基于GC-MS代谢组学技术对叶酸调控鸡原代肝细胞代谢的研究

doi:10.11843/j.issn.0366-6964.2018.09.013

Abstract

Abstract:

The study was conducted to investigate the effect of folic acid on metabolism of chicken primary hepatocytes, aiming to provide a reference for exploring regulation mechanism of folic acid on hepatic metabolism and also provide theoretical and scientific basis for its application in poultry. New-born chickens were used for separating primary liver cells. When the confluence reached to about 80%, hepatocytes were exposed to treatment medium. The control group and folic acid addition group were contained in the study. There were 6 replicates for each group. After 12 h treatment, cells were collected to detect triglyceride, NEFA and cholesterols contents. Metabolomics analysis was carried out by GC-MS method. The OPLS-DA model was established to identify differential metabolites, and pathways enriched based on differential metabolites were analyzed through KEGG system. The results showed that folic acid significantly decreased triglyceride and cholesterol contents in hepatocytes compared with the control group (P<0.05). NEFA level was higher in folic acid addition group (P<0.05). Twenty three up-regulation and 9 down-regulation metabolites were found between two groups, which mainly included amino acid, small molecule sugar and acid. Thirty one pathways were enriched based on differential metabolites containing amino acid metabolism, carbohydrate metabolism, lipid metabolism, and so on. The results indicated that folic acid could promote lipid hydrolysis in chicken primary hepatocytes, and was involved in regulating sugar, lipid and amino acid metabolism, which would provide a reference for exploring regulatory mechanism of folic acid on hepatic metabolism in future.

Key words: folic acid, metabolomics, hepatocytes, differential metabolites

CLC Number:

S831.5

LIU Yan-li, SUN Wen-qiang, HAN Di, DUAN Yu-lan, YANG Xin, YANG Xiao-jun. The Study on Folic Acid Regulating Metabolism of Chicken Primary Hepatocytes by GC-MS Metabolomics[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1919-1927.

