Steinberg Jay, Halter Jeffrey, Schiller Henry J, Dasilva Monica, Landas Steve, Gatto Louis A, Maisi Paivi, Sorsa Timo, Rajamaki Minna, Lee Hsi-Ming, Nieman Gary F
Department of Surgery, SUNY Upsate Medical University, Syracuse, NY 13210, USA.
J Surg Res. 2003 May 15;111(2):185-95. doi: 10.1016/s0022-4804(03)00089-1.
Neutrophil activation with concomitant matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) release has been implicated in the development of sepsis-induced acute lung injury. We hypothesized that COL-3, a chemically modified tetracycline known to inhibit MMP-2 and MMP-9, would reduce lung injury and improve survival in rats following cecal ligation and puncture (CLP).
Sprague-Dawley rats were separated into five groups: 1) sham CLP+ carboxymethylcellulose (CMC; vehicle for COL-3, n = 6); 2) sham CLP + COL-3 (n = 6); 3) CLP + CMC (n = 10); 4) CLP + single-dose (SD) COL-3 administered concomitant with CLP (n = 9); and 5) CLP + multiple-dose (MD) COL-3 administered concomitant with CLP and at 24 h after CLP (n = 15). Rats were sacrificed at 168 h (7 days) or immediately after death, with survival defined as hours after CLP. Histological lung assessment was made based on neutrophil infiltration, alveolar wall thickening, and intraalveolar edema fluid. Lung MMP-2 and MMP-9 levels were assessed by immunohistochemistry. MMP-2 and MMP-9 levels were correlated with survival by simple regression analysis.
The mortality of rats in the cecal ligation and puncture without treatment group (CLP + CMC) was 70% at 168 h. A single dose of COL-3 in the CLP + COL-3 (SD) group significantly reduced mortality to 54%. Furthermore, with a repeat dose of COL-3 at 24 h after CLP, mortality was significantly reduced to 33%. Pathologic lung changes seen histologically in the CLP + CMC group were significantly reduced by COL-3. A significant reduction in lung tissue levels of MMP-2 and MMP-9 was noted in both groups treated with COL-3. Reduction of MMP-2 and MMP-9 levels correlated with improved survival.
Inhibition of MMP-2 and MMP-9 by COL-3 in a clinically relevant model of sepsis-induced acute lung injury reduces pulmonary injury and improves survival in a dose-dependent fashion. Our results suggest that prophylactic treatment with COL-3 in high-risk patients may reduce the morbidity and mortality associated with sepsis-induced acute respiratory distress syndrome.
中性粒细胞激活并伴随基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）释放与脓毒症诱导的急性肺损伤的发生有关。我们推测，COL-3，一种已知可抑制MMP-2和MMP-9的化学修饰四环素，将减轻盲肠结扎和穿刺（CLP）后大鼠的肺损伤并提高其存活率。
将Sprague-Dawley大鼠分为五组：1）假手术CLP+羧甲基纤维素（CMC；COL-3的载体，n = 6）；2）假手术CLP + COL-3（n = 6）；3）CLP + CMC（n = 10）；4）CLP +与CLP同时给予单剂量（SD）COL-3（n = 9）；5）CLP +与CLP同时及CLP后24小时给予多剂量（MD）COL-3（n = 15）。在168小时（7天）或死亡后立即处死大鼠，存活时间定义为CLP后的小时数。基于中性粒细胞浸润、肺泡壁增厚和肺泡内水肿液进行肺组织学评估。通过免疫组织化学评估肺MMP-2和MMP-9水平。通过简单回归分析将MMP-2和MMP-9水平与存活率相关联。
未治疗的盲肠结扎和穿刺组（CLP + CMC）大鼠在168小时时的死亡率为70%。CLP + COL-3（SD）组中的单剂量COL-3显著降低死亡率至54%。此外，在CLP后24小时重复给予COL-3，死亡率显著降低至33%。COL-3显著减轻了CLP + CMC组组织学上可见的病理性肺改变。在接受COL-3治疗的两组中均观察到肺组织中MMP-2和MMP-9水平显著降低。MMP-2和MMP-9水平的降低与存活率提高相关。
在脓毒症诱导的急性肺损伤的临床相关模型中，COL-3对MMP-2和MMP-9的抑制以剂量依赖的方式减轻肺损伤并提高存活率。我们的结果表明，对高危患者进行COL-3预防性治疗可能降低与脓毒症诱导的急性呼吸窘迫综合征相关的发病率和死亡率。
