猪肌源性前体脂肪细胞的分离培养和成脂诱导分化研究
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摘要
本研究旨在建立猪肌源性前体脂肪细胞的体外培养体系,并探究成脂诱导分化过程中相关基因的表达,为进一步研究调控猪肌内脂肪沉积的分子机制提供试验材料。采集15日龄仔猪的背最长肌组织,使用Ⅱ型胶原酶进行消化,分离获得前体脂肪细胞,进行原代、传代培养以及成脂诱导,观察细胞形态、测定生长曲线、油红O染色鉴定以及检测成脂关键基因PPARγ、FABP4、C/EBPα、C/EBPβ、ZFP423、SREBP1、PRDM16和HSL在诱导分化过程中的表达量。结果表明,分离的猪肌源性前体脂肪细胞在6～7 h,发现少量细胞开始贴壁,贴壁的细胞呈似圆型、小梭型以及其它不规则形状,经传代后细胞形态均一,呈典型的成纤维细胞样,细胞的生长曲线呈S型。添加分化培养基进行诱导后,所有基因在细胞分化的早期均有表达,FABP4、C/EBPα、SREBP1与PPARγ基因在分化早期表达量极显著升高(P <0. 01),在后期表达量极显著下降(P <0. 01),C/EBPβ、ZFP423和PRMD16基因的表达量在分化过程中未见显著变化,HSL的表达量随着分化进行极显著降低(P <0. 01)。在诱导分化12 d左右,油红O染色呈现红色。综上表明,本研究成功建立了猪肌源性前体脂肪细胞的分离与培养体系,进行成脂诱导分化,揭示分化过程中关键基因的表达规律,为进一步研究猪肌内脂肪沉积的分子机制和改善肉质品质奠定基础。
The aim of this study was to establish the culture system of porcine myogenic adipocytes cell in vitro,and to understand the mRNA expression regulation of adipogenesis related genes during adipogenic differentiation process and to provide experimental materials for further studying on the mechanism of intramuscular fat deposition in pig. Samples of longissimus dorsi muscle of the 15 days old piglets were collected and the precursor adipocytes cell was obtained by digested with type II collagenase,and the cell were separated,primary and passage cultured and adipogenic induction. The morphology of cell were observed and the growth curve was determined. The adipogenesis was measured by Oil red O staining and the mRNA expression of adipogenesis related genes,such as PPARγ,FABP4,C/EBPα,C/EBPβ,ZFP423,SREBP1,PRMD16 and HSL were detected by qRT-PCR during induced differentiation. The results showed that a small number of myogenic preadipocytes cells began to adhere to the cell wall atabout 6-7 h,and the shape of the cells were round,small shuttle and irregular,and became uniform in fibroblast-like appearance after cell passage. The growth curve was typical " S" shaped. All genes detected in this study were expressed at early stage of adipogenic differentiation after adding differentiation medium,in which the mRNA expression of FABP4,C/EBPα,SREBP1 and PPARγ at early stage was very significantly higher( P < 0. 01) and then was decreased significantly( P < 0. 01). There was no significant change for the mRNA expression of C/EBPβ,ZFP423 and PRMD16 genes during different differentiation stages. The mRNA expression of HSL was significantly reduced with the differentiation process( P< 0. 01). Oil red O staining was red after induction for 12 days. In conclusion,The system of the isolation,culture and adipogenic differentiation of porcine myogenic adipocytes cell was successfully established,and the expression and regulation of key genes involved in adipogenesis was investigated during cell differentiation process,which provide the foundation for further studying on the mechanism of intramuscular fat deposition and improvement of meat quality in pig production.
