柠檬酸对小鼠脂肪细胞甘油三酯及其代谢产物含量的影响
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摘要
本试验旨在研究柠檬酸对小鼠脂肪细胞甘油三酯(TG)及其代谢产物含量的影响。将小鼠3T3-L1前脂细胞诱导成成熟的脂肪细胞作为研究对象,在诱导培养第14天,加入含0(对照组)、20(试验Ⅰ组)、50(试验Ⅱ组)、200μmol/L的柠檬酸完全培养基(试验Ⅲ组)培养细胞,于0、36、72 h收集细胞检测TG合成和分解代谢中关键限速酶及代谢产物的含量。结果表明:在36和72 h,试验Ⅰ、Ⅱ、Ⅲ组TG含量显著高于对照组(P<0.05);各试验组醛缩酶(FDA)、乙酰辅酶A羧化酶(ACC)和脂肪酸合成酶(FAS)含量均显著或极显著高于对照组(P<0.05或P<0.01);各试验组激素敏感脂肪酶(HSL)、肉碱脂酰转移酶1(CPT1)、游离脂肪酸(FFA)及乙酰辅酶A含量均极显著低于对照组(P<0.01)。结果提示,柠檬酸能够促进小鼠脂肪细胞TG的合成和沉积,能显著或极显著提高TG合成代谢中FDA、ACC、FAS产物的含量,极显著降低TG分解代谢中HSL、CPT1、FFA及乙酰辅酶A产物的含量,20μmol/L为最佳剂量。
The purpose of this study was to investigate the effects of citric acid on contents of triglyceride and its metabolites of mouse adipocytes. Firstly inoculated the mouse 3 T3-L1 cells into mature adipocytes as the test subject,the cells were cultured in complete mediums containing 0( CK group),20( test group Ⅰ),50( test group Ⅱ) and 200 μmol/L citric acid( test group Ⅲ) on the 14 th day,respectively. The cells were collected at 0,36 and 72 h for the detection of key rate-limiting enzyme contents in TG synthesis and catabolism,and TG metabolite contents,respectively. The results showed as follows: at 36 an 72 h,TG content in test groupsⅠ,Ⅱ and Ⅲ was significantly higher than that in CK group( P<0.05). The contents of aldolase( FDA),acetyl-Co A carboxylase( ACC) and fatty acid synthetase( FAS) in test groups were significantly higher than those in CK group( P< 0.05 or P< 0.01). The contents of hormone-sensitive lipase( HSL),carnitine palmityl transferase 1( CPT1),free fatty acids( FFA) and acetyl Co A in test groups were significantly lower than those in control group( P< 0.01). In conclusion,citric acid can promote the synthesis and deposition of TG of mouse adipocytes,significantly increase the contents of FDA,ACC,and FAS in TG synthesis,and significantly reduce the contents of HSL,CPT1,FFA,acetyl Co A in TG disintegration; the optimal dosage of citric acid is 20 μmol/L.
The purpose of this study was to investigate the effects of citric acid on contents of triglyceride and its metabolites of mouse adipocytes. Firstly inoculated the mouse 3 T3-L1 cells into mature adipocytes as the test subject,the cells were cultured in complete mediums containing 0( CK group),20( test group Ⅰ),50( test group Ⅱ) and 200 μmol/L citric acid( test group Ⅲ) on the 14 th day,respectively. The cells were collected at 0,36 and 72 h for the detection of key rate-limiting enzyme contents in TG synthesis and catabolism,and TG metabolite contents,respectively. The results showed as follows: at 36 an 72 h,TG content in test groupsⅠ,Ⅱ and Ⅲ was significantly higher than that in CK group( P<0.05). The contents of aldolase( FDA),acetyl-Co A carboxylase( ACC) and fatty acid synthetase( FAS) in test groups were significantly higher than those in CK group( P< 0.05 or P< 0.01). The contents of hormone-sensitive lipase( HSL),carnitine palmityl transferase 1( CPT1),free fatty acids( FFA) and acetyl Co A in test groups were significantly lower than those in control group( P< 0.01). In conclusion,citric acid can promote the synthesis and deposition of TG of mouse adipocytes,significantly increase the contents of FDA,ACC,and FAS in TG synthesis,and significantly reduce the contents of HSL,CPT1,FFA,acetyl Co A in TG disintegration; the optimal dosage of citric acid is 20 μmol/L.
引文
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[19]董婧.肉碱对鲤鱼幼鱼脂肪酸代谢调控作用的研究[D].博士学位论文.长春:吉林农业大学,2014:2-6.
[20]ABU-ELHEIGA L,MATZUK M M,ABO-HASHEM A K A,et al.Continuous fatty acid oxidation and reduce fat storage in m ice lacking acetyl-Co A carboxylase 2[J].Science,2001,291:2613-2616.
[21]童晋.油菜柠檬酸合酶与柠檬酸裂解酶基因克隆及功能研究[D].博士学位论文.北京:中国农业科学院,2009:3-21.
[22]杨竹青,瞿明仁,赵向辉,等.烟酸对反刍动物脂类代谢和肉质的影响及其作用机制[J].动物营养学报,2013,25(6):1150-1157.
[23]WANG Y X,JONES V B,URS S,et al.The human fatty acid synthase gene and de novo lipogenesis are coordinately regulated in hum an adipose tissue[J].The Journal of Nutrition,2004,134(5):1032-1038.
