CAJ | 학술논문

目的探讨胃肠道血管畸形( GIVM)的发病机制及沙利度胺治疗GIVM所致消化道出血的作用机制.方法收集10例因GIVM导致消化道大出血而行部分肠切除术患者的病变组织、非病变组织以及非GIVM患者的正常肠黏膜组织,分别进行免疫组化染色检测血管生成素2(Ang-2)、Notch1、Dll4的表达.体外培养人脐静脉内皮细胞(HUVEC)至对数生长期,同步化后分别用4种浓度的沙利度胺(25、50、100、200 mg/L)刺激24 h或48 h,另设溶剂对照组.实时定量PCR法测定沙利度胺刺激24 h后各组Ang-2、Notch1、Dll4 mRNA的表达水平,Western blot法测定沙利度胺作用48 h后各组Ang-2、Notch1、Dll4蛋白的表达水平.结果免疫组化染色显示GIVM病变肠段血管扩张、扭曲,与GIVM患者的非病变组织及非GIVM患者的正常肠黏膜组织相比,其Ang-2、Notch1、Dll4的表达明显增强,且Ang-2的表达与Notch1、Dll4的表达水平呈正相关(r值均为0.732,P值均为0 016),Dll4与Notch1的表达呈完全正相关(r=1.000,P=0.000).实时定量PCR和Western blot结果提示,沙利度胺能明显抑制Ang-2、Notch1及Dll4的mRNA和蛋白表达,且这种抑制作用呈剂量相关.结论 Ang-2、Notch1、Dll4可能与GIVM的发病机制相关,沙利度胺可抑制其表达,且具有剂量依赖性.这一作用可能是沙利度胺抑制血管生成,从而有效治疗GIVM所致消化道出血的机制之一.
목적 탐토위장도혈관기형( GIVM)적발병궤제급사리도알치료GIVM소치소화도출혈적작용궤제.방법 수집10례인GIVM도치소화도대출혈이행부분장절제술환자적병변조직、비병변조직이급비GIVM환자적정상장점막조직,분별진행면역조화염색검측혈관생성소2(Ang-2)、Notch1、Dll4적표체.체외배양인제정맥내피세포(HUVEC)지대수생장기,동보화후분별용4충농도적사리도알(25、50、100、200 mg/L)자격24 h혹48 h,령설용제대조조.실시정량PCR법측정사리도알자격24 h후각조Ang-2、Notch1、Dll4 mRNA적표체수평,Western blot법측정사리도알작용48 h후각조Ang-2、Notch1、Dll4단백적표체수평.결과 면역조화염색현시GIVM병변장단혈관확장、뉴곡,여GIVM환자적비병변조직급비GIVM환자적정상장점막조직상비,기Ang-2、Notch1、Dll4적표체명현증강,차Ang-2적표체여Notch1、Dll4적표체수평정정상관(r치균위0.732,P치균위0 016),Dll4여Notch1적표체정완전정상관(r=1.000,P=0.000).실시정량PCR화Western blot결과제시,사리도알능명현억제Ang-2、Notch1급Dll4적mRNA화단백표체,차저충억제작용정제량상관.결론 Ang-2、Notch1、Dll4가능여GIVM적발병궤제상관,사리도알가억제기표체,차구유제량의뢰성.저일작용가능시사리도알억제혈관생성,종이유효치료GIVM소치소화도출혈적궤제지일.
Objective To study the pathogcncsis of gastrointestinal vascular malformation (GIVM) and the potential mechanism of thalidomide in the treatment of gastrointestinal bleeding due to GIVM.Methods We collected the surgical intestinal specimens from 10 patients who suffered from massive hemorrhage of gastrointestinal tract owning to GIVM and the normal intestinal mucosa around the lesions,as well as normal intestinal mucosa from healthy subjects.Immunohistochemical(IHC) staining was carried out to investigate the differences of angiopoietin 2 ( Ang2 ),Notch1 and delta like ligand 4 (Dll4) in the above three intestinal mucosa to find the relationship with the pathogenesis of GIVM. Human umbilical vein endothelial cells(HUVECs) were cultured with 0,25,50,100 and 200 mg/L thalidomide for 24 or 48 hours to observe their mRNA and protein expressions of Ang2,Notch1,Dll4 by real-time PCR and Western blot.Results By IHC staining,more expressions of Ang2,Notch1 and Dll4 in the lesions were detected than those in the normal intestinal mucosa around the lesions and the normal intestinal mucosa in healthy people.The expressions of Ang2,Notch1 and Dll4 were significantly correlated (P =0.016,r =0.732),and the expressions of Notch1 and Dll4 were absolutely correlated ( P =0.000,r =1.000).Real-time PCR and Western blot showed that thalidomide could down-regulate the expressions of them,which were in a concentration-dependent manner.Conclusion Ang2,Notch1 and Dll4 may correlate with the pathogenesis of GIVM,while thalidomide can concentration-dependently down-regulate the expression of Ang2,Notch1 and Dll4,which may be one of the mechanism that thalidomide play a therapeutic role in GIVM.