染料木黄酮对大鼠体内N-羟乙酰神经氨酸含量的影响及其与唾液酸转移酶相互作用的探讨
|
摘要
本研究旨在研究染料木黄酮(Genistein,Gen)对大鼠体内N-羟乙酰神经氨酸(N-glycolylneuraminic acid,Neu5Gc)生物合成的影响。选取80只4周龄SD雄性大鼠,随机平均分为对照组和Gen组,分别灌胃5%的乙醇溶液和300 mg/(kg·d)的Gen溶液。利用荧光高效液相色谱(HPLC-FLD)检测大鼠后腿肌肉、肾脏、肝脏组织中Neu5Gc的含量,并采用Gen与唾液酸转移酶(Sialyltransferase,ST)分子对接,初步探讨了其抑制Neu5Gc合成的机理。结果表明:灌胃15 d时,后腿肌肉和肝脏组织中的Neu5Gc的含量分别降低了13.77%和15.45%,而肾脏组织中Neu5Gc的含量变化差异不显著;30 d时,在肌肉组织中未检出Neu5Gc,在肝脏组织中的Neu5Gc的含量降低了13.35%,肾脏组织中Neu5Gc的含量没有显著的变化;45 d时,在后腿肌肉、肾脏组织和肝脏组织中的Neu5Gc含量分别降低了32.65%、16.80%和32.78%;60d时,在后腿肌肉、肾脏组织和肝脏组织中Neu5Gc含量降低了12.72%、12.30%和11.42%。Gen与ST活性位点残基His319、Ser151、Gly293、Thr328形成氢键,且与残基His302、His301、Trp300、Ser271、Phe292、Thr328、Ser325、Ile274形成疏水作用。因此分子间弱相互作用是导致Gen抑制ST活性的主要原因。该研究结果为后续开展宰前降低红肉中Neu5Gc的方法提供了基础实验方法支撑。
To investigate the effects of genistein(Gen) on the biosynthesis of N-glycolylneuraminic acid(Neu5 Gc) in rats, 80 4-week-old male SD rats were randomly equally into the control and genistein groups. The rats of control and genistein groups were fed 5% ethanol and 300 mg/(kg·d) genistein respectively by gavage. The contents of Neu5 Gc in hind leg muscle, kidney and liver tissues of rats were measured by using high performance liquid chromatography coupled with fluorescence detector(HPLC/FLD), and the mechanism of inhibition of Neu5 Gc synthesis was investigated by using the molecular docking of Gen and sialyltransferase. On the 15 th day, the content of Neu5 Gc in hind leg muscle and liver tissues decreased 13.77% and 15.45%, respectively, and there was no significant change in the content of Neu5 Gc in kidney tissues. On the 30 th day, the content of Neu5 Gc in liver tissues decreased 13.35%, however, there was no significant change in the content of Neu5 Gc in kidney tissues and Neu5 Gc was not detected in hind leg muscle. The content of Neu5 Gc in hind leg muscle, kidney and liver tissues decreased respectively 32.65%, 32.78%, 16.80% and 12.72%, 11.42%, 12.30% while rats fed on the 45 th and the 60 th days. Genistein has formed the hydrogen bond with sialyltransferase activity site residues His319, Ser151, Gly293, Thr328 and formed a hydrophobic interactions with the residues His302, His301, Trp300, Ser271, Phe292, Thr328, Ser325 and Ile274. The results of molecular docking indicated that the weak intermolecular interaction was the main cause of genistein inhibiting sialyltransferase activity. The research results provided an experimental basis for the subsequent reduction of Neu5 Gc in red meat before slaughter.
