巨噬细胞abca1对ox-ldl诱导的炎症因子 的调节及其意义

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1、巨噬细胞ABCA1对Ox-LDL诱导的炎症因子的调节及其意义作者：郭志刚，吴平生，李建华，赖文岩【摘要】　　【目的】研究在氧化低密度脂蛋白（Ox-LDL）诱导作用下，ATP结合盒转运子A1（ABCA1）对巨噬细胞中细胞间黏附分子－1（ICAM-1）、单核细胞化学趋向蛋白－1（MCP-1）mRNA和蛋白质及白介素－1β（IL-1β）蛋白质表达的影响，在细胞水平证实ABCA1对动脉粥样硬化的影响。【方法】用氟波酯(PMA)刺激THP-1细胞使之转变为巨噬细胞，Ox-LDL(30?滋g/mL)刺激3、6、12、24h后，以荧光定量RT－PCR和Wes

2、tern蛋白印迹法及酶联免疫吸附法（ELISA）检测ABCA1、ICAM-1、MCP-1及IL-1βmRNA和蛋白质表达量；用反义寡核苷酸（100nmol/L）抑制ABCA1的表达，观察Ox-LDL刺激下上述指标的改变。【结果】给予Ox-LDL刺激后，巨噬细胞ABCA1、ICAM-1、MCP-1的mRNA和蛋白及IL-1β蛋白质表达均增高；给予反义寡核苷酸转染后Ox-LDL刺激3、6h，上述指标的mRNA表达降低(P<0.01)，12、24h蛋白质表达降低(P<0.01)。【结论】在巨噬细胞，ABCA1可增加Ox-LDL诱导的炎症因

3、子表达，参与动脉粥样硬化的发生。【关键词】ATP结合盒转运子A1；氧化低密度脂蛋白；巨噬细胞；动脉粥样硬化；炎症因子14Abstract:【Objective】ToevaluatetheeffectsofATPbindingcassettetransporterA1(ABCA1)onmRNAandproteinexpressionofintercellularadhesionmolecule-1(ICAM-1)andmonocytechemoattractantprotein-1(MCP-1)andproteinexpressionofinte

4、rleukin-1β(IL-1β)inTHP-1macrophageinducedbyoxidizedlowdensitylipoprotein(Ox-LDL)inordertoinvestigatethemechanismthatABCA1contributestoatherosclerogenesis.【Methods】MonocyticTHP-1cellswereculturedwith100nmol/Lphorbolmyristateacetate(PMA)for72hourstoleadcellsintoTHP-1macrophage

5、.Ox-LDL(30?滋g/mL)wasaddedintoculturemediaandTHP-1macrophagecellswereharvestedat3,6,12,and24hours,respectively.ThemRNAandproteinlevelsofABCA1,ICAM-1,MCP-1andIL-1βwereinvestigatedbyreal-timefluorescentquantitativeRT-PCR,Westernblot,andELISAmethods.Afterphosphorothioateoligonuc

6、leotidesofABCA1mixturewereaddtoculturemediaatfinalconcentrationof100nmol/L,thesameexperimentswererepeated.【Results】ThemRNAandproteinamountsofABCA1,ICAM-1,andMCP-1andalsoproteinamountofIL-1βwereincreasedaftermacrophageincubatedwithOx-LDL.Transfectedwithantisenseoligonucleotid

7、esofABCA1,theexpressivelevels14ofmRNAweredecreasedat3and6hours(P<0.01),andproteinat12and24hours(P<0.01).【Conclusion】ABCA1couldincreasetheexpressionofinflammatorycytokinesinmacrophageinducedbyOx-LDLandcontributedtoatherosclerogenesis.Keyword:ATP-bindingcassetteA1(ABCA1)

8、;oxidizedlowdensitylipoprotein;THP-1macrophage;atherosclerosis;inflammatory