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1、TNF-α影响胰岛素对原代培养大鼠脂肪细胞增殖及生脂基因的转录表达动物52(1):123～129,2006ActaZoologicaSinicaTNF.I]'影响胰岛素对原代培养大鼠脂肪细胞增殖及生脂基因的转录表达*卢建雄,2杨公社H陈粉粉1.西北农林科技大学动物脂肪沉积和肌肉发育实验室,陕西杨凌7121002.西北民族大学生命科学与工程学院,兰州730030摘要采用RT—PCR,MTT比色法和油红O染色提取法,分别测定外源TNF—a胰岛素对原代培养大鼠前体脂肪细胞增殖及脂肪生成和碳水化合物反应元件结合蛋白(ChREBP),固醇调控
2、元件结合蛋白(SREBP)一lc,脂肪酸合酶(FAs)及乙酰辅酶A羧化酶(ACC)1基因转录表达的影响.结果表明,TNF-a可有效抑制胰岛素对原代培养大鼠前体脂肪细胞增殖的促进作用,并通过抑制胰岛素对转录因子SREBP—lc和脂肪酸合酶FAS及ACC1基因转录表达的促进作用,减少脂肪酸的生物合成,抑制成熟脂肪细胞的脂肪生成,证明TNF-a是动物脂肪形成的有效抑制因子.此外,TNF-a下调ChREBPmRNA表达,但胰岛素在低糖条件下对其表达没有明显影响[动物52(1):123—129,2006].关键词肿瘤坏死因子一a胰岛素脂肪细胞
3、增殖脂肪生成转录表达TNF—ID'affectsproliferationandtranscriptionoflipogenic—relatedgenesstimulatedbyinsulininprimaryculturedratadipocytesLUJian—Xiong,YANGGong—She,CHENFen—Fen1.LaboratoryofAnimalFatDepositionandMuscleDevelopment,NorthwestSci—TechUniversityofAgricultureandForestry,
4、Yangling712100,Shaanxi,China2.CollegeofLifeScienceandEngineering,NorthwestMinoritiesUniversity,Lanzhou730030,ChinaAbstractTNF—aiSapleiotropiccytokinesecretedinadiposetissue.TostudytheeffectsofTNF-aonadipogenesisstimu—latedbyinsulininvitro,ratpreadipocyteswereisolatedby
5、collagenasetreatmentofadiposetissuefrommaleSDrats(20d-old),andculturedinmediawithTNF-a,insulinandTNF-a+insulin,respectively.PreadipocyteproliferationwasmeasureelwithMTT.Thelipogenesisandrelativelevelsoflipogenic—relatedgenemRNAsinmatureadipocytesweremea—suredthroughoil
6、Red0stainingandsemiquantitativeRT—PCR.respectively.TheresultsshowedthatTNF—aeffectivelyinhibitedpreadipocyteproliferationstimulatedbyinsulinanddecreasedlipogenesisthroughinhibitingstimulationofinsulinontranscriptionalexpressionsoflipogenicgenessuchasacetyl—CoAcarboxyla
7、sea(ACC1),fattyacidsynthase(FAS),sterolregulatoryelementbindingprotein(SREBP)一lcinadipocytes,suggestingthatTNF-aisaeffectiveinhibitorofadipogenesisinadiposetissue.Inaddition,mRNAexpressionofthecarbohydrateresponseelementbindingprotein(ChREBP)wassignificantlyinhibitedby
8、TNF-a,whileitwasnotinfluencedbyinsulininadipocytesin1oWglucosemedia【ActaZoologicaSinica52(1):123—129,20061.KeywordsTN
