叶酸和维生素B_(12)对五龙鹅肝脏中磷脂酸磷酸酯酶1基因表达量的影响及其与血清脂类代谢、脂肪沉积和肉品质指标的相关性分析
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摘要
本试验旨在探究饲粮中添加不同水平叶酸和维生素B12对五龙鹅肝脏中磷脂酸磷酸酯酶1(Lpin1)基因表达量的影响及在不同组织中的表达差异性,并分析Lpin1基因表达量与血清脂类代谢、脂肪沉积、肉品质指标之间的相关性。试验选用5周龄五龙鹅420只,随机分为7个组,每个组6个重复,每个重复10只鹅。试验采用2×3两因素(叶酸添加水平×维生素B12添加水平)交叉等重复的析因设计,空白对照组饲喂基础饲粮,不添加叶酸和维生素B12;试验组饲粮叶酸添加水平分别为0.25、2.00 mg/kg,维生素B12添加水平分别为0.003、0.009、0.018 mg/kg。试验期为4周。结果表明:1)不同添加水平的叶酸和维生素B12的互作对五龙鹅的末重影响显著(P<0.05)。2)不同添加水平的叶酸和维生素B12的互作对皮脂率、腹脂率、肌间脂带宽、胸肌肌内脂肪率和腿肌肌内脂肪率影响显著(P<0.05)。3)不同添加水平的叶酸和维生素B12及其互作对五龙鹅肝脏中Lpin1基因表达量影响显著(P<0.05)。4) Lpin1基因在五龙鹅的心脏、肝脏、肾脏、腹脂、肺脏、肌胃、腺胃、胸肌、腿肌、脾脏、胰腺11个组织中均有表达,其中Lpin1基因表达量表达量最高的为腹脂,显著高于其他组织(P<0.05);其次为肌胃、胰腺、肺脏,显著高于腺胃、心脏、肾脏、腿肌、胸肌(P<0.05)。5)五龙鹅肝脏中Lpin1基因表达量与血清甘油三酯、高密度脂蛋白胆固醇含量呈显著负相关(P<0.05),与血清葡萄糖、总胆固醇、低密度脂蛋白胆固醇含量呈负相关(P>0.05)。6)五龙鹅肝脏中Lpin1基因表达量与腹脂率、腿肌肌内脂肪率呈显著正相关(P<0.05),与胸肌肌内脂肪率、肌间脂带宽呈极显著正相关(P<0.01),与皮脂率呈负相关(P>0.05)。7)五龙鹅肝脏中Lpin1基因表达量与胸肌和腿肌pH、肉色、剪切力、失水率均无显著相关性(P>0.05),但与腿肌滴水损失呈显著正相关(P<0.05)。由此可见,不同添加水平的叶酸和维生素B12及其互作对五龙鹅肝脏中Lpin1基因表达量有显著影响,且Lpin1基因表达量与血清脂类代谢、脂肪沉积和肉品质指标有一定的相关性。
Abstract:This study was conducted to investigated the effect of dietary different supplemental levels of folic acid and vitamin B_(12) on liver lipid phosphate phosphohydrolase 1(Lpin1) gene expression of Wulong geese and its expression difference in different tissues,and to further analysis the correlation of Lpin1 gene expression with serum lipid metabolism,fat deposition and meat quality indices.A total of 420 five-week-old Wulong geese were randomly divided into 7 groups with 6 replicates per group and 10 geese per replicate.The trial used a 2×3 two factors(folic acid supplemental level×vitamin B_(12) supplemental level) across-equal factorial design,geese in the blank control group were fed a basal diet without folic acid and vitamin B_(12);geese in experimental groups were fed experiment diets,which folic acid supplemental levels were 0.25,2.00 mg/kg,respectively,and vitamin B_(12) supplemental levels were 0.003,0.009,0.018 mg/kg,respectively.The experiment lasted for 4 weeks.The results showed as follow :1) the interaction of different supplemental levels of folic acid and vitamin B_(12) had significant effect on the final weight of Wulong geese(P<0.05).2) The interaction of different supplemental levels of folic acid and vitamin B_(12) had significant effects on the percentage of subcutaneous fat,percentage of abdominal fat,intermuscular fat bandwidth,intramuscular fat rate of breast muscle,intramuscular fat rate of leg muscle of Wulong geese(P<0.05).3) Different supplemental levels of folic acid and vitamin B_(12) and their interaction had significant effect on the liver Lpin1 gene expression of Wulong geese(P<0.05).4) The Lpin1 gene expression was detected in 11 tissues of heart,liver,kidney,abdominal fat,lung,gizzard,glandular stomach,breast muscle,leg muscle,spleen and pancreas;the Lpin1 gene expression in abdominal fat was the highest,and it significantly higher than that in other tissues(P<0.05);secondly was in gizzard,pancreas and lung,significantly higher than that in glandular stomach,heart,kidney,leg muscle and breast muscle(P<0.05).5) The liver Lpin1 gene expression of Wulong geese had significant negative correlations with the contents of triglyceride and high density lipoprotein cholesterol in