VEGF单克隆抗体在胰腺缺血再灌注损伤中的保护作用

Effect of VEGF MCAb in ischemia-reperfusion injury in pancreas of rats

目的:探讨VEGF单克隆抗体对胰腺缺血再灌注(ischemia-reperfusion,I/R)损伤的保护作用及其机制。方法:采用钳闭大鼠腹腔干、肠系膜上动脉制备胰腺缺血再灌注损伤模型。应用ELISA试剂检测VEGF,TNF-α表达,并应用TUNEL法检测胰腺细胞凋亡,RT-PCR,蛋白质印迹法和免疫组织化学染色检测胰腺组织中Fas,FasLmRNA和蛋白的表达情况。结果:I/R组血清中的TNF-α,VEGF明显高于VEGF单抗组(P<0.01)。I/R组胰腺组织中TUNEL染色见典型的细胞核固缩及凋亡小体形成。应用VEGF单克隆抗体后,胰腺组织中胰腺细胞凋亡指数明显下降。和I/R组相比,胰腺组织中Fas,FasL mRNA和蛋白表达水平均明显降低,免疫组化亦显示Fas,FasL在VEGF单抗组中显色弱于I/R组。结论:VEGF单克隆抗体可以降低I/R过程中TNF-α,VEGF表达水平,并且抑制Fas的表达,抑制过度凋亡,减轻缺血再灌注对胰腺的损伤。

Objective： To study the effect and mechanism of VEGF MCAb in preventing ischemia-reperfusion injury.Methods： Ischemia-reperfusion models were made by ligated both the anterior menseneric artery and the celiac artery of 70 rats.The expression of TNF-α,VEGF was detected by ELISA.Apoptosis of pancreatic acinar cells was examined by terminal deoxynucleotidyl transferase mediated dUTPbiot in nick end labeling（TUNEL） method.The expression of Fas,FasL mRNA and protein were detected by Real-Time PCR,Western Blot and immunohistochemistry in rat pancreatic samples.Results： Compared with ischemia-reperfusion group,VEGFMCAb can significantly decrease the level of TNF-α,VEGF in serum and apoptosis index.The expression level of Fas,FasL mRNA and protein in I/R group was significant higher than pancreatic tissue in group treating with VEGFMCAb.In I/R group,Fas,FasL immunoreactivity was more intense than in corresponding VEGFMCAb treated tissue.Conclusion： VEGFMCAb can protect pancreas against ischemia-reperfusion injury and can inhibit excessive apoptosis through down-regulation of Fas/FasL.