目的 探讨停灌时间、再灌注时间对心肌缺血再灌注损伤程度的影响,为防止缺血再灌注损伤的发生或减轻其损伤程度提供依据.方法 2015年9月—2017年5月,选取健康成年雄性实验树鼩,采用随机数字表法分为5组,保证每组采用Langendorff离体心脏灌注系统成功建立心肌缺血再灌注损伤模型10只.A组稳定灌注30 min后取5只实验树鼩观察心肌梗死面积,其余继续灌注直至90 min.B组稳定灌注30 min,停灌15 min,再灌注30 min.C组稳定灌注30 min,停灌15 min,再灌注60 min.D组稳定灌注30 min,停灌30 min,再灌注30 min.E组稳定灌注30 min,停灌30 min,再灌注60 min.B~E组分别在停灌后取5只实验树鼩观察心肌梗死面积,其余继续再灌注.灌注结束,沿心脏冠状面切片,计算梗死面积.取灌注液、心肌组织,检测丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)水平.结果 A组、D组、E组停灌与再灌注后实验树鼩心肌梗死面积比较,差异无统计学意义(P>0.05);B组、C组再灌注后实验树鼩心肌梗死面积较停灌后增加(P<0.05).B~E组再灌注后心肌梗死面积大于A组,D、E组心肌梗死面积大于B、C组(P<0.05).双因素方差分析显示,停灌时间对灌注液、心肌组织的ALT、AST,以及对灌注液CK-MB、LDH的主效应显著,而再灌注时间对灌注液、心肌组织的ALT、AST、CK-MB、LDH的主效应均显著(P<0.05).结论 缺血时间和再灌注时间是影响心肌缺血再灌注损伤程度的关键因素,其中以再灌注时间更为重要.%Objective To investigate the effect of suspension time and perfusion time on myocardial ischemia-reperfusion injury,in order to provide experimental evidence for the prevention or reduction of ischemia-reperfusion injury. Methods This study was conducted between September 2015 and May 2017.Healthy adult male tree shrews were divided into five groups by random number table,and there were 10 tree shrews in each group,which were successfully established myocardial ischemia-reperfusion injury models through Langendorff technique.Treatment in group A was stable perfusion for 30 min,and 5 tree shrews were uesd to observed infarction areas,and the others were continue perfusing for 60 min.In group B,treatment included stable perfusion for 30 min,suspension for 15 min and reperfusion for 30 min;in group C,treatment was stable perfusion for 30 min,suspension for 15 min and reperfusion for 60 min.In group D,stable perfusion,suspension and reperfusion were undertaken for 30,30 and 30 min,respectively;in group E,stable perfusion,suspension and reperfusion were undertaken for 30,30 and 60 min,respectively.In B-E groups,5 tree shrews were uesd to observed infarction areas after suspension,and the others were continue reperfusing.The infarction area was calculated based on heart coronal sections.Perfusion fluid and myocardial tissues were obtained to determine the levels of alanine aminotransferase(ALT),aspartate transaminase (AST),creatine kinase-MB(CK-MB) and lactate dehydrogenase(LDH).Results There was no statistically significant difference in myocardial infarction area between suspension and reperfusion in group A,D and E(P>0.05).The myocardial infarction area were increased after reperfusion than that after suspension in group B and C(P<0.05).The infarction areas in group B-E were significantly larger than that in group A,and group D and E had a larger infarction area than group B and C (P<0.05).Two-factor analysis of variance showed that the main effect of suspension time was significant for the ALT and AST level in perfusion fluid and myocardial tissues,as well as for the level of CK-MB and LDH in perfusion fluid;while the main effect of reperfusion time was significant for the ALT,AST,CK-MB and LDH levels in both perfusion fluid and myocardial tissues(P<0.05).Conclusion Both suspension time and reperfusion time are key factors in affecting myocardial ischemia-reperfusion injury,and reperfusion time is more important.
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