臭氧预处理对家兔全心缺血-再灌注损伤的影响

Effects of ozone pretreatment on heart ischemia-reperfusion injury in rabbits

目的研究臭氧预处理对家兔离体心脏全心缺血-再灌注损伤的影响,并探讨其作用机制。方法健康成年家兔24只,雌雄不拘,体重2.25~2.5 kg。随机分为正常对照组(C组)、缺血-再灌注组(IR组)、氧气预处理组(O2P组)和臭氧预处理组(O3P组),每组6只。所有动物常规饲养,C组和IR组不做术前处理;于术前5 d O2P组给予O210 ml,O3P组给予40μg/ml O3 10 ml腹腔内注射,固定时间注射,每日一次。制备离体心脏缺血-再灌注损伤模型,IR组、O2P组和O3P组平衡20 min,停灌30 min,再灌注60 min。C组单纯灌注110 min。于平衡20 min(T0)、再灌注5 min(T1)、再灌注60 min(T2)时记录左室发展压(LVDP),并取冠脉流出液,采用ELISA法测定肌酸激酶同工酶MB(CK-MB)、肌钙蛋白T(cTnT)、IL-6、IL-10浓度。于再灌注60 min末,取心肌组织,用TTC染色法测定心肌梗死面积,HE染色观察病理学结果。结果与C组比较,T1-T2时IR组、O2P组和O3P组HR明显减慢(P<0.05),LVDP明显降低(P<0.05),CK-MB、cTnT、IL-6浓度明显升高(P<0.05),心肌梗死面积百分比明显增大(P<0.05),心肌细胞损伤加重。与IR组比较,T1-T2时O3P组HR明显增快(P<0.05),LVDP明显升高(P<0.05),CK-MB、cTnT、IL-6浓度明显降低(P<0.05),IL-10浓度明显升高(P<0.05),心肌梗死面积明显减小(P<0.05),心肌细胞损伤减轻。IR组和O2P组HR,LVDP,CK-MB,IL-6,IL-10浓度和心肌梗死面积差异均无统计学意义。结论臭氧预处理减轻家兔离体心脏缺血-再灌注损伤,其保护机制可能与抑制促炎因子释放和促进抗炎因子表达有关。

Objective To study the effect of ozone pretreatment on heart ischemia-reperfusion injury in isolated heart of rabbits, and to explore its mechanism. Methods Twenty-four healthy adult rabbits, male and female, weighting 2.25-2.5 kg, were randomly divided into four groups: normal control group(group C), ischemia-reperfusion group(group IR), oxygen pretreatment group(group O2P) and ozone pretreatment group(group O3P). All animals were fedconventionally and no preoperative treatment was performed in the group C and group IR. Group O2P and group O3P were injected intraperitoneally with O2 10 ml and 40 μg/ml O3 10 ml respectively, 5 days before surgery at fixed time, once daily. The ischemia-reperfusion injury model of isolated heart was prepared, which was balanced for 20 min, stopped for 30 min, and reperfused for 60 min. Simple perfusion was performed for 110 min in group C. Left ventricular developmental pressure(LVDP)was recorded at 20 min of equilibration(T0), 5 min of reperfusion(T1) and 60 min of reperfusion(T2). Coronary effluent was taken for measurement of creatine kinase isoenzyme MB(CK-MB), troponin T(cTnT), IL-6 and IL-10 concentrations by ELISA. At the end of 60 min of reperfusion, myocardial tissue was taken, the infarct size was determined by TTC staining, and the pathological results were observed by HE staining. Results Compared with group C, the HR and LVDP of group IR, group O2P and group O3P were significantly decreased(P < 0.05), and the concentrations of CK-MB, cTnT, and IL-6 were significantly increased at T1-T2. The percentage of myocardial infarct size(P < 0.05) and myocardial cell injury were significantly increased in group IR, group O2P and group O3P. Compared with the group IR, the HR and LVDP in group O3P were significantly increased(P < 0.05), and the concentrations of CK-MB, cTnT, and IL-6 were significantly decreased(P < 0.05). The concentration of IL-10 was significantly increased(P < 0.05) at T1-T2. The area of myocardial infarction was significantly reduced(P < 0.05), and myocardial cell damage was reduced in group O3P. There was no significant difference in HR, LVDP, CK-MB, IL-6, IL-10 and myocardial infarction area between group IR and group O2P. Conclusion Ozone pretreatment attenuates ischemia-reperfusion injury in isolated rabbit hearts, and the protective mechanism may be related to inhibiting the release of pro-inflammatory factors and promoting the expression of anti-inflammatory factors.