不同时长冷暴露下C2C12细胞中O-GlcNAc和IL-6表达的变化

doi:10.11843/j.issn.0366-6964.2021.023

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (1): 219-225.doi: 10.11843/j.issn.0366-6964.2021.023

不同时长冷暴露下C2C12细胞中O-GlcNAc和IL-6表达的变化

胡亚婕, 刘洋, 姚睿智, 史宏昭, 刘鹏, 徐彬, 吕红明, 李士泽*

黑龙江八一农垦大学动物科技学院, 动物医学基础国家级实验教学示范中心;黑龙江省牛病防治重点实验室, 大庆 163319

收稿日期:2020-05-06 出版日期:2021-01-23 发布日期:2021-01-19
通讯作者: 李士泽,主要从事动物应激生物学、应激分子调控机制研究,E-mail:byndlsz@163.com
作者简介:胡亚婕(1996-),女,山东临沂人,硕士生,主要从事动物应激分子调控机制研究,E-mail:huyajiess@163.com
基金资助:
国家自然科学基金面上项目（31972637；31772695）；黑龙江省自然科学基金重点项目（ZD2019C004）

Variation of O-GlcNAc and IL-6 Expression in C2C12 Cells to Different Durations of Cold Exposure

HU Yajie, LIU Yang, YAO Ruizhi, SHI Hongzhao, LIU Peng, XU Bin, LÜ Hongming, LI Shize*

Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, The National Experimental Teaching Demonstration Center of Veterinary Medcine, College of Animal Science & Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China

Received:2020-05-06 Online:2021-01-23 Published:2021-01-19

摘要/Abstract

摘要： 冷暴露下骨骼肌中O-GlcNAc（O-linked β-N-acetylglucosamine）糖基化修饰水平显著升高，作为"应激感受器"调控细胞信号转导及基因转录等；而骨骼肌作为分泌器官通过收缩产生的肌源性IL-6具有免疫作用，更能够调节骨骼肌中糖、脂代谢。本试验旨在检测不同时长冷暴露下小鼠成肌细胞系C2C12中O-GlcNAc相关蛋白和IL-6表达变化，初步探究冷暴露下O-GlcNAc与IL-6之间的潜在联系。本试验通过体外培养小鼠成肌细胞系C2C12，并随机分为不做冷暴露处理的对照组（cold 0 h组，也称为control组，37℃±0.5℃）和冷暴露（32℃±0.5℃）3、6、9、12 h处理组，采用Western blot方法检测不同时长冷暴露下O-GlcNAc糖基化水平和OGT、OGA及IL-6表达水平。结果显示，与对照组相比，冷暴露3 h，OGT表达水平显著升高（P<0.05），冷暴露6 h，OGA表达水平显著升高（P<0.05）；冷暴露3 h，O-GlcNAc糖基化修饰水平和IL-6表达水平极显著升高（P<0.01）。不同时长冷暴露下，C2C12细胞中O-GlcNAc糖基化修饰水平及IL-6表达水平显著升高，并且，不同时长冷暴露下，O-GlcNAc糖基化修饰和IL-6表达变化趋势高度相似，提示冷暴露下O-GlcNAc糖基化修饰可能参与IL-6表达的调控。

