trichostatin-a and Acute-Lung-Injury

Acute lung disease is characterized by inflammation. This research aimed to investigate effect of trichostatin A (TSA) on microRNA-146a (miR-146a) and tumor necrosis factor α (TNF-α) in lipopolysaccharide (LPS)-induced alveolar macrophage injury model.. Rat alveolar macrophage, NR8383, was cultured and induced using LPS to establish acute lung injury model in vitro level. Cell Counting Kit-8 (CCK-8) assay was used to determine cell viability of NR8383 cells. TSA was administrated to LPS-induced NR8383 cells. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) assay was utilized to evaluate TNF-α and miR-146a mRNA expression in LPS and/or TSA treated NR8383 cells. Enzyme-link immunosorbent assay (ELISA) was used to examine TNF-α levels.. This study selected 1 ng/ml and 10 ng/ml TSA as the optimal concentrations for treating NR8383 cells. LPS-induced acute lung injury model was successfully established. TSA administration significantly enhanced accounts of LPS-stimulated NR8383 cells. LPS induction significantly increased miR-146a mRNA expression in NR8383 cells compared to NR8383 cells (p<0.05). TSA administration significantly reduced the levels of TNF-α in LPS-induced NR8383 cells compared to those in LPS-induced NR8383 cells (p<0.05). TSA administration significantly enhanced miR-146a expression in LPS-induced NR8383 cells compared to that in LPS-induced NR8383 cells (p<0.05).. TSA administration exerted anti-inflammation functions in LPS-induced acute lung injury model in vitro, which might be triggered by inhibiting TNF-α molecule and upregulating miR-146a expression. The present data hint that TSA could be considered as a potential therapeutic agent for treating acute lung injury.

Topics: Acute Lung Injury; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Survival; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Hydroxamic Acids; Lipopolysaccharides; MicroRNAs; Rats; Tumor Necrosis Factor-alpha; Up-Regulation

Pulmonary infections remain the most common cause of Acute Respiratory Distress Syndrome (ARDS), a pulmonary inflammatory disease with high mortality, for which no targeted therapy currently exists. We have previously demonstrated an ameliorated syndrome with early, broad spectrum Histone Deacetylase (HDAC) inhibition in a murine model of gram-negative pneumonia-induced Acute Lung Injury (ALI), the underlying pulmonary pathologic phenotype leading to ARDS. With the current project we aim to determine if selective inhibition of a specific HDAC leads to a similar pro-survival phenotype, potentially pointing to a future therapeutic target.. C57Bl/6 mice underwent endotracheal instillation of 30×10Escherichia coli (strain 19138) versus saline (n = 24). Half the infected mice were administered Trichostatin A (TSA) 30 min later. All animals were sacrificed 6 h later for tissue sampling and HDAC quantification, while another set of animals (n = 24) was followed to determine survival. Experiments were repeated with selective siRNA inhibition of the HDAC demonstrating the greatest inhibition versus scrambled siRNA (n = 24).. TSA significantly ameliorated the inflammatory phenotype and improved survival in infected-ALI mice, and HDAC7 was the HDAC with the greatest transcription and protein translation suppression. Similar results were obtained with selective HDAC7 siRNA inhibition compared with scrambled siRNA.. HDAC7 appears to play a key role in the inflammatory response that leads to ALI after gram-negative pneumonia in mice.

Topics: Acute Lung Injury; Animals; Disease Models, Animal; Escherichia coli; Escherichia coli Infections; Histone Deacetylase Inhibitors; Hydroxamic Acids; Male; Mice; Mice, Inbred C57BL; Pneumonia, Bacterial

Acute lung injury (ALI) and idiopathic pulmonary fibrosis (IPF) are both serious public health problems with high incidence and mortality rate in adults, and with few drugs available for the efficient treatment in clinic. In this study, we identified that two known histone deacetylase (HDAC) inhibitors, suberanilohydroxamic acid (SAHA, 1) and its analogue 4-(dimethylamino)-N-[7-(hydroxyamino)-7-oxoheptyl]benzamide (2), are effective inhibitors of Leukotriene A4 hydrolase (LTA4H), a key enzyme in the biosynthesis of leukotriene B4 (LTB4), across a panel of 18 HDAC inhibitors, using enzymatic assay, thermofluor assay, and X-ray crystallographic investigation. Importantly, both 1 and 2 markedly diminish early neutrophilic inflammation in mouse models of ALI and IPF under a clinical safety dose. Detailed mechanisms of down-regulation of proinflammatory cytokines by 1 or 2 were determined in vivo. Collectively, 1 and 2 would provide promising agents with well-known clinical safety for potential treatment in patients with ALI and IPF via pharmacologically inhibiting LAT4H and blocking LTB4 biosynthesis.

