不同高精料耐受时间对泌乳期山羊肝脏糖脂代谢的影响
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摘要
为了研究饲喂高精料日粮对泌乳期山羊肝脏糖脂代谢的影响,试验选用16只健康经产泌乳中期关中奶山羊,随机分为3组,低精料组(LC,n=5)、长期高精料组(HL,n=7)和短期高精料组(HS,n=4)。实验结束后采集血液和肝脏组织。结果显示,与LC组相比,HL和HS组奶山羊血糖和肝糖原水平显著升高(P<0.05),血液中甘油三酯(TG)水平显著降低(P<0.05),胆固醇(Tch)含量没有显著变化(P>0.05);HS组山羊肝脏TG和Tch水平显著增加(P<0.05),HL组无显著变化(P>0.05)。HL组肝脏中丙酮酸羧化酶(PC)和葡萄糖转运蛋白2(GLUT2)mRNA表达水平显著上调;HS组肝脏PC、固醇调节元件结合蛋白1(SREBP1)、硬脂酰辅酶A去饱和酶(SCD)、二酰基甘油酰基转移酶(DGAT)和3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)mRNA表达水平显著升高(P<0.05),SREBP-2有升高的趋势(P=0.76),肉碱棕榈酰转移酶-1(CPT-1)和胆固醇7α-羟化酶(CYP7α1)mRNA表达显著下降(P<0.05);线粒体型磷酸烯醇式丙酮酸羧激酶(PCK2)蛋白表达水平显著升高(P<0.05)。以上结果表明,短期饲喂高精料日粮能增强肝脏糖异生能力,促进肝脏胆固醇和脂肪合成;而长期饲喂高精料日粮能增强肝脏糖异生作用,但对胆固醇和脂肪代谢相关基因的表达没有影响。
The aim of this study was to investigate the effects of short-term and long-term feeding with a high concentrate diet on glucose and lipid metabolism in the liver of lactating goats.Sixteen mid-lactating goats were randomly divided into three groups:one group was fed with a high concentrate diet for 4 weeks(HS;65%concentrate diet,n=4);one group was fed with a high concentrate diet for 19 weeks(HL;65%concentration diet,n=7);and the control group was fed with a low concentrate diet during the whole experimental period(LC;35% concentrate diet,n=5).Compared with the LC group,the HS and HL groups showed significantly increased levels of plasma glucose and hepatic glycogen and markedly decreased triglyceride(TG)levels in plasma,but total cholesterol(Tch)in plasma showed no significant difference(P>0.05).The concentrations of hepatic TG and Tch were significantly increased in the HS group(P<0.05)but not in the HL group(P<0.05).Compared with the LC group,the HL group showed significantly upregulated transcript levels of the genes encoding pyruvic carboxylase and glucose transporter 2 in the liver(P <0.05).In the HS group,the transcript levels of genes encoding pyruvic carboxylase,sterol regulation element-binding protein-1,stearoylCoA desaturase,diacylgycerol acyltransferase,and 3-hydroxy-3-methyl glutaryl coenzyme A reductase in the liver were markedly increased,as was the protein level of phosphoenol pyruvate carboxykinase 2.The transcript levels of the gene encodingsterol regulation element-binding protein-2 tended to be higher(0.05
The aim of this study was to investigate the effects of short-term and long-term feeding with a high concentrate diet on glucose and lipid metabolism in the liver of lactating goats.Sixteen mid-lactating goats were randomly divided into three groups:one group was fed with a high concentrate diet for 4 weeks(HS;65%concentrate diet,n=4);one group was fed with a high concentrate diet for 19 weeks(HL;65%concentration diet,n=7);and the control group was fed with a low concentrate diet during the whole experimental period(LC;35% concentrate diet,n=5).Compared with the LC group,the HS and HL groups showed significantly increased levels of plasma glucose and hepatic glycogen and markedly decreased triglyceride(TG)levels in plasma,but total cholesterol(Tch)in plasma showed no significant difference(P>0.05).The concentrations of hepatic TG and Tch were significantly increased in the HS group(P<0.05)but not in the HL group(P<0.05).Compared with the LC group,the HL group showed significantly upregulated transcript levels of the genes encoding pyruvic carboxylase and glucose transporter 2 in the liver(P <0.05).In the HS group,the transcript levels of genes encoding pyruvic carboxylase,sterol regulation element-binding protein-1,stearoylCoA desaturase,diacylgycerol acyltransferase,and 3-hydroxy-3-methyl glutaryl coenzyme A reductase in the liver were markedly increased,as was the protein level of phosphoenol pyruvate carboxykinase 2.The transcript levels of the gene encodingsterol regulation element-binding protein-2 tended to be higher(0.05
引文
[1]Plaizier J C,Krause D O,Gozho G N,et al.Subacute ruminal acidosis in dairy cows:The physiological causes,incidence and consequences.Veterinary Journal,2008,176(1):21-31.