To investigate the effects of genistein(Gen) on the biosynthesis of N-glycolylneuraminic acid(Neu5 Gc) in rats, 80 4-week-old male SD rats were randomly equally into the control and genistein groups. The rats of control and genistein groups were fed 5% ethanol and 300 mg/(kg·d) genistein respectively by gavage. The contents of Neu5 Gc in hind leg muscle, kidney and liver tissues of rats were measured by using high performance liquid chromatography coupled with fluorescence detector(HPLC/FLD), and the mechanism of inhibition of Neu5 Gc synthesis was investigated by using the molecular docking of Gen and sialyltransferase. On the 15 th day, the content of Neu5 Gc in hind leg muscle and liver tissues decreased 13.77% and 15.45%, respectively, and there was no significant change in the content of Neu5 Gc in kidney tissues. On the 30 th day, the content of Neu5 Gc in liver tissues decreased 13.35%, however, there was no significant change in the content of Neu5 Gc in kidney tissues and Neu5 Gc was not detected in hind leg muscle. The content of Neu5 Gc in hind leg muscle, kidney and liver tissues decreased respectively 32.65%, 32.78%, 16.80% and 12.72%, 11.42%, 12.30% while rats fed on the 45 th and the 60 th days. Genistein has formed the hydrogen bond with sialyltransferase activity site residues His319, Ser151, Gly293, Thr328 and formed a hydrophobic interactions with the residues His302, His301, Trp300, Ser271, Phe292, Thr328, Ser325 and Ile274. The results of molecular docking indicated that the weak intermolecular interaction was the main cause of genistein inhibiting sialyltransferase activity. The research results provided an experimental basis for the subsequent reduction of Neu5 Gc in red meat before slaughter.
引文
[1]Liang ML,Zhu QJ,Liu CL,et al.Dissociation effect of different treatments on N-Glycolylneuraminic acid in beef.Food Sci,2018,39(10):28-34(in Chinese).梁美莲,朱秋劲,刘春丽,等.不同加工处理方式对牛肉中Neu5Gc解离的影响.食品科学,2018,39(10):28-34.
[2]Malykh YN,Schauer R,Shaw L.N-Glycolylneuraminic acid in human tumours.Biochimie,2001,83(7):623-634.
[3]Fan XD,Li HW.Progress in studies on relationship between red meat intake,N-Glycolylneuraminic acid and cancer risk.Food Sci,2014,35(15):326-329(in Chinese).范杏丹,李红卫.红肉、N-羟乙酰神经氨酸与癌症关系的研究进展.食品科学,2014,35(15):326-329.
[4]Alisson-Silva F,Kawanishi K,Varki A.Human risk of diseases associated with red meat intake:analysis of current theories and proposed role for metabolic incorporation of a non-human sialic acid.Mol Aspects Med,2016,51:16-30.
[5]Zarogoulidis K,Eleftheriou P,Domvri K,et al.Determination of antibodies against the red meat derived antigen Neu5Gc in patients with lung cancer.Probable association with disease development and progress.Eur Respir J,2015,46(S59):PA533.
[6]Samraj AN,L?ubli H,Varki N,et al.Involvement of a non-human sialic acid in human cancer.Front Oncol,2014,4:1-13.
[7]Endo T,Seeberger PH,Hart GW,et al.Glycoscience:Biology and Medicine.Tokyo:Springer,2015:1-8.
[8]Wang MM.Establishment and optimization of method for sialic acid detection from animal-originated foods and the high-throughput method for sialylated conjunction extraction[D].Nanjing:Nanjing Agricultural University,2015(in Chinese).王毛毛.动物性食品中唾液酸测定及唾液酸复合物高通量提取方法研究[D].南京:南京农业大学,2015.
[9]Ji S,Wang F,Chen Y,et al.Developmental changes in the level of free and conjugated sialic acids,Neu5Ac,Neu5Gc and KDN in different organs of pig:a LC-MS/MS quantitative analyses.Glycoconj J,2017,34(1):21-30.
[10]Iwasaki M,Inoue S,Kitajima K,et al.Novel oligosaccharide chains on polysialoglycoproteins isolated from rainbow trout eggs.A unique carbohydrate sequence with a sialidase-resistant sialyl group,DGalNAc.beta1 leads to 4(NeuGc2leads to 3)DGalNAc.Biochemistry,1984,23(2):305-310.
