限饲对妊娠中期母羊血液生化指标及内脏脂肪组织脂肪代谢的影响
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摘要
本试验旨在研究限饲对妊娠中期母羊血液生化指标、内脏脂肪组织(VAT)脂肪酸组成及脂肪代谢相关基因表达的影响。选取妊娠湘东黑山羊16只,妊娠(45±3)d,体重(29.86±3.07)kg,随机分配至对照组(C组,100%妊娠营养需求)和限饲组(R组,60%妊娠营养需求),每组8只,试验期为妊娠45~100 d。妊娠101 d时,检测血液生化指标以及瘤胃网膜、肠系膜和肾周脂肪组织中脂肪酸组成、脂肪代谢和能量感知相关基因的表达。结果表明,R组与C组相比:1)血液胰高血糖素、游离脂肪酸含量显著升高(P<0.05),血液瘦素、脂联素和高密度脂蛋白胆固醇含量显著降低(P<0.05)。2)网膜脂花生四烯酸、棕榈油酸、亚油酸、二十碳三烯酸和多不饱和脂肪酸含量显著降低(P<0.05);系膜脂豆蔻酸含量显著降低(P<0.05);肾周脂亚油酸、花生四烯酸、二十碳三烯酸和多不饱和脂肪酸含量显著降低(P<0.05)。3)网膜脂腺苷酸活化蛋白激酶的催化亚基α2(AMPKα2)基因表达有上调的趋势(0.05≤P≤0.10);系膜脂脂肪酸合成酶(FASN)、硬脂酰辅酶A脱氢酶1(SCD1)、解偶联蛋白2(UCP2)基因表达有下调的趋势(0.05≤P≤0.10),过氧化物酶体增殖激活受体γ(PPARγ)基因表达有上调的趋势(0.05≤P≤0.10);肾周脂FASN、UCP2基因表达显著下调(P<0.05),腺苷酸活化蛋白激酶的非催化亚基β1(AMPKβ1)基因表达显著上调(P<0.05),肉碱脂酰转移酶1A(CPT1A)基因表达有上调的趋势(0.05≤P≤0.10)。综上所述,妊娠中期限饲导致母羊血液中脂肪代谢调控因子(瘦素和脂联素)含量下降,脂肪代谢产物(游离脂肪酸)含量上升,导致VAT的脂肪酸组成(PUFA含量下降),并经下调脂肪合成基因(FASN和SCD1)、上调脂肪动员基因(CPT1A)及下调基础能量代谢基因(UCP2)表达的途径,使母体VAT脂肪合成作用减弱、脂质分解作用增强来调节机体能量稳态。
This experiment was conducted to investigate the effects of feeding restriction on blood biochemical indexes,fatty acid composition and gene expressions related to lipid metabolism of visceral adipose tissue( VAT) in mid-pregnancy ewes. Sixteen Xiangdong black goats [gestation time of( 45±3) d],body weight of( 29.86 ±3.07) kg]were chosen and randomly assigned to control group( C group,100% of pregnancy nutritional requirements)and restricted group( R group,40% of pregnancy nutritional requirements),and each group comprised of8 dams. The experiment period was 45 to 100 d of gestation. At day 101 of gestation,blood biochemical indexes,fatty acid composition and gene expressions related to lipid metabolism and energy sensing in omental,mesenteric and perirenal adipose tissues were detected. The results showed that R group compared with C group: 1) blood contents of glucagon and free fatty acid were significantly increased( P<0.05),and blood contents of leptin,adiponectin and high-density lipoprotein cholesterol were significantly decreased( P<0.05). 2) The contents of arachidonic acid( C20∶4 n6),palmitoleic acid( C16∶1),linoleic acid( C18∶2 n6 c),epoxyeicosatrienoic acids( C20∶3 n6)and polyunsaturated fatty acid( PUFA) in omental adipose tissue were significantly decreased( P<0.05); the content of myristic acid in mesenteric adipose tissue was significantly decreased( P<0.05); the contents of linoleic acid,arachidonic acid,epoxyeicosatrienoic acids and PUFA in perirenal adipose tissue were significantly decreased( P<0.05). 3) The expression of adenosine 5'-monophosphate-activated protein kinase α 2( AMPKα2) gene in omental adipose tissue tended to be increased( 0.05≤P≤0.10); the expressions of fatty acid synthetase( FASN),stearoyl-CoA desaturase-1( SCD1) and uncoupling protein 2( UCP2) genes in mesenteric adipose tissue tended to be decreased( 0.05≤P≤0.10),and the expression of peroxisome proliferator-activated receptor γ( PPARγ) gene tended to be increased( 0.05≤P≤0.10); in perirenal adipose tissue,the expressions of FASN and UCP2 genes were significantly decreased( P < 0. 05),while that of adenosine 5'-monophosphate-activated protein kinase β1( AMPKβ1) gene was significantly increased( P<0.05),and that of carnitine palmitoyltransferase 1 A( CPT1 A)gene tended to be increased( 0.05≤P≤0.10). These results indicate that feeding restriction during mid-gestation decreases contents of lipid metabolism regulator factors( leptin and adiponectin),increases contents of lipid metabolites( free fatty acids),and affects fatty acid composition( decreases PUFA content),down-regulates gene( FASN and SCD1) expressions related to lipid synthesis,up-regulates gene( CPT1 A) expression related to lipid mobilization,and down-regulates gene( UCP2) expression related to basal energy metabolism in VAT of dams,thereby attenuates lipid anabolism and enhances lipid mobilization in VAT to regulate body energy balance.
