Tissue samples were homo genized and the cytoplasmic and nuclear proteins were extracted respectively according to instructions of Nuclear and Cytoplasmic Protein Extraction Kit. Protein concentrations were determined by BCA protein assay kit. Samples were separated on a denaturing 12% pol yacrylamide gel ARQ197 cost and transferred to a nitrocellulose membrane. NF B p65 protein was detected by chemi luminescence using a rabbit polyclonal antibody accord ing to the manufacturers instructions. Statistical analyses Data were entered into a database and analyzed using SPSS software, and expressed as means SEM. Statisti cally significant differences between groups were deter mined by ANOVA followed by Students t test. Significance was accepted when p 0. 05.

Results Effect of Butyrate on MPO Activity and NO Concentrations in Lung Tissues of Mice with ALI After LPS administration, the MPO activity in lung tis sues was significantly and continuously increased com pared with the control and butyrate groups from 1 to 24 hours. In addition, the concentrations of NO were significantly increased at 1 hour and peaked at 3 hours after LPS administration. However, in LPS butyrate group, butyrate pretreatment markedly decreased the MPO activity and NO concentrations at different time point. Effect of Butyrate on the Concentrations of TNF a and IL 1b in BALF of Mice with ALI The concentrations of TNF a and IL 1b in BALF were significantly increased at 1 hour and peaked at 3 hours after LPS administration. Butyrate pre treatment efficiently reduced the production of TNF a and IL 1b at different time point.

Effect of Butyrate on the Lung edema of Mice with ALI Compared with the control and butyrate groups, the lung wet dry weight ratios were significantly and con tinuously increased from 1 to 24 hours after LPS admin istration. The increase of the lung wet dry weight ratios was significantly reduced by butyrate administration at different time point. Effect of Butyrate on the Pulmonary Histopathological Changes of Mice with ALI Lung tissues from the control and butyrate groups showed a normal structure and no histopathological changes under a light microscope. In LPS group, the lungs stained with hematoxylin eosin indicated widespread alveolar wall thickness caused by of expression of NF B p65 in cytoplasm and nucleus, same as control group, were observed with administration of butyrate alone at each time point.

Discussion The different results were observed in previous studies in vitro in terms of HDAC inhibitors anti inflammatory effects. ITF2357, a HDAC inhibitor, reduced IL 1, TNF a and interferon g expression in LPS stimulated human peripheral blood mononuclear cells. Butyrate also reduced IL 12 production by human blood monocytes, and inhibited NO production in RAW macrophage cells. However, pharmacological Carfilzomib inhibition of HDAC edema, severe hemorrhage in the alveolus, alveolus col lapse and obvious inflammatory cells infiltration.