CAJ | 학술논문

目的检测血管内皮生长因子(vascular endothelial growth factor,VEGF)在门静脉高压症(portal hypertension,PHT)内脏血管中的表达变化,探讨其在内脏高动力循环中的作用及机制.方法临床检测肝硬化PHT患者脾脏动脉内VEGF通路相关蛋白的表达.动物实验观察正常组(N组)7只大鼠和四氯化碳(CCl4)诱导的肝硬化门静脉高压症组(PHT组)7只大鼠的门静脉直径、门静脉血流速度(portal vein blood flow velocity,PBV)、门静脉血流量(portal vein blood flow,PBF)和门静脉压力(portal vein pressure,PP)以及离体肠系膜微动脉对去甲肾上腺素(norepinephrine,NE)的反应性变化.通过SU5416选择性抑制VEGF通路后,对比离体肠系膜微动脉收缩反应性的变化以及肠系膜动脉内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)活化程度的变化.细胞实验中通过原代培养动脉内皮细胞进行体外实验,验证VEGF对eNOS活化的影响.结果 ①肝硬化PHT患者脾动脉VEGF表达明显升高.②动物实验中门静脉直径在N组和PHT组之间差异无统计学意义,PHT组PBV和PBF明显低于N组,SU5416对PHT组PBV及PBF无明显改善作用.③PHT组PP水平明显高于N组,SU5416对PHT组PP无明显降低作用.④PHT组肠系膜微动脉对NE的反应性明显降低,EC50值明显增大,SU5416能部分改善这种低反应性.⑤PHT组肠系膜动脉VEGF、VEGFR-2、eNOS和p-eNOS的蛋白表达较N组明显上调,SU5416能明显降低VEGFR-2与表达和eNOS的活化.⑥体外实验证实,VEGF可促进eNOS的活化.结论肝硬化PHT内脏动脉中过度生成的VEGF部分通过促进eNOS表达和活化的方式降低内脏动脉对NE的反应性,参与内脏高动力循环的形成.
목적 검측혈관내피생장인자(vascular endothelial growth factor,VEGF)재문정맥고압증(portal hypertension,PHT)내장혈관중적표체변화,탐토기재내장고동력순배중적작용급궤제.방법 림상검측간경화PHT환자비장동맥내VEGF통로상관단백적표체.동물실험관찰정상조(N조)7지대서화사록화탄(CCl4)유도적간경화문정맥고압증조(PHT조)7지대서적문정맥직경、문정맥혈류속도(portal vein blood flow velocity,PBV)、문정맥혈류량(portal vein blood flow,PBF)화문정맥압력(portal vein pressure,PP)이급리체장계막미동맥대거갑신상선소(norepinephrine,NE)적반응성변화.통과SU5416선택성억제VEGF통로후,대비리체장계막미동맥수축반응성적변화이급장계막동맥내피형일양화담합매(endothelial nitric oxide synthase,eNOS)활화정도적변화.세포실험중통과원대배양동맥내피세포진행체외실험,험증VEGF대eNOS활화적영향.결과 ①간경화PHT환자비동맥VEGF표체명현승고.②동물실험중문정맥직경재N조화PHT조지간차이무통계학의의,PHT조PBV화PBF명현저우N조,SU5416대PHT조PBV급PBF무명현개선작용.③PHT조PP수평명현고우N조,SU5416대PHT조PP무명현강저작용.④PHT조장계막미동맥대NE적반응성명현강저,EC50치명현증대,SU5416능부분개선저충저반응성.⑤PHT조장계막동맥VEGF、VEGFR-2、eNOS화p-eNOS적단백표체교N조명현상조,SU5416능명현강저VEGFR-2여표체화eNOS적활화.⑥체외실험증실,VEGF가촉진eNOS적활화.결론 간경화PHT내장동맥중과도생성적VEGF부분통과촉진eNOS표체화활화적방식강저내장동맥대NE적반응성,삼여내장고동력순배적형성.
Objective To detect the variation of vascular endothelial growth factor (VEGF) in the splanchnic vessels under portal hypertension (PHT) and explore its mechanism and influence on the process of hyperdynamic circulation.Methods In humans,the level of VEGF pathway related proteins were detected in the splenic artery of PHT patients in clinical trials.In animal experiments,the following parameters were observed for rats in the control group (group N,n =7) and the CCl4 induced portal hypertension group (group PHT,n=7):portal vein diameter,portal vein blood flow velocity (PBV),portal vein blood flow (PBF),portal vein pressure (PP),norepinephrine (NE) reactivity in the isolated mesenteric artery microcirculation,contractile reactivity and degree of endothelial ni tric oxide synthase (eNOS) activation in the mesenteric artery by selectively inhibiting the VEGF signal pathway with SU5416.In cell experiments,primary culturing of arterial endothelial cells in vitro were used to verify the effects of VEGF on eNOS activation.Results The results showed that VEGF expression levels in the splenic artery of PHT patients significantly increased.In animal experiments,there was not a significant difference in portal vein diameter between group N and group PHT.How ever,the PBV and PBF of group PHT were lower than those of group N,and SU5416 had no clear effect on PBV and PBF in group PHT.PP of group PHT was much higher than that of group N,and SU5416 had little influence on reducing PP in group PHT.The contractile response of mesenteric artery microcirculation to NE in group PHT decreased significantly,EC50 increased a lot,and SU5416 improved this hypoergia partially.The protein levels of VEGF,VEGFR-2,eNOS,and p-eNOS in the mesentery artery of group PHT raised quite a lot compared to group N,and SU5416 decreased the protein level of VEGFR-2 and activation of eNOS significantly.Experiments in vitro confirmed that VEGF could promote the activation of eNOS.Conclusion The excessive VEGF produced in visceral arteries under PHT may participate in the process of hyperdynamic circulation partially through promoting the synthesis and activation of eNOS and then reducing visceral arteries' response to NE.