[11]Muralikrishna G,Reuter G,Peter-Katalini?J,et al.Identification of a new ganglioside from the starfish Asterias rubens.Carbohydr Res,1992,236:321-326.
[12]Chou HH,Takematsu H,Diaz S,et al.A mutation in human CMP-sialic acid hydroxylase occurred after the Homo-pan divergence.Proc Natl Acad Sci USA,1998,95(20):11751-11756.
[13]Hayakawa T,Aki I,Varki A,et al.Fixation of the human-specific CMP-N-acetylneuraminic acid hydroxylase pseudogene and implications of haplotype diversity for human evolution.Genetics,2006,172(2):1139-1146.
[14]Chen HJ,Ding C.Progress on Neu5Gc.Prog Veterin Med,2009,30(4):64-68(in Chinese).陈鸿军,丁铲.N-羟乙酰神经氨酸研究进展.动物医学进展,2009,30(4):64-68.
[15]Banda K,Gregg CJ,Chow R,et al.Metabolism of vertebrate amino sugars with N-glycolyl groups:mechanisms underlying gastrointestinal incorporation of the non-human sialic acid xeno-autoantigen N-glycolylneuraminic acid.J Biol Chem,2012,287(34):28852-28864.
[16]Glass CK,Olefsky JM.Inflammation and lipid signaling in the etiology of insulin resistance.Cell Metab,2012,15(5):635-645.
[17]Rocha VZ,Libby P.Obesity,inflammation,and atherosclerosis.Nat Rev Cardiol,2009,6(6):399-409.
[18]L?fling JC,Paton AW,Varki NM,et al.A dietary non-human sialic acid may facilitate hemolytic-uremic syndrome.Kidney Int,2009,76(2):140-144.
[19]Serna IV A,Boedeker EC.Pathogenesis and treatment of Shiga toxin-producing Escherichia coli infections.Curr Opin Gastroenterol,2008,24(1):38-47.
[20]Melton-Celsa A,Mohawk K,Teel L,et al.Pathogenesis of Shiga-toxin producing Escherichia coli.Curr Top Microbiol Immunol,2012,357:67-103.
[21]Jiang Y,Fu H,Feng CY,et al.Effects of different treatments on N-glycolylneuraminic acid dissociation in red meat.Meat Res,2015,29(12):52-57(in Chinese).蒋芸,付浩,冯昌雨,等.不同处理方式对红肉中N-羟乙酰神经氨酸解离的影响.肉类研究,2015,29(12):52-57.
[22]Bardor M,Nguyen DH,Diaz S,et al.Mechanism of uptake and incorporation of the non-human sialic acid N-glycolylneuraminic acid into human cells.J Biol Chem,2005,280(6):4228-4237.
[23]Wang F,Zhang HL,Guang CE,et al.Mechanism of interaction between sialyltransferase and its inhibitory soyasaponin I.J Food Sci Biotechnol,2015,34(4):355-360(in Chinese).王棐,张海玲,光翠娥,等.大豆皂苷Ⅰ抑制唾液酸转移酶的分子机理研究.食品与生物技术学报,2015,34(4):355-360.
[24]Sun MM,Wang P,Li ZY,et al.Research progress of soybean active ingredients.Soyb Sci,2018,37(6):975-983(in Chinese).孙明明,王萍,李智媛,等.大豆活性成分研究进展.大豆科学,2018,37(6):975-983.
[25]Akiyama T,Ishida J,Nakagawa S,et al.Genistein,a specific inhibitor of tyrosine-specific protein kinases.J Biol Chem,1987,262(12):5592-5595.
[26]McClain RM,Wolz E,Davidovich A,et al.Acute,subchronic and chronic safety studies with genistein in rats.Food Chem Toxicol,2006,44(1):56-80.
[27]Sun JX,Zhang YZ,Gao F.The research progress on physiological function of genistein.Food Res Dev,2005,26(3):92-95(in Chinese).孙洁心,张永忠,高锋.染料木黄酮生理功能的研究进展.食品研究与开发,2005,26(3):92-95.
