血管内皮细胞对平滑肌细胞表型转化的影响

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1、万方数据ResearchArtides78基础研究n管内皮鱼车晓聪刘水潘妻凰是国§【捕2l目的分析共培养时太鼠血管内皮细胞(VECs)对大鼠血管平滑肌细胞(vSMCs)表型转化的影响．方法VSMCs直接种植于培养板表面，VECs删种植于与VSMCs相对的浮胶底面。免疫荧光方挂观察和鍪窟VECs和VSMCs，RT-PCR和共聚焦E傲镜分析丧型相关#园的表达。结果搋代培养的大鼠VECs呈铺路石样形志．vwF巍色阳性。原代培养的大鼠VSMCs免疫最光垡色d-SMA呈阳性。RT-PCR椅测结果表明．48h共培养组VSMCs的合成表型相

2、关基因CRBP-l、sm哪b的表达水平显著高干单独培葬组．分别为I4倍，l5倍-72h选到峰值．分别为I7倍、2I倍．96h开始下降，搀培养组中收缩表!标记物Smoothelfa—B和SM-MHC的基园表达木平在48h、72h＆著低于单拙培养组．96hSm∞thelin—B却高于单独培养维。单抽培养组上进备g因的变化趋势T变或保持稳定。免癌荧光结果显示SM—MHC*白表达在共培养组中96h后从下降转为升高(P<005)。结论在共培养体系中，血管内皮细胞对缸管平滑肌细胞表型转化的作用表现为先促进向合成型转化，96h后促进向性缩型

3、转化。!*望i加表型-细胞共培养一血管内皮细胞t血管平滑肌细胞t基目表达Eff*lofVaKularEndotheliAlCeosoBne呻帅icTrlnsifionofSmithMn"leCellsLiXiao-cong’,￡ⅢShuiiPanLuan女ng’WuUuo—qa蝌ILaboratoryofMoleculaBiologY．ShanghaiMedicalCollegelucianUniversimShanghai,2000MChina；。D口一“fof№^ⅫⅡandEngin∞Hng,Fndan¨一1口Shangh

4、ai,200433．ChinaE^-’x”a“，Obj∞dveToobesetheinflue【tccofv雠u[arendothelialcetls(VECs)oRphenolypealtemad⋯ofscu】Ⅱsmithmusclecells(VSMCs)MethedsVECsandVSMCsweRisolatedfromthoracica0衄ofmt—gcoll89咖∞digestion∞dtissuesectJon“1m力7％∞slmetivelyAsinglelayerofVSMCswm“lmredoBthedown

5、sideoffl∞tinggelofTypeIcollag∞flora㈣tailsdfacedasinglelayerofVECsattachingoNthesuff嘲ofcellcultureplateThecrosstalkbetweenVECs曲dVSMCsw％allowedIhrou曲cul呲mediaVSMCs如dVECswemidentifiedbyimmBBofluoTcscentstainingTheCXpTgSSIOOSofSM—MHCSmoothelin-BSmembandCRBP-1byVSMCseult

6、urcdonthefloatinggel⋯analyzedbyRT-PCRandconfocalm⋯⋯‰uIbVECsshowedatypicalcobblestoneⅢorphologyandwc∞positiveforvWFstainingVSMCswemconfi皿edbyd—SMAcxp”sslonInthefloatinggel⋯u】mKSyStem．theexpTessionofCRBP-1锄dSmembthemosts血g吼tm出Bforthesyntheticphe

7、loq坤ofVSMCsincrustedl4

8、-foldⅫd15-foldbythe⋯1n肿withVECsatthe48hi／mepoint，peakedm72hwithI7一foldand2I-foldI一％⋯aⅢwithculturca]oDcaccordingtoRT-PCRanalysisThenthetmndoftheexp”sslonofCRBPI∞dSmemb⋯rscdtheexpTesslonat96hisl⋯hⅫthoseat72hOttthecOn”awthe∞n怕cⅢcphenotyp。ITlarkerSmoothelin—BandSM—MHCe“

9、pmssl0⋯significantlylowerthanthosecuItⅢtdaloneat48hand72hIlOW,VeT,thecxDTcssIonofSmoothelin—Brevveda崎96hco-col【IIresignifie札tlyhigherthant