摘要
目的:探讨缺血预适应(ischemic preconditioning,IPC)对青年与老年大鼠缺血/再灌注(ischemia/reperfusion,IR)心肌损伤的影响。方法:雄性3月龄(青年)与20月龄(老年)Wistar大鼠,应用离体心脏灌流方法复制心肌IR与IPC模型。实验分为青年缺血/再灌注(YIR)组、青年缺血预适应(YPC)组、老年缺血再灌注(OIR)组与老年缺血预适应(OPC)组。透射电镜观察心肌及心肌线粒体超微结构变化;TTC染色测定心肌梗死面积;比色法测定冠脉流出液中乳酸脱氢酶(LDH)活性、心肌组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;酶联免疫吸附法测定心肌组织硝基化与羰基化蛋白质含量,TUNEL方法检测心肌细胞凋亡;氧电极法测定线粒体呼吸功能及钙诱导的线粒体渗透性转运孔开放情况。结果:与YIR组比较,YPC组心肌梗死面积明显减少,冠脉流出液中LDH活性降低,心肌组织的SOD活性增加,MDA含量降低,心肌硝基化与羰基化蛋白含量降低。电镜下可见YPC组心肌及分离的心肌线粒体膜结构完整、基质致密。YIR组心肌线粒体呼吸控制率与Ⅲ态耗氧量及P/O比值均明显增加,质子漏出减少,钙诱导的线粒体肿胀明显减轻,心肌细胞凋亡率下降。而与OIR组比较,OPC组上述指标均无显著统计学差异。与YPC组比较,OPC组心肌超微结构损伤明显增加,心肌氧化应激水平增加,线粒体呼吸功能下降,心肌细胞凋亡与坏死增多。结论:缺血预适应能够保护青年大鼠心肌缺血/再灌注损伤;而老年大鼠心脏对预适应刺激减敏,导致缺血预适应心肌保护作用钝化,这可能与老龄IPC心脏氧化应激水平增加导致线粒体损伤、细胞凋亡有关。
AIM： To study the protective effect of ischemia preconditioning （ IPC ） on ischemia/reperfusion （ IR）-damaged myocardium in young and old rats .METHODS： Male Wistar rats aged at 3 months （ young ） and 20 months （ old） were used to establish myocardial IPC model and IR model with the method of Langendorff heart perfusion . The rats were divided into young ischemia/reperfusion （ YIR） group, young ischemic preconditioning （ YPC） group, old ischemia/reperfusion （ OIR） group and old ischemic preconditioning （ OPC） group.Transmission electron microscopy was used to observe the ultrastructural changes of myocardial tissue and myocardial mitochondria .The myocardial infarction area was determined by TTC staining .The lactate dehydrogenase （ LDH） content in coronary effluent fluid and the levels of superoxide dismutase （ SOD） and malondialdehyde （ MDA） in myocardial tissues were detected by the method of colorimetry . The levels of nitrated and carbonylated proteins in myocardial tissue were measured by ELISA .The myocardial cell apoptosis was analyzed by TUNEL assay .The mitochondrial respiratory function and mitochondrial permeability transition pore opening induced by calcium load were evaluated by oxygen electrode method .RESULTS： Compared with YIR group , the myocardial infarction area in YPC group was obviously smaller , SOD activity in myocardial tissues increased , LDH activity in coronary effluent fluid and the content of MDA decreased , and the levels of nitrated and carbonylated proteins in the cardiac tissues reduced .In YPC group, the mitochondrial membrane structure appeared intact , cristae of the mitochondria showed close arrangement , and the matrix was compressed under the electron microscope .Myocardial mitochondrial respiratory control rate , state Ⅲoxygen consumption and the P/O ratio in YIR group all significantly increased , proton leak decreased, mitochondrial swelling induced by calcium distinctly reduced , and myocardial apoptosis rate declined .No significant difference of the above indexes between OIR group and OPC group was observed .Compared with YPC group , myocardial ultrastructural damage increased clearly , cardiac oxidative stress increased , mitochondrial respiratory function declined, and cell apoptosis and necrosis increased in OPC group .CONCLUSION：Ischemic preconditioning has protective effect against myocardial IR injury in young rat hearts , while old rat hearts were less sensitive to ischemic preconditioning , leading to bluntness of cardioprotection with IPC in aging hearts .This may be related to mitochondrial injury and severe cellular apoptosis caused by increase of cardiac oxidative stress levels in the aging ischemic preconditioning heart .
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第10期1737-1743,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81170178)
关键词
缺血/再灌注
缺血预适应
氧化应激
线粒体功能
心肌
Ischemia/reperfusion
Ischemic preconditioning
Oxidative stress
Mitochondrial function
Myocardium
