Active site inhibited factor VIIa attenuates myocardial ischemia/reperfusion injury in mice
Active site inhibited factor VIIa attenuates myocardial ischemia/reperfusion injury in mice
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DOI：
10.1111/j.1538-7836.2008.03232.x
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发表时间：
2009-02-01
影响因子：
10.4
通讯作者：
Ten Cate, H.
中科院分区：
文献类型：
--
作者：
Loubele, S. T. B. G.;Spek, C. A.;Ten Cate, H.
背景：抑制特定的凝血途径，如因子VIIa-组织因子复合体，已被证明可以减轻缺血/再灌注(I/R)损伤，但其细胞机制尚未探讨。目的：探讨活性部位抑制因子VIIa(ASIS)抗心肌I/R损伤作用的细胞机制。方法：在小鼠心肌I/R损伤模型中，观察特定的小鼠重组蛋白的作用。缺血1h后再灌流2、6、24 h。在再灌流诱导前后给予小鼠ASIS或安慰剂。结果：ASIS可使心肌I/R损伤在3次再灌流时减少40%以上。多重连接依赖的探针扩增(MLPA)分析显示，给予ASIS后，缺血心肌中CD14、TLR-4、IL-1受体相关激酶(IRAK)和I-kappa Bα的mRNA表达减少，表明Toll样受体-4(TLR-4)和随后的核因子-kappa B(NF-kappa B)介导的细胞信号转导受到抑制。ASIS可降低I/R后核活化的核因子-kappaB和受核因子-kappaB通路影响的蛋白水平，包括组织因子(Tf)和IL-6。再灌流6h和2 4h后，给予ASIS治疗后，梗死区中性粒细胞浸润减少。然而，没有证据表明ASIS对细胞凋亡有影响(TUNEL染色和MLPA分析)。结论：ASIS给药后心肌I/R损伤减轻主要是由于炎症相关致死性I/R损伤减轻，可能是通过核因子-kappaB机制介导的。
Background: Inhibition of specific coagulation pathways such as the factor VIIa-tissue factor complex has been shown to attenuate ischemia/reperfusion (I/R) injury, but the cellular mechanisms have not been explored. Objectives: To determine the cellular mechanisms involved in the working mechanism of active site inhibited factor VIIa (ASIS) in the protection against myocardial I/R injury. Methods: We investigated the effects of a specific mouse recombinant in a mouse model of myocardial I/R injury. One hour of ischemia was followed by 2, 6 or 24 h of reperfusion. Mouse ASIS or placebo was administered before and after induction of reperfusion. Results: ASIS administration reduced myocardial I/R injury by more than 40% at three reperfusion times. Multiplex ligation dependent probe amplification (MLPA) analysis showed reduced mRNA expression in the ischemic myocardium of CD14, TLR-4, interleukin-1 (IL-1) receptor-associated kinase (IRAK) and I kappa B alpha upon ASIS administration, indicative of inhibition of toll-like receptor-4 (TLR-4) and subsequent nuclear factor-kappa B (NF-kappa B) mediated cell signaling. Levels of nuclear activated NF-kappa B and proteins influenced by the NF-kappa B pathway including tissue factor (TF) and IL-6 that were increased after I/R, were attenuated upon ASIS administration. After 6 and 24 h of reperfusion, neutrophil infiltration into the area of infarction was decreased upon ASIS administration. There was, however, no evidence of an effect of ASIS on apoptosis (Tunel staining and MLPA analysis). Conclusions: We conclude that the diminished amount of myocardial I/R injury after ASIS administration is primarily due to attenuated inflammation-related lethal I/R injury, probably mediated through the NF-kappa B mechanism.