血红素加氧酶-1对大鼠体外循环术后肺组织抗炎性损伤的研究  

Research on the anti-inflammation effect and mechanism of heme oxygenase-1 on lung injury after cardiopulmonary bypass

作　　者：生伟[1] 潘丽华[2] 池一凡[1] 孙龙[1] 牛兆倬[1] 林明山[1] 

机构地区：[1]青岛大学附属青岛市市立医院心脏中心,山东省青岛市266071 [2]威海市立医院心内科

出　　处：《中国心血管病研究》2017年第8期764-767,I0002,共5页Chinese Journal of Cardiovascular Research

基　　金：青岛市卫计委课题（项目编号：2013-WSZD012）

摘　　要：目的 观察钴原卟啉诱导肺组织血红素加氧酶-1（HO-1）表达在大鼠体外循环术后肺组织损伤中发挥的抗炎性损伤作用,并对其作用机制进行分析.方法 雄性Wistar大鼠144只,随机分为3组：A组（对照组）、B组（钴原卟啉,CoPP）、C组[钴原卟啉（CoPP）＋锌原卟啉（ZnPP）].建立改良大鼠体外循环肺损伤模型.制模前24 h,A组大鼠腹腔注射生理盐水,B组腹腔注射CoPP（5 mg/kg）,C组腹腔注射CoPP（5 mg/kg）和ZnPP（5 mg/kg）.制模成功后采用断颈法分别于体外循环前（T0）,体外循环结束即刻（T1）,体外循环后2h（T2）、6 h（T3）、12 h（T4）、24 h（T5）将大鼠处死,每个时间点8只.取肺组织检测相应的指标.用多聚甲醛固定部分右肺组织,然后用石蜡包埋,以用于HO-1蛋白表达的免疫组化方法测定.剩余肺组织置于液氮中冻存备用,用于检测肺组织匀浆胆红素的含量,酶联免疫吸附（ELISA）法检测肿瘤坏死因子α（TNF-α）表达.结果 B组大鼠肺组织中HO-1蛋白表达在体外循环（CPB）开始前后均有明显增加,其活性测定结果与蛋白表达结果一致,在各时间点均强于A组和C组（P＜0.05）;各组HO-1蛋白表达在T2期最高,A、B、C三组分别为0.1852±0.070、0.2873±0.091、0.1917±0.082.B组大鼠肺组织TNF-α含量在CPB后各时间点均低于A组和C组（p＜0.05）;三组TNF-α的含量在T2期最高,分别为（480.42±41.67）pg/ml、（358.19±32.35）pg/ml、（466.53±39.42）pg/ml.结论 CoPP可以诱导肺组织HO-1的高表达,而且经过CPB后仍保持较高的表达.内源性HO-1高表达在体外循环肺损伤中具有抗炎作用,可有效抑制TNF-α炎性细胞因子分泌,从而减轻体外循环术后肺的炎性损伤.Objective The aim of this study is to determine the anti-inflammation effects of heme oxygenase-1 on lung injury after cardiopulmonary bypass,and to investigate its probable mechanisms.Methods 144 male Wistar rats were divided into 3 groups：group A （control group）,group B （cobalt protoporphyrin,CoPP）,group C [CoPP and zinc protoporphyrin（ZnPP）] randomly.A modified rat model of CPB-induced lung injury was established.Normal saline was injected 24 h before modified rat model establishment in group A intraperitoneally,and CoPP（5 mg/kg） in Group B,CoPP（5 mg/kg） and ZnPP（5 mg/kg） in group C respectively.After the modified rat model was established,the lung tissues were taken at different time for the relevant indicators test：before CPB（T0）,0 min after CPB（T1）,2 h after CPB（T2）,6 h（T3）,12 h（T4） and 24 h（T5）.Partial right lung tissue of rats in each group was fixed by paraformaldehyde as soon as excision,and then it was embedded in paraffin,then used for the test the expression of HO-1 protein in each group by immunohistochemistry.The rest of the lung tissue was placed in liquid nitrogen for detection of lung tissue homogenate bilirubin content,and the determination of TNF-α expression by ELISA method.Results The HO-1 protein expressions in lung tissue in group B were higher than those in group A and group C in any given time interval,and the same as HO-1 activity （P＜0.05）,the expressions of HO-1 protein of group A,B and C at T2 stage were 0.1852±0.070,0.2873±0.091,0.1917±0.082 respectively.TNF-α content in group B at all time point after bypass were lower than those of group A and group C （P＜0.05）,the TNF-α contents of group A,B and C at T2 stage were （480.42±41.67）pg/ml,（358.19±32.35）pg/ml,（466.53±39.42）pg/ml respectively.Conclusion CoPP can induce a large amount of HO1 expression in lung tissue,and it is still highly expressed after CPB,so as to play an important part in anti-inflammation,and reduce lung injury.

关 键 词：血红素加氧酶-1 体外循环 肺损伤 炎症反应 

分 类 号：Q95-33[生物学—动物学] R654.2[医药卫生—外科学]