体外循环系统下加压灌注改善模型猪下肢血运

Enhancement-perfusion therapy by extracorporeal circulation system ameliorates the blood supply of ischemic lower limbs

背景:课题组前期应用体外循环系统模拟体内生理环境对离体断肢进行灌注保存,并取得了一定研究成果,达到了延长体外断肢寄养时间的目的。目的:研究体外循环系统下加压灌注疗法对改善缺血下肢血运的作用。方法:选取健康成年雄性巴马小型猪18只,随机分为3组(n=6)。A组为正常对照组,不做任何缺血处理;B、C两组通过结扎右后肢股动脉建立下肢缺血模型;缺血4周后,C组进行加压灌注疗法,1 h/(次·d)。1周后应用数字化X光机行右后肢股动脉造影,观察各组股动脉形态及远端侧支循环变化;取各组右后肢胫前肌标本,行苏木精-伊红染色测定毛细血管密度;并用Western blot法检测血管内皮生长因子蛋白相对表达量;采用ELISA法检测血清标本中血管内皮生长因子水平。结果与结论:(1)与B组相比,C组右后肢股动脉造影直径明显增加,远端侧支循环明显丰富;(2)C组毛细血管密度较B组明显增加(P<0.05),但A、C两组差异无显著性意义(P>0.05);(3)Western blot法显示,与A组相比,B、C组血管内皮生长因子蛋白量均显著升高(P<0.05),但以C组为最高(P<0.05);(4)ELISA法检测发现,与A组相比,B、C组血管内皮生长因子血清含量均显著升高(P<0.05),且C组最高(P<0.05);(5)可见,体外循环系统下加压灌注疗法可以明显改善缺血下肢血运,可能为临床治疗下肢缺血提供新选择。

BACKGROUND:In the preliminary study,extracorporeal circulation system simulated physiological environment has been applied in limb preservation,significantly prolonging the severed limb preserving time.OBJECTIVE:To study the effect of enhancement-perfusion by extracorporeal circulation system on the blood supply of ischemic lower limbs.METHODS:Eighteen adult Bama mini pigs were equally randomized into three groups:group A(no intervention),groups B and C(establishing the ischemic model by arteria cruralis ligation at the right posterior lower limb).At 4 weeks after modeling,the model pigs in the group C received enhancement-perfusion therapy 1 hour per day.One week later,the situations of angiography were detected by digital radiography.The tibialis anterior separated from each pig was used to test the microvessel density by hematoxylin-eosin staining.The expression levels of vascular endothelial growth factor in the skeletal muscle and serum samples were monitored by western blot and ELISA,respectively.RESULTS AND CONCLUSION:Both diameter and distal collateral circulation of the arteria cruralis in the group C were much better than those in the group B.The microvessel density in the group C was significantly higher than that in the group B(P<0.05),but the insignificant difference between groups A and C was found(P>0.05).Both histionic and serum levels of vascular endothelial growth factor were significantly increased in the groups B and C compared with the group A,especially in the group C(P<0.05).In summary,the enhancement-perfusion therapy by extracorporeal circulation system can considerably ameliorate blood supply of lower limbs,providing a promising method for treatment of ischemic lower limbs.