The aim of this study was to establish the culture system of porcine myogenic adipocytes cell in vitro,and to understand the mRNA expression regulation of adipogenesis related genes during adipogenic differentiation process and to provide experimental materials for further studying on the mechanism of intramuscular fat deposition in pig. Samples of longissimus dorsi muscle of the 15 days old piglets were collected and the precursor adipocytes cell was obtained by digested with type II collagenase,and the cell were separated,primary and passage cultured and adipogenic induction. The morphology of cell were observed and the growth curve was determined. The adipogenesis was measured by Oil red O staining and the mRNA expression of adipogenesis related genes,such as PPARγ,FABP4,C/EBPα,C/EBPβ,ZFP423,SREBP1,PRMD16 and HSL were detected by qRT-PCR during induced differentiation. The results showed that a small number of myogenic preadipocytes cells began to adhere to the cell wall atabout 6-7 h,and the shape of the cells were round,small shuttle and irregular,and became uniform in fibroblast-like appearance after cell passage. The growth curve was typical " S" shaped. All genes detected in this study were expressed at early stage of adipogenic differentiation after adding differentiation medium,in which the mRNA expression of FABP4,C/EBPα,SREBP1 and PPARγ at early stage was very significantly higher( P < 0. 01) and then was decreased significantly( P < 0. 01). There was no significant change for the mRNA expression of C/EBPβ,ZFP423 and PRMD16 genes during different differentiation stages. The mRNA expression of HSL was significantly reduced with the differentiation process( P< 0. 01). Oil red O staining was red after induction for 12 days. In conclusion,The system of the isolation,culture and adipogenic differentiation of porcine myogenic adipocytes cell was successfully established,and the expression and regulation of key genes involved in adipogenesis was investigated during cell differentiation process,which provide the foundation for further studying on the mechanism of intramuscular fat deposition and improvement of meat quality in pig production.
引文
[1]BREWER M S,ZHU L G,MCKEITH F K. Marbling effects on quality characteristics of pork loin chops:consumer purchase intent,visual and sensory characteristics[J]. Meat Sci,2001,59(2):153-163.
[2]ALTMANN M,PLIQUETT U. Prediction of intramuscular fat by impedance spectroscopy[J]. Meat Sci,2006,72(4):666-671.
[3]TANG Q Q,LANE M D. Adipogenesis:from stem cell to adipocyte[J]. Annu Rev Biochem,2012,81:715-736.
[4]FARMER S R. Transcriptional control of adipocyte formation[J]. Cell Metab,2006,4(4):263-273.
[5]RAMJI D P,FOKA P. CCAAT/enhancer-binding proteins:structure,function and regulation[J]. Biochem J,2002,365(3):561-575.
[6]GERBENS F,VERBURG F J,VAN MOERKERK H T B,et al. Associations of heart and adipocyte fatty acidbinding protein gene expression with intramuscular fat content in pigs[J]. J Anim Sci,2001,79(2):347-354.
[7]SEALE P,CONROE H M,ESTALL J,et al. Prdm16 determines the thermogenic program of subcutaneous white adipose tissue in mice[J]. J Clin Ivest,2011,121(1):96-105.
[8]鞠大鹏,詹丽杏.脂肪细胞分化及其调控的研究进展[J].中国细胞生物学学报,2010,32(5):690-695.JU D P,ZHAN L X. Developments in regulation of adipocytes differentiation[J]. Chinese Journal of Cell Biology,2010,32(5):690-695.(in Chinese)
[9]YEAMAN S J. Hormone-sensitive lipase-a multipurpose enzyme in lipid metabolism[J]. BBA-Mol Cell Res,1990,1052(1):128-132.
[10]GUPTA R K,ARANY Z,SEALE P,et al. Transcriptional control of preadipocyte determination by Zfp423[J]. Nature,2010,464(7288):619-623.
[11]孙成娟,许厚强,赵佳福,等.从江香猪肌内和皮下脂肪前体细胞分化过程中相关基因的表达[J].农业生物技术学报,2017,25(12):1979-1988.SUN C J,XU H Q,ZHAO J F,et al. The expression of related genes on intramuscular and subcutaneous preadipocytes during differentiation in Congjiang Xiang pig(Sus scrofa)[J]. Journal of Agricultural Biotechnology,2017,25(12):1979-1988.(in Chinese)
[12]林森,林亚秋,朱江江,等. Wnt10b对山羊前体脂肪细胞分化相关基因表达的影响[J].畜牧兽医学报,2018,49(4):685-692.LIN S,LIN Y Q,ZHU J J,et al. Effect of Wnt10b on the expression of precursor adipocytes differentiation related genes in goat[J]. Acta Veterinaria et Zootechnica Sinica,2018,49(4):685-692.(in Chinese)
[13]RUIZ-OJEDA F J,RUPéREZ A I,GOMEZ-LLORENTE C,et al. Cell models and their application for studying adipogenic differentiation in relation to obesity:a review[J]. Int J Mol Sci,2016,17(7):1040.
[14]CORDANI N,PISA V,POZZI L,et al. Nitric oxide controls fat deposition in dystrophic skeletal muscle by regulating fibro-adipogenic precursor differentiation[J]. Stem Cells,2014,32(4),874-885.