[2]Alzahal O,Rustomo B,Odongo N E,et al.Technical note:A system for continuous recording of ruminal pH in cattle.Journal of Animal Science,2007,85(1):213-217.
[3]Kleen J L,Hooijer G A,Rehage J,et al.Subacute ruminal acidosis in Dutch dairy herds.Veterinary Record,2009,164(22):681-684.
[4]Aschenbach J R,Kristensen N B,Donkin S S,et al.Gluconeogenesis in dairy cows:The secret of making sweet milk from sour dough.Iubmb Life,2010,62(12):869-877.
[5]Li H M.The Effect of Different Energy Intake During the Dry Period on Glyconeogenesis in Periparturient Dairy Cows.Harbin:Northeast Agricultural University,2014.李红梅.干奶期不同能量摄食对奶牛糖异生的影响.哈尔滨:东北农业大学,2006.
[6]Hurtaud C,Lemosquet S,Rulquin H.Effect of graded duodenal infusions of glucose on yield and composition of milk from dairy cows.2.Diets based on grass silage.Journal of Dairy Science,2000,83(12):2952.
[7]Kristensen N B.Splanchnic metabolism of volatile fatty acids in the dairy cow.Animal Science An International Journal of Fundamental&Applied Research,2005,80(80):3-10.
[8]Tagari H,Jr W K,Theurer B,et al.Mammary uptake,portal-drained visceral flux,and hepatic metabolism of free and peptide-bound amino acids in cows fed steam-flaked or dry-rolled sorghum grain diets.Journal of Dairy Science,2008,91(2):679-697.
[9]Dong H B,Wang S Q,Jia Y Y,et al.Effect of long-term feeding of high concentrate diet on plasma biochemical parameters and expression of genes involved in glucose metabolism in lactating goats.Acta Prataculturae Sinica,2014,23(5):295-302.董海波,王绍庆,贾媛媛,等.长期饲喂高精料日粮对泌乳期山羊血液生化参数及组织中糖代谢相关基因表达的影响.草业学报,2014,23(5):295-302.
[10]Seifter S,Dayton S S,Seifter S,et al.The estimation of glycogen with the anthrone reagent.Archives of Biochemistry and Biophysics,1950,25:191-200.
[11]Dong J I,Xin S J.Development and data analysis of real-time fluorescent quantitative PCR.Letters in Biotechnology,2009,20(4):598-600.
[12]Ai X J,Wu X L,Zhu Y Q,et al.Characteristics of some plasma biochemical parameter during perinatal period in cow.China Dairy Cattle,2004,(1):23-25.艾晓杰,吴晓林,朱勇琪,等.初胎奶牛围产期某些血液生化成分的特点.中国奶牛,2004,(1):23-25.
[13]Zhang S K.Effect of Different Forage Ratio Diets on Milk Fat/milk Protein and Its Mechanism in Lactating Goats.Nanjing:Nanjing Agricultural University,2012.张树坤.不同精粗比日粮对泌乳奶山羊乳脂肪/乳蛋白的影响及其机制.南京:南京农业大学,2012.
[14]Zhou C P.Effect of Different Dietary Carbohydrate Levels on Growth Performance,Immunity and Carbohydrate Metabolic Enzymes in Wuchang Bream(Megalobrama amblycephala).Nanjing:Nanjing Agricultural University,2012.周传朋.不同水平碳水化合物日粮对团头鲂的生长、免疫及相关糖代谢酶的影响.南京:南京农业大学,2012.
