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1、糖基化终产物对THP-1巨噬细胞酰基辅酶A:胆固醇酰基转移酶-1表达的影响作者：杨启红,徐强,张红,司良毅单位：1第三军医大学西南医院老年病科,重庆400038;2解放军第113医院老年病科,浙江宁波 摘 要:目的 观察糖基化终产物(advancedglycosylationend-products,AGEs)对THP-1巨噬细胞酰基辅酶A:胆固醇酰基转移酶-1(acylcoenzymeA:cholesterolacyltransferase-1,ACAT-1)表达的影响。方法 将糖基化-牛血清白蛋白(AGE-modifiedbovineserumalbumin,
2、AGE-BSA)与佛波酯(PMA)诱导分化72h的THP-1巨噬细胞共同孵育,运用RT-PCR和Westernblot分别检测巨噬细胞ACAT-1mRNA和蛋白的表达水平。结果 PMA诱导72h后,THP-1细胞停止增殖,由单核细胞分化成为巨噬细胞。用50、100、200和400mg/LAGE-BSA处理巨噬细胞后,细胞ACAT-1mRNA相对表达量逐渐增加;ACAT-1蛋白表达的平均积分光密度值也逐渐增加,二者较对照牛血清白蛋白组明显增加(P<0.05)。用200mg/LAGE-BSA作用巨噬细胞0、12、24、36h和48h后,细胞ACAT-1mRNA相对
3、表达量逐渐升;ACAT-1蛋白表达的平均积分光密度值也逐渐升高,较0h明显增加(P<0.05)。结论 AGEs可上调THP-1巨噬细胞ACAT-1mRNA和蛋白的表达,呈浓度和时间依赖性,这可能与糖尿病动脉粥样硬化有关。 关键词:糖尿病;糖基化终产物;酰基辅酶A:胆固醇酰基转移酶-1;基因表达 中图法分类号:R392.12;R392.6;R587.102 文献标识码:AEffectsofadvancedglycationend-productsonexpressionofacylcoenzymeA:cho-lesterolacyltransferase
4、-1inTHP-1macrophagesYANGQi-hong1,XUQiang1,ZHANGHong2,SILiang-yi1(1DepartmentofGeriatrics,SouthwestHospital,ThirdMilitaryMedi-calUniversity,Chongqing400038;2DepartmentofGeriatrics,No.113HospitalofPLA,NingboZhejiang315021,China) Abstract:Objective Toinvestigatetheeffectsofadvancedglycos
5、ylationend-products(AGEs)ontheexpressionofacylcoenzymeA:cholesterolacyltransferase-1(ACAT-1)inculturedTHP-1macrophages.Methods THP-1cellsweredifferentiatedintomacrophagesafterculturedinRPMI1640mediacontaining0.1μmol/LPMAfor72h.THP-1macrophageswerethenexposedtoAGE-modifiedbovineserum
6、albumin(AGE-BSA,atconcentrationof50,100,200mg/L)for24horto200mg/LAGE-BSAfor0,12,24,36h.Ex-pressionofACAT-1mRNAandproteininTHP-1macrophageswasmeasuredbyRT-PCRandWesternblot,re-pectively.Results Afterinducedby0.1μmol/LPMA,THP-1cellsstoppedproliferationanddifferentiatedintomacrophages.
7、TreatmentofTHP-1macrophageswithAGE-BSAresultedinanincreaseinthemRNAandproteinlevelsforACAT-1inadose-andtime-dependentmanner.Conclusion AGEsupregulatetheexpressionlevelsofmRNAandproteinforACAT-1inculturedTHP-1macrophages,whichmightbepartlyinvolvedintheatherogenesisindiabeticpatients.