关键词: 冷暴露, O-GlcNAc, IL-6, OGT, OGA

Abstract: Under cold exposure, O-linked β-N-acetylglucosamine (O-GlcNAc) in skeletal muscle increased significantly, and O-GlcNAc, as a "stress receptor", regulated cell signal transduction and gene transcription. Skeletal muscle, as a secretory organ, produces myogenic IL-6 through contraction, which not only plays an immune role but also can regulate the metabolism of glucose and lipid in skeletal muscle. Therefore, the purpose of this study was to detect the expression of O-GlcNAc related protein and cytokine IL-6 in C2C12 under different durations of cold exposure. To explore the potential relationship between O-GlcNAc and IL-6 under of cold exposure. In this experiment, C2C12 was cultured in vitro and randomly divided into control group (37℃±0.5℃) and cold exposure (32℃±0.5℃) 3, 6, 9, and 12 hours treatment group. Western blot was used to detect the glycosylation level of O-GlcNAc and the expression level of OGT, OGA, and IL-6 under different durations of exposure. The results showed that compared with the normal temperature control group, the expression level of OGT was significantly increased (P<0.05) after 3 hours of cold exposure and the expression level of OGA was significantly increased (P<0.05) after 6 hours of cold exposure; the expression level of O-GlcNAc glycosylation and IL-6 were significantly increased (P<0.01) after 3 hours of cold exposure. The expression levels of O-GlcNAc glycosylation and IL-6 in C2C12 cells were significantly increased under different durations of cold exposure. At the same time, the trend of O-GlcNAc glycosylation and IL-6 expression under different durations of cold exposure was highly similar, suggesting that O-GlcNAc glycosylation participated in the regulation of IL-6 expression under cold exposure.

Key words: cold exposure, O-GlcNAc, IL-6, OGT, OGA

中图分类号:

S852.2

胡亚婕, 刘洋, 姚睿智, 史宏昭, 刘鹏, 徐彬, 吕红明, 李士泽. 不同时长冷暴露下C2C12细胞中O-GlcNAc和IL-6表达的变化[J]. 畜牧兽医学报, 2021, 52(1): 219-225.

HU Yajie, LIU Yang, YAO Ruizhi, SHI Hongzhao, LIU Peng, XU Bin, LÜ Hongming, LI Shize. Variation of O-GlcNAc and IL-6 Expression in C2C12 Cells to Different Durations of Cold Exposure[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 219-225.