This experiment was conducted to investigate the effects of feeding restriction on blood biochemical indexes,fatty acid composition and gene expressions related to lipid metabolism of visceral adipose tissue( VAT) in mid-pregnancy ewes. Sixteen Xiangdong black goats [gestation time of( 45±3) d],body weight of( 29.86 ±3.07) kg]were chosen and randomly assigned to control group( C group,100% of pregnancy nutritional requirements)and restricted group( R group,40% of pregnancy nutritional requirements),and each group comprised of8 dams. The experiment period was 45 to 100 d of gestation. At day 101 of gestation,blood biochemical indexes,fatty acid composition and gene expressions related to lipid metabolism and energy sensing in omental,mesenteric and perirenal adipose tissues were detected. The results showed that R group compared with C group: 1) blood contents of glucagon and free fatty acid were significantly increased( P<0.05),and blood contents of leptin,adiponectin and high-density lipoprotein cholesterol were significantly decreased( P<0.05). 2) The contents of arachidonic acid( C20∶4 n6),palmitoleic acid( C16∶1),linoleic acid( C18∶2 n6 c),epoxyeicosatrienoic acids( C20∶3 n6)and polyunsaturated fatty acid( PUFA) in omental adipose tissue were significantly decreased( P<0.05); the content of myristic acid in mesenteric adipose tissue was significantly decreased( P<0.05); the contents of linoleic acid,arachidonic acid,epoxyeicosatrienoic acids and PUFA in perirenal adipose tissue were significantly decreased( P<0.05). 3) The expression of adenosine 5'-monophosphate-activated protein kinase α 2( AMPKα2) gene in omental adipose tissue tended to be increased( 0.05≤P≤0.10); the expressions of fatty acid synthetase( FASN),stearoyl-CoA desaturase-1( SCD1) and uncoupling protein 2( UCP2) genes in mesenteric adipose tissue tended to be decreased( 0.05≤P≤0.10),and the expression of peroxisome proliferator-activated receptor γ( PPARγ) gene tended to be increased( 0.05≤P≤0.10); in perirenal adipose tissue,the expressions of FASN and UCP2 genes were significantly decreased( P < 0. 05),while that of adenosine 5'-monophosphate-activated protein kinase β1( AMPKβ1) gene was significantly increased( P<0.05),and that of carnitine palmitoyltransferase 1 A( CPT1 A)gene tended to be increased( 0.05≤P≤0.10). These results indicate that feeding restriction during mid-gestation decreases contents of lipid metabolism regulator factors( leptin and adiponectin),increases contents of lipid metabolites( free fatty acids),and affects fatty acid composition( decreases PUFA content),down-regulates gene( FASN and SCD1) expressions related to lipid synthesis,up-regulates gene( CPT1 A) expression related to lipid mobilization,and down-regulates gene( UCP2) expression related to basal energy metabolism in VAT of dams,thereby attenuates lipid anabolism and enhances lipid mobilization in VAT to regulate body energy balance.
