Effects and Regulating Mechanism of Betaine on Lipid Metabolism and DNA Methylation in Primary Germinal Hepatocytes of Geese

  • YANG Zhi ,
  • YANG Yu ,
  • YANG Jingjing ,
  • WANG Zhiyue ,
  • YANG Haiming
Expand
  • 1. Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China;
    2. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

Received date: 2020-11-06

  Online published: 2021-05-14

Supported by

 

Abstract

This experiment was conducted to study the effects and regulating mechanism of betaine on lipid metabolism and DNA methylation in primary germinal hepatocytes of geese. Geese primary germinal hepatocytes were used as the research material in this experiment. Experiment 1:this experiment was studied the effects of different concentrations [0 (control), 2.5, 5.0 and 10.0 mmol/L] of betaine on DNA methylation related enzyme activity and DNA methylation in promoter region of VNN-1 (pantetheinase) in geese primary germinal hepatocytes. Experiment 2:this experiment was investigated the effects of different treatments [normal culture medium (control), 5.0 μmol/L DNA methylation enzyme inhibitor (AZA), 5.0 mmol/L betaine, 5.0 μmol/L AZA+5.0 mmol/L betaine] on the expression of VNN-1 and fatty acid synthase (FAS) genes in geese primary germinal hepatocytes, and explored the molecular mechanism of VNN-1 promoter region methylation regulates the molecular mechanism of gene expression. The results showed as follows:1) the content of low-density lipoprotein (LDL) in betaine culture medium containing 2.5, 5.0 and 10.0 mmol/L was significantly lower than that in the control group (P<0.05). 2) The S-adenosylmethionine/S-adenosylhomocysteine (SAM/SAH) ratio showed an increasing trend with increasing betaine concentration, with a significant primary linear relationship (P<0.05). 3)The expression of VNN-1 gene in hepatocytes of 5.0 and 10.0 mmol/L betaine groups was significantly lower than that of control group and 2.5 mmol/L betaine group (P<0.05); DNA methylation in VNN-1 promoter region of hepatocytes of 5.0 and 10.0 mmol/L betaine groups was significantly higher than that of control group (P<0.05). 4) The expression of VNN-1 gene in hepatocytes of AZA group and AZA+betaine group was significantly higher than that of the control group at 96 h of AZA treatment (P<0.05). AZA treatment significantly increased the expression of FAS gene in hepatocytes at 48 h of treatment compared with the control group (P<0.05). In conclusion, adding betaine to the primary germinal hepatocyte culture medium of goose can inhibit the lipid accumulation of hepatocytes and improve the overall DNA methylation level of the cells. In addition, negative feedback of methylation in the promoter region of VNN-1 gene regulates gene expression. AZA inhibite DNA methylation and find that the high expression of VNN-1 and FAS are associated with DNA demethylation.

Cite this article

YANG Zhi , YANG Yu , YANG Jingjing , WANG Zhiyue , YANG Haiming . Effects and Regulating Mechanism of Betaine on Lipid Metabolism and DNA Methylation in Primary Germinal Hepatocytes of Geese[J]. Chinese Journal of Animal Nutrition, 2021 , 33(5) : 2918 -2926 . DOI: 10.3969/j.issn.1006-267x.2021.05.049

