Differential cell count bloodLeukocytes were quantified by flowcytometry using TruCount-Tubes and differentiated according to their side-scatter/forward-scatter properties and CD45 and Gr-1 expression.Quantification of cytokinesCytokines were quantified from total protein of flushed homogenized left lungs and compound libraries blood samples (BioRad, Hercules, CA, USA).Measurement of Alanine transaminase levelsAlanine transaminase (ALT) levels were measured by routine laboratory test at the Institute of Laboratory Medicine and Pathobiochemistry of the Charit�� – Universit?tsmedizin Berlin.Statistic analysesGroups were compared using One-Way-ANOVA following Newman-Keuls post test. For comparison of two groups Mann-Whitney U-Test was applied. P-values < 0.05 were considered significant. Data are represented as mean +/- SEM.

ResultsSimvastatin prevented oxygenation failure in VILIThe decline of the peripheral oxygen saturation (SpO2) observed in ventilated mice was prevented by Simvastatin treatment (Figure (Figure1a).1a). At the termination of the experiment, blood gas analysis was performed in arterial blood samples. The P/F ratio was higher in simvastatin treated mice (Figure (Figure1b1b).Figure 1Simvastatin improved oxygenation in VILI. (a) Peripheral Oxygen Saturation (SpO2) was monitored continuously and (b) P/F ratio was assessed at the end of the 6 h ventilation period in simvastatin (6 h Vent. + Simva) or sham (6 h Vent.) treated mice. Simvastatin …Simvastatin reduced VILI-associated pulmonary vascular leakageMV induced a marked increase of pulmonary microvascular permeability in mice, indicated by an elevated HSA BAL/plasma ratio.

Pulmonary hyperpermeability was decreased by Simvastatin treatment (Figure (Figure22).Figure 2Simvastatin reduced VILI-associated lung hyperpermeability. Human serum albumin (HSA; 1 mg) was injected 90 minutes prior to termination of the experiment. In non-ventilated simvastatin (NV + Simva) or sham (NV) treated mice, and in ventilated and simvastatin …Simvastatin attenuated endothelial injury in ventilated miceNon-ventilated, untreated or simvastatin treated mice exhibited intact alveolar epithelium and capillary endothelium (Figure 3a-d). Capillary endothelial cells of ventilated and untreated mice were swollen and showed loss of intracellular vesicles and caveolae (Figure 3e, f).

In ventilated and simvastatin treated lungs, endothelial cells displayed fewer signs of injury as compared to ventilated and untreated mice. Swelling of endothelial cells occurred only sporadically, and normal distribution of vesicles and caveolae was preserved by Cilengitide simvastatin (Figure 3g, h).Figure 3Simvastatin reduced VILI-associated endothelial injury. In lung sections of non ventilated, sham treated mice (NV) (a, b) and non-ventilated, simvastatin treated mice (NV + Simva) (c, d), structurally intact capillaries containing numerous caveolae and …