References

[1] WIS'NIEWSKI J R,VILDHEDE A,NORÉN A,et al.In-depth quantitative analysis and comparison of the human hepatocyte and hepatoma cell line HepG2 proteomes[J].J Proteomics,2016,136:234-247.
[2] TEDESCO S,BAYAT N,DANIELSSON G,et al.Proteomic and lipidomic analysis of primary mouse hepatocytes exposed to metal and metal oxide nanoparticles[J].J Integr OMICS,2015,184:1-14.
[3] FAN Y H,HE C H,LIU G F,et al.Optimization of the isolation and cultivation of Cyprinus carpio primary hepatocytes[J].Cytotechnology,2008,58(2):85-92.
[4] 刘腾飞,田静,耿智霞,等.抗菌药物对LPS诱导鸡肝细胞损伤的影响及复方甘草酸单胺的修复效应[J].畜牧兽医学报,2015,46(2):309-316.
LIU T F,TIAN J,GENG Z X,et al.Effects of antibiotics on LPS induced liver cell injury and hepatoprotective activity of compound ammonium glycyrrhizin[J].Acta Veterinaria et Zootechnica Sinica,2015,46(2):309-316.(in Chinese)
[5] 刘丽,季辉,彭麟,等.鸡肝原代细胞药物代谢模型的建立与优化[J].南京农业大学学报,2015,38(1):127-133.
LIU L,JI H,PENG L,et al.The establishment and optimization of the drug metabolic model of adult chicken primary hepatocytes[J]. Journal of Nanjing Agricultural University,2015,38(1):127-133.(in Chinese)
[6] HOU D X,KUNITAKE T,KUSUDA J,et al.Primary culture of chicken hepatocytes as an in vitro model for determining the influence of dioxin[J].Biosci Biotechnol Biochem,2001,65(1):218-221.
[7] LIU Y L,ZHI L H,SHEN J,et al.Effect of in ovo folic acid injection on hepatic IGF2 expression and embryo growth of broilers[J].J Anim Sci Biotechnol,2016,7:40.
[8] LI S Z,ZHI L H,LIU Y L,et al.Effect of in ovo feeding of folic acid on the folate metabolism,immune function and epigenetic modification of immune effector molecules of broiler[J].Br J Nutr,2016,115(3):411-421.
[9] ALLAH E S H A,BADARY D M.Folic acid protects against lead acetate-induced hepatotoxicity by decreasing NF-κB,IL-1β production and lipid peroxidation mediataed cell injury[J].Pathophysiology,2017,24(1):39-44.
[10] 刘艳利,党燕娜,段玉兰,等.鸡原代肝细胞培养及叶酸对脂质代谢相关基因表达的影响[J].中国农业科学,2017,50(21):4205-4211.
LIU Y L,DANG Y N,DUAN Y L,et al.Effect of folic acid on lipid metabolism associated gene expression in primarily cultured chickens hepatocytes[J].Scientia Agricultura Sinica,2017,50(21):4205-4211.(in Chinese)
[11] 刘雪莲,张盼盼,田方圆,等.基于GC-MS代谢组学技术对内脂素调控雏鸡采食机制的研究[J].畜牧兽医学报,2017,48(7):1251-1259.
LIU X L,ZHANG P P,TIAN F Y,et al.Research of visfatin regulating chicks feed intake based on GC-MS metabolomics[J].Acta Veterinaria et Zootechnica Sinica,2017,48(7):1251-1259.(in Chinese)
[12] PILKIS S J,CLAUS T H.Hepatic gluconeogenesis/glycolysis:regulation and structure/function relationships of substrate cycle enzymes[J].Annu Rev Nutr,1991,11:465-515.
[13] BROSNAN M E,BROSNAN J T.Formate:the neglected member of one-carbon metabolism[J].Annu Rev Nutr,2016,36:369-388.
[14] 孙玲伟,包凯,李影,等.奶牛临床和亚临床酮病的血浆代谢组学研究[J].中国农业科学,2014,47(8):1588-1599.
SUN L W,BAO K,LI Y,et al.Plasma metabolomics study of dairy cows with clinical and subclinical ketosis[J].Scientia Agricultura Sinica,2014,47(8):1588-1599.(in Chinese)
[15] YOSHINARI O,IGARASHI K.Anti-diabetic effect of pyroglutamic acid in type 2 diabetic Goto-Kakizaki rats and KK-Ay mice[J].Br J Nutr,2011,106(7):995-1004.
[16] WU G Y,FANG Y Z,YANG S,et al.Glutathione metabolism and its implications for health[J].J Nutr,2004,134(3):489-492.
[17] MERRY T L,TRAN M,STATHOPOULOS M,et al.High-fat-fed obese glutathione peroxidase 1-deficient mice exhibit defective insulin secretion but protection from hepatic steatosis and liver damage[J].Antioxid Redox Sign,2014,20(14):2114-2129.
[18] GLISZCZYNSKA-SWIGLO A.Folates as antioxidants[J].Food Chem,2007,101(4):1480-1483.
[19] JOSHI R,ADHIKARI S,PATRO B S,et al.Free radical scavenging behavior of folic acid:evidence for possible antioxidant activity[J].Free Radic Biol Med,2001,30(12):1390-1399.
[20] CUI S S,LI W,LV X,et al.Folic acid attenuates homocysteine and enhances antioxidative capacity in atherosclerotic rats[J].Appl Physiol Nutr Meta,2017,42(10):1015-1022
[21] LEVEILLE G A,ROMSOS D R,YEH Y Y,et al.Lipid biosynthesis in the chick.A consideration of site of synthesis,influence of diet and possible regulatory mechanisms[J].Poult Sci,1975,54(4):1075-1093.
[22] DA SILVA R P,KELLY K B,AL RAJABI A,et al.Novel insights on interactions between folate and lipid metabolism[J].Biofactors,2014,40(3):277-283.
[23] 李吕木,PEEBLES D.孵化期间鸡胚、肝脏、卵黄及蛋白组成的变化及其相关关系[J].畜牧兽医学报,2005,36(6):574-577.
LI L M,PEEBLES D.Chick embryo,liver,yolk and albumen compositional changes and their relationships during incubation[J].Acta Veterinaria et Zootechnica Sinica,2005,36(6):574-577.(in Chinese)
[24] NOBLE RC,COCCHI M.Lipid metabolism and the neonatal chicken[J].Prog Lipid Res,1990,29(2):107-140.
[25] MCNEIL C J,HAY S M,RUCKLIDGE G J,et al.Maternal diets deficient in folic acid and related methyl donors modify mechanisms associated with lipid metabolism in the fetal liver of the rat[J].Br J Nutr,2009,102(10):1445-1452.

The Study on Folic Acid Regulating Metabolism of Chicken Primary Hepatocytes by GC-MS Metabolomics

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