Buerke Michael, Pruefer Diethard, Sankat Dennis, Carter Justin M, Buerke Ute, Russ Martin, Schlitt Axel, Friedrich Ivar, Börgermann Jochen, Vahl Christian F, Werdan Karl
Department of Internal Medicine III, Martin Luther University Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle/Saale, Germany.
Circulation. 2007 Sep 11;116(11 Suppl):I121-6. doi: 10.1161/CIRCULATIONAHA.106.680249.
Reperfusion injury of ischemic myocardium has been attributed to neutrophil infiltration, inflammatory activation and cardiac necrosis/apoptosis. Serine protease inhibition with aprotinin is cardioprotective, but the mechanism is unknown.
We studied aprotinin in a rat model of myocardial ischemia for 20 minutes and reperfusion for 20 minutes, 8 hours or 24 hours. Aprotinin (20,000 IU/kg) given 5 minutes before reperfusion significantly reduced leukocyte accumulation (P<0.01), myocardial injury (determined by CK depletion, P<0.01) and myocyte apoptosis (P<0.05) compared with vehicle treated rats. Differential gene expression analysis showed myocardial ischemia plus reperfusion increased expression of proinflammatory genes like P-selectin, E-selectin, intercellular adhesion molecule, tumor necrosis factor-alpha, tumor necrosis factor-alpha receptor, interleukin-6, monocyte chemoattractant protein-1, p53, and Fas (CD59). Aprotinin before reperfusion suppressed expression of these inflammatory genes. Finally, differential protein expression analysis demonstrated increased intercellular adhesion molecule-1, tumor necrosis factor-alpha, and p53 after myocardial ischemia plus reperfusion, and this effect was diminished by aprotinin.
We demonstrated myocardial ischemia plus reperfusion induced leukocyte accumulation, inflammation, gene expression, protein expression and finally tissue injury and showed aprotinin limiting reperfusion injury through each of these stages, even after 24 hours of reperfusion. This effect seems partly attributable to suppression of proinflammatory genes and leukocyte accumulation. This work casts further light on the complex signaling of ischemia and reperfusion.
缺血心肌的再灌注损伤被认为与中性粒细胞浸润、炎症激活及心脏坏死/凋亡有关。抑肽酶抑制丝氨酸蛋白酶具有心脏保护作用，但其机制尚不清楚。
我们在大鼠心肌缺血20分钟并再灌注20分钟、8小时或24小时的模型中研究了抑肽酶。与给予赋形剂处理的大鼠相比，再灌注前5分钟给予抑肽酶（20,000 IU/kg）可显著减少白细胞聚集（P<0.01）、心肌损伤（通过肌酸激酶消耗测定，P<0.01）和心肌细胞凋亡（P<0.05）。差异基因表达分析显示，心肌缺血加再灌注可增加促炎基因如P-选择素、E-选择素、细胞间黏附分子、肿瘤坏死因子-α、肿瘤坏死因子-α受体、白细胞介素-6、单核细胞趋化蛋白-1、p53和Fas（CD59）的表达。再灌注前给予抑肽酶可抑制这些炎症基因的表达。最后，差异蛋白质表达分析表明，心肌缺血加再灌注后细胞间黏附分子-1、肿瘤坏死因子-α和p53增加，而抑肽酶可减弱这种作用。
我们证明心肌缺血加再灌注可诱导白细胞聚集、炎症、基因表达、蛋白质表达，最终导致组织损伤，并表明抑肽酶可在再灌注的各个阶段限制再灌注损伤，即使在再灌注24小时后也是如此。这种作用似乎部分归因于对促炎基因和白细胞聚集的抑制。这项工作进一步揭示了缺血和再灌注的复杂信号传导。