[15]Dy N M,Gostout C J,Balm R K.Bleeding from the endoscopically-identified Dieulafoy lesion of the proximal small intestine and colon.The American Journal of Gastroenterology,1995,90(1):108-111.
[16]Guo Y Q,Zhou Y,Xu X F,et al.Effect of finely ground wheat induced subacute ruminal acidosis on milk fat synthesis and expression of key lipogenic enzyme genes in liver tissue in dairy cows.Acta Veterinaria et Zootechnica Sinica,2014,45(7):1120-1128.郭勇庆,邹杨,徐晓锋,等.细粉碎小麦诱导亚急性瘤胃酸中毒对奶牛乳脂合成和肝脂肪合成关键酶基因表达量的影响.畜牧兽医学报,2014,45(7):1120-1128.
[17]Van Nevel C J,Demeyer D I.Influence of pH on lipolysis and biohydrogenation of soybean oil by rumen contents in vitro.Reproduction,Nutrition,Development,1996,36(1):53-63.
[18]Goto M,Yoshioka T,Battelino T,et al.TNF|[agr]|decreases gluconeogenesis in hepatocytes isolated from 10-day-old rats.Pediatric Research,2001,49(4):552.
[19]Lu D X.Regulatory theory system of glucose nutrition and its application in ruminant.Animal Husbandry and Feed Science,2010,31(6):402-409.卢德勋.反刍动物葡萄糖营养调控理论体系及其应用.畜牧与饲料科学,2010,31(6):402-409.
[20]Nijland M J,Mitsuya K,Li C,et al.Epigenetic modification of fetal baboon hepatic phosphoenolpyruvate carboxykinase following exposure to moderately reduced nutrient availability.Journal of Physiology,2010,588(8):1349-1359.
[21]Modaressi S,Brechtel K,Christ B,et al.Human mitochondrial phosphoenolpyruvate carboxykinase 2gene.Structure,chromosomal localization and tissue-specific expression.The Biochemical Journal,1998,333(Pt 2):359-366.
[22]Wood I S,Trayhurn P.Glucose transporters(GLUT and SGLT):expanded families of sugar transport proteins.The British Journal of Nutrition,2003,89(1):3-9.
[23]Leturque A,Brot-Laroche E,Gall M L,et al.The role of GLUT2in dietary sugar handling.Journal of Physiology and Biochemistry,2005,61(4):529-537.
[24]Chung M,Ha S,Jeong S,et al.Cloning and characterization of bovine stearoyl CoA desaturasel cDNA from adipose tissues.Bioscience Biotechnology&Biochemistry,2000,64(7):1526-1530.
[25]Miyazaki M,Kim Y C,Ntambi J M.A lipogenic diet in mice with a disruption of the stearoyl-CoA desaturase 1gene reveals a stringent requirement of endogenous monounsaturated fatty acids for triglyceride synthesis.Journal of Lipid Research,2001,42(7):1018-1024.
[26]Listenberger L L,Han X,Lewis S E,et al.Triglyceride accumulation protects against fatty acid-induced lipotoxicity.Proceedings of The National Academy of Sciences of the United States of America,2003,100(6):3077-3082.
[27]Yu Y H,Ginsberg H N.The role of acyl-CoA:diacylglycerol acyltransferase(DGAT)in energy metabolism.Annals of Medicine,2004,36(4):252-261.
[28]Yokoyama C,Wang X,Briggs M R,et al.SREBP-1,a basic-helix-loop-helix-leucine zipper protein that controls transcription of the low density lipoprotein receptor gene.Cell,1993,75(1):187-197.
[29]MascaróC,Acosta E,Ortiz J A,et al.Control of human muscle-type carnitine palmitoyltransferase I gene transcription by peroxisome proliferator-activated receptor.Journal of Biological Chemistry,1998,273(15):8560-8563.
[30]Jia Y Y.Effect of Subacute Ruminal Acidosis on Cortisol Concentration and Hepatie Lipid Metabolism and Its Mechanism in Dairy Goats.Nanjing:Nanjing Agricultural University,2013.贾媛媛.亚急性瘤胃酸中毒对山羊血液皮质醇浓度和肝脏脂代谢的影响及其机制研究.南京:南京农业大学,2013.