引文
[1]GUZMAN C,CABRERA R,CARDENAS M,et al.Protein restriction during fetal and neonatal developm ent in the rat alters reproductive function and accelerates reproductive ageing in fem ale progeny[J].Journal of Physiology,2006,572(1):97-108.
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[3]ROSARIO F J,JANSSON N,KANAI Y,et al.Maternal protein restriction in the rat inhibits placental insulin,m TOR,and STAT3 signaling and dow n-regulates placental am ino acid transporters[J].Endocrinology,2011,152(3):1119-1129.
[4]BARCROFT J.Researches on pre-natal life[M].Oxford:Blackw ell Scientific Publications,1946.
[5]VIRTUE S,VIDAL-PUIG A.Adipose tissue expandability,lipotoxicity and the m etabolic syndrom e—an allostatic perspective[J].Genes&Nutrition,2007,2(1):41-45.
[6]NIELSEN M O,KONGSTED A H,THYGESEN M P,et al.Late gestation undernutrition can predispose for visceral adiposity by altering fat distribution patterns and increasing the preference for a high-fat diet in early postnatal life[J].British Journal of Nutrition,2013,109(11):2098-2110.
[7]DE BLASIO M J,GATFORD K L,ROBINSON J S,et al.Placental restriction of fetal grow th reduces size at birth and alters postnatal grow th,feeding activity,and adiposity in the young lam b[J].Am erican Journal of Physiology:Regulatory,Integrative and Com parative Physiology,2007,292(2):R875-R886.
[8]KHANAL P,HUSTED S V,AXEL A M,et al.Late gestation over-and undernutrition predispose for visceral adiposity in response to a post-natal obesogenic diet,but w ith differential im pacts on glucose-insulin adaptations during fasting in lam bs[J].Acta Physiologica,2013,210(1):110-126.
[9]张宏福.动物营养参数与饲养标准[M].2版.北京:中国农业出版社,2010.
[10]张丽英.饲料分析及饲料质量检测技术[M].2版.北京:中国农业大学出版社,2003.
[11]ICHIHARA K,SHIBAHARA A,YAMAMOTO K,et al.An im proved m ethod for rapid analysis of the fatty acids of glycerolipids[J].Lipids,1996,31(5):535-539.
[12]WAJCHENBERG B L.Subcutaneous and visceral adipose tissue:their relation to the m etabolic syndrom e[J].Endocrine Reviews,2000,21(6):697-738.
[13]HABEGGER K M,HEPPNER K M,GEARY N,et al.The m etabolic actions of glucagon revisited[J].Nature Review s Endocrinology,2010,6(12):689-697.
[14]EHRHARDT R A,SLEPETIS R M,BELL A W,et al.M aternal leptin is elevated during pregnancy in sheep[J].Dom est Anim Endocrinol,2001,21(2):85-96.
[15]NEDVIDKOVAJ,SMITKA K,KOPSK Y'V,et al.Adiponectin,an adipocyte-derived protein[J].Physiological Research,2005,54(2):133-140.
[16]VASSEUR F,LEPRETRE F,LACQUEMANT C,et al.The genetics of adiponectin[J].Current Diabetes Reports,2003,3(2):151-158.
[17]DIEZ J J,IGLESIAS P.The role of the novel adipocyte-derived horm one adiponectin in hum an disease[J].European Journal of Endocrinology,2003,148(3):293-300.
[18]HARRIS R B S,APOLZAN J W.Changes in glucose tolerance and leptin responsiveness of rats offered a choice of lard,sucrose,and chow[J].Am erican Journal of Physiology:Regulatory,Integrative and Comparative Physiology,2012,302(11):R1327-R1339.
