催乳素对奶牛脂肪细胞脂代谢关键基因的作用
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摘要
催乳素(prolactin,PRL)是启动和维持泌乳的重要激素,在泌乳期间能调节机体将营养物质和能量优先流向乳腺,用于乳脂和乳蛋白的合成。本试验采用体外分离培养奶牛前脂肪细胞,诱导分化为成熟脂肪细胞,加入PRL探讨其对脂肪细胞脂代谢相关基因的作用。结果显示,奶牛白色脂肪细胞存在催乳素受体(prolactin receptor,PRLR),且在PRL质量浓度为75μg/L,作用时间为9h时,PRLR表达量达到最高,此时调节脂合成的核转录因子SREBPE-1c及下游控制脂合成的基因FAS和LPL均显著降低,75μg/L组与对照组相比差异极显著(P<0.01);控制脂分解的基因HSL显著升高,75μg/L组与对照组相比差异显著(P<0.05);细胞培养上清液中的甘油含量显著升高,75μg/L组与对照组相比差异显著(P<0.05)。结果表明,PRL能直接作用于奶牛白色脂肪细胞,降低脂合成作用并促进脂分解作用。
Prolactin(PRL)is an important hormone which stimulates and maintains milk production.It regulates metabolism and transfers nutrition to breast from other tissue during lactation.Pre-adipocytes have been isolated from white adipose tissue of dairy cows and then differentiated into mature adipocytes in this study.Different concentrations of PRL were added to the mature adipocytes to study the effect of PRL on lipid metabolism.The results demonstrated the PRLR was expressed in bovine adipocytes and mRNA levels of PRLR reached the maximum at 9hafter75μg/L PRLtreatment in adipocytes.Meanwhile,the mRNA levels of lipogenesis regulator SREBP-1cand its target genes FAS and LPL were decreased significantly whereas lipolysis regulator HSL was increased significantly.Furthermore,75μg/L PRL treatment significantly increased the glycerinum in supernatant.Taken together,these results demonstrate that PRL increases the lipolysis and decreases the lipogenesis in bovine adipocytes directly through PRLR.
Prolactin(PRL)is an important hormone which stimulates and maintains milk production.It regulates metabolism and transfers nutrition to breast from other tissue during lactation.Pre-adipocytes have been isolated from white adipose tissue of dairy cows and then differentiated into mature adipocytes in this study.Different concentrations of PRL were added to the mature adipocytes to study the effect of PRL on lipid metabolism.The results demonstrated the PRLR was expressed in bovine adipocytes and mRNA levels of PRLR reached the maximum at 9hafter75μg/L PRLtreatment in adipocytes.Meanwhile,the mRNA levels of lipogenesis regulator SREBP-1cand its target genes FAS and LPL were decreased significantly whereas lipolysis regulator HSL was increased significantly.Furthermore,75μg/L PRL treatment significantly increased the glycerinum in supernatant.Taken together,these results demonstrate that PRL increases the lipolysis and decreases the lipogenesis in bovine adipocytes directly through PRLR.
引文
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[16]Bertile F,Raclot T.mRNA levels of SREBP-1cdo not coincide with the changes in adipose lipogenic gene expression[J].Biochem Biophys Res Commun,2004,325(3):827-834.
[17]Graugnard D E,Bionaz M,Trevisi E,et al.Blood immunometabolic indices and polymorphonuclear neutrophil function in peripartum dairy cows are altered by level of dietary energy prepartum[J].J Dairy Sci,2012,95(4):1749-1758.
[18]张克春,谭勋.围产期奶牛NEFA的营养调控研究进展[J].乳业科学与技术,2007,3(1):49-52.
[19]史晓霞,王建国,等.非酯化脂肪酸对体外培养犊牛原代肝细胞中炎性因子表达及其活性的影响[J].中国兽医学报,2014,34(1):92-96.
[20]Fajas L,Schoonjans K,Gelman L,et al.Regulation of peroxisome proliferator-activated receptorγexpression by adipocyte differentiation and determination factor 1/sterol regulatory element binding protein 1:implications for adipocyte differentiation and metabolism[J].Mol Cell Biol,1999,19(8):5495-5503.