[31]Liu J M.Effect of long-term phychological stress on plasma corticosterone,catecholamine level and immunological function in rats.Acta Academiae Medicinae Qingdao,2005,41(3):226-228.刘继民.长期心理应激对大鼠血浆皮质酮、儿茶酚胺水平及免疫功能的影响.青岛大学医学院学报,2005,41(3):226-228.
[32]Sen S,Makkar H P,Becker K.Alfalfa saponins and their implication in animal nutrition.Journal of Agricultural and Food Chemistry,1998,46(1):131-140.
[33]Jurevics H,Hostettler J,Barrett C,et al.Diurnal and dietary-induced changes in cholesterol synthesis correlate with levels of mRNA for HMG-CoA reductase.Journal of Lipid Research,2000,41(7):1048-1054.
[34]Liu H.Changes in Expression of Genes Involved in Liver Cholesterol Metabolism of Insulin Resistance and Diabetic Rats.Shijiazhuang:Hebei Medical University,2010.刘赫.单纯胰岛素抵抗及糖尿病大鼠肝胆固醇代谢相关基因mRNA表达变化.石家庄:河北医科大学,2010.
[35]Qiang Y,Chen Y,Han L,et al.Inflammatory stress increases unmodified LDL uptake via LDL receptor:an alternative pathway for macrophage foam-cell formation.Inflammation Research,2009,58(11):809-818.
[36]Joo K M,Lee J H,Jeon H Y,et al.Pharmacokinetic study of ginsenoside Re with pure ginsenoside Re and ginseng berry extracts in mouse using ultra performance liquid chromatography/mass spectrometric method.Journal of Pharmaceutical and Biomedical Analysis,2010,51(1):278-283.
[37]Jones R D,Repa J J,Russell D W,et al.Delineation of biochemical,molecular,and physiological changes accompanying bile acid pool size restoration in Cyp7a1(-/-)mice fed low levels of cholic acid.The American Journal of Physiology Gastrointestinal and Liver Physiology,2012,303(2):263-274.
[38]Wu L,Zhao Y,Gong J B,et al.Effect of chronic restraint stress of serum glucose and lipid in healthy rats.International Journal of Endocrinology and Metabolism,2010,30(2):80-82.武磊,赵云,弓景波,等.慢性束缚应激对健康大鼠血糖及血脂的影响.国际内分泌代谢杂志,2010,30(2):80-82.
[2]Alzahal O,Rustomo B,Odongo N E,et al.Technical note:A system for continuous recording of ruminal pH in cattle.Journal of Animal Science,2007,85(1):213-217.
[3]Kleen J L,Hooijer G A,Rehage J,et al.Subacute ruminal acidosis in Dutch dairy herds.Veterinary Record,2009,164(22):681-684.
[4]Aschenbach J R,Kristensen N B,Donkin S S,et al.Gluconeogenesis in dairy cows:The secret of making sweet milk from sour dough.Iubmb Life,2010,62(12):869-877.
[5]Li H M.The Effect of Different Energy Intake During the Dry Period on Glyconeogenesis in Periparturient Dairy Cows.Harbin:Northeast Agricultural University,2014.李红梅.干奶期不同能量摄食对奶牛糖异生的影响.哈尔滨:东北农业大学,2006.
[6]Hurtaud C,Lemosquet S,Rulquin H.Effect of graded duodenal infusions of glucose on yield and composition of milk from dairy cows.2.Diets based on grass silage.Journal of Dairy Science,2000,83(12):2952.
[7]Kristensen N B.Splanchnic metabolism of volatile fatty acids in the dairy cow.Animal Science An International Journal of Fundamental&Applied Research,2005,80(80):3-10.
[8]Tagari H,Jr W K,Theurer B,et al.Mammary uptake,portal-drained visceral flux,and hepatic metabolism of free and peptide-bound amino acids in cows fed steam-flaked or dry-rolled sorghum grain diets.Journal of Dairy Science,2008,91(2):679-697.
