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1、ISSN100727626中国生物化学与分子生物学报2003年4月CN1123870PQChineseJournalofBiochemistryandMolecularBiology19(2):250～255血清饥饿可诱导人血管平滑肌细胞再分化3韩 梅, 温进坤, 郑 斌, 聂 磊(河北医科大学基础医学研究所,河北省医学生物技术重点实验室,石家庄 050017)摘要 体外培养的分化型血管平滑肌细胞(vascularsmoothmusclecells,VSMC)以特异性标志基因表达、长梭形外观及对兴奋剂刺激产生收缩反应为其表型特征.以
2、血清饥饿法培养处于超汇合(overconfluence)状态的人VSMC,观察其分化型标志基因表达活性及其与细胞形态特征和收缩反应性之间的关系,探讨细胞生存环境对VSMC基因表达及表型的影响.研究显示,生长至超汇合的VSMC由含血清培养转为血清饥饿后,收缩蛋白如SMα肌动蛋白(SMα2actin)、SM22α、h12calponin、肌球蛋白重链(MHC)SM1和SM2亚型的表达活性明显上调,证实血清饥饿诱导的收缩蛋白基因表达和血清应答因子(serumresponsefactor,SRF)与CArG顺式元件结合活性的增强有关.同时,
3、血清饥饿还可激活参与VSMC分化调节的转录调控因子SmLIM、Gax和分化相关蛋白HRG21基因的转录.随着血清饥饿培养时间的延长,VSMC逐渐形成多层、束状、成极性排列的形式,对兴奋剂刺激产生的收缩反应明显增强.结果表明,超汇合状态的去分化型VSMC脱离血清刺激后,可以再分化成熟并重新获得收缩能力.关键词 血管平滑肌细胞,再分化,标志基因,收缩,细胞形态+中图分类号 R329128RedifferentiationofCulturedHumanVascularSmoothMuscleCellsafterSerumDeprivati
4、on3HANMei,WENJin2kun,ZHENGBin,NIELei(InstituteofBasicMedicine,HebeiMedicalUniversity,HebeiLaboratoryofMedicalBiotechnology,Shijiazhuang050017,China)AbstractDifferentiatedvascularsmoothmusclecells(VSMC)arecharacterizedasexpressionofspecificmarkergenes,elongatedandspindl
5、e2shapedmorphologyandtheabilitytocontractinresponsetoagoniststimulation.TodeterminewhetherhumanVSMCphenotypecouldrevertfromdedifferentiationtodifferentiation,overconfluentVSMCwasincubatedinserum2freeM199,andexpressionofdifferentiationmarkergeneswasdetectedbyRT2PCR,Nort
6、hernblottingandWesternblotting.Therelationshipsbetweenattainmentofacontractilephenotypeandexpressionofdifferentiationmarkergeneswereinvestigated.TheresultsindicatedthatoverconfluentVSMCafterserumwithdrawalsequentiallyexpressedspecificcontractileproteins,includingSMα2ac
7、tin,SM2MHCisoformsSM1andSM2,h12calponinandSM22α.Duringprolongedperiodsof“starvation”,theincreasesintheexpressionofmarkergeneswereassociatedwithelevationofSRFbindingtoCArGcis2elements.Onserumwithdrawal,VSMCexpressedthetranscriptionfactorsthatarenecessaryforregulatingVSM
8、Cdifferentiation,suchasSmLIM,GaxandHRG21,whichwererestrictedindifferentiatedVSMCexpressionandundetectablebeforeserumw
