叶酸添加水平对肉仔鸡肝脏中脂肪代谢相关基因表达和甲基化的影响
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摘要
本试验旨在考察叶酸添加水平对肉仔鸡血清中脂蛋白脂肪酶(LPL)和脂肪酸合成酶(FAS)活性、过氧化物酶体增殖物激活受体γ(PPARγ)含量以及肝脏中脂肪代谢相关基因表达和甲基化的影响。选择1日龄爱拔益加(AA)肉仔鸡300只,随机分成3组,每组5个重复,每个重复20只。对照组饲喂基础饲粮,试验组分别饲喂在基础饲粮中添加5和10 mg/kg叶酸的试验饲粮。试验期为42 d。结果显示:1)与对照组相比,饲粮中添加10 mg/kg叶酸显著降低了21日龄肉仔鸡血清中LPL活性(P<0.05);饲粮中添加5和10 mg/kg叶酸极显著降低了42日龄肉仔鸡血清中LPL活性(P<0.01),极显著提高了21和42日龄肉仔鸡血清中PPARγ含量(P<0.01),极显著降低了21和42日龄肉仔鸡血清中FAS活性(P<0.01)。2)实时荧光定量PCR结果发现,饲粮中添加5和10 mg/kg叶酸均未显著影响42日龄肉仔鸡肝脏中LPL和PPARγ基因的相对表达量(P>0.05),但均显著降低了FAS基因的相对表达量(P <0.05)。3)甲基化分析显示,饲粮中添加5和10 mg/kg叶酸均未显著影响42日龄肉仔鸡肝脏中LPL和PPARγ基因的甲基化比率(P>0.05),但饲粮中添加5 mg/kg叶酸极显著提高了42日龄肉仔鸡肝脏中FAS基因的甲基化比率(P<0.01)。由此得出,饲粮中添加5和10 mg/kg叶酸极显著降低了42日龄肉仔鸡血清中LPL和FAS的活性,极显著增加了PPARγ的含量,显著降低了肝脏中FAS基因的表达;此外,饲粮中添加5 mg/kg叶酸还极显著提高了42日龄肉仔鸡肝脏中FAS基因的甲基化水平。
The aims of this study were to investigate the effects of different folate supplemental levels on the activities of lipoprotein lipase( LPL),fatty acid synthase( FAS) and peroxisome proliferator-activated receptor gamma( PPARγ) content in serum and the expression and methylation of lipid metabolism related genes in liver of broilers. A total of 300 one-day-old Arbor Acres( AA) broilers were randomly allotted to 3 groups with 5 replicates of 20 broilers each. Broilers in control group were fed a basal diet,and broilers in trial groups were fed the basal diet supplemented with 5 and 10 mg/kg folate,respectively. The experiment lasted for 42 days. The results showed as follows: 1) compared with the control group,diet supplemented with 10 mg/kg folate significantly reduced the activity of LPL in serum of 21-day-old broilers( P<0.05); diet supplemented with 5 and 10 mg/kg folate significantly reduced the activity of LPL in serum of 42-day-old broilers( P <0.01),significantly increased the content of PPARγ in serum of 21-and 42-day-old broilers( P<0.01),and significantly reduced the activity of FAS in serum of 21-and 42-day-old broilers( P<0.01). 2) The results of real-time quantitative PCR indicated that the relative expression levels of LPL and PPARγ genes in liver of 42-day-old broilers were not significantly affected by diet supplemented with 5 and 10 mg/kg folate( P>0.05).Compared with the control group,the relative expression levels of FAS gene in liver of 42-day-old broilers was significantly decreased by diet supplemented with 5 and 10 mg/kg folate( P<0.05). 3) The results of methylation analysis showed that the methylation ratios of LPL and PPARγ genes in liver of 42-day-old broilers were not significantly affected by diet supplemented with 5 and 10 mg/kg folate( P>0.05). Compared with the control group,diet supplemented with 5 mg/kg folate significantly improved the methylation ratio of FAS gene in liver of 42-day-old broilers( P<0.01). In conclusion,diet supplemented with 5 and 10 mg/kg folate can significantly decrease serum LPL and FAS activities,significantly increase serum PPARγ content,significantly reduce the expression of FAS gene in liver of 42-day-old broilers. Moreover,diets supplemented with 5 mg/kg folate can also significantly increase the methylation level of FAS gene in liver of 42-day-old broilers.[Chinese Journal of Animal Nutrition,2019,31( 5) : 2098-2106]
