑制因子对肉鸡心肌脂肪酸代谢的影响

The Effects of Macrophage Migration Inhibitory on Metabolism of Free-Fatty-Acid in Broiler Chicken Cardiocytes in Hypoxia

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