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1、at-RA抑制大鼠骨髓間充質干細胞成脂分化張蔓,舒秀蘭,李文迪,劉晶晶,陳潔,劉友學(400014重慶,重慶醫(yī)科大學附屬兒童醫(yī)院兒童發(fā)育疾病研究省部共建教育部重點實驗室,兒科學重慶市重點實驗室,重慶市(兒童發(fā)育重大疾病診治與預防)國際科技合作基地)[摘要]目的探討高濃度全反式視黃酸(all-transretinoicacid,at-RA)抑制大鼠骨髓間充質干細胞(ratbonemarrowmesenchymalstemcells,rBMSCs)向脂肪細胞分化的機制。方法體外分離、培養(yǎng)和誘導rBMSCs;在成脂誘導液中加入0.1、
2、0.5、1.0、5.0?mol/L不同濃度(0.1~5.0?mol/L)的at-RA,光學顯微鏡下觀察細胞形態(tài)及油紅O染色變化;油紅O染色提取法分析不同濃度at-RA對BMSCs成脂分化的影響;real-timePCR測定細胞視黃酸受體(RAR?、RAR?)、CCAAT增強子結合蛋白?(CEBP?)及過氧化物酶體增殖激活受體?2(PPAR?2)mRNA水平的變化。結果細胞形態(tài)及油紅O染色觀察發(fā)現(xiàn),在0.1、0.5、1.0、5.00.1~5.0?mol/Lat-RA作用下rBMSCs向脂肪細胞分化受到抑制,且呈濃度依賴性;油紅O染
3、色提取法半定量測定結果顯示,隨著at-RA濃度升高,分化細胞內(nèi)脂肪含量也隨著降低(P<0.01);real-timePCR檢測顯示與血清組(未加at-RA)相比,加入at-RA后,rBMSCs的CEBP?和PPAR?2表達顯著降低(P<0.05),RAR?表達無顯著差異,但RAR?表達顯著增加(P<0.01)。結論0.1、0.5、1.0、5.00.1~5.0?mol/Lat-RA可抑制rBMSCs向脂肪細胞分化,其機制可能與RAR?的上調及CEBP?、PPAR?2的下調有關。[關鍵詞]全反式視黃酸;骨髓間充質干細胞;成脂分化;視
4、黃酸受體[中圖法分類號][文獻標志碼]Aat-RAinhibitsitsadipogenicdifferentiationofratbonemarrowmesenchymalstemcellsZhangMan,ShuXiulan,LiWendi,LiuJingjing,ChenJie,LiuYouxue(MinistryofEducationKeyLaboratoryofChildDevelopmentandDisorders,KeyLaboratoryofPediatricsinChongqing,ChongqingInter
5、nationalScienceandTechnologyCooperatioCenterforChildDevelopmentandDisorder,Children’sHospitalofChongqingMedicalUniversity,Chongqing,400014,China)[Abstract]ObjectiveToinvestigatethemechanismofall-transretinoicacid(at-RA)toinhibittheadipogenicdifferentiationofratbonema
6、rrowmesenchymalstemcells(rBMSCs).MethodsRatBMSCswereisolated,incubatedandinducedtodifferentiateintoadipocytes.At-RAathighconcentrations(0.1?5.0?mol/L)wasaddedtotheadipogenicinducedmedium.ThemorphologicchangesofrBMSCsduringdifferentiationwereobservedundermicroscope.Th
7、eeffectofat-RAontheadipogenicdifferentiationwasanalyzedbyoilredOstaining.Theexpressionofretinoicacidreceptors?and??RAR?,RAR??????CCAAT/enhancerbindingprotein??(CEBP?)andperoxisomeproliferators-activatedreceptor?2?PPAR?2??wasanalyzedbyreal-timequantitativepolymerasech
8、ainreaction(real-timePCR).ResultsAttheconcentrationof0.1?5.0?mol/L,at-RAinhibitedrBMSCstodifferentiateintoadipocytes.Andtheeffectwa