The aims of this study were to investigate the effects of different folate supplemental levels on the activities of lipoprotein lipase( LPL),fatty acid synthase( FAS) and peroxisome proliferator-activated receptor gamma( PPARγ) content in serum and the expression and methylation of lipid metabolism related genes in liver of broilers. A total of 300 one-day-old Arbor Acres( AA) broilers were randomly allotted to 3 groups with 5 replicates of 20 broilers each. Broilers in control group were fed a basal diet,and broilers in trial groups were fed the basal diet supplemented with 5 and 10 mg/kg folate,respectively. The experiment lasted for 42 days. The results showed as follows: 1) compared with the control group,diet supplemented with 10 mg/kg folate significantly reduced the activity of LPL in serum of 21-day-old broilers( P<0.05); diet supplemented with 5 and 10 mg/kg folate significantly reduced the activity of LPL in serum of 42-day-old broilers( P <0.01),significantly increased the content of PPARγ in serum of 21-and 42-day-old broilers( P<0.01),and significantly reduced the activity of FAS in serum of 21-and 42-day-old broilers( P<0.01). 2) The results of real-time quantitative PCR indicated that the relative expression levels of LPL and PPARγ genes in liver of 42-day-old broilers were not significantly affected by diet supplemented with 5 and 10 mg/kg folate( P>0.05).Compared with the control group,the relative expression levels of FAS gene in liver of 42-day-old broilers was significantly decreased by diet supplemented with 5 and 10 mg/kg folate( P<0.05). 3) The results of methylation analysis showed that the methylation ratios of LPL and PPARγ genes in liver of 42-day-old broilers were not significantly affected by diet supplemented with 5 and 10 mg/kg folate( P>0.05). Compared with the control group,diet supplemented with 5 mg/kg folate significantly improved the methylation ratio of FAS gene in liver of 42-day-old broilers( P<0.01). In conclusion,diet supplemented with 5 and 10 mg/kg folate can significantly decrease serum LPL and FAS activities,significantly increase serum PPARγ content,significantly reduce the expression of FAS gene in liver of 42-day-old broilers. Moreover,diets supplemented with 5 mg/kg folate can also significantly increase the methylation level of FAS gene in liver of 42-day-old broilers.[Chinese Journal of Animal Nutrition,2019,31( 5) : 2098-2106]
引文
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[21] SIE K K Y,LI J,LY A,et al.Effect of maternal and postw eaning folic acid supplementation on global and gene-specific DNA methylation in the liver of the rat offspring[J]. M olecular Nutrition&Food Research,2013,57(4):677-685.
[22]赵欣,曹丁丁,李媛媛,等.叶酸对人脂肪间充质干细胞成脂分化的影响[J].北京医学,2016,38(4):316-318.
[23] YU X Q,LIU R R,ZHAO G P,et al. Folate supplementation modifies CCAAT/enhancer-binding proteinαmethylation to mediate differentiation of preadipocytes in chickens[J]. Poultry Science,2014,93(10):2596-2603.
[24] MULLEN G E,YET L.Progress in the development of fatty acid synthase inhibitors as anticancer targets[J].Bioorganic&M edicinal Chemistry Letters,2015,25(20):4363-4369.
[25]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.
[26]喻小琼.叶酸对鸡脂肪沉积的影响及表观遗传调控机制研究[D].硕士学位论文.北京:中国农业科学院,2014.
[2] CRIDER K S,YANG T P,BERRY R J,et al. Folate and DNA methylation:a review of molecular mechanisms and the evidence for folate’s role[J].Advances Nutrition,2012,3(1):21-38.
[3] LUCOCK M.Folic acid:nutritional biochemistry,molecular biology,and role in disease processes[J]. M olecular Genetics and M etabolism,2000,71(1/2):121-138.
[4] NAZKI F H,SAMEER A S,GANAIE B A. Folate:metabolism,genes,polymorphisms and the associated diseases[J].Gene,2014,533(1):11-20.
[5] DE-REGIL L M,PENA-ROSAS J P,FERNANDEZGAXIOLA A C,et al. Effects and safety of periconceptional oral folate supplementation for preventing birth defects[J].Cochrane Database of Systematic Review s,2015,14(12):CD007950.
[6] LAM T Y,SETO S W,AU A L S,et al. Folic acid supplementation modifiesβ-adrenoceptor-mediated in vitro lipolysis of obese/diabetic(+db/+db)mice[J].Experimental Biology and M edicine,2009,234(9):1047-1055.
