高糖诱导山羊骨骼肌卫星细胞成脂分化过程中相关基因表达的变化

doi:10.11843/j.issn.0366-6964.2014.05.005

摘要/Abstract

摘要：

本研究采用高糖培养基诱导山羊骨骼肌卫星细胞（Skeletal muscle satellite cells，SMSCs）成脂分化，检测诱导分化过程中脂肪酸合成及成脂分化相关基因表达量，为进一步揭示SMSCs的成脂分化机理提供依据。采用含25 mmol•L^-1葡萄糖的F12培养基诱导山羊SMSCs成脂分化，油红O染色观察脂肪滴形成，qPCR检测诱导前（0 d）和分别在高糖、低糖培养基中培养2 、4 、6 、8 、10 d时脂肪酸合成及成脂分化相关基因ACC、FAS、C/EBPα、PPARγ、SREBP-1、AdipoQ mRNA的相对表达量。结果表明：油红O染色观察到山羊SMSCs高糖培养6 d即形成较多脂肪滴，而低糖培养至10 d也无明显脂肪滴形成。ACC、C/EBPα表达量在高糖培养2 d后显著升高并维持较高值（P<0.05）；SREBP-1、FAS表达量在高糖培养6 d后显著升高（P<0.05）；PPARγ表达量在高糖培养10 d后显著升高（P<0.05）；AdipoQ表达量在高糖培养2、8、10 d时显著升高（P<0.05）。本试验发现高浓度葡萄糖能直接或间接上调脂肪酸合成及成脂肪分化相关基因的表达，促进山羊SMSCs向类脂肪细胞转化。

Abstract:

This study aimed to induce skeletal muscle satellite cells (SMSCs) to differentiate into adipocytes by using high concentration glucose，and detect the relative expressions of adipogenic related and adipogenic differentiation related genes，and lay a foundation for futher studying the mechanism of SMSCs adipogenic differentiation.High concentration glucose (25 mmol•L^-1) F12 culture medium was used for inducing，and cells were stained with Oil Red O，qPCR was employed to detect the relative expressions of ACC，FAS，C/EBPα，PPARγ，SREBP-1 and AdipoQ at 0 d and five time points (2，4，6，8 and 10 d) under the condition of high and low concentration glucose.The results showed that lots of lipid droplets were observed in cells induced by high concentration glucose at the 6 d.In high concentration glucose group，the mRNA expressions of ACC and C/EBPα significantly increased at the 2 d (P<0.05) and maintained relative high expression levels at later stages，and the expressions of SREBP-1 and FAS were both significantly up-regulated at the 6 d (P<0.05)，while the expression of PPARγ significantly increased at the 10 d (P<0.05).The expression of AdipoQ significantly ascended at the 2，8 and 10 d (P<0.05).In this study，we found that high concentration glucose up-regulated the expressions of the adipogenic-related genes and promoted the SMSCs differentiating into adipocyte-like cells.

中图分类号:

S827
S813.3

薛科，王林杰，陈利，王薏琳，仲涛，李利，张红平. 高糖诱导山羊骨骼肌卫星细胞成脂分化过程中相关基因表达的变化[J]. 畜牧兽医学报, 2014, 45(5): 706-713.

XUE Ke， WANG Lin-jie, CHEN Li， WANG Yi-lin， ZHONG Tao， LI Li， ZHANG Hong-ping. Adipogenic-related Gene Expressions in Goat Skeletal Muscle Satellite Cells Treated with High Concentration Glucose[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(5): 706-713.

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https://www.xmsyxb.com/CN/Y2014/V45/I5/706

