Differential contribution of necrosis and apoptosis in myocardial ischemia-reperfusion injury
Differential contribution of necrosis and apoptosis in myocardial ischemia-reperfusion injury
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DOI：
10.1152/ajpheart.00935.2003
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发表时间：
2004-05-01
影响因子：
4.8
通讯作者：
Levitsky, S
中科院分区：
文献类型：
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作者：
McCully, JD;Wakiyama, H;Levitsky, S
坏死和凋亡对心肌损伤的作用不同。确定这些过程在缺血-再灌注损伤中的作用将有助于心肌组织的保护。在Langendorff灌注的兔心脏（n = 47）中研究了全脑缺血（GI）0（对照组）、5（GI-5）、10（GI-10）、15（GI-15）、20（GI-20）、25（GI-25）和30 min（GI-30）和再灌注120 min的坏死和凋亡。通过氯化三苯基四氮唑（TTC）染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）、bax、bcl 2、聚（ADP）核糖聚合酶（PARP）切割、半胱天冬酶-3、-8和-9切割和活性、Fas配体（FasL）和Fas激活的死亡结构域（FADD）来确定心肌损伤。使用不可逆半胱天冬酶-3、-8和-9抑制剂分别测定细胞凋亡的贡献（n = 42）。在GI-20、GI-25和GI-30，左心室峰发展压（LVPDP）和收缩期缩短（SS）显著降低，梗死面积和TUNEL阳性细胞显著增加（与对照组相比P < 0.05）。在GI-5至GI-30时，促凋亡bax、PARP裂解以及半胱天冬酶-3和-9裂解和活性明显。Fas、FADD和caspase-8的裂解和活性未发生改变。在GI-25和GI-30，不可逆地抑制caspase-3和caspase- 9活性可显著减少梗死面积（P < 0.05），但对LVPDP和SS无影响。心肌损伤是由于坏死和凋亡的显著增加（与对照组相比P < 0.05），这通过TUNEL、TTC染色和GI-20的半胱天冬酶活性来证明。内在的促凋亡激活在缺血早期是明显的，但在GI-25之前并不显著影响梗死面积。在GI-20、GI-25和GI-30，坏死对梗死面积的贡献显著大于凋亡。在早期再灌注过程中，半胱天冬酶抑制剂可显著降低细胞凋亡，但这种保护作用并不能改善缺血后即刻的功能恢复。
Necrosis and apoptosis differentially contribute to myocardial injury. Determination of the contribution of these processes in ischemia-reperfusion injury would allow for the preservation of myocardial tissue. Necrosis and apoptosis were investigated in Langendorff-perfused rabbit hearts (n = 47) subjected to 0 ( Control group), 5 (GI-5), 10 (GI-10), 15 (GI-15), 20 (GI-20), 25 (GI-25), and 30 min (GI-30) of global ischemia (GI) and 120 min of reperfusion. Myocardial injury was determined by triphenyltetrazolium chloride (TTC) staining, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling ( TUNEL), bax, bcl2, poly( ADP) ribose polymerase ( PARP) cleavage, caspase-3, -8, and -9 cleavage and activity, Fas ligand (FasL), and Fas-activated death domain ( FADD). The contribution of apoptosis was determined separately ( n = 42) using irreversible caspase-3, - 8, and - 9 inhibitors. Left ventricular peak developed pressure (LVPDP) and systolic shortening ( SS) were significantly decreased and infarct size and TUNEL-positive cells were significantly increased ( P < 0.05 vs. Control group) at GI-20, GI-25, and GI-30. Proapoptotic bax, PARP cleavage, and caspase-3 and - 9 cleavage and activity were apparent at GI-5 to GI-30. Fas, FADD, and caspase-8 cleavage and activity were unaltered. Irreversible inhibition of caspase-3 and - 9 activity significantly decreased ( P < 0.05) infarct size at GI-25 and GI-30 but had no effect on LVPDP or SS. Myocardial injury results from a significant increase in both necrosis and apoptosis ( P < 0.05 vs. Control group) evident by TUNEL, TTC staining, and caspase activity at GI-20. Intrinsic proapoptotic activation is evident early during ischemia but does not significantly contribute to infarct size before GI-25. The contribution of necrosis to infarct size at GI-20, GI-25, and GI-30 is significantly greater than that of apoptosis. Apoptosis is significantly decreased by caspase inhibition during early reperfusion, but this protection does not improve immediate postischemic functional recovery.