McDonald M C, Mota-Filipe H, Paul A, Cuzzocrea S, Abdelrahman M, Harwood S, Plevin R, Chatterjee P K, Yaqoob M M, Thiemermann C
Department of Experimental Medicine and Nephrology, William Harvey Research Institute, St. Bartholomew's and The Royal London School of Medicine and Dentistry, London EC1M 6BQ, UK.
FASEB J. 2001 Jan;15(1):171-186. doi: 10.1096/fj.99-0645com.
There is limited evidence that inhibition of the activity of the cytosolic cysteine protease calpain reduces ischemia/reperfusion injury. The multiple organ injury associated with hemorrhagic shock is due at least in part to ischemia (during hemorrhage) and reperfusion (during resuscitation) of target organs. Here we investigate the effects of calpain inhibitor I on the organ injury (kidney, liver, pancreas, lung, intestine) and dysfunction (kidney) associated with hemorrhagic shock in the anesthetized rat. Hemorrhage and resuscitation with shed blood resulted in an increase in calpain activity (heart), activation of NF-kappaB (kidney), expression of iNOS and COX-2 (kidney), and the development of multiple organ injury and dysfunction, all of which were attenuated by calpain inhibitor I (10 mg/kg i.p.), administered 30 min prior to hemorrhage. Chymostatin, a serine protease inhibitor that does not prevent the activation of NF-kappaB, had no effect on the organ injury/failure caused by hemorrhagic shock. Pretreatment (for 1 h) of murine macrophages or rat aortic smooth muscle cells (activated with endotoxin) with calpain inhibitor I attenuated the binding of activated NF-kappaB to DNA and the degradation of IkappaBalpha, IkappaBbeta, and IkappaBvarepsilon. Selective inhibition of iNOS activity with L-NIL reduced the circulatory failure and liver injury, while selective inhibition of COX-2 activity with SC58635 reduced the renal dysfunction and liver injury caused by hemorrhagic shock. Thus, we provide evidence that the mechanisms by which calpain inhibitor I reduces the circulatory failure as well as the organ injury and dysfunction in hemorrhagic shock include 1) inhibition of calpain activity, 2) inhibition of the activation of NF-kappaB and thus prevention of the expression of NFkappaB-dependent genes, 3) prevention of the expression of iNOS, and 4) prevention of the expression of COX-2. Inhibition of calpain activity may represent a novel therapeutic approach for the therapy of hemorrhagic shock.
仅有有限的证据表明，抑制胞质半胱氨酸蛋白酶钙蛋白酶的活性可减轻缺血/再灌注损伤。与失血性休克相关的多器官损伤至少部分归因于靶器官的缺血（出血期间）和再灌注（复苏期间）。在此，我们研究钙蛋白酶抑制剂I对麻醉大鼠失血性休克相关的器官损伤（肾脏、肝脏、胰腺、肺、肠道）和功能障碍（肾脏）的影响。用失血进行出血和复苏导致钙蛋白酶活性增加（心脏）、NF-κB激活（肾脏）、诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2）表达（肾脏）以及多器官损伤和功能障碍的发生，所有这些均被在出血前30分钟腹腔注射的钙蛋白酶抑制剂I（10毫克/千克）所减轻。抑肽酶是一种丝氨酸蛋白酶抑制剂，不能阻止NF-κB的激活，对失血性休克引起的器官损伤/衰竭没有影响。用钙蛋白酶抑制剂I预处理（1小时）小鼠巨噬细胞或大鼠主动脉平滑肌细胞（用内毒素激活）可减弱激活的NF-κB与DNA的结合以及IκBα、IκBβ和IκBε的降解。用L-NIL选择性抑制iNOS活性可减轻循环衰竭和肝脏损伤，而用SC58635选择性抑制COX-2活性可减轻失血性休克引起的肾功能障碍和肝脏损伤。因此，我们提供的证据表明，钙蛋白酶抑制剂I减轻失血性休克中循环衰竭以及器官损伤和功能障碍的机制包括：1）抑制钙蛋白酶活性；2）抑制NF-κB的激活，从而防止NF-κB依赖性基因的表达；3）防止iNOS的表达；4）防止COX-2的表达。抑制钙蛋白酶活性可能代表一种治疗失血性休克的新方法。
