心肌缺血预适应作用对老年大鼠心肌的影响及其机制的研究

摘要

目的：本研究探索心肌缺血预适应（IPC）对老年大鼠心肌缺血再灌注（I/R）后的影响及机制。　　方法：取21~23月龄老年Wistar大鼠32只，建立离体心脏Langendorff灌注模型，分为4组（每组8只）：对照组、I/R组、IPC组、强化IPC组。对照组采用全心灌流180 min，不做任何处理。I/R组采用心脏平衡灌流30 min后，缺血30 min，再复灌120 min。IPC组采用心脏平衡灌流10 min，经两次缺血5 min再灌注5 min后，缺血30 min，再复灌120 min。强化IPC组采用心脏平衡灌流10 min，经四次缺血5 min再灌注5 min后，缺血30 min ，再复灌120 min。比较各组复灌30 min、60 min、90 min、120 min后心排血量的恢复率以及左心室发展压、左心室内压最大上升和下降速率（±dp/dtmax）的恢复率；检测缺血前及复灌120 min后冠状动脉流出液中肌酸激酶MB同工酶（CK-MB）和乳酸脱氢酶活性，心肌组织中丙二醛含量和超氧化物歧化酶的活性。各组取心尖肌做冰冻切片行过氧化物酶体增生激活受体γ协同刺激因子1α（PGC-1α）免疫组织化学染色，计算出平均积分吸光度。　　结果：IPC组与I/R组丙二醛含量、CK-MB及超氧化物歧化酶活性，左心室发展压、±dp/dtmax恢复率比较差异无统计学意义（P>0.05），而强化IPC组与I/R组比较，CK-MB和超氧化物歧化酶活性明显减少（P<0.01），丙二醛含量明显降低（P<0.05），超氧化物歧化酶活性明显增加（P<0.01），心排血量、左心室发展压、±dp/dtmax恢复率明显增加（P<0.01），差异有统计学意义。PGC-1α的表达：IPC组与I/R组比较差异无统计学意义（P>0.05），而强化IPC组表达明显增加，与I/R组比较差异有统计学意义（P<0.01）。　　结论：老年大鼠心肌IPC的保护作用减弱，可能机制是老年心肌PGC-1α蛋白表达降低，使老年心肌调节超氧化物歧化酶活性的作用下降，进而影响老年心肌对应激反应的防护能力，使IPC对老年心肌不能发挥保护作用。而强化IPC使PGC-1α表达增加，恢复了对超氧化物歧化酶活性的调节能力，进而恢复了IPC对老年心肌的保护作用。%Objective: To explore the impact of Ischemic preconditioning (IPC) in aged experimental rats after myocardial ischemia-reperfusion (I/R) with its mechanism. Methods: A total of 32 Wistar rats at the age of (21-23) months were divided into 4 groups, n=8 in each group.①Control group, the rats received cardiac perfusion for 180 min. ②I/R group, the rats received cardiac perfusion for 30 min, followed by ischemia for 30 min, then reperfusion for 120min.③IPC group, the rats received cardiac perfusion for 10 min, followed by ischemia and reperfusion 2 times (5 min in each time), then ischemia 30 min and reperfusion 120 min. ④ Enhanced IPC group, rats received cardiac perfusion for 10 min, followed by ischemia and reperfusion 4 times (5 min in each time), then ischemia 30 min and reperfusion 120 min. The recovery rate of cardiac output (CO), left ventricular developed pressure (LVDP) and the recovery rate of maximum rise and fall of left ventricular pressure (±dp/dtmax) at (30, 60, 90, 120) min after reperfusion were recorded respectively. The creatine kinase (CK-MB), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were examined before ischemia and 120 min after reperfusion. The apical peroxisome proliferator-activated receptorγco-stimulatory factor 1α(PGC-1α) was examined by immuno-histochemistry. Results: The MDA content, CK-MB, SOD activities LVDP and (±dp/dtmax) recovery were similar between IPC group and I/R group, P>0.05. While compared with I/R group, the Enhanced IPC group showed decreased CK-MB activity and MDA content, increased SOD activity and CO, LVDP and (±dp/dtmax) recovery rate, all P<0.01. The PGC-1αexpression was similar between IPC group and I/R group, P>0.05. While compared with I/R group, the Enhanced IPC group had increased PGC-1αexpression, P<0.01. Conclusion: The cardiac IPC was weakened in aged rats which might be because of decreased PGC-1αexpression, the enhanced IPC may up-regulate PGC-1αexpression and therefore, protect the cardiac tissue in aged experimental rats.