参考文献 24

[1]	FEBBRAIO M A, PEDERSEN B K. Contraction-induced myokine production and release:is skeletal muscle an endocrine organ?[J]. Exerc Sport Sci Rev, 2005, 33(3):114-119.
[2]	STEENSBERG A. The role of IL-6 in exercise-induced immune changes and metabolism[J]. Exerc Immunol Rev, 2003, 9:40-47.
[3]	PEDERSEN B K, STEENSBERG A, KELLER P, et al. Muscle-derived interleukin-6:lipolytic, anti-inflammatory and immune regulatory effects[J]. Pflügers Arch, 2003, 446(1):9-16.
[4]	HART G W, HOUSLEY M P, SLAWSON C. Cycling of O-linked β-N-acetylglucosamine on nucleocytoplasmic proteins[J]. Nature, 2007, 446(7139):1017-1022.
[5]	TORRES C R, HART G W. Topography and polypeptide distribution of terminal N-acetylglucosamine residues on the surfaces of intact lymphocytes. Evidence for O-linked GlcNAc[J]. J Biol Chem, 1984, 259(5):3308-3317.
[6]	HALTIWANGER R S, HOLT G D, HART G W. Enzymatic addition of O-GlcNAc to nuclear and cytoplasmic proteins. Identification of a uridine diphospho-N-acetylglucosamine:peptide β-N-acetylgluco-saminyltransferase[J]. J Biol Chem, 1990, 265(5):2563-2568.
[7]	DONG L Y, HART G W. Purification and characterization of an O-GlcNAc selective N-acetyl-β-D-glucosaminidase from rat spleen cytosol[J]. J Biol Chem, 1994, 269(30):19321-19330.
[8]	LIU S, SHENG H X, YU Z, et al. O-linked β-N-acetylglucosamine modification of proteins is activated in post-ischemic brains of young but not aged mice:implications for impaired functional recovery from ischemic stress[J]. J Cereb Blood Flow Metab, 2016, 36(2):393-398.
[9]	HART G W. Minireview series on the thirtieth anniversary of research on O-GlcNAcylation of nuclear and cytoplasmic proteins:nutrient regulation of cellular metabolism and physiology by O-GlcNAcylation[J]. J Biol Chem, 2014, 289(50):34422-34423.
[10]	ZACHARA N E, O'DONNELL N, CHEUNG W D, et al. Dynamic O-GlcNAc modification of nucleocytoplasmic proteins in response to stress:a survival response of mammalian cells[J]. J Biol Chem, 2004, 279(29):30133-30142.
[11]	NGOH G A, HAMID T, PRABHU S D, et al. O-GlcNAc signaling attenuates ER stress-induced cardiomyocyte death[J]. Am J Physiol Heart Circ Physiol, 2009, 297(5):H1711-H1719.
[12]	YAO R Z, YANG Y Y, LIAN S, et al. Effects of acute cold stress on liver O-GlcNAcylation and glycometabolism in mice[J]. Int J Mol Sci, 2018, 19(9):2815.
[13]	LIU Y L, BI H, CHI S M, et al. The effect of compound nutrients on stress-induced changes in serum IL-2, IL-6 and TNF-α levels in rats[J]. Cytokine, 2007, 37(1):14-21.
[14]	ZHOU D, KUSNECOV A W, SHURIN M R, et al. Exposure to physical and psychological stressors elevates plasma interleukin 6:relationship to the activation of hypothalamic-pituitary-adrenal axis[J]. Endocrinology, 1993, 133(6):2523-2530.
[15]	STEPTOE A, HAMER M, CHIDA Y. The effects of acute psychological stress on circulating inflammatory factors in humans:a review and meta-analysis[J]. Brain, Behav, Immun, 2007, 21(7):901-912.
[16]	NUKINA H, SUDO N, AIBA Y, et al. Restraint stress elevates the plasma interleukin-6 levels in germ-free mice[J]. J Neuroimmunol, 2001, 115(1-2):46-52.
[17]	YILDIRIM N C, YUREKLI M. The effect of adrenomedullin and cold stress on interleukin-6 levels in some rat tissues[J]. Clin Exp Immunol, 2010, 161(1):171-175.
[18]	GIROTTI M, DONEGAN J J, MORILAK D A. Chronic intermittent cold stress sensitizes neuro-immune reactivity in the rat brain[J]. Psychoneuroendocrinology, 2011, 36(8):1164-1174.
[19]	WERNSTEDT I, EDGLEY A, BERNDTSSON A, et al. Reduced stress- and cold-induced increase in energy expenditure in interleukin-6-deficient mice[J]. Am J Physiol Regul Integr Comp Physiol, 2006, 291(3):R551-R557.
[20]	FEBBRAIO M A, HISCOCK N, SACCHETTI M, et al. Interleukin-6 is a novel factor mediating glucose homeostasis during skeletal muscle contraction[J]. Diabetes, 2004, 53(7):1643-1648.
[21]	KNUDSEN J G, BERTHOLDT L, JOENSEN E, et al. Skeletal muscle interleukin-6 regulates metabolic factors in iWAT during HFD and exercise training[J]. Obesity, 2015, 23(8):1616-1624.
[22]	GOLKS A, TRAN T T T, GOETSCHY J F, et al. Requirement for O-linked N-acetylglucosaminyltrans-ferase in lymphocytes activation[J]. EMBO J, 2007, 26(20):4368-4379.
[23]	RAMAKRISHNAN P, CLARK P M, MASON D E, et al. Activation of the transcriptional function of the NF-κB protein C-rel by O-GlcNAc glycosylation[J]. Sci Signal, 2013, 27(290):ra75.
[24]	YANG W H, PARK S Y, NAM H W, et al. NFκB activation is associated with its O-GlcNAcylation state under hyperglycemic conditions[J]. Procee Natl Acad Sci U S A, 2008, 105(45):17345-17350.

不同时长冷暴露下C2C12细胞中O-GlcNAc和IL-6表达的变化

Variation of O-GlcNAc and IL-6 Expression in C2C12 Cells to Different Durations of Cold Exposure

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