[15]赵存真,李黄琨,马云,等.腺病毒介导的MAT2B过表达促进猪肌内脂肪细胞分化[J].畜牧兽医学报,2017,48(5):802-809.ZHAO C Z,LI H K,MA Y,et al. MAT2B overexprssion mediated by adenovirus promoting porcine intramuscular preadipocyte differentiation[J]. Acta Veterinaria et Zootechnica Sinica,2017,48(5):802-809.(in Chinese)
[16]张萌萌,魏胜娟,王艺如,等.牛肌内前体脂肪细胞的分离培养及分化相关基因的表达规律研究[J].畜牧与兽医,2018,50(5):1-6.ZHANG M M,WEI S J,WANG Y R,et al. Primary culture and expression of differentiation-related genes in bovine intramuscular preadipocytes[J]. Animal Husbandry&Veterinary Medicine,2018,50(5):1-6.(in Chinese)
[17]耿晓晖,富俊才,韩凯,等.绵羊肌内脂肪前体细胞的体外培养及分选[J].中国农业大学学报,2014,19(5):110-115.GENG X H,FU J C,HAN K,et al. In vitro culture and purification of ovine intramuscular preadipocyte[J].Journal of China Agricultural University,2014,19(5):110-115.(in Chinese)
[18]屈长青,张国华,陈粉粉,等.猪前体脂肪细胞的原代培养[J].农业生物技术学报,2005,13(5):649-653.QU C Q,ZHANG G H,CHEN F F,et al. Primary culture of porcine preadipocyte[J]. Journal of Agricultural Biotechnology,2005,13(5):649-653.(in Chinese)
[19]杜琛,付绍印,韩志玲,等.绒山羊肌内前体脂肪细胞的基因表达分析[J].畜牧兽医学报,2013,44(10):1532-1538.DU C,FU S Y,HAN Z L,et al. Gene expression analysis on intramuscular preadipocytes in the cashmere goats[J]. Acta Veterinaria et Zootechnica Sinica,2013,44(10):1532-1538.(in Chinese)
[20]陈妍,张国华,陆会宁,等.八眉猪肌内前体脂肪细胞的分离培养及诱导分化[J].广东农业科学,2014,41(13):110-113,123.CHEN Y,ZHANG G H,LU H N,et al. Culture and induced differentiation of intramuscular preadipocytes from Bamei pigs in vitro[J]. Guangdong Agricultural Sciences,2014,41(13):110-113,123.(in Chinese)
[21]SATOH A,STEIN L,IMAI S. The role of mammalian sirtuins in the regulation of metabolism,aging,and longevity[M]//Histone Deacetylases:the Biology and Clinical Implication. Springer Berlin,Heidelberg,2011:125-162.
[22]LEFTEROVA M I,ZHANG Y,STEGER D J,et al.PPARγand C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale[J]. Gene Dev,2008,22(21):2941-2952.
[23]LEE J E,GE K. Transcriptional and epigenetic regulation of PPARγexpression during adipogenesis[J]. Cell Biosci,2014,4(1):29.
[24]马晶晶,章涛. PPARγ功能与疾病关系研究进展[J].中国药理学通报,2012,28(5):601-604.MA J J,ZHANG T. Function of PPARγand its relationship with diseases:a present review[J]. Chinese Pharmacological Bulletin,2012,28(5):601-604.(in Chinese)
[25]NOH J R,KIM Y H,HWANG J H,et al. Scoparone inhibits adipocyte differentiation through down-regulation of peroxisome proliferators-activated receptorγin 3T3-L1 preadipocytes[J]. Food Chem,2013,141(2):723-730.
[26]ROSEN E D,MACDOUGALD O A. Adipocyte differentiation from the inside out[J]. Nat Rev Mol Cell Biol,2006,7(12):885-896.
[27]TANAKA T,YOSHIDA N,KISHIMOTO T,et al. Defective adipocyte differentiation in mice lacking the C/EBPβand or C/EBPδgene[J]. EMBO J,1997,16(24):7432-7443.
[28]NTAMBI J M,YOUNG-CHEUL K. Adipocyte differentiation and gene expression[J]. J Nutr,2000,130(12):3122S-3126S.
[29]KIM J B,SPIEGELMAN B M. ADD1/SREBP1 promotes adipocyte differentiation and gene expression linked to fatty acid metabolism[J]. Gene Dev,1996,10(9):1096-1107.
