体外循环法对犬骨骼肌缺血再灌注损伤的保护作用

Protective effects of ECC on ischemia-reperfusion injury in canine skeletal muscle

目的探讨体外循环法(extracorporeal circulation,ECC)对犬骨骼肌缺血再灌注损伤的保护作用。方法首先建立犬骨骼肌缺血再灌注损伤模型,通过HE染色法观察组织炎性细胞浸润,利用TUNEL法观察组织凋亡的变化,特异性试剂盒检测LD、SOD等酶的变化,ELISA法检测IL-10和TNF-α等细胞因子的变化,评价ECC法再灌注模式对犬骨骼肌缺血再灌注损伤的保护作用。结果模型组动物的组织炎性细胞浸润明显,诱导细胞凋亡发生,血清中LD[(2.6±0.3)mmol/ml vs.(5.2±0.1)mmol/ml]、SOD[(15.2±1.2)U/ml vs.(31.1±2.1)U/ml]等酶的水平显著增加,同时血清中IL-10[(60.4±2.3)pg/ml vs.(89.2±1.5)pg/ml]和TNF-α[(172.3±8.7)ng/l vs.(273.4±12.5)ng/l]等细胞因子的含量显著增加(P<0.01)。ECC法再灌注处理的动物,其组织炎性反应和细胞凋亡程度显著低于模型组动物,血清中上述酶和细胞因子的水平也显著降低,分别为:LD[(3.5±0.1)mmol/ml vs.(5.2±0.1)mmol/ml],SOD[(21.3±1.3)U/ml vs.(31.1±2.1)U/ml],IL-10[(69.4±2.6)pg/ml vs.(89.2±1.5)pg/ml]和TNF-α[(190.8±12.1)ng/l vs.(273.4±12.5)ng/l](P<0.05)。结论ECC法再灌注模式,对犬骨骼肌缺血再灌注损伤具有一定的保护作用。

Objective To investigate the protective effect of ECC on ischemia-reperfusion injury in canine skeletal muscle.Methods The inflammatory cell infiltration was observed by HE staining.The changes of tissue apoptosis were observed using the TUNEL method.At the same time,LD and SOD were detected by the specific kit.Changes in such cytokines as TNF-αand IL-10 were detected by ELISA.Results In the model group,inflammatory cell infiltration was significantly increased accompanied by obvious apoptosis.In addition,levels of LD[(2.6±0.3)mmol/ml vs(5.2±0.1)mmol/ml]and SOD[(15.2±1.2)U/ml vs(31.1±2.1)U/ml]were significantly increased(P<0.01).Furthermore,the levels of such cytokines as IL-10[(60.4±2.3)vs(89.2±1.5)pg/ml]and TNF-α[(172.3±8.7)ng/l vs(273.4±12.5)ng/l]were increased significantly(P<0.01).In the ECC group,the tissue inflammatory response was reduced significantly.The levels of the above enzymes and cytokines were also reduced obviously,LD[(3.5±0.1)mmol/ml vs(5.2±0.1)mmol/ml],SOD[(21.3±1.3)U/ml vs(31.1±2.1)U/ml],IL-10[(69.4±2.6)pg/ml vs(89.2±1.5)pg/ml],TNF-α[(190.8±12.1)ng/l vs 273.4±12.5 ng/l](P<0.05).Conclusions The protective effect on canine skeletal muscle ischemia-reperfusion injury is observed after pretreatment with the ECC method.