参考文献

［1］JANG Y C，SINHA M，CERLETTI M，et al.Skeletal muscle stem cells: effects of aging and metabolism on muscle regenerative function ［J］.Cold Spring Harb Symp Quant Biol，2011，76：101-111.
［2］MAURO A.Satellite cell of skeletal muscle fibers ［J］.J Biophys Biochem Cytol，1961，9：493-495.
［3］WOZNIAK A C，KONG J，BOCK E，et al.Signaling satellite-cell activation in skeletal muscle: markers，models，stretch，and potential alternate pathways ［J］.Muscle Nerve，2005，31：283-300.
［4］SIEGEL A L，KUHLMANN P K，COMELISON D D.Muscle satellite cell proliferation and association: new insights from myofiber time-lapse imaging ［J］.Skelet Muscle，2011，1（1）：7.
［5］SAITO Y，NONAKA I.Initiation of satellite cell replication in bupivacaine-induced myonecrosis ［J］.Acta Neuropathol，1994，88：252-257.
［6］KATAGIRI T，YAMAGUCHI A，KOMAKI M，et al.Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage ［J］.J Cell Biol，1994，127：1755-1766.
［7］NISHIMURA R，KATO Y，CHEN D，et al.Smad5 and DPC4 are key molecules in mediating BMP-2-induced osteoblastic differentiation of the pluripotent mesenchymal precursor cell line C2C12 ［J］.J Biol Chem，1998，273：1872-1879.
［8］LEE E J，KAMLI M R，BHAT A R，et al.Effect of porcine placenta steroid extract on myogenic satellite cell proliferation，transdifferentiation，and lipid accumulation ［J］.In vitro Cell Dev Biol Anim，2012，48：326-333.
［9］BELOOR J，KANG H K，MOON Y S.Serum lipids can convert bovine myogenic satellite cells to adipocytes ［J］.Asian Aust J Anim Sci，2010，23：1519-1526.
［10］WU H，REN Y，LI S，et al.In vitro culture and induced differentiation of sheep skeletal muscle satellite cells ［J］.Cell Biol Int，2012，36：579-587.
［11］TEBOUL L，GAILLARD D，STACCINI L，et al.Thiazolidinediones and fatty acids convert myogenic cells into adipose-like cells ［J］.J Biol Chem，1995，270：28183-28187.
［12］GUILLET-DENIAU I，PICHARD A L，KONE A，et al.Glucose induces de novo lipogenesis in rat muscle satellite cells through a sterol-regulatory-element-binding-protein-1c-dependent pathway ［J］.J Cell Sci，2004，117：1937-1944.
［13］YUE T，YIN J，LI F，et al.High glucose induces differentiation and adipogenesis in porcine muscle satellite cells via mTOR ［J］.BMB Rep，2010，43：140-145.
［14］LOPEZ-CASILLAS F，BAI D H，LUO X C，et al.Structure of the coding sequence and primary amino acid sequence of acetyl-coenzyme A carboxylase ［J］.Proc Natl Acad Sci U S A，1988，85：5784-5788.
［15］CHAKRAVARTY B，GU Z，CHIRALA S S，et al.Human fatty acid synthase: structure and substrate selectivity of the thioesterase domain ［J］.Proc Natl Acad Sci U S A，2004，101：15567-15572
［16］MULLER C，ALUNNI-FABBRONI M，KOWENZ-LEUTZ E，et al.Separation of C/EBPalpha-mediated proliferation arrest and differentiation pathways ［J］.Proc Natl Acad Sci U S A，1999，96：7276-7281.
［17］WANG H，MAECHLER P，ANTINOZZI P A，et al.The transcription factor SREBP-1c is instrumental in the development of beta-cell dysfunction ［J］.J Biol Chem，2003，278：16622-16629.
［18］BUGGE A，MANDRUP S.Molecular mechanisms and genome-wide aspects of PPAR subtype specific transactivation ［J］.PPAR Res，2010，2010：1-12.
［19］HUANG Z G，XIONG L，LIU Z S，et al.The development changes and effect on IMF content of H-FABP and PPAR mRNA expression in sheep muscle ［J］.Acta Genetica Sinica，2006，33：507-514.
［20］RODEN M，PRICE T B，PERSEGHIN G，et al.Mechanism of free fatty acid-induced insulin resistance in humans ［J］.J Clin Invest，1996，97：2859-2865.
［21］BLACHNIO-ZABIELSKA A U，BARANOWSKI M，HIRNLE T，et al.Increased bioactive lipids content in human subcutaneous and epicardial fat tissue correlates with insulin resistance ［J］.Lipids，2012，47：1131-1141.
［22］LIU S，WANG L，WANG N，et al.Oleate induces transdifferentiation of chicken fibroblasts into adipocyte-like cells ［J］.Comp Biochem Physiol A Mol Integr Physiol，2009，154：135-141.
［23］MATSUBARA Y，SATO K，ISHII H，et al.Changes in mRNA expression of regulatory factors involved in adipocyte differentiation during fatty acid induced adipogenesis in chicken ［J］.Comp Biochem Physiol A Mol Integr Physiol，2005，141：108-115.
［24］LONG Y C，ZIERATH J R.AMP-activated protein kinase signaling in metabolic regulation ［J］.J Clin Invest，2006，116：1776-1783.
［25］MENG X，YANG J M，CAO Y J，et al.Increasing fatty acid production in E.coli by simulating the lipid accumulation of oleaginous microorganisms ［J］.J Ind Microbiol Biotechnol，2011，38：919-925.
［26］KIM C W，MOON Y A，PARK S W，et al.Induced polymerization of mammalian acetyl-CoA carboxylase by MIG12 provides a tertiary level of regulation of fatty acid synthesis ［J］.Proc Natl Acad Sci USA，2010，107：9626-9631.
［27］SMITH S，WITKOWSKI A，JOSHI A K.Structural and functional organization of the animal fatty acid synthase ［J］.Prog Lipid Res，2003，42：289-317.
［28］HUANG G C，ZHANG J S，TANG Q Q.Involvement of C/EBP-alpha gene in in vitro activation of rat hepatic stellate cells ［J］.Biochem Biophys Res Commun，2004，324：1309-1318.
［29］RODEN M，PRICE T B，PERSEGHIN G，et al.Mechanism of free fatty acid-induced insulin resistance in humans ［J］.J Clin Invest，1996，97：2859-2865.
［30］XU H，LUO J，LI F，et al.Effects of SREBP-1 over-expression on fatty acid metabolism related genes expression in goats ［J］.Chinese Journal of Biotechnology，2012，28：1306-1316.
［31］FARMER S R.Transcriptional control of adipocyte formation ［J］.Cell Metab，2006，4：263-273.
［32］TONTONOZ P，HU E，SPIEGELMAN B M.Stimulation of adipogenesis in fibroblasts by PPAR gamma 2，a lipid-activated transcription factor ［J］.Cell，1994，79：1147-1156.
［33］ZIEMKE F，MANTZOROS C S.Adiponectin in insulin resistance: lessons from translational research ［J］.Am J Clin Nutr，2010，91：258S-261S.

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