[30]YELLATURU C R,DENG X,PARK E A,et al. Insulin enhances the biogenesis of nuclear sterol regulatory element-binding protein(SREBP)-1c by posttranscriptional down-regulation of Insig-2A and its dissociation from SREBP cleavage-activating protein(SCAP)·SREBP-1c complex[J]. J Biol Chem,2009,284(46):31726-31734.
[31]SHIMOMURA I,SHIMANO H,KORN B S,et al. Nuclear sterol regulatory element-binding proteins activate genes responsible for the entire program of unsaturated fatty acid biosynthesis in transgenic mouse liver[J]. J Biol Chem,1998,273(52):35299-35306.
[32]TAPPY L,LK A. Metabolic effects of fructose and the worldwide increase in obesity[J]. Physiol Rev,2010,90(1):23-46.
[33]HARADA K,SHEN W J,PATEL S,et al. Resistance to high-fat diet-induced obesity and altered expression of adipose-specific genes in HSL-deficient mice[J]. Am J Physiol-Endoc M,2003,285(6):E1182-E1195.
[34]XU A M,WANG Y,XU J Y,et al. Adipocyte fatty acid–binding protein is a plasma biomarker closely associated with obesity and metabolic syndrome[J]. Clin Chem,2006,52(3):405-413.
[35]BERNLOHR D A,BOLANOWSKI M A,KELLY T J Jr,et al. Evidence for an increase in transcription of specific m RNAs during differentiation of 3T3-L1 preadipocytes[J]. J Biol Chem,1985,260(9):5563-5567.
[36]CHRISTY R J,YANG V W,NTAMBI J M,et al. Differentiation-induced gene expression in 3T3-L1 preadipocytes:CCAAT/enhancer binding protein interacts with and activates the promoters of two adipocyte-specific genes[J]. Gene Dev,1989,3(9):1323-1335.
[37]ZHAO C P,TIAN F,YU Y,et al. Muscle transcriptomic analyses in Angus cattle with divergent tenderness[J].Mol Biol Rep,2012,39(4):4185-4193.
[38]WEI S J,ZHANG L F,ZHOU X,et al. Emerging roles of zinc finger proteins in regulating adipogenesis[J]. Cell Mol Life Sci,2013,70(23):4569-4584.
[39]SEALE P,KAJIMURA S,YANG W L,et al. Transcriptional control of brown fat determination by PRDM16[J]. Cell Metab,2007,6(1):38-54.
[40]SEALE P,BJORK B,YANG W L,et al. PRDM16 controls a brown fat/skeletal muscle switch[J]. Nature,2008,454(7207):961-967.
[41]KAJIMURA S,SEALE P,TOMARU T,et al. Regulation of the brown and white fat gene programs through a PRDM16/Ct BP transcriptional complex[J]. Gene Dev,2008,22(10):1397-1409.
[2]ALTMANN M,PLIQUETT U. Prediction of intramuscular fat by impedance spectroscopy[J]. Meat Sci,2006,72(4):666-671.
[3]TANG Q Q,LANE M D. Adipogenesis:from stem cell to adipocyte[J]. Annu Rev Biochem,2012,81:715-736.
[4]FARMER S R. Transcriptional control of adipocyte formation[J]. Cell Metab,2006,4(4):263-273.
[5]RAMJI D P,FOKA P. CCAAT/enhancer-binding proteins:structure,function and regulation[J]. Biochem J,2002,365(3):561-575.
[6]GERBENS F,VERBURG F J,VAN MOERKERK H T B,et al. Associations of heart and adipocyte fatty acidbinding protein gene expression with intramuscular fat content in pigs[J]. J Anim Sci,2001,79(2):347-354.
[7]SEALE P,CONROE H M,ESTALL J,et al. Prdm16 determines the thermogenic program of subcutaneous white adipose tissue in mice[J]. J Clin Ivest,2011,121(1):96-105.
[8]鞠大鹏,詹丽杏.脂肪细胞分化及其调控的研究进展[J].中国细胞生物学学报,2010,32(5):690-695.JU D P,ZHAN L X. Developments in regulation of adipocytes differentiation[J]. Chinese Journal of Cell Biology,2010,32(5):690-695.(in Chinese)
[9]YEAMAN S J. Hormone-sensitive lipase-a multipurpose enzyme in lipid metabolism[J]. BBA-Mol Cell Res,1990,1052(1):128-132.
