Anderson Dana R, Taylor Stephanie L, Fetterer David P, Holmes Wesley W
Medical Toxicology Branch, Analytical Toxicology Division, U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD 21010, USA.
Toxicology. 2009 Sep 1;263(1):41-6. doi: 10.1016/j.tox.2008.08.025. Epub 2008 Sep 21.
Sulfur mustard (SM)-induced lung injury has been associated with protease activation, oxidative injury and inflammatory response culminating in tissue necrosis. The protease inhibitors aprotinin and ilomastat and the antioxidant trolox were evaluated for efficacy in ameliorating SM-induced lung injury. Anesthetized spontaneously breathing rats (N=6-8/group) were intratracheally intubated and exposed to 1.4 mg/kg SM (0.35 mg SM in 0.1 ml of ethanol) or ethanol alone by vapor inhalation for 50 min. At 1 min before the exposure rats were treated with one of the following: intravenous aprotinin, 4.4 mg/kg; intraperitoneal (ip) ilomastat, 25mg/kg; or ip trolox, 500 microg/kg. Aprotinin-treated animals received supplemental 2.2mg/kg doses at 1 min and 6h post-exposure (PE). A whole body plethysmograph system was used to monitor pulmonary function (PF) parameters for 1h before exposure (baseline), and from 5-6 and 23-24h post-exposure. SM inhalation caused significant increases in several PF parameters, including tidal volume, peak inspiratory flow, peak expiratory flow, end expiratory pause and enhanced pause. Consistent with the reported development of SM-induced pathology, these changes were minimal at the 5-6-h time and significant at the 23-24-h timepoint. At the later time it is known from previous work that airways are becoming obstructed with loose cellular debris, damaged cells and exudate, which contributed to the changes in PF parameters. Treatment with aprotinin or ilomastat eliminated these PF changes, yielding results comparable with controls for each of these parameters. Lung lavage fluid analysis showed that SM caused a significant increase in total protein (TP) and in the cytokines IL-1alpha and IL-13. Aprotinin treatment prevented the increases in TP and IL-1alpha production, ilomastat prevented the increased production of IL-13, and trolox treatment did not significantly prevent the SM-related increases in TP, IL-1alpha or IL-13. Histopathologic examination of lung tissue 24h post-exposure showed minimal alveolar effects caused by SM, while damage to bronchiolar regions was much more severe due to the highly reactive nature of SM. While aprotinin and ilomastat both alleviated the PF perturbations, surprisingly only aprotinin reduced the observed pathology, both grossly and histologically. These early results indicate that treatment with aprotinin and to a lesser extent ilomastat reduces some of the direct inflammatory response and damage associated with SM-induced lung injury. This research was supported by the Defense Threat Reduction Agency - Joint Science and Technology Office, Medical S&T Division.
硫芥（SM）所致肺损伤与蛋白酶激活、氧化损伤及炎症反应相关，最终导致组织坏死。对蛋白酶抑制剂抑肽酶和伊洛马司他以及抗氧化剂曲洛昔芬在改善SM所致肺损伤方面的疗效进行了评估。将麻醉状态下自主呼吸的大鼠（每组N = 6 - 8只）经气管插管，通过蒸汽吸入暴露于1.4 mg/kg SM（0.1 ml乙醇中含0.35 mg SM）或仅暴露于乙醇中50分钟。在暴露前1分钟，用以下药物之一对大鼠进行治疗：静脉注射抑肽酶，4.4 mg/kg；腹腔注射（ip）伊洛马司他，25 mg/kg；或腹腔注射曲洛昔芬，500 μg/kg。接受抑肽酶治疗的动物在暴露后1分钟和6小时（PE）接受补充剂量2.2 mg/kg。使用全身体积描记系统在暴露前1小时（基线）以及暴露后5 - 6小时和23 - 24小时监测肺功能（PF）参数。吸入SM导致多个PF参数显著增加，包括潮气量、吸气峰流速、呼气峰流速、呼气末停顿和增强停顿。与报道的SM所致病理学发展一致，这些变化在5 - 6小时时最小，在23 - 24小时时间点显著。在较晚时间，根据先前的研究可知气道被松散的细胞碎片、受损细胞和渗出物阻塞，这导致了PF参数的变化。用抑肽酶或伊洛马司他治疗可消除这些PF变化，这些参数的结果与对照组相当。肺灌洗流体分析表明，SM导致总蛋白（TP）以及细胞因子IL - 1α和IL - 13显著增加。抑肽酶治疗可防止TP和IL - 1α产生增加，伊洛马司他可防止IL - 13产生增加，而曲洛昔芬治疗未显著防止与SM相关的TP、IL - 1α或IL - 13增加。暴露后24小时对肺组织进行组织病理学检查显示，SM对肺泡的影响最小，而由于SM的高反应性，细支气管区域的损伤更为严重。虽然抑肽酶和伊洛马司他均减轻了PF紊乱，但令人惊讶的是，只有抑肽酶在大体和组织学上均减轻了观察到的病理学变化。这些早期结果表明，抑肽酶治疗以及程度较轻的伊洛马司他治疗可减轻一些与SM所致肺损伤相关的直接炎症反应和损伤。本研究得到了国防威胁降低局 - 联合科学与技术办公室医学科技司的支持。