[19]HAGHIAC M,BASU S,PRESLEY L,et al.Patterns of adiponectin expression in term pregnancy:im pact of obesity[J].The Journal of Clinical Endocrinology&M etabolism,2014,99(9):3427-3434.
[20]茅慧玲,刘建新.反刍动物肌肉脂肪酸营养调控研究进展[J].饲料工业,2010,31(23):30-34.
[21]BANSKALIEVA V V,SAHLU T,GOETSCH A L.Fatty acid com position of goat m uscles and fat depots:a review[J].Sm all Rum inant Research,2000,37(3):255-268.
[22]双金,敖力格日玛,敖长金.苏尼特羊体脂脂肪酸组成的研究[J].畜牧兽医学报,2015,46(8):1363-1374.
[23]DERVISHI E,SERRANO C,JOY M,et al.Effect of the feeding system on the fatty acid com position,expression of theΔ9-desaturase,peroxisom e proliferatoractivated receptor alpha,gam m a,and sterol regulatory elem ent binding protein 1 genes in the sem itendinous m uscle of light lam bs of the Rasa aragonesa breed[J].BMC Veterinary Research,2010,6:40.
[24]DERVISHI E,SERRANO C,JOY M,et al.The effect of feeding system in the expression of genes related w ith fat m etabolism in sem itendinous m uscle in sheep[J].Meat Science,2011,89(1):91-97.
[25]蒋金航,马云,王新庄.PPARγ基因调控脂肪细胞分化的研究进展[J].中国畜牧杂志,2014,50(9):91-95.
[26]YEON S H,LEE S H,CHOI B H,et al.Genetic variation of FASN is associated w ith fatty acid com position of Hanw oo[J].M eat Science,2013,94(1):133-138.
[27]MENENDEZ J A,LUPU R.Fatty acid synthase-catalyzed de novo fatty acid biosynthesis:from anabolicenergy-storage pathw ay in norm al tissues to jack-ofall-trades in cancer cells[J].Archivum Im m unologiae et Therapiae Experim entalis,2004,52(6):414-426.
[28]PATON C M,NTAMBI J M.Biochemical and physiological function of stearoyl-Co A desaturase[J].Am erican Journal of Physiology Endocrinology and M etabolism,2009,297(1):E28-E37.
[29]HOLM C.Molecular mechanisms regulating hormonesensitive lipase and lipolysis[J].Biochem ical Society Transactions,2003,31(6):1120-1124.
[30]PUIGSERVER P,WU Z D,PARK C W,et al.A coldinducible coactivator of nuclear receptors linked to adaptive therm ogenesis[J].Cell,1998,92(6):829-839.
[31]孙亮,朱小泉,王沥,等.核辅激活因子PGC-1表达的分子调控机制[J].中国生物化学与分子生物学报,2005,21(4):431-439.
[32]张艳芳.CPT1基因对猪脂肪沉积的影响及其调控机制的研究[D].博士学位论文.杭州:浙江大学,2010.
[33]BARTELDS B,TAKENS J,SMID G B,et al.Myocardial carnitine palm itoyltransferaseⅠexpression and long-chain fatty acid oxidation in fetal and new born lam bs[J].Am erican Journal of Physiology:Heart and Circulatory Physiology,2004,286(6):H2243-H2248.
[34]MCINNES K J,BROWN K A,HUNGER N I,et al.Regulation of LKB1 expression by sex horm ones in adipocytes[J].International Journal of Obesity,2012,36(7):982-985.
[35]WU B,DU Y,LIU C,et al.Effect of repeated fasting/refeeding on body w eight control and energy balance regulation in rats[J].Journal of Hygiene Research,2010,39(5):601-605.
[36]MOON J S,LEE S,PARK M A,et al.UCP2-induced fatty acid synthase prom otes NLRP3 inflam m asom e activation during sepsis[J].Journal of Clinical Investigation,2015,125(2):665-680.