serum(P<0.05),and had negative correlations with the contents of glucose,total cholesterol,low density lipoprotein cholesterol in serum(P>0.05).6) The liver Lpin1 gene expression of Wulong geese had significant positive correlations with the percentage of abdominal fat and intramuscular fat rate of leg muscle(P<0.05),had significant positive correlations with the intermuscular fat bandwidth,intramuscular fat rate of breast muscle(P<0.01),and had a negative correlation with the percentage of subcutaneous fat(P>0.05).7) The liver Lpin1 gene expression of Wulong geese had no significant correlation with the pH,meat color,shearing force and water loss rate in breast muscle and leg muscle(P>0.05),but had a significant positive correlation with the drip loss of leg muscle(P<0.05).In conclusion,different supplemental levels of folic acid and vitamin B_(12) and their interaction have significant effects on the liver Lpin1 gene expression of Wulong geese,and the Lpin1 gene expression has a certain correlation with serum lipid metabolism,fat deposition and meat quality indices.
Abstract:This study was conducted to investigated the effect of dietary different supplemental levels of folic acid and vitamin B_(12) on liver lipid phosphate phosphohydrolase 1(Lpin1) gene expression of Wulong geese and its expression difference in different tissues,and to further analysis the correlation of Lpin1 gene expression with serum lipid metabolism,fat deposition and meat quality indices.A total of 420 five-week-old Wulong geese were randomly divided into 7 groups with 6 replicates per group and 10 geese per replicate.The trial used a 2×3 two factors(folic acid supplemental level×vitamin B_(12) supplemental level) across-equal factorial design,geese in the blank control group were fed a basal diet without folic acid and vitamin B_(12);geese in experimental groups were fed experiment diets,which folic acid supplemental levels were 0.25,2.00 mg/kg,respectively,and vitamin B_(12) supplemental levels were 0.003,0.009,0.018 mg/kg,respectively.The experiment lasted for 4 weeks.The results showed as follow :1) the interaction of different supplemental levels of folic acid and vitamin B_(12) had significant effect on the final weight of Wulong geese(P<0.05).2) The interaction of different supplemental levels of folic acid and vitamin B_(12) had significant effects on the percentage of subcutaneous fat,percentage of abdominal fat,intermuscular fat bandwidth,intramuscular fat rate of breast muscle,intramuscular fat rate of leg muscle of Wulong geese(P<0.05).3) Different supplemental levels of folic acid and vitamin B_(12) and their interaction had significant effect on the liver Lpin1 gene expression of Wulong geese(P<0.05).4) The Lpin1 gene expression was detected in 11 tissues of heart,liver,kidney,abdominal fat,lung,gizzard,glandular stomach,breast muscle,leg muscle,spleen and pancreas;the Lpin1 gene expression in abdominal fat was the highest,and it significantly higher than that in other tissues(P<0.05);secondly was in gizzard,pancreas and lung,significantly higher than that in glandular stomach,heart,kidney,leg muscle and breast muscle(P<0.05).5) The liver Lpin1 gene expression of Wulong geese had significant negative correlations with the contents of triglyceride and high