Carbon Monoxide-Releasing Molecule-2 Reduces Intestinal Epithelial Tight-Junction Damage and Mortality in Septic Rats.
Carbon Monoxide-Releasing Molecule-2 Reduces Intestinal Epithelial Tight-Junction Damage and Mortality in Septic Rats.
复制标题
一氧化碳释放分子 2 减少脓毒症大鼠肠上皮紧密连接损伤和死亡率
DOI：
10.1371/journal.pone.0145988
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Ji Z
中科院分区：
文献类型：
--
作者：
Zhang S;Zheng S;Wang X;Shi Q;Wang X;Yuan S;Wang G;Ji Z
肠上皮细胞紧密连接的损伤在脓毒症中起重要作用。最近我们发现一氧化碳释放分子-2（CORM-2）能够保护LPS诱导的肠上皮细胞紧密连接损伤，本研究将探讨CORM-2是否能够保护大鼠盲肠结扎穿孔（CLP）模型的肠上皮细胞紧密连接。根据标准程序使用雄性Sprague-Dawley（SD）大鼠产生CLP模型，并且在CLP诱导后立即静脉内用CORM-2或无活性CORM-2（iCORM-2）8 mg/kg处理，并在24小时或72小时后安乐死（仅用于死亡率）。形态学变化进行了研究，使用透射电子显微镜和共聚焦显微镜。采用Western blotting检测肌球蛋白轻链（MLC）磷酸化水平和重要TJ蛋白水平。采用ELISA试剂盒检测细胞因子、IL-1β和TNF-α。使用FD-4作为标记物评价总体肠上皮通透性。CORM-2而不是iCORM-2显著减少脓毒症诱导的肠粘膜损伤（包括TJ破坏）、TJ蛋白减少（包括闭锁小带-I（ZO-1）、密蛋白-1和闭锁蛋白）、MLC磷酸化和促炎细胞因子释放。总体结果显示，CORM-2抑制脓毒症诱导的肠上皮通透性变化，降低脓毒症大鼠的死亡率。我们的数据强烈表明CORM-2可能是一种潜在的治疗剂，通过抑制炎症，恢复肠上皮屏障和降低死亡率。
Damage to intestinal epithelial tight junctions plays an important role in sepsis. Recently we found that Carbon Monoxide-Releasing Molecule-2 (CORM-2) is able to protect LPS-induced intestinal epithelial tight junction damage and in this study we will investigate if CORM-2 could protect intestinal epithelial tight junctions in the rat cecal ligation and puncture (CLP) model. The CLP model was generated using male Sprague-Dawley (SD) rats according to standard procedure and treated with CORM-2 or inactive CORM-2 (iCORM-2), 8 mg/kg, i.v. immediately after CLP induction and euthanized after 24h or 72h (for mortality rate only). Morphological changes were investigated using both transmission electron and confocal microscopy. The levels of important TJ proteins and phosphorylation of myosin light chain (MLC) were examined using Western blotting. Cytokines, IL-1β and TNF-α were measured using ELISA kits. The overall intestinal epithelial permeability was evaluated using FD-4 as a marker. CORM-2, but not iCORM-2, significantly reduced sepsis-induced damage of intestinal mucosa (including TJ disruption), TJ protein reduction (including zonula occludens-l (ZO-1), claudin-1 and occludin), MLC phosphorylation and proinflammatory cytokine release. The overall outcomes showed that CORM-2 suppressed sepsis-induced intestinal epithelial permeability changes and reduced mortality rate of those septic rats. Our data strongly suggest that CORM-2 could be a potential therapeutic reagent for sepsis by suppressing inflammation, restoring intestinal epithelial barrier and reducing mortality.