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1、生物工程学报ChinJBiotech2009,May25;25(5):739-744journals.im.ac.cnChineseJournalofBiotechnologyISSN1000-3061cjb@im.ac.cn©2009InstituteofMicrobiology,CAS&CSM,Allrightsreserved组织工程与细胞培养钙信号调节小鼠前体脂肪细胞分化和脂质蓄积的机制112王丽,孙超,康靖全1西北农林科技大学动物科技学院,杨凌7121002西北农林科技大学生命科学学院,杨凌712100摘要:本试验用醋酸钙、p38丝裂原激活蛋白激酶(p38MAPK)
2、抑制剂SB203580及钙通道阻滞剂和激动剂刺激小鼠前体脂肪细胞。通过实时定量PCR技术检测前体脂肪细胞分化标志基因和钙信号相关受体基因表达水平,用油红O染色2+2+提取法和Fura-2/AM荧光法测定胞内脂质蓄积情况及胞浆游离Ca浓度([Ca]i)变化,以探讨钙信号调节前体脂肪细胞分化的潜在机制。结果表明：钙通道阻滞剂和激动剂显著改变了脂蛋白脂酶(LPL),过氧化物增殖激活受体γ(PPARγ)、脂肪酸合成酶(FAS)的表达水平，且影响细胞内的脂质蓄积。与降低外钙摄入相比，降低内钙释放能促进前体脂肪细胞分化(P<0.01)，而提高外钙摄入与提高内钙释放相比，提高外钙摄入显著抑
3、制前体脂肪细胞分化(P<0.01)。2+SB203580可降低胞浆[Ca]i浓度，促进前体细胞分化和脂质蓄积(P<0.01)。但钙信号并未影响维生素D受体(VDR)和细胞外钙敏感受体(CaSR)的表达水平。提示钙信号可能通过p38MAPK通路影响前体脂肪细胞分化和脂质蓄积。关键词:钙信号,p38MAPK,小鼠,前体脂肪细胞分化,脂质蓄积Themechanismofcalciumsignalregulatepreadipocytedifferentiationandlipidaccumulationinmice112LiWang,ChaoSun,andJingquanKang1
4、CollegeofAnimalScience&Technology,NorthwestAgricultureandForestryUniversity,Yangling712100,China2CollegeofLifeSciences,NorthwestAgricultureandForestryUniversity,Yangling712100,ChinaAbstract:Westimulatedpreadipocyteofmicewithcalciumacetate,p38mitogen-activatedproteinkinase(p38MAPK)inhibitorS
5、B203580,theparalysorsandexcitomotorsofcalciumchannel.Thenwedetectedexpressionlevelofpreadipocytedifferentiation’s2+markergenesandcalciumsignalrelatedacceptorgenesbyreal-timePCR,anddeterminedintracellularfreeCaconcentration2+([Ca]i])withFura-2/AM,intracellularlipidaccumulationbyoilredOstaini
6、ng.Ouraimwastoinvestigatethepotentialmechanismbetweencalciumsignalandpreadipocytedifferentiation.Theresultsindicatedthattheparalysorsandexcitomotorsofcalciumchannelchangedtheexpressionleveloflipoproteinlipase(LPL),peroxisomeproliferators-activatedreceptorgamma(PPARγ),fatty2+acidsynthetase(F
7、AS),andthelipidaccumulation,markedly.ComparedwithexocellularCa’sdecrease,inhibitedintracellular2+2+Ca’sliberationcanpromotedpreadipocytedifferentiation(P<0.01),andcomparedwithintracellularCa’sincrease,promoted2+2+exocellularCa’singestinhibitedpreadipocyt