density lipoprotein cholesterol in serum(P<0.05),and had negative correlations with the contents of glucose,total cholesterol,low density lipoprotein cholesterol in serum(P>0.05).6) The liver Lpin1 gene expression of Wulong geese had significant positive correlations with the percentage of abdominal fat and intramuscular fat rate of leg muscle(P<0.05),had significant positive correlations with the intermuscular fat bandwidth,intramuscular fat rate of breast muscle(P<0.01),and had a negative correlation with the percentage of subcutaneous fat(P>0.05).7) The liver Lpin1 gene expression of Wulong geese had no significant correlation with the pH,meat color,shearing force and water loss rate in breast muscle and leg muscle(P>0.05),but had a significant positive correlation with the drip loss of leg muscle(P<0.05).In conclusion,different supplemental levels of folic acid and vitamin B_(12) and their interaction have significant effects on the liver Lpin1 gene expression of Wulong geese,and the Lpin1 gene expression has a certain correlation with serum lipid metabolism,fat deposition and meat quality indices.
引文
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[14]ESECELI H,DEGIRMENCIOGLU N,BILGIC M.The effect of inclusion of chromium yeast(Co-FatorⅡ,Alltech Inc.)and folic acid to the rations of laying hens on performance,egg quality,egg yolk cholesterol,f olic acid and chromium levels[J].Journal of Animal and Veterinary Advances,2010,9(2):384-391.
[15]BURDGE G C,LILLYCROP K A,JACKSON A A,et al.The nature of the growth pattern and of the metabolic response to fasting in the rat are dependent upon the dietary protein and folic acid intakes of their pregnant dams and post-weaning fat consumption[J].British Journal of Nutrition,2008,99(3):540-549.
[16]PHAN J,PTERFY M,REUE K.Lipin expression preceding peroxisome proliferator-activated receptor-γis critical for adipogenesis,in vivo and in vitro[J].The Journal of Biological Chemistry,2004,279(28):29558-29564.
[17]REUE K,ZHANG P X.The lipin protein family:dual roles in lipid biosynthesis and gene expression[J].FEBS Letters,2008,582(1):90-96.
[18]DOUGLAS D S,MORAN J L,BERMINGHAM J R,et al.Concurrent Lpin1 and Nrcam mouse mutations result in severe peripheral neuropathy with transitory hindlimb paralysis[J].Journal of Neuroscience,2009,29(39):12089-12100.
[19]MICHOT C,HUBERT L,BRIVET M,et al.LPIN1gene mutations:a major cause of severe rhabdomyolysis in early childhood[J].Human Mutation,2010,31(7):E1564-E1573.
[20]唐修君,陈宽雄,高玉时,等.我国地方鹅种肉品质研究进展[J].中国家禽,2008,30(24):48-50.
[21]WANG R,WANG T,LU W,et al.Three indel variants in chicken LPIN1 exon 6/flanking region are associated with performance and carcass traits[J].British Poultry Science,2015,56(6):621-630.
[22]LI S Y,CHEN W,KANG X T,et al.Distinct tissue expression profiles of chicken Lpin1-α/βisoforms and the effect of the variation on muscle fiber traits[J].Gene,2013,515(2):281-290.
[2]刘欢.叶酸、维生素B12、B6与神经退行性疾病[J].天津医科大学学报,2005,11(4):650-653.
[3]STEKOL J A,WEISS S,WEISS K W.Vitamin B12and folic acid in the synthesis of choline in the rat[J].Archives of Biochemistry and Biophysics,1952,36(1):5-10.
[4]ERKURT M A,AYDOGDU I,DIKILITA爦M,et al.Effects of cyanocobalamin on immunity in patients with pernicious anemia[J].Medical Principles and Practice,2008,17(2):131-135.