[9]Dong H B,Wang S Q,Jia Y Y,et al.Effect of long-term feeding of high concentrate diet on plasma biochemical parameters and expression of genes involved in glucose metabolism in lactating goats.Acta Prataculturae Sinica,2014,23(5):295-302.董海波,王绍庆,贾媛媛,等.长期饲喂高精料日粮对泌乳期山羊血液生化参数及组织中糖代谢相关基因表达的影响.草业学报,2014,23(5):295-302.
[10]Seifter S,Dayton S S,Seifter S,et al.The estimation of glycogen with the anthrone reagent.Archives of Biochemistry and Biophysics,1950,25:191-200.
[11]Dong J I,Xin S J.Development and data analysis of real-time fluorescent quantitative PCR.Letters in Biotechnology,2009,20(4):598-600.
[12]Ai X J,Wu X L,Zhu Y Q,et al.Characteristics of some plasma biochemical parameter during perinatal period in cow.China Dairy Cattle,2004,(1):23-25.艾晓杰,吴晓林,朱勇琪,等.初胎奶牛围产期某些血液生化成分的特点.中国奶牛,2004,(1):23-25.
[13]Zhang S K.Effect of Different Forage Ratio Diets on Milk Fat/milk Protein and Its Mechanism in Lactating Goats.Nanjing:Nanjing Agricultural University,2012.张树坤.不同精粗比日粮对泌乳奶山羊乳脂肪/乳蛋白的影响及其机制.南京:南京农业大学,2012.
[14]Zhou C P.Effect of Different Dietary Carbohydrate Levels on Growth Performance,Immunity and Carbohydrate Metabolic Enzymes in Wuchang Bream(Megalobrama amblycephala).Nanjing:Nanjing Agricultural University,2012.周传朋.不同水平碳水化合物日粮对团头鲂的生长、免疫及相关糖代谢酶的影响.南京:南京农业大学,2012.
[15]Dy N M,Gostout C J,Balm R K.Bleeding from the endoscopically-identified Dieulafoy lesion of the proximal small intestine and colon.The American Journal of Gastroenterology,1995,90(1):108-111.
[16]Guo Y Q,Zhou Y,Xu X F,et al.Effect of finely ground wheat induced subacute ruminal acidosis on milk fat synthesis and expression of key lipogenic enzyme genes in liver tissue in dairy cows.Acta Veterinaria et Zootechnica Sinica,2014,45(7):1120-1128.郭勇庆,邹杨,徐晓锋,等.细粉碎小麦诱导亚急性瘤胃酸中毒对奶牛乳脂合成和肝脂肪合成关键酶基因表达量的影响.畜牧兽医学报,2014,45(7):1120-1128.
[17]Van Nevel C J,Demeyer D I.Influence of pH on lipolysis and biohydrogenation of soybean oil by rumen contents in vitro.Reproduction,Nutrition,Development,1996,36(1):53-63.
[18]Goto M,Yoshioka T,Battelino T,et al.TNF|[agr]|decreases gluconeogenesis in hepatocytes isolated from 10-day-old rats.Pediatric Research,2001,49(4):552.
[19]Lu D X.Regulatory theory system of glucose nutrition and its application in ruminant.Animal Husbandry and Feed Science,2010,31(6):402-409.卢德勋.反刍动物葡萄糖营养调控理论体系及其应用.畜牧与饲料科学,2010,31(6):402-409.
[20]Nijland M J,Mitsuya K,Li C,et al.Epigenetic modification of fetal baboon hepatic phosphoenolpyruvate carboxykinase following exposure to moderately reduced nutrient availability.Journal of Physiology,2010,588(8):1349-1359.
[21]Modaressi S,Brechtel K,Christ B,et al.Human mitochondrial phosphoenolpyruvate carboxykinase 2gene.Structure,chromosomal localization and tissue-specific expression.The Biochemical Journal,1998,333(Pt 2):359-366.
[22]Wood I S,Trayhurn P.Glucose transporters(GLUT and SGLT):expanded families of sugar transport proteins.The British Journal of Nutrition,2003,89(1):3-9.
[23]Leturque A,Brot-Laroche E,Gall M L,et al.The role of GLUT2in dietary sugar handling.Journal of Physiology and Biochemistry,2005,61(4):529-537.