[7] KUMAR K A,LALITHA A,PAVITHRA D,et al.M aternal dietary folate and/or vitamin B12restrictions alter body composition(adiposity)and lipid metabolism in Wistar rat offspring[J]. Journal of Nutritional Biochemistry,2013,24(1):25-31.
[8] SINCLAIR K D,ALLEGRUCCI C,SINGH R,et al.DNA methylation,insulin resistance,and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status[J]. Proceedings of the National Academy of Sciences of the United States of America,2007,104(49):19351-19356.
[9] DOMINGUEZ-SALAS P,MOORE S E,BAKER M S,et al. M aternal nutrition at conception modulates DNA methylation of human metastable epialleles[J].Nature Communication,2014,5:3746.
[10] LAMBROT R,XU C,SAINT-PHAR S,et al.Low paternal dietary folate alters the mouse sperm epigenome and is associated w ith negative pregnancy outcomes[J].Nature Communication,2013,4(1):2889.
[11] TAOUIS M,DRIDI S,CASSY S,et al. Chicken leptin:properties and actions[J].Domestic Animal Endocrinology,2001,21(4):319-327.
[12] YANG X,HUANG Y F,SUN C H,et al. Maternal prenatal folic acid supplementation programs offspring lipid metabolism by aberrant dna methylation in hepatic ATGL and adipose LPL in rats[J].Nutrients,2017,9(9):935,doi:10.3390/nu9090935.
[13] LEE A H,GLIMCHER L H.Intersection of the unfolded protein response and hepatic lipid metabolism[J].Cellular and M olecular Life Sciences,2009,66(17):2835-2850.
[14] CHMURZYNSKA A,STACHOWIAK M,GAWECKI J,et al.Protein and folic acid content in the maternal diet determine lipid metabolism and response to high-fat feeding in rat progeny in an age-dependent manner[J].Genes&Nutrition,2012,7:223-234.
[15] ZECHNER R,STRAUSS J,FRANK S,et al.The role of lipoprotein lipase in adipose tissue development and metabolism[J]. International Journal of Obesity,2000,24(Suppl.4):S53-S56.
[16] FEOLI-FONSECA J C,LEVY E,GODARD M,et al.Familial lipoprotein lipase deficiency in infancy:clinical,biochemical,and molecular study[J]. The Journal of Pediatrics,1998,133(3):417-423.
[17] LAPERROUSAZ E,MOULLEV S,DENIS R G,et al.Lipoprotein lipase in hypothalamus is a key regulator of body w eight gain and glucose homeostasis in mice[J].Diabetologia,2017,60(7):1314-1324.
[18] XING J Y,KANG L,JIANG Y L. Effect of dietary betaine supplementation on lipogenesis gene expression and CpG methylation of lipoprotein lipase gene in broilers[J].M olecular Biology Reports,2011,38(3):1975-1981.
[19] MICHALIK L,AUWERX J,BERGER J P,et al.International union of pharmacology. LXI. Peroxisome proliferator-activated receptors[J]. Pharmacological Review s,2006,58(4):726-741.
[20] JONES J R,BARRICK C,KIM K A,et al.Deletion of PPAR gamma in adipose tissues of mice protects against high fat diet-induced obesity and insulin resistance[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(17):6207-6212.
[21] SIE K K Y,LI J,LY A,et al.Effect of maternal and postw eaning folic acid supplementation on global and gene-specific DNA methylation in the liver of the rat offspring[J]. M olecular Nutrition&Food Research,2013,57(4):677-685.
[22]赵欣,曹丁丁,李媛媛,等.叶酸对人脂肪间充质干细胞成脂分化的影响[J].北京医学,2016,38(4):316-318.
[23] YU X Q,LIU R R,ZHAO G P,et al. Folate supplementation modifies CCAAT/enhancer-binding proteinαmethylation to mediate differentiation of preadipocytes in chickens[J]. Poultry Science,2014,93(10):2596-2603.
[24] MULLEN G E,YET L.Progress in the development of fatty acid synthase inhibitors as anticancer targets[J].Bioorganic&M edicinal Chemistry Letters,2015,25(20):4363-4369.
[25]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.
[26]喻小琼.叶酸对鸡脂肪沉积的影响及表观遗传调控机制研究[D].硕士学位论文.北京:中国农业科学院,2014.