[5]HAN G S,WU W I,CARMAN G M.The Saccharomyces cerevisiae Lipin homolog is a Mg2+-dependent phosphatidate phosphatase enzyme[J].The Journal of Biological Chemistry,2006,281(14):9210-9218.
[6]PHAN J,REUE K,et al.Lipin,a lipodystrophy and obesity gene[J].Cell Metabolism,2005,1(1):73-83.
[7]中华人民共和国农业部.家禽生产性能名词术语和度量统计方法NY/T 823-2004[S]北京:中国标准出版社,2008.
[8]中华人民共和国农业部.肉与肉制品GB/T 9695-2008[S].北京:中国标准出版社,2008.
[9]王宝维,程漫漫,孔敏,等.饲粮中添加叶酸和维生素B12对五龙鹅生长性能、屠宰性能、脂肪沉积及极长链脂肪酸延长酶7基因表达量的影响[J].动物营养学报,2018,30(11):4433-4443.
[10]YAO-BORENGASSER A W,RASOULI N,VARMAV,et al.Lipin expression is attenuated in adipose tissue of insulin-resistant human subjects and increases with peroxisome proliferator-activated receptorγactivation[J].Diabetes,2006,55(10):2811-2818.
[11]DONKOR J,SARIAHMETOGLU M,DEWALD J,et al.Three mammalian lipins act as phosphatidate phosphatases with distinct tissue expression patterns[J].Journal of Biological Chemistry,2007,282(6):3450-3457.
[12]HE X P,XU X W,ZHAO S H,et al.Investigation of Lpin1 as a candidate gene for fat deposition in pigs[J].Molecular Biology Reports,2009,36(5):1175-1180.
[13]李家驹.应激对肉仔鸡小肽吸收的影响及小肽调控脂肪代谢的机理研究[D].博士学位论文.北京:中国农业科学院,2010.
[14]ESECELI H,DEGIRMENCIOGLU N,BILGIC M.The effect of inclusion of chromium yeast(Co-FatorⅡ,Alltech Inc.)and folic acid to the rations of laying hens on performance,egg quality,egg yolk cholesterol,f olic acid and chromium levels[J].Journal of Animal and Veterinary Advances,2010,9(2):384-391.
[15]BURDGE G C,LILLYCROP K A,JACKSON A A,et al.The nature of the growth pattern and of the metabolic response to fasting in the rat are dependent upon the dietary protein and folic acid intakes of their pregnant dams and post-weaning fat consumption[J].British Journal of Nutrition,2008,99(3):540-549.
[16]PHAN J,PTERFY M,REUE K.Lipin expression preceding peroxisome proliferator-activated receptor-γis critical for adipogenesis,in vivo and in vitro[J].The Journal of Biological Chemistry,2004,279(28):29558-29564.
[17]REUE K,ZHANG P X.The lipin protein family:dual roles in lipid biosynthesis and gene expression[J].FEBS Letters,2008,582(1):90-96.
[18]DOUGLAS D S,MORAN J L,BERMINGHAM J R,et al.Concurrent Lpin1 and Nrcam mouse mutations result in severe peripheral neuropathy with transitory hindlimb paralysis[J].Journal of Neuroscience,2009,29(39):12089-12100.
[19]MICHOT C,HUBERT L,BRIVET M,et al.LPIN1gene mutations:a major cause of severe rhabdomyolysis in early childhood[J].Human Mutation,2010,31(7):E1564-E1573.
[20]唐修君,陈宽雄,高玉时,等.我国地方鹅种肉品质研究进展[J].中国家禽,2008,30(24):48-50.
[21]WANG R,WANG T,LU W,et al.Three indel variants in chicken LPIN1 exon 6/flanking region are associated with performance and carcass traits[J].British Poultry Science,2015,56(6):621-630.
[22]LI S Y,CHEN W,KANG X T,et al.Distinct tissue expression profiles of chicken Lpin1-α/βisoforms and the effect of the variation on muscle fiber traits[J].Gene,2013,515(2):281-290.