[10]GUPTA R K,ARANY Z,SEALE P,et al. Transcriptional control of preadipocyte determination by Zfp423[J]. Nature,2010,464(7288):619-623.
[11]孙成娟,许厚强,赵佳福,等.从江香猪肌内和皮下脂肪前体细胞分化过程中相关基因的表达[J].农业生物技术学报,2017,25(12):1979-1988.SUN C J,XU H Q,ZHAO J F,et al. The expression of related genes on intramuscular and subcutaneous preadipocytes during differentiation in Congjiang Xiang pig(Sus scrofa)[J]. Journal of Agricultural Biotechnology,2017,25(12):1979-1988.(in Chinese)
[12]林森,林亚秋,朱江江,等. Wnt10b对山羊前体脂肪细胞分化相关基因表达的影响[J].畜牧兽医学报,2018,49(4):685-692.LIN S,LIN Y Q,ZHU J J,et al. Effect of Wnt10b on the expression of precursor adipocytes differentiation related genes in goat[J]. Acta Veterinaria et Zootechnica Sinica,2018,49(4):685-692.(in Chinese)
[13]RUIZ-OJEDA F J,RUPéREZ A I,GOMEZ-LLORENTE C,et al. Cell models and their application for studying adipogenic differentiation in relation to obesity:a review[J]. Int J Mol Sci,2016,17(7):1040.
[14]CORDANI N,PISA V,POZZI L,et al. Nitric oxide controls fat deposition in dystrophic skeletal muscle by regulating fibro-adipogenic precursor differentiation[J]. Stem Cells,2014,32(4),874-885.
[15]赵存真,李黄琨,马云,等.腺病毒介导的MAT2B过表达促进猪肌内脂肪细胞分化[J].畜牧兽医学报,2017,48(5):802-809.ZHAO C Z,LI H K,MA Y,et al. MAT2B overexprssion mediated by adenovirus promoting porcine intramuscular preadipocyte differentiation[J]. Acta Veterinaria et Zootechnica Sinica,2017,48(5):802-809.(in Chinese)
[16]张萌萌,魏胜娟,王艺如,等.牛肌内前体脂肪细胞的分离培养及分化相关基因的表达规律研究[J].畜牧与兽医,2018,50(5):1-6.ZHANG M M,WEI S J,WANG Y R,et al. Primary culture and expression of differentiation-related genes in bovine intramuscular preadipocytes[J]. Animal Husbandry&Veterinary Medicine,2018,50(5):1-6.(in Chinese)
[17]耿晓晖,富俊才,韩凯,等.绵羊肌内脂肪前体细胞的体外培养及分选[J].中国农业大学学报,2014,19(5):110-115.GENG X H,FU J C,HAN K,et al. In vitro culture and purification of ovine intramuscular preadipocyte[J].Journal of China Agricultural University,2014,19(5):110-115.(in Chinese)
[18]屈长青,张国华,陈粉粉,等.猪前体脂肪细胞的原代培养[J].农业生物技术学报,2005,13(5):649-653.QU C Q,ZHANG G H,CHEN F F,et al. Primary culture of porcine preadipocyte[J]. Journal of Agricultural Biotechnology,2005,13(5):649-653.(in Chinese)
[19]杜琛,付绍印,韩志玲,等.绒山羊肌内前体脂肪细胞的基因表达分析[J].畜牧兽医学报,2013,44(10):1532-1538.DU C,FU S Y,HAN Z L,et al. Gene expression analysis on intramuscular preadipocytes in the cashmere goats[J]. Acta Veterinaria et Zootechnica Sinica,2013,44(10):1532-1538.(in Chinese)
[20]陈妍,张国华,陆会宁,等.八眉猪肌内前体脂肪细胞的分离培养及诱导分化[J].广东农业科学,2014,41(13):110-113,123.CHEN Y,ZHANG G H,LU H N,et al. Culture and induced differentiation of intramuscular preadipocytes from Bamei pigs in vitro[J]. Guangdong Agricultural Sciences,2014,41(13):110-113,123.(in Chinese)
[21]SATOH A,STEIN L,IMAI S. The role of mammalian sirtuins in the regulation of metabolism,aging,and longevity[M]//Histone Deacetylases:the Biology and Clinical Implication. Springer Berlin,Heidelberg,2011:125-162.
[22]LEFTEROVA M I,ZHANG Y,STEGER D J,et al.PPARγand C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale[J]. Gene Dev,2008,22(21):2941-2952.