[37]DE QUEIROZ K B,GUIMARAES J B,COIMBRA C C,et al.Endurance training increases leptin expression in the retroperitoneal adipose tissue of rats fed w ith a high-sugar diet[J].Lipids,2014,49(1):85-96.
[2]ROOS S,LAGERLOF O,WENNERGREN M,et al.Regulation of am ino acid transporters by glucose and grow th factors in cultured prim ary hum an trophoblast cells is m ediated by m TOR signaling[J].Am erican Journal of Physiology Cell Physiology,2009,297(3):C723-C731.
[3]ROSARIO F J,JANSSON N,KANAI Y,et al.Maternal protein restriction in the rat inhibits placental insulin,m TOR,and STAT3 signaling and dow n-regulates placental am ino acid transporters[J].Endocrinology,2011,152(3):1119-1129.
[4]BARCROFT J.Researches on pre-natal life[M].Oxford:Blackw ell Scientific Publications,1946.
[5]VIRTUE S,VIDAL-PUIG A.Adipose tissue expandability,lipotoxicity and the m etabolic syndrom e—an allostatic perspective[J].Genes&Nutrition,2007,2(1):41-45.
[6]NIELSEN M O,KONGSTED A H,THYGESEN M P,et al.Late gestation undernutrition can predispose for visceral adiposity by altering fat distribution patterns and increasing the preference for a high-fat diet in early postnatal life[J].British Journal of Nutrition,2013,109(11):2098-2110.
[7]DE BLASIO M J,GATFORD K L,ROBINSON J S,et al.Placental restriction of fetal grow th reduces size at birth and alters postnatal grow th,feeding activity,and adiposity in the young lam b[J].Am erican Journal of Physiology:Regulatory,Integrative and Com parative Physiology,2007,292(2):R875-R886.
[8]KHANAL P,HUSTED S V,AXEL A M,et al.Late gestation over-and undernutrition predispose for visceral adiposity in response to a post-natal obesogenic diet,but w ith differential im pacts on glucose-insulin adaptations during fasting in lam bs[J].Acta Physiologica,2013,210(1):110-126.
[9]张宏福.动物营养参数与饲养标准[M].2版.北京:中国农业出版社,2010.
[10]张丽英.饲料分析及饲料质量检测技术[M].2版.北京:中国农业大学出版社,2003.
[11]ICHIHARA K,SHIBAHARA A,YAMAMOTO K,et al.An im proved m ethod for rapid analysis of the fatty acids of glycerolipids[J].Lipids,1996,31(5):535-539.
[12]WAJCHENBERG B L.Subcutaneous and visceral adipose tissue:their relation to the m etabolic syndrom e[J].Endocrine Reviews,2000,21(6):697-738.
[13]HABEGGER K M,HEPPNER K M,GEARY N,et al.The m etabolic actions of glucagon revisited[J].Nature Review s Endocrinology,2010,6(12):689-697.
[14]EHRHARDT R A,SLEPETIS R M,BELL A W,et al.M aternal leptin is elevated during pregnancy in sheep[J].Dom est Anim Endocrinol,2001,21(2):85-96.
[15]NEDVIDKOVAJ,SMITKA K,KOPSK Y'V,et al.Adiponectin,an adipocyte-derived protein[J].Physiological Research,2005,54(2):133-140.
[16]VASSEUR F,LEPRETRE F,LACQUEMANT C,et al.The genetics of adiponectin[J].Current Diabetes Reports,2003,3(2):151-158.
[17]DIEZ J J,IGLESIAS P.The role of the novel adipocyte-derived horm one adiponectin in hum an disease[J].European Journal of Endocrinology,2003,148(3):293-300.
[18]HARRIS R B S,APOLZAN J W.Changes in glucose tolerance and leptin responsiveness of rats offered a choice of lard,sucrose,and chow[J].Am erican Journal of Physiology:Regulatory,Integrative and Comparative Physiology,2012,302(11):R1327-R1339.
