吸入氢气对大鼠心肌缺血再灌注损伤的影响

Impact of inhaling hydrogen on rats with myocardial ischemia reperfusion injury

目的探讨吸入氢气对大鼠心肌缺血再灌注损伤的影响。方法 40只雄性SD大鼠随机分为4组,每组10只,分别为:假手术组(SH组),缺血再灌注组(I/R组),1%氢气组(1%H2组),2%氢气组(2%H_2组)。除SH组外,其他各组通过结扎左冠状动脉前降支30 min后恢复血流并持续120 min,制作大鼠缺血再灌注损伤模型。1%H_2组、2%H_2组在再灌注过程中持续吸入相应浓度的氢气。观察并记录缺血前(T_0)、再灌注即刻(T_1)、再灌注30 min(T_2)、再灌注60 min(T_3)、再灌注120 min(T_4)各时点的心率(HR)、平均动脉压(MAP),并在再灌注120 min时从尾静脉抽取5 ml静脉血,进行血清肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)检测。随后处死大鼠取心脏,计算心肌梗死的面积,并采用酶联免疫吸附法(ELISA)测定心肌8-羟基脱氧鸟苷(8-OHdG)、高迁移率族蛋白1(HMGB1)、核因子-κB(NF-κB)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)的浓度。结果各组HR在各时点的比较差异均无统计学意义(P>0.05)。与SH组比较,I/R组、1%H_2组、2%H_2组的梗死面积、CK-MB、cTnI、8-OHdG、HMGB1、NFκB、TNF-α、IL-6显著升高,而MAP水平降低(P<0.05)。与I/R组比较,1%H_2组、2%H_2组的梗死面积、CK-MB、cTnI、8-OHdG、HMGB1、NF-κB、TNF-α、IL-6显著降低,MAP水平升高(P<0.05)。1%H_2组和2%H_2组组间比较各观察指标差异无统计学意义(P>0.05)。结论吸入氢气能减轻大鼠心肌缺血再灌注损伤,其机制可能与抑制氧化应激损伤、炎性反应有关。

Objective To explore the impact of inhaling hydrogen on rats with myocardial ischemia reperfusion injury rat. Methods 40 male rats were randomly divided into 4 groups： sham-operated group （SH group）,ischemia reperfusion group （I/R group）,1% hydrogen group （1%H2 group） and 2% hydrogen group （2%H2 group）,each group had 10 rats.Apart from SH group,ischemia reperfusion injury rats model were made by ligation left anterior descending coronary artery for 30 min and then restored blood flow for continuing 120 min.Appropriate concentration of hydrogen were continuous in- haled in 1%H2 group and 2%H2 group.Heart rate （HR） before ischemia （T0）,immediate reperfusion （T1）,reperfusion for 30 min （T2）,reperfusion for 60 min （T3） reperfusion for 120 min （T4） and mean arterial pressure （MAP） were observed and recorded.5 ml venous blood was extracted at 120 min after reperfusion,Creatine kinase-MB （CK-MB） and cardiac troponin I （cTnI） were tested.Then the rats were killed and hearts were removed.The area of myocardial infarction area was calculated and the levels of 8-hydroxy-2＇-deoxyguanosine （8-OHdG）,high mobility group box 1 （HMGB1）,nuclear factor-kappa B （NF-KB）,tumor necrosis factor α （TNF-α） and interleukin-6 （IL-6） were tested by enzyme linked im- munosorbent assay （ELISA）. Results The HR had no significant difference at different time among 4 groups.Compared with SH group,the infarction area,lelves of CK-MB,cTnI,8-OHdG,HMGB1,NF-κB,TNF-α and IL-6 in I/R group, 1%H2 group and 2%H2 group significantly increased,while the level of MAP decreased （P〈0.05）.Compared with I/R group,the infarction area,levels of CK-MB,cTnI,8-OHdG,HMGB1,NF-KB,TNF-α and IL-6 in 1%H2 group and 2%H2 group sig- nificantly decreased,while the level of MAP increased （P〈0.05）.The index between 1%H2 group and 2%H2 group had no significant difference （≥0.05）. Conclusion Inhaling hydrogen can alleviate myocardial ischemia reperfusion injury in rat.The mechanism may be related to inflammatory reaction and inhibit oxidative stress injury of hydrogen.