[24]Chung M,Ha S,Jeong S,et al.Cloning and characterization of bovine stearoyl CoA desaturasel cDNA from adipose tissues.Bioscience Biotechnology&Biochemistry,2000,64(7):1526-1530.
[25]Miyazaki M,Kim Y C,Ntambi J M.A lipogenic diet in mice with a disruption of the stearoyl-CoA desaturase 1gene reveals a stringent requirement of endogenous monounsaturated fatty acids for triglyceride synthesis.Journal of Lipid Research,2001,42(7):1018-1024.
[26]Listenberger L L,Han X,Lewis S E,et al.Triglyceride accumulation protects against fatty acid-induced lipotoxicity.Proceedings of The National Academy of Sciences of the United States of America,2003,100(6):3077-3082.
[27]Yu Y H,Ginsberg H N.The role of acyl-CoA:diacylglycerol acyltransferase(DGAT)in energy metabolism.Annals of Medicine,2004,36(4):252-261.
[28]Yokoyama C,Wang X,Briggs M R,et al.SREBP-1,a basic-helix-loop-helix-leucine zipper protein that controls transcription of the low density lipoprotein receptor gene.Cell,1993,75(1):187-197.
[29]MascaróC,Acosta E,Ortiz J A,et al.Control of human muscle-type carnitine palmitoyltransferase I gene transcription by peroxisome proliferator-activated receptor.Journal of Biological Chemistry,1998,273(15):8560-8563.
[30]Jia Y Y.Effect of Subacute Ruminal Acidosis on Cortisol Concentration and Hepatie Lipid Metabolism and Its Mechanism in Dairy Goats.Nanjing:Nanjing Agricultural University,2013.贾媛媛.亚急性瘤胃酸中毒对山羊血液皮质醇浓度和肝脏脂代谢的影响及其机制研究.南京:南京农业大学,2013.
[31]Liu J M.Effect of long-term phychological stress on plasma corticosterone,catecholamine level and immunological function in rats.Acta Academiae Medicinae Qingdao,2005,41(3):226-228.刘继民.长期心理应激对大鼠血浆皮质酮、儿茶酚胺水平及免疫功能的影响.青岛大学医学院学报,2005,41(3):226-228.
[32]Sen S,Makkar H P,Becker K.Alfalfa saponins and their implication in animal nutrition.Journal of Agricultural and Food Chemistry,1998,46(1):131-140.
[33]Jurevics H,Hostettler J,Barrett C,et al.Diurnal and dietary-induced changes in cholesterol synthesis correlate with levels of mRNA for HMG-CoA reductase.Journal of Lipid Research,2000,41(7):1048-1054.
[34]Liu H.Changes in Expression of Genes Involved in Liver Cholesterol Metabolism of Insulin Resistance and Diabetic Rats.Shijiazhuang:Hebei Medical University,2010.刘赫.单纯胰岛素抵抗及糖尿病大鼠肝胆固醇代谢相关基因mRNA表达变化.石家庄:河北医科大学,2010.
[35]Qiang Y,Chen Y,Han L,et al.Inflammatory stress increases unmodified LDL uptake via LDL receptor:an alternative pathway for macrophage foam-cell formation.Inflammation Research,2009,58(11):809-818.
[36]Joo K M,Lee J H,Jeon H Y,et al.Pharmacokinetic study of ginsenoside Re with pure ginsenoside Re and ginseng berry extracts in mouse using ultra performance liquid chromatography/mass spectrometric method.Journal of Pharmaceutical and Biomedical Analysis,2010,51(1):278-283.
[37]Jones R D,Repa J J,Russell D W,et al.Delineation of biochemical,molecular,and physiological changes accompanying bile acid pool size restoration in Cyp7a1(-/-)mice fed low levels of cholic acid.The American Journal of Physiology Gastrointestinal and Liver Physiology,2012,303(2):263-274.
[38]Wu L,Zhao Y,Gong J B,et al.Effect of chronic restraint stress of serum glucose and lipid in healthy rats.International Journal of Endocrinology and Metabolism,2010,30(2):80-82.武磊,赵云,弓景波,等.慢性束缚应激对健康大鼠血糖及血脂的影响.国际内分泌代谢杂志,2010,30(2):80-82.