门静脉高压大鼠腹膜血管内皮生长因子的表达及意义

目的探讨门静脉高压大鼠腹膜血管内皮生长因子(VEGF)的表达及意义。方法雄性SD大鼠随机分为3组:门静脉结扎(PVL)组32只,假手术(SO)组16只,门静脉结扎联合沙利度安治疗(PVL-T)组8只。为诱导腹水形成,于手术后第1,3,5,7天分批给予每只大鼠腹腔注射33.33%葡萄糖溶液1mL,30min后收集腹水标本并测总量,同时采用ELISA测定腹水中VEGF含量;留取腹膜组织做VEGF免疫组化染色和实时RT-PCR。结果术后第1、3天PVL组大鼠腹水总量和VEGF含量显著大于SO组(P<0.05),术后第5、7天大鼠腹水总量和VEGF含量在PVL组和SO组之间无显著性差异(P>0.05),术后第3天PVL-T组腹水总量显著低于PVL组(3.4±0.27mLvs4.92±1.53mL,P<0.05)。免疫组化结果显示PVL组VEGF呈强阳性表达,而SO组VEGF则呈弱阳性表达,且表达部位多在腹膜间皮细胞、巨噬细胞胞浆;实时RT-PCR结果表明PVL组腹膜VEGFmRNA表达显著高于SO组(P<0.05)。结论门静脉高压可增加由于渗透性改变所致大鼠腹水形成,腹膜VEGF高表达可能与大鼠腹水形成有关,抑制VEGF活性可能是治疗门静脉高压所致腹水的一种新的治疗策略。

Expression of vascular endothelial growth factor in peritoneum of portal hypertensive rats and their significance

【Objective】 To investigate expression of vascular endothelial growth factor in peritoneum of portal hypertensive rats and their significance.【Methods】 Male S-D Rats were randomly divided into three groups: portal vein ligation(PVL) group with 32 rats,sham-operated(SO) group with 16 rats,Portal vein ligation combined with thalidomide treatment(PVL-T) group with 8 rats.To induce the formation of ascites,1.0 mL of 33.33% glucose was injected into the abdomen of every rat on day1,3,5,and 7 after operation.30 minutes later,ascites was collected and the total capacity was measured.VEGF levels in ascites were measured by ELISA.Expression of VEGF in peritoneum tissues was studied using real-time RT-PCR and immunohistochemistry.【Results】 On day 1,3 after operation,the amount of ascites and VEGF level in PVL rats was significantly higher than that in SO rats(P <0.05).Whereas,on day 5,7 after operation,the amount of ascites and VEGF level was no significant difference between the PVL group and the SO group(P >0.05).On day 3 after operation,the amount of ascites in PVL-T group was significantly lower than that in PVL group(3.4±0.27 mL vs 4.92±1.53 mL,P <0.05).Histological results showed that expression of VEGF was strongly positive in PVL group,while the SO group was weakly positive.The positive parts mainly located in peritoneal mesothelial cells and cytoplasm of macrophages.Real-time RT-PCR results also showed that VEGF-mRNA expression of peritoneum in PVL group was significantly higher than that in SO group(P <0.05).【Conclusion】 Portal hypertension increases ascites formation which is caused by a difference of osmolality.Increased expression of VEGF in peritoneum may be associated with ascites formation in rats.Inhibition of VEGF activity maybe a new strategy for the treatment of ascites,which is caused by portal hypertensive.