Supplementary MaterialsAdditional File 1: Materials, Supplementary Tables and Figures. mice by modulating glucose uptake, adenosine triphosphate and lactate content. Furthermore upregulation of peroxisome proliferator-activated receptor- coactivator-1, nuclear respiratory factor 1 and mitochondrial transcription factor A genes in cardiac tissue were revealed in septic mice treated with carbon monoxide. Taken together, the results indicate that exogenous carbon monoxide effectively modulated mitochondrial energetic metabolisms by interfering with expression of peroxisome proliferator-activated receptor- coactivator-1, nuclear respiratory factor 1 and mitochondrial transcription factor A genes, consequently exerted an important improvement in sepsis-induced cardiac dysfunction. i.vt 0.05 was considered to be statistically significant. Results Effect of CORM-2 on survival of septic mice Survival of mice was analyzed during 72 h after CLP procedures. As shown in Fig ?Fig1,1, no deaths occurred in the sham group. Most of mice challenged with CLP died between 12 h and 24 h. CORM-2 administration dramatically increased the survival rate of septic mice from 25% to 63%. iCORM-2, which did not release carbon monoxide, had no effect on survival of septic mice. Open in a separate window Fig 1 Effect of carbon monoxide releasing molecule-2 (CORM-2) on survival of septic mice. Mice were challenged with cecal ligation and puncture (CLP) and treated Rabbit polyclonal to RFP2 with CORM-2 or AP24534 enzyme inhibitor inactive CORM-2 (iCORM-2). All mice had normal access to water and food. Animal survival was monitored 4 times a day for up to 72 h after CLP. Most of mice challenged with CLP died between 12 h and 24 h. CORM-2 dramatically increased the survival of septic mice from 25% to 63%. ** 0.01 compared with sham, ## 0.01 compared with CLP. Effect of CORM-2 on cardiac injury of septic mice In septic mice, significant increases of myocardial enzymes in serum were seen at 6, 12 and 24 h after CLP surgery ( 0.001. Fig ?Fig2A).2A). The administration of CORM-2 significantly abolished this elevation ( 0.001). After administration of CORM-2, the elevation of IL-1 and TNF- levels were effectively abolished ( 0.05). Open in a AP24534 enzyme inhibitor separate window Fig 2 Effect of carbon monoxide releasing molecule-2 (CORM-2) on cardiac injury of septic mice. Mice were challenged with CLP and treated with CORM-2 or iCORM-2. A, Myocardial enzymes, including LDH, AST, CK and CK-MB, were detected at 6, 12 and 24 h after CLP surgery. Myocardial enzymes were increased at 6, 12 and 24 h after CLP. administration of CORM-2 significantly abolished this elevation. B, 24 h following CLP surgery, apoptosis of cardiac cells was assessed by Annexin V/PI staining. Representative flow cytometry (FC) images were shown. C, FC results showed a high apoptotic rate at 24 h after CLP surgery and then a significant decrease in AP24534 enzyme inhibitor the presence of CORM-2. D, At 24 h after CLP, the levels of interleukin (IL)-1 and tumor necrosis factor (TNF-) in serum were markedly increased compared with those in sham mice. After administration of CORM-2, the elevation of TNF- and IL-1 levels were effectively abolished. Data were shown as mean SD, * 0.001). The elevation was abolished after administration of CORM-2 ( 0.05). Consistent results were found in MMP alternations. As shown in Fig ?Fig3B,3B, mitochondrial depolarization was observed 24 h after surgery ( 0.001) and this decline was prevented by CORM-2 ( 0.01) (Fig ?(Fig3C).3C). Treatment with CORM-2 significantly inhibited the alterations ( 0.05). Representative electron microscopic images (Fig ?(Fig3D)3D) showed some early changes of mitochondria like breakage of inner and outer membranes, variable swelling and lose of matrix density in CLP mice compared with sham mice. CORM-2 effectively inhibited these structural destructions. Open in a separate window Fig 3 Effect of carbon monoxide releasing molecule-2 (CORM-2) on mitochondrial damage in the heart of septic mice. Mice were challenged with.