戊乙奎醚对肺缺血再灌注损伤大鼠的拮抗作用

Antagonistic effect of penehyclidine on lung ischemia reperfusion injury rats

目的研究戊乙奎醚对肺缺血再灌注损伤的拮抗作用及其与高迁移率族蛋白B1(HMGB1)-晚期糖基化终末产物受体(RAGE)信号通路的关系。方法将120只Wistar雄性大鼠随机分为4组:假手术组、模型组(肺缺血再灌注组)和低、高剂量实验组(0.1,1 mg·kg^(-1)戊乙奎醚),每组30只。术毕心脏放血法处死大鼠对标本进行处理,以酶联免疫吸附(ELISA)法测定肺匀浆髓过氧化物酶(MPO)活性、肿瘤坏死因子-α(TNF-α)含量及血清中白细胞介素-1β(IL^(-1)β)、白细胞介素-6(IL-6)的含量;以蛋白质印迹法检测肺泡灌洗液中肺表面活性蛋白(SP-A)及肺组织中RAGE的水平;以免疫组化法测定肺组织HMGB1的表达。结果假手术组、模型组和低、高剂量实验组MPO分别为(3.67±0.53),(8.86±0.42),(7.91±0.44),(6.21±0.41)pg·mL^(-1),TNF-α分别为(5.34±0.41),(10.49±0.39),(8.49±0.44),(7.19±0.43)pg·mL^(-1),IL^(-1)β分别为(4.52±0.42),(12.39±0.42),(8.71±0.41),(6.35±0.45)pg·mL^(-1),IL-6分别为(38.58±0.42),(58.42±0.41),(43.92±0.42),(30.65±0.39)pg·mL^(-1),与假手术组比较,模型组和低、高剂量实验组的MPO、TNF-α、IL^(-1)β、IL-6的表达均有增加(均P<0.05)。与模型组比较,低、高剂量实验组肺组织炎症因子及HMGB1、RAGE的表达均降低(均P<0.05)。结论HMGB1、RAGE参与肺缺血再灌注损伤的发生过程;戊乙奎醚对肺缺血再灌注损伤起到保护作用,其机制可能是通过对HMGB1-RAGE信号通路的拮抗作用来实现的。

Objective To study the antagonistic effect of penehyclidine on lung ischemia-reperfusion injury and its relationship with high mobility group protein B1(HMGB1)-advanced glycation end product receptor(RAGE)signaling pathway.Methods Totally 120 male Wistar rats were randomly divided into 4 groups:sham operation group,model group(pulmonary ischemia-reperfusion group),low dose experimental group and high dose experimental group(0.1,1 mg·kg^(-1) penehyclidine),with 30 rats in each group.The rats were sacrificed by cardiac exsanguination after operation,and the activity of myeloperoxidase(MPO)in lung homogenate,the content of tumor necrosis factor-α(TNF-α)and the content of interleukin-1β(IL^(-1)β)and interleukin-6(IL-6)in serum were determined by enzyme linked immunosorbent assay(ELISA).Western blotting was used to detect the levels of lung surface active protein(SP-A)in alveolar lavage fluid and RAGE in lung tissue.The expression of HMGB1 in lung tissue was determined by immunohistochemical method.Results In sham operation group,model group and low,high dose experimental groups,MPO were(3.67±0.53),(8.86±0.42),(7.91±0.44),(6.21±0.41)pg·mL^(-1);TNF-αwere(5.34±0.41),(10.49±0.39),(8.49±0.44),(7.19±0.43)pg·mL^(-1);IL^(-1)βwere(4.52±0.42),(12.39±0.42),(8.71±0.41),(6.35±0.45)pg·mL^(-1);IL-6 were(38.58±0.42),(58.42±0.41),(43.92±0.42),(30.65±0.39)pg·mL^(-1).Compared with sham operation group,the expression of MPO,TNF-α,IL^(-1)βand IL-6 in model group and low,high dose experimental groups were increased(all P<0.05).Compared with model group,the expressions of inflammatory factors,HMGB1 and RAGE in lung tissues of low,high dose experimental groups were decreased(all P<0.05).Conclusion HMGB1 and RAGE are involved in the occurrence of lung ischemia-reperfusion injury.Penehyclidine has a protective effect on lung ischemia-reperfusion injury,which may be achieved through antagonistic effect on HMGB1-RAGE signaling pathway.