References

[1] ALIREZAEI M,GHEISARI R H,RANJBAR R V.et al.Betaine:a promising antioxidant agent for enhancement of broiler meat quality[J].British Poultry Science,2012,53(5):699-707.  
[2] YANG S,ZHAO N N,SUN B,et al.Grandmaternal betaine supplementation enhances hepatic IGF2 expression in F2 rat offspring through modification of promoter DNA methylation[J].Journal of the Science of Food and Agriculture,2020,100(4):1486-1494.  
[3] PITARI G,MAURIZI G,FLATI V,et al.Enzymatic synthesis of S-aminoethyl-L-cysteine from pantetheine[J].Biochimica et Biophysica Acta (BBA):General Subjects,1992,1116(1):27-33.  
[4] ROMMELAERE S,MILLET V,MANH T P V,et al.Sox17 regulates liver lipid metabolism and adaptation to fasting[J].PLOS One,2014,9(8):e104925.
[5] NAQUET P,PITARI G,DUPRÈ S,et al.Role of the Vnn1 pantetheinase in tissue tolerance to stress[J].Biochemical Society Transactions,2014,42(4):1094-1100.  
[6] JELTSCH A.Beyond Watson and Crick:DNA methylation and molecular enzymology of DNA methyltransferases[J].ChemBioChem,2002,3(4):275-293.
[7] 洪胜辉,张军,张蕊,等.鹅原代肝细胞的简易、高纯分离及培养[J].江苏农业科学,2012,40(4):56-58. HONG S H,ZHANG J,ZHANG R,et al.Simple and high purity isolation and culture of goose primary hepatocytes[J].Jiangsu Agricultural Science,2012,40(4):56-58.(in Chinese)
[8] 杨雨,阳金金,万雨,等.饲粮中添加甜菜碱对仔鹅生长性能及其调控因子的影响[J].动物营养学报,2020,32(9):4140-4147. YANG Y,YANG J J,WAN Y,et al.Effects of dietary betaine on growth performance,and its regulating factors of geese[J].Chinese Journal of Animal Nutrition,2020,32(9):4140-4147.(in Chinese)
[9] WU X, OUYANG Y, WANG B,et al.Hypermethylation of the IRAK3-activated MAPK signaling pathway to promote the development of glioma[J].Cancer Management and Research,2020,7(12):7043-7059.
[10] YANG Z,WANG Z Y,YANG H M,et al.Effects of dietary methionine and betaine on slaughter performance,biochemical and enzymatic parameters in goose liver and hepatic composition[J].Animal Feed Science and Technology,2017,228:48-58.
[11] ZHANG L F,QI Y L,ALUO Z,et al.Betaine increases mitochondrial content and improves hepatic lipid metabolism[J].Food & Function,2018,10(1):216-223.
[12] ZHAN X A,LI J X,XU Z R,et al.Effects of methionine and betaine supplementation on growth performance,carcase composition and metabolism of lipids in male broilers[J].British Poultry Science,2006,47(5):576-580.  
[13] SAUNDERSON C L,MACKINLAY J.Changes in body-weight,composition and hepatic enzyme activities in response to dietary methionine,betaine and choline levels in growing chicks[J].British Journal of Nutrition,1990,63(2):339-349.  
[14] SINGHAL N K,STERNBACH S,FLEMING S,et al.Betaine restores epigenetic control and supports neuronal mitochondria in the cuprizone mouse model of multiple sclerosis[J].Epigenetics,2020,15(8):871-886.  
[15] WEICKHMANN A K,KELLER H,WURM J P,et al.The structure of the SAM/SAH-binding riboswitch[J].Nucleic Acids Research,2019,47(5):2654-2665.  
[16] ABOBAKER H,HU Y,OMER N A,et al.Maternal betaine suppresses adrenal expression of cholesterol trafficking genes and decreases plasma corticosterone concentration in offspring pullets[J].Journal of Animal Science and Biotechnology,2019,10(1):87-97.  
[17] JONES P A,LIANG G N.Rethinking how DNA methylation patterns are maintained[J].Nature Reviews Genetics,2009,10(11):805-811.  
[18] CHEN S Y,ZHANG W X,SUN C,et al.Systemic nanoparticle-mediated delivery of pantetheinase Vanin-1 regulates lipolysis and adiposity in abdominal white adipose tissue[J].Advanced Science,2020,7(14):2000542.
[19] 李洁,周浩,郁建锋,等.鹅VNN1基因的克隆及组织表达分析[J].南京农业大学学报,2018,41(1):140-146. LI J,ZHOU H,YU J F,et al.Cloning and tissue expression analysis of VNN1 gene in geese[J].Journal of Nanjing Agricultural University,2018,41(1):140-146.(in Chinese)
[20] VAN DIEPEN J A,JANSEN P A,BALLAK D B,et al.PPAR-alpha dependent regulation of vanin-1 mediates hepatic lipid metabolism[J].Journal of Hepatology,2014,61(2):366-372.  
[21] CHEN S Y,ZHANG W X,TANG C Q,et al.Vanin-1 is a key activator for hepatic gluconeogenesis[J].Diabetes,2014,63(6):2073-2085.  
[22] ROMMELAERE S,MILLET V,GENSOLLEN T,et al.PPARalpha regulates the production of serum Vanin-1 by liver[J].FEBS Letters,2013,587(22):3742-3748.  
[23] CHEN X,XING M.Effects of 5-Aza-2'-deoxycytidine on hormone secretion and epigenetic regulation in sika deer ovarian granulosa cells[J].Reproduction In Domestic Animals,2021,56(2):360-369.  
[24] CARVALHO C D S,FERNANDES M N.Effect of copper on liver key enzymes of anaerobic glucose metabolism from freshwater tropical fish Prochilodus lineatus[J].Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology,2008,151(3):437-442.  
[25] 王昕源,张雨,杨楠,等.DNMT3a通过提升基因内部甲基化介导紫杉醇诱导的LINE-1异常表达[J].遗传,2020,42(1):100-111. WANG X Y,ZHANG Y,YANG N,et al.DNMT3a mediates paclitaxel-induced aberrant LINE-1 expression through elevated internal gene methylation[J].Genetics,2020,42(1):100-111.(in Chinese)
[26] SHEN J,LIU Y L,REN X C,et al.Changes in DNA methylation and chromatin structure of pro-inflammatory cytokines stimulated by LPS in broiler peripheral blood mononuclear cells[J].Poultry Science,2016,95(7):1636-1645.  
[27] LI X,SUN Q W,LI X,et al.Dietary betaine supplementation to gestational sows enhances hippocampal IGF2 expression in newborn piglets with modified DNA methylation of the differentially methylated regions[J].European Journal of Nutrition,2015,54(7):1201-1210.  
[28] LING L,CHEN L B,ZHANG C N,et al.High glucose induces podocyte epithelial-to-mesenchymal transition by demethylation-mediated enhancement of MMP9 expression[J].Molecular Medicine Reports,2018,17(4):5642-5651.
[29] NISHIZAWA Y,IKEDA R,YAMAMOTO M,et al.5-aza-2-deoxycytidine enhances the sensitivity of 5-fluorouracil by demethylation of the thymidine phosphorylase promoter[J].Anticancer Research,2019,39(8):4129-4136.  
Outlines

/