[2]Li X,Li X,Bai G,et al.Effects of non-esterified fatty acids on the gluconeogenesis in bovine hepatocytes[J].Mol Cell Biochem,2012,359(1/2):385-388.
[3]Kelley K W,Weigent D A,Kooijman R.Protein hormones and immunity[J].Brain Behav Immun,2007,21(4):384-392.
[4]Nagano M,Kelly P A.Tissue distribution and regulation of rat prolactin receptor gene expression.Quantitative analysis by polymerase chain reaction[J].J Biol Chem,1994,269(18):37-45.
[5]Jensen D R,Gavigan S,Sawicki V,et al.Regulation of lipoprotein lipase activity and mRNA in the mammary gland of the lactating mouse[J].Biochem J,1994,298:321-327.
[6]Zechner R,Strauss J,Frank S,et al.The role of lipoprotein lipase in adipose tissue development and metabolism[J].Int J Obesity Relat Metabolic Disorders:J Int Assoc Stud Obesity,2000,24:53-60.
[7]Ling C,Svensson L,Oden B,et al.Identification of functional prolactin(PRL)receptor gene expression:PRL inhibits lipoprotein lipase activity in human white adipose tissue[J].J Clin Endocrinol Metabolism,2003,88(4):1804-1808.
[8]Ling C,Hellgren G,Gebre-Medhin M,et al.Prolactin(PRL)receptor gene expression in mouse adipose tissue:Increases during lactation and in PRL-transgenic mice 1[J].Endocrinology,2000,141(10):3564-3572.
[9]Wang X,Huang M,Wang Y.The effect of insulin,TNFαand DHA on the proliferation,differentiation and lipolysis of preadipocytes isolated from large yellow croaker(Pseudosciaena Crocea R.)[J].PloS One,2012,7(10):48-69.
[10]Rosen E D,Walkey C J,Puigserver P,et al.Transcriptional regulation of adipogenesis[J].Genes&development,2000,14(11):1293-1307.
[11]Gurnell M.Peroxisome proliferator-activated receptorγand the regulation of adipocyte function:lessons from human genetic studies[J].Best Practice Res Clin Endocrinol Metabolism,2005,19(4):501-523.
[12]Mueller E,Drori S,Aiyer A,et al.Genetic analysis of adipogenesis through peroxisome proliferator-activated receptorγisoforms[J].J Biol Chem,2002,277(44):25-30.
[13]Ren D,Collingwood T N,Rebar E J,et al.PPARγknockdown by engineered transcription factors:exogenous PPARγ2but not PPARγ1reactivates adipogenesis[J].Gene Dev,2002,16(1):27-32.
[14]Sekiya M,Yahagi N,Matsuzaka T,et al.SREBP-1-independent regulation of lipogenic gene expression in adipocytes[J].J Lipid Res,2007,48(7):1581-1591.
[15]Palmer D G,Rutter G A,TavaréJ M.Insulin-stimulated fatty acid synthase gene expression does not require increased sterol response element binding protein 1transcription in primary adipocytes[J].Biochem Biophys Res Commun,2002,291(3):439-443.
[16]Bertile F,Raclot T.mRNA levels of SREBP-1cdo not coincide with the changes in adipose lipogenic gene expression[J].Biochem Biophys Res Commun,2004,325(3):827-834.
[17]Graugnard D E,Bionaz M,Trevisi E,et al.Blood immunometabolic indices and polymorphonuclear neutrophil function in peripartum dairy cows are altered by level of dietary energy prepartum[J].J Dairy Sci,2012,95(4):1749-1758.
[18]张克春,谭勋.围产期奶牛NEFA的营养调控研究进展[J].乳业科学与技术,2007,3(1):49-52.
[19]史晓霞,王建国,等.非酯化脂肪酸对体外培养犊牛原代肝细胞中炎性因子表达及其活性的影响[J].中国兽医学报,2014,34(1):92-96.
[20]Fajas L,Schoonjans K,Gelman L,et al.Regulation of peroxisome proliferator-activated receptorγexpression by adipocyte differentiation and determination factor 1/sterol regulatory element binding protein 1:implications for adipocyte differentiation and metabolism[J].Mol Cell Biol,1999,19(8):5495-5503.