[23]LEE J E,GE K. Transcriptional and epigenetic regulation of PPARγexpression during adipogenesis[J]. Cell Biosci,2014,4(1):29.
[24]马晶晶,章涛. PPARγ功能与疾病关系研究进展[J].中国药理学通报,2012,28(5):601-604.MA J J,ZHANG T. Function of PPARγand its relationship with diseases:a present review[J]. Chinese Pharmacological Bulletin,2012,28(5):601-604.(in Chinese)
[25]NOH J R,KIM Y H,HWANG J H,et al. Scoparone inhibits adipocyte differentiation through down-regulation of peroxisome proliferators-activated receptorγin 3T3-L1 preadipocytes[J]. Food Chem,2013,141(2):723-730.
[26]ROSEN E D,MACDOUGALD O A. Adipocyte differentiation from the inside out[J]. Nat Rev Mol Cell Biol,2006,7(12):885-896.
[27]TANAKA T,YOSHIDA N,KISHIMOTO T,et al. Defective adipocyte differentiation in mice lacking the C/EBPβand or C/EBPδgene[J]. EMBO J,1997,16(24):7432-7443.
[28]NTAMBI J M,YOUNG-CHEUL K. Adipocyte differentiation and gene expression[J]. J Nutr,2000,130(12):3122S-3126S.
[29]KIM J B,SPIEGELMAN B M. ADD1/SREBP1 promotes adipocyte differentiation and gene expression linked to fatty acid metabolism[J]. Gene Dev,1996,10(9):1096-1107.
[30]YELLATURU C R,DENG X,PARK E A,et al. Insulin enhances the biogenesis of nuclear sterol regulatory element-binding protein(SREBP)-1c by posttranscriptional down-regulation of Insig-2A and its dissociation from SREBP cleavage-activating protein(SCAP)·SREBP-1c complex[J]. J Biol Chem,2009,284(46):31726-31734.
[31]SHIMOMURA I,SHIMANO H,KORN B S,et al. Nuclear sterol regulatory element-binding proteins activate genes responsible for the entire program of unsaturated fatty acid biosynthesis in transgenic mouse liver[J]. J Biol Chem,1998,273(52):35299-35306.
[32]TAPPY L,LK A. Metabolic effects of fructose and the worldwide increase in obesity[J]. Physiol Rev,2010,90(1):23-46.
[33]HARADA K,SHEN W J,PATEL S,et al. Resistance to high-fat diet-induced obesity and altered expression of adipose-specific genes in HSL-deficient mice[J]. Am J Physiol-Endoc M,2003,285(6):E1182-E1195.
[34]XU A M,WANG Y,XU J Y,et al. Adipocyte fatty acid–binding protein is a plasma biomarker closely associated with obesity and metabolic syndrome[J]. Clin Chem,2006,52(3):405-413.
[35]BERNLOHR D A,BOLANOWSKI M A,KELLY T J Jr,et al. Evidence for an increase in transcription of specific m RNAs during differentiation of 3T3-L1 preadipocytes[J]. J Biol Chem,1985,260(9):5563-5567.
[36]CHRISTY R J,YANG V W,NTAMBI J M,et al. Differentiation-induced gene expression in 3T3-L1 preadipocytes:CCAAT/enhancer binding protein interacts with and activates the promoters of two adipocyte-specific genes[J]. Gene Dev,1989,3(9):1323-1335.
[37]ZHAO C P,TIAN F,YU Y,et al. Muscle transcriptomic analyses in Angus cattle with divergent tenderness[J].Mol Biol Rep,2012,39(4):4185-4193.
[38]WEI S J,ZHANG L F,ZHOU X,et al. Emerging roles of zinc finger proteins in regulating adipogenesis[J]. Cell Mol Life Sci,2013,70(23):4569-4584.
[39]SEALE P,KAJIMURA S,YANG W L,et al. Transcriptional control of brown fat determination by PRDM16[J]. Cell Metab,2007,6(1):38-54.
[40]SEALE P,BJORK B,YANG W L,et al. PRDM16 controls a brown fat/skeletal muscle switch[J]. Nature,2008,454(7207):961-967.
[41]KAJIMURA S,SEALE P,TOMARU T,et al. Regulation of the brown and white fat gene programs through a PRDM16/Ct BP transcriptional complex[J]. Gene Dev,2008,22(10):1397-1409.