Topics: Acute Lung Injury; Animals; Epoxide Hydrolases; Female; Histone Deacetylase Inhibitors; Idiopathic Pulmonary Fibrosis; Leukotriene B4; Mice; Mice, Inbred C57BL; Neutrophils

Mechanical ventilation (MV) used in patients with acute respiratory distress syndrome (ARDS) can cause diffuse lung inflammation, an effect termed ventilator-induced lung injury, which may produce profound pulmonary fibrogenesis. Histone deacetylases (HDACs) and serine/threonine kinase/protein kinase B (Akt) are crucial in modulating the epithelial-mesenchymal transition (EMT) during the reparative phase of ARDS; however, the mechanisms regulating the interactions among MV, EMT, HDACs, and Akt remain unclear. We hypothesized that trichostatin A (TSA), a HDAC inhibitor, can reduce MV-augmented bleomycin-induced EMT by inhibiting the HDAC4 and Akt pathways.. Five days after bleomycin treatment to mimic acute lung injury (ALI), wild-type or Akt-deficient C57BL/6 mice were exposed to low-tidal-volume (low-VT, 6 mL/kg) or high-VT (30 mL/kg) MV with room air for 5 h after receiving 2 mg/kg TSA. Nonventilated mice were examined as controls.. Following bleomycin exposure in wild-type mice, high-VT MV induced substantial increases in microvascular leaks; matrix metalloproteinase-9 (MMP-9) and plasminogen activator inhibitor-1 proteins; free radical production; Masson's trichrome staining; fibronectin, MMP-9, and collagen 1a1 gene expression; EMT (identified by increased localized staining of α-smooth muscle actin and decreased staining of E-cadherin); total HDAC activity; and HDAC4 and Akt activation (P < 0.05). In Akt-deficient mice, the MV-augmented lung inflammation, profibrotic mediators, EMT profiles, Akt activation, and pathological fibrotic scores were reduced and pharmacologic inhibition of HDAC4 expression was triggered by TSA (P < 0.05).. Our data indicate that TSA treatment attenuates high-VT MV-augmented EMT after bleomycin-induced ALI, in part by inhibiting the HDAC4 and Akt pathways.

Topics: Actins; Acute Lung Injury; Animals; Bleomycin; Cadherins; Collagen Type I; Epithelial-Mesenchymal Transition; Free Radicals; Gene Expression Regulation; Histone Deacetylase Inhibitors; Histone Deacetylases; Humans; Hydroxamic Acids; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Mice, Knockout; Proto-Oncogene Proteins c-akt; Respiration, Artificial; Serpin E2; Signal Transduction; Tidal Volume; Ventilator-Induced Lung Injury

Trichostatin A (TSA) is a histone deacetylase inhibitor with anti-inflammatory effects. Nonetheless, little information is available about the effect of TSA in ischemia-reperfusion (IR)-induced lung injury. In a perfused rat lung model, IR was induced by 40min of ischemia followed by 60min of reperfusion. The rat lungs were randomly divided into several groups including control, control+TSA (0.1mg/kg), IR, and IR+various dosages of TSA (0.05, 0.075, 0.1mg/kg). Bronchoalveolar lavage fluids and lung tissues were obtained and examined at the end of the experiment. TSA dose-dependently diminished IR-induced increased vascular permeability and edema, pulmonary artery pressure, and histological changes in the lungs. Additionally, TSA suppressed lavage tumor necrosis factor-α and cytokine-induced neutrophil chemoattractant concentrations, cell infiltration, and myeloperoxidase-positive cells in the lung tissue. Furthermore, TSA attenuated the phosphorylation of extracellular signal-regulated kinase, p38, and c-Jun N-terminal kinase, degradation of the inhibitor of nuclear factor (NF)-κB, and nuclear NF-κB levels. TSA also decreased poly (ADP-ribose) polymerase but enhanced acetylated histone H3 acetylation, Bcl-2, and mitogen-activated protein kinase phosphatase-1 (MKP-1) expression in IR lung tissue. Therefore, TSA exerted a protective effect on IR-induced lung injury via increasing histone acetylation and MKP-1 protein expression, repressing NF-κB, mitogen-activated protein kinase, and apoptosis signaling pathways.

Topics: Acetylation; Acute Lung Injury; Animals; Anti-Inflammatory Agents; Apoptosis; Arterial Pressure; Bronchoalveolar Lavage Fluid; Chemokine CXCL1; Dual Specificity Phosphatase 1; Histone Deacetylase Inhibitors; Histones; Hydroxamic Acids; Male; Mitogen-Activated Protein Kinases; NF-kappa B; Pulmonary Artery; Rats, Sprague-Dawley; Reperfusion Injury; Signal Transduction; Tumor Necrosis Factor-alpha

Acute lung injury (ALI) during sepsis is characterized by bilateral alveolar infiltrates, lung edema and respiratory failure. Here, we examined the efficacy the DNA methyl transferase (DNMT) inhibitor 5-Aza 2-deoxycytidine (Aza), the histone deacetylase (HDAC) inhibitor Trichostatin A (TSA), as well as the combination therapy of Aza and TSA (Aza+TSA) provides in the protection of ALI. In LPS-induced mouse ALI, post-treatment with a single dose of Aza+TSA showed substantial attenuation of adverse lung