[24]王倩倩,杨彪,夏丽丽,等.鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化[J].畜牧兽医学报,2016,47(4):700-708.
[2]赵俭.不同真菌来源的ATP-柠檬酸裂解酶基因的克隆与表达[D].硕士学位论文.武汉:华中农业大学,2011:2-9.
[3]杨文洲,薛永常,农晓帆,等.柠檬酸对啤酒酵母TCA循环中有机酸的影响[J].大连轻工业学院学报,2007,26(4):313-315.
[4]田静.中国西门塔尔牛生产性能测定及CS基因多态性与肉质性状的关联分析[D].博士学位论文.长春:吉林大学,2012:3-21.
[5]郭秀玲,徐民岗,张秀丽等.小鼠3T3-L1前脂肪细胞培养与诱导分化方法的建立[J].中国药物与临床,2013,12(13):1542-1545,1662.
[6]闫莉,许言午,王晓梁,等.双抗体夹心法检测3-硝基酪氨酸方法的建立及应用[J].中国应用生理学杂志,2009,25(4):569-572.
[7]KHU L Y,STOREY K B,RUBSTOV A M,et al.Regulation of glucokinase activity in liver of hibernating ground squirrel Spermophilus undulatus[J].Biokhim iya,2014,79(7):727-732.
[8]门丽媛,刘帅,宋士一,等.达乌尔黄鼠育肥过程和冬眠期白色脂肪组织糖代谢相关基因的差异表达[J].兽类学报,2015,35(4):422-430.
[9]程钰蓉,孙志杰,崔球.丙酮酸脱氢酶竞争性抑制剂调控裂殖壶菌脂肪酸合成的研究[J].食品工业科技,2016,37(16):161-166.
[10]MAO J Q,DEMAYO F J,LI H G,et al.Liver-specific deletion of acetyl-Co A carboxylase 1 reduces hepatic triglyceride accum ulation w ithout affecting glucose hom eostasis[J].Proceedings of the National Academ y of Sciences of the United States of Am erica,2006,103(22):8552-8557.
[11]BARBER M C,PRICE N T,TRAVERS M T.Structure and regulation of acetyl-Co A carboxylase genes of m etazoa[J].Biochim ica et Biophysica Acta:M olecular and Cell Biology of Lipids,2005,1733(1):1-28.
[12]DENTIN R,BENHARMED F,PEGORIER J P,et al.Polyunsaturated fatty acids suppress glycolytic and lipogenic genes through the inhibition of ChREBP nuclear protein translocation[J].Journal of Clinical Investigation,2005,115(10):2843-2854.
[13]LORENTE-CEBRIN S,KULYTA,HEDN P,et al.Relationship betw een site-spcific HSL phosphorylation and adipocyte lipocyte in obese w om en[J].Obesity Facts,2011,4(5):365-371.
[14]CHONG X,HE J H,JIANG H F,et al.Direct effect of glucocorticoids on lipolysis in adipocytes[J].M olecular Endocrinology,2009,23(8):1161-1170.
[15]CORNACIU I,BOESZOERMENYI A,LINDERM UTH H,et al.The m inim al dom ain of adipose triglyceride(ATGL)ranges until leucine 254 and can be activated and inhibited by cgi-58 and GS respectively[J].PLo S One,2011,6(10):26349.
[16]CHAKRABARTI P,KANDROR K V.Fox O1 controls insulin-dependent adipose triglyceride lipase(ATGL)expression and lipolysis in adipocytes[J].Journal of Biological Chem istry,2009,284(20):13296-13300.
[17]SERR J,SUH Y,LEE K.Regulation of adipose triglyceride lipase by fasting and refeeding in avian species[J].Poultry Science,2009,88(12):2585-2591.
[18]袁禹惠.饲料中脂肪及花生四烯酸水平对半滑舌鳎(Cynoglossus semilaevis)幼鱼生长、脂肪酸组成及代谢相关基因表达的影响[D].硕士学位论文.青岛:中国海洋大学,2015:2-9.
[19]董婧.肉碱对鲤鱼幼鱼脂肪酸代谢调控作用的研究[D].博士学位论文.长春:吉林农业大学,2014:2-6.
[20]ABU-ELHEIGA L,MATZUK M M,ABO-HASHEM A K A,et al.Continuous fatty acid oxidation and reduce fat storage in m ice lacking acetyl-Co A carboxylase 2[J].Science,2001,291:2613-2616.
[21]童晋.油菜柠檬酸合酶与柠檬酸裂解酶基因克隆及功能研究[D].博士学位论文.北京:中国农业科学院,2009:3-21.
[22]杨竹青,瞿明仁,赵向辉,等.烟酸对反刍动物脂类代谢和肉质的影响及其作用机制[J].动物营养学报,2013,25(6):1150-1157.
[23]WANG Y X,JONES V B,URS S,et al.The human fatty acid synthase gene and de novo lipogenesis are coordinately regulated in hum an adipose tissue[J].The Journal of Nutrition,2004,134(5):1032-1038.
[24]王倩倩,杨彪,夏丽丽,等.鹅乙酰辅酶A酰基转移酶2基因的克隆及其在鹅肥肝形成过程中的表达变化[J].畜牧兽医学报,2016,47(4):700-708.