[19]HAGHIAC M,BASU S,PRESLEY L,et al.Patterns of adiponectin expression in term pregnancy:im pact of obesity[J].The Journal of Clinical Endocrinology&M etabolism,2014,99(9):3427-3434.
[20]茅慧玲,刘建新.反刍动物肌肉脂肪酸营养调控研究进展[J].饲料工业,2010,31(23):30-34.
[21]BANSKALIEVA V V,SAHLU T,GOETSCH A L.Fatty acid com position of goat m uscles and fat depots:a review[J].Sm all Rum inant Research,2000,37(3):255-268.
[22]双金,敖力格日玛,敖长金.苏尼特羊体脂脂肪酸组成的研究[J].畜牧兽医学报,2015,46(8):1363-1374.
[23]DERVISHI E,SERRANO C,JOY M,et al.Effect of the feeding system on the fatty acid com position,expression of theΔ9-desaturase,peroxisom e proliferatoractivated receptor alpha,gam m a,and sterol regulatory elem ent binding protein 1 genes in the sem itendinous m uscle of light lam bs of the Rasa aragonesa breed[J].BMC Veterinary Research,2010,6:40.
[24]DERVISHI E,SERRANO C,JOY M,et al.The effect of feeding system in the expression of genes related w ith fat m etabolism in sem itendinous m uscle in sheep[J].Meat Science,2011,89(1):91-97.
[25]蒋金航,马云,王新庄.PPARγ基因调控脂肪细胞分化的研究进展[J].中国畜牧杂志,2014,50(9):91-95.
[26]YEON S H,LEE S H,CHOI B H,et al.Genetic variation of FASN is associated w ith fatty acid com position of Hanw oo[J].M eat Science,2013,94(1):133-138.
[27]MENENDEZ J A,LUPU R.Fatty acid synthase-catalyzed de novo fatty acid biosynthesis:from anabolicenergy-storage pathw ay in norm al tissues to jack-ofall-trades in cancer cells[J].Archivum Im m unologiae et Therapiae Experim entalis,2004,52(6):414-426.
[28]PATON C M,NTAMBI J M.Biochemical and physiological function of stearoyl-Co A desaturase[J].Am erican Journal of Physiology Endocrinology and M etabolism,2009,297(1):E28-E37.
[29]HOLM C.Molecular mechanisms regulating hormonesensitive lipase and lipolysis[J].Biochem ical Society Transactions,2003,31(6):1120-1124.
[30]PUIGSERVER P,WU Z D,PARK C W,et al.A coldinducible coactivator of nuclear receptors linked to adaptive therm ogenesis[J].Cell,1998,92(6):829-839.
[31]孙亮,朱小泉,王沥,等.核辅激活因子PGC-1表达的分子调控机制[J].中国生物化学与分子生物学报,2005,21(4):431-439.
[32]张艳芳.CPT1基因对猪脂肪沉积的影响及其调控机制的研究[D].博士学位论文.杭州:浙江大学,2010.
[33]BARTELDS B,TAKENS J,SMID G B,et al.Myocardial carnitine palm itoyltransferaseⅠexpression and long-chain fatty acid oxidation in fetal and new born lam bs[J].Am erican Journal of Physiology:Heart and Circulatory Physiology,2004,286(6):H2243-H2248.
[34]MCINNES K J,BROWN K A,HUNGER N I,et al.Regulation of LKB1 expression by sex horm ones in adipocytes[J].International Journal of Obesity,2012,36(7):982-985.
[35]WU B,DU Y,LIU C,et al.Effect of repeated fasting/refeeding on body w eight control and energy balance regulation in rats[J].Journal of Hygiene Research,2010,39(5):601-605.
[36]MOON J S,LEE S,PARK M A,et al.UCP2-induced fatty acid synthase prom otes NLRP3 inflam m asom e activation during sepsis[J].Journal of Clinical Investigation,2015,125(2):665-680.
[37]DE QUEIROZ K B,GUIMARAES J B,COIMBRA C C,et al.Endurance training increases leptin expression in the retroperitoneal adipose tissue of rats fed w ith a high-sugar diet[J].Lipids,2014,49(1):85-96.