[28]Naaz A,Yellayi S,Zakroczymski MA,et al.The soy isoflavone genistein decreases adipose deposition in mice.Endocrinology,2003,144(8):3315-3320.
[29]Nielsen ILF,Williamson G.Review of the factors affecting bioavailability of soy isoflavones in humans.Nutr Cancer,2007,57(1):1-10.
[30]Penza M,Montani C,Romani A,et al.Genistein affects adipose tissue deposition in a dose-dependent and gender-specific manner.Endocrinology,2006,147(12):5740-5751.
[31]Zhou YZ,Zhu QJ,Chang R,et al.Effects of Kaempferol and Quercetin on the Synthesis of Neu5GcinRats and its Molecular Mechanism.Chin JBiochem Mol Biol,2019,35(1):101-111.周樱子,朱秋劲,常瑞,等.山奈酚和槲皮素对大鼠体内Neu5Gc合成的影响及分子机制探究.中国生物化学与分子生物学报,2019,35(1):101-111.
[32]Xiu JC.Research progress of soy isoflavones in breed of pig,cow and sheep.Sci Technol Innovat,2018,(11):140-142(in Chinese).修建成.大豆异黄酮在猪牛羊养殖中的应用进展.科学技术创新,2018,(11):140-142.
[33]Rao FV,Rich JR,Raki?B,et al.Structural insight into mammalian sialyltransferases.Nat Struct Mol Biol,2009,16(11):1186-1188.
[34]Duan YB,Yin Y,Meng FL,et al.Design,synthesis and biological evaluation of benzothiazoles as highly potent rock inhibitors through molecular docking and free energy calculations.Chem J Chin Univ,2017,38(9):1568-1577(in Chinese).段永斌,殷燕,孟凡丽,等.基于分子对接和自由能计算的高活性苯并噻唑类ROCK抑制剂的设计、合成和生物学评价.高等学校化学学报,2017,38(9):1568-1577.
[35]Chen P,Li LK,Chen XL,et al.Effects of soy isoflavone on hepatic fatty acid metabolism of rats.JSichuan Agric Univ,2018,36(2):267-272(in Chinese).陈苹,李立科,陈晓林,等.大豆异黄酮对大鼠肝脏酸代谢的影响.四川农业大学学报,2018,36(2):267-272.
[36]Sohn HY,Keller M,Gloe T,et al.The small G-protein Rac mediates depolarization-induced superoxide formation in human endothelial cells.JBiol Chem,2000,275(25):18745-18750.
[37]Zhu GC.Experimental study on genistein prevention and treatment of transplant arteriosclerosis in arotic transplants of rat.Huazhong Univ Sci and Technol,2006(in Chinese).朱国超.Gen防治大鼠移植动脉硬化的实验研究华中科技大学,2006.
[2]Malykh YN,Schauer R,Shaw L.N-Glycolylneuraminic acid in human tumours.Biochimie,2001,83(7):623-634.
[3]Fan XD,Li HW.Progress in studies on relationship between red meat intake,N-Glycolylneuraminic acid and cancer risk.Food Sci,2014,35(15):326-329(in Chinese).范杏丹,李红卫.红肉、N-羟乙酰神经氨酸与癌症关系的研究进展.食品科学,2014,35(15):326-329.
[4]Alisson-Silva F,Kawanishi K,Varki A.Human risk of diseases associated with red meat intake:analysis of current theories and proposed role for metabolic incorporation of a non-human sialic acid.Mol Aspects Med,2016,51:16-30.
[5]Zarogoulidis K,Eleftheriou P,Domvri K,et al.Determination of antibodies against the red meat derived antigen Neu5Gc in patients with lung cancer.Probable association with disease development and progress.Eur Respir J,2015,46(S59):PA533.
[6]Samraj AN,L?ubli H,Varki N,et al.Involvement of a non-human sialic acid in human cancer.Front Oncol,2014,4:1-13.
[7]Endo T,Seeberger PH,Hart GW,et al.Glycoscience:Biology and Medicine.Tokyo:Springer,2015:1-8.
[8]Wang MM.Establishment and optimization of method for sialic acid detection from animal-originated foods and the high-throughput method for sialylated conjunction extraction[D].Nanjing:Nanjing Agricultural University,2015(in Chinese).王毛毛.动物性食品中唾液酸测定及唾液酸复合物高通量提取方法研究[D].南京:南京农业大学,2015.
[9]Ji S,Wang F,Chen Y,et al.Developmental changes in the level of free and conjugated sialic acids,Neu5Ac,Neu5Gc and KDN in different organs of pig:a LC-MS/MS quantitative analyses.Glycoconj J,2017,34(1):21-30.
[10]Iwasaki M,Inoue S,Kitajima K,et al.Novel oligosaccharide chains on polysialoglycoproteins isolated from rainbow trout eggs.A unique carbohydrate sequence with a sialidase-resistant sialyl group,DGalNAc.beta1 leads to 4(NeuGc2leads to 3)DGalNAc.Biochemistry,1984,23(2):305-310.
[11]Muralikrishna G,Reuter G,Peter-Katalini?J,et al.Identification of a new ganglioside from the starfish Asterias rubens.Carbohydr Res,1992,236:321-326.
[12]Chou HH,Takematsu H,Diaz S,et al.A mutation in human CMP-sialic acid hydroxylase occurred after the Homo-pan divergence.Proc Natl Acad Sci USA,1998,95(20):11751-11756.
[13]Hayakawa T,Aki I,Varki A,et al.Fixation of the human-specific CMP-N-acetylneuraminic acid hydroxylase pseudogene and implications of haplotype diversity for human evolution.Genetics,2006,172(2):1139-1146.
[14]Chen HJ,Ding C.Progress on Neu5Gc.Prog Veterin Med,2009,30(4):64-68(in Chinese).陈鸿军,丁铲.N-羟乙酰神经氨酸研究进展.动物医学进展,2009,30(4):64-68.
[15]Banda K,Gregg CJ,Chow R,et al.Metabolism of vertebrate amino sugars with N-glycolyl groups:mechanisms underlying gastrointestinal incorporation of the non-human sialic acid xeno-autoantigen N-glycolylneuraminic acid.J Biol Chem,2012,287(34):28852-28864.
[16]Glass CK,Olefsky JM.Inflammation and lipid signaling in the etiology of insulin resistance.Cell Metab,2012,15(5):635-645.
[17]Rocha VZ,Libby P.Obesity,inflammation,and atherosclerosis.Nat Rev Cardiol,2009,6(6):399-409.
[18]L?fling JC,Paton AW,Varki NM,et al.A dietary non-human sialic acid may facilitate hemolytic-uremic syndrome.Kidney Int,2009,76(2):140-144.
[19]Serna IV A,Boedeker EC.Pathogenesis and treatment of Shiga toxin-producing Escherichia coli infections.Curr Opin Gastroenterol,2008,24(1):38-47.
[20]Melton-Celsa A,Mohawk K,Teel L,et al.Pathogenesis of Shiga-toxin producing Escherichia coli.Curr Top Microbiol Immunol,2012,357:67-103.
[21]Jiang Y,Fu H,Feng CY,et al.Effects of different treatments on N-glycolylneuraminic acid dissociation in red meat.Meat Res,2015,29(12):52-57(in Chinese).蒋芸,付浩,冯昌雨,等.不同处理方式对红肉中N-羟乙酰神经氨酸解离的影响.肉类研究,2015,29(12):52-57.
[22]Bardor M,Nguyen DH,Diaz S,et al.Mechanism of uptake and incorporation of the non-human sialic acid N-glycolylneuraminic acid into human cells.J Biol Chem,2005,280(6):4228-4237.
[23]Wang F,Zhang HL,Guang CE,et al.Mechanism of interaction between sialyltransferase and its inhibitory soyasaponin I.J Food Sci Biotechnol,2015,34(4):355-360(in Chinese).王棐,张海玲,光翠娥,等.大豆皂苷Ⅰ抑制唾液酸转移酶的分子机理研究.食品与生物技术学报,2015,34(4):355-360.
[24]Sun MM,Wang P,Li ZY,et al.Research progress of soybean active ingredients.Soyb Sci,2018,37(6):975-983(in Chinese).孙明明,王萍,李智媛,等.大豆活性成分研究进展.大豆科学,2018,37(6):975-983.
[25]Akiyama T,Ishida J,Nakagawa S,et al.Genistein,a specific inhibitor of tyrosine-specific protein kinases.J Biol Chem,1987,262(12):5592-5595.
[26]McClain RM,Wolz E,Davidovich A,et al.Acute,subchronic and chronic safety studies with genistein in rats.Food Chem Toxicol,2006,44(1):56-80.
[27]Sun JX,Zhang YZ,Gao F.The research progress on physiological function of genistein.Food Res Dev,2005,26(3):92-95(in Chinese).孙洁心,张永忠,高锋.染料木黄酮生理功能的研究进展.食品研究与开发,2005,26(3):92-95.
[28]Naaz A,Yellayi S,Zakroczymski MA,et al.The soy isoflavone genistein decreases adipose deposition in mice.Endocrinology,2003,144(8):3315-3320.
[29]Nielsen ILF,Williamson G.Review of the factors affecting bioavailability of soy isoflavones in humans.Nutr Cancer,2007,57(1):1-10.
[30]Penza M,Montani C,Romani A,et al.Genistein affects adipose tissue deposition in a dose-dependent and gender-specific manner.Endocrinology,2006,147(12):5740-5751.
[31]Zhou YZ,Zhu QJ,Chang R,et al.Effects of Kaempferol and Quercetin on the Synthesis of Neu5GcinRats and its Molecular Mechanism.Chin JBiochem Mol Biol,2019,35(1):101-111.周樱子,朱秋劲,常瑞,等.山奈酚和槲皮素对大鼠体内Neu5Gc合成的影响及分子机制探究.中国生物化学与分子生物学报,2019,35(1):101-111.
[32]Xiu JC.Research progress of soy isoflavones in breed of pig,cow and sheep.Sci Technol Innovat,2018,(11):140-142(in Chinese).修建成.大豆异黄酮在猪牛羊养殖中的应用进展.科学技术创新,2018,(11):140-142.
[33]Rao FV,Rich JR,Raki?B,et al.Structural insight into mammalian sialyltransferases.Nat Struct Mol Biol,2009,16(11):1186-1188.
[34]Duan YB,Yin Y,Meng FL,et al.Design,synthesis and biological evaluation of benzothiazoles as highly potent rock inhibitors through molecular docking and free energy calculations.Chem J Chin Univ,2017,38(9):1568-1577(in Chinese).段永斌,殷燕,孟凡丽,等.基于分子对接和自由能计算的高活性苯并噻唑类ROCK抑制剂的设计、合成和生物学评价.高等学校化学学报,2017,38(9):1568-1577.
[35]Chen P,Li LK,Chen XL,et al.Effects of soy isoflavone on hepatic fatty acid metabolism of rats.JSichuan Agric Univ,2018,36(2):267-272(in Chinese).陈苹,李立科,陈晓林,等.大豆异黄酮对大鼠肝脏酸代谢的影响.四川农业大学学报,2018,36(2):267-272.
[36]Sohn HY,Keller M,Gloe T,et al.The small G-protein Rac mediates depolarization-induced superoxide formation in human endothelial cells.JBiol Chem,2000,275(25):18745-18750.
[37]Zhu GC.Experimental study on genistein prevention and treatment of transplant arteriosclerosis in arotic transplants of rat.Huazhong Univ Sci and Technol,2006(in Chinese).朱国超.Gen防治大鼠移植动脉硬化的实验研究华中科技大学,2006.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |