Bihari Aurelia, Cepinskas Gediminas, Forbes Thomas L, Potter Richard F, Lawendy Abdel-Rahman
Division of Orthopaedic Surgery, University of Western Ontario, London, Ontario, Canada; Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada.
Centre for Critical Illness Research, Lawson Health Research Institute, London, Ontario, Canada; Department of Medical Biophysics, University of Western Ontario, London, Ontario, Canada.
J Vasc Surg. 2017 Dec;66(6):1864-1871. doi: 10.1016/j.jvs.2016.11.065. Epub 2017 Feb 17.
Ischemia-reperfusion (IR) is a limb- and life-threatening complication of acute limb ischemia and musculoskeletal trauma. Carbon monoxide-releasing molecules (CORMs) have recently been shown to protect microvascular perfusion and to reduce inflammation and injury in various ischemic animal models. The purpose of this study was to examine the effects of water-soluble CORM-3 on the extent of IR-induced muscle injury.
Wistar rats were randomized into three groups: sham (no ischemia), IR + CORM-3 (10 mg/kg intraperitoneally), and IR + inactive CORM-3 (iCORM-3; 10 mg/kg intraperitoneally). No-flow ischemia was induced by the application of a tourniquet to the hind limb for 2 hours; tourniquet release commenced the reperfusion phase. Both CORM-3 and iCORM-3 were injected immediately after tourniquet release. Temporal changes in microvascular perfusion, cellular tissue injury (ethidium bromide and bisbenzimide staining), and inflammatory response (leukocyte recruitment) within the extensor digitorum longus muscle were assessed using intravital video microscopy every 15 minutes for a total of 90 minutes after initiation of reperfusion. Systemic levels of tumor necrosis factor-α were also measured.
Hind limb IR resulted in (1) a significant no-reflow phenomenon followed by progressive increase in microvascular perfusion deficit (21% ± 2% continuously perfused capillaries in IR vs 76% ± 4% in sham [P < .001]; 52% ± 8% nonperfused capillaries in IR vs 13% ± 2% in sham at 90 minutes of reperfusion [P < .001]), (2) tissue injury (ethidium bromide and bisbenzimide staining of 0.52 ± 0.07 in IR vs 0.05 ± 0.03 in sham at 90 minutes of reperfusion [P < .001]), (3) leukocyte recruitment (13.7 ± 0.9 adherent leukocytes/30 seconds/1000 μm in IR vs 1.8 ± 0.5 adherent leukocytes/30 seconds/1000 μm in sham at 90 minutes of reperfusion [P < .001]), and (4) an increase in circulating tumor necrosis factor-α levels. Systemic administration of CORM-3 (but not of iCORM-3) effectively reduced the IR-associated skeletal muscle perfusion deficits, tissue injury, and inflammatory activation.
CORM-3 displays potent protective and anti-inflammatory effects in an experimental model of hind limb IR, suggesting a potential therapeutic application of CORMs in treatment of ischemic conditions.
缺血再灌注（IR）是急性肢体缺血和肌肉骨骼创伤中一种危及肢体和生命的并发症。最近研究表明，一氧化碳释放分子（CORMs）可保护微血管灌注，并减少各种缺血动物模型中的炎症和损伤。本研究旨在探讨水溶性CORM-3对IR诱导的肌肉损伤程度的影响。
将Wistar大鼠随机分为三组：假手术组（无缺血）、IR + CORM-3组（腹腔注射10 mg/kg）和IR + 无活性CORM-3组（iCORM-3；腹腔注射10 mg/kg）。通过在后肢应用止血带2小时诱导无血流缺血；松开止血带开始再灌注阶段。在松开止血带后立即注射CORM-3和iCORM-3。在再灌注开始后的90分钟内，每隔15分钟使用活体视频显微镜评估趾长伸肌内微血管灌注、细胞组织损伤（溴化乙锭和双苯甲酰亚胺染色）和炎症反应（白细胞募集）的时间变化。还测量了肿瘤坏死因子-α的全身水平。
后肢IR导致（1）明显的无复流现象，随后微血管灌注缺损逐渐增加（IR组持续灌注的毛细血管为21% ± 2%，假手术组为76% ± 4% [P <.001]；再灌注90分钟时，IR组非灌注毛细血管为52% ± 8%，假手术组为13% ± 2% [P <.001]），（2）组织损伤（再灌注90分钟时，IR组溴化乙锭和双苯甲酰亚胺染色为0.52 ± 0.07，假手术组为0.05 ± 0.03 [P <.001]），（3）白细胞募集（再灌注90分钟时，IR组每30秒每1000μm有13.7 ± 0.9个黏附白细胞，假手术组为1.8 ± 0.5个黏附白细胞/30秒/1000μm [P <.001]），以及（4）循环肿瘤坏死因子-α水平升高。全身给予CORM-3（而非iCORM-3）可有效减少与IR相关的骨骼肌灌注缺损、组织损伤和炎症激活。
CORM-3在大鼠后肢IR实验模型中显示出强大的保护和抗炎作用，提示CORMs在缺血性疾病治疗中具有潜在的治疗应用价值。
