CAJ | 학술논문

目的探讨自体静脉移植后血管内皮生长因子(VEGF)对产生促红细胞生成素的肝癌细胞B4受体(EphB4)表达的调控及其抑制移植静脉内膜增生的机制.方法建立小鼠自体颈静脉移植重建颈总动脉的模型,采用实时定量聚合酶链反应(Real-time PCR)方法,检测术后不同时间点VEGF和EphB4 mRNA水平;利用小干扰RNA (siRNA)阻断内源性VEGF的产生,比较处理组与非处理组内膜增生情况;以VEGF刺激鼠静脉内皮细胞观察EphB4表达;分别用VEGF传导通路的阻断剂预处理细胞,再以VEGF刺激细胞,检测EphB4表达.结果阻断移植静脉VEGF的表达后,EphB4表达显著减少(处理组0.52±0.14比非处理组2.10 ±0.79,P＜0.05),且内膜增生程度明显高于未处理组[处理组(29.8±2.09) μm比非处理组(14.7 ±0.05) μm,P＜0.05].VEGF下调EphB4呈时间及剂量依赖性.VEGF通过VEGF受体2(VEGF-R2)及其下游信号传导通路细胞外调节激酶(ERK) 1/2抑制EphB4表达.结论 VEGF可能通过负性调控EphB4的表达而抑制移植静脉内膜增生,此调节作用通过VEGF-R2-ERK1/2途径实现.
목적 탐토자체정맥이식후혈관내피생장인자(VEGF)대산생촉홍세포생성소적간암세포B4수체(EphB4)표체적조공급기억제이식정맥내막증생적궤제.방법 건립소서자체경정맥이식중건경총동맥적모형,채용실시정량취합매련반응(Real-time PCR)방법,검측술후불동시간점VEGF화EphB4 mRNA수평;이용소간우RNA (siRNA)조단내원성VEGF적산생,비교처리조여비처리조내막증생정황;이VEGF자격서정맥내피세포관찰EphB4표체;분별용VEGF전도통로적조단제예처리세포,재이VEGF자격세포,검측EphB4표체.결과 조단이식정맥VEGF적표체후,EphB4표체현저감소(처리조0.52±0.14비비처리조2.10 ±0.79,P＜0.05),차내막증생정도명현고우미처리조[처리조(29.8±2.09) μm비비처리조(14.7 ±0.05) μm,P＜0.05].VEGF하조EphB4정시간급제량의뢰성.VEGF통과VEGF수체2(VEGF-R2)급기하유신호전도통로세포외조절격매(ERK) 1/2억제EphB4표체.결론 VEGF가능통과부성조공EphB4적표체이억제이식정맥내막증생,차조절작용통과VEGF-R2-ERK1/2도경실현.
Objective To observe the relationship between vasculaendothelial growth facto(VEGF) and venoufingerprinEPH receptoB4 (EphB4) expression in vein grafadaptation, so ato discusthe mechanism of VEGF in inhibiting vein grafintimal hyperplasiby regulating EphB4.MethodEstablish raauto-jugulavein grafmodel to determine VEGF mRNA, EphB4 mRNlevel in early and late phase awell acompare vasculawall thickening in vein grafadaptation;knockdown endogenouVEGF in early phase by using small interfering RN(siRNA) and observe EphB4 expression and the tendency of intima-medithickening in latephase.Mouse endothelial cellwere stimulated with VEGF, opretreated with VEGF-receptor2 (VEGF-R2), extracellulasignal-regulated kinase (ERK) 1/2, oprotein kinase (Akt) inhibitors, and then EphB4 expression wadetermined by real-time quantitative polymerase chain reaction (Real-time PCR).ResultKnockdown endogenouVEGF lead to decrease of EphB4 expression (knockdown VEGF vs.wide type : 0.52 ±0.14 vs.2.10 ± 0.79, P ＜ 0.05, GAPDH =1) awell aincrease of intima-medithickening [knockdown VEGF vs.wide type : (29.8 ± 2.09) μm vs.(14.7 ± 0.05) μm, P ＜ 0.05];VEGF down-regulated EphB4 in time-and dosage-dependenfashion.VEGF suppression of EphB4 expression wadependent, aleasin part, on VEGF-R2 and itdownstream ERK1/2 pathway bunothe Akpathway.Conclusion VEGF irequired fodiminished EphB4 expression in vein graft.The mechanism of VEGF inhibiintimal hyperplasimay vidown-regulation of EphB4 through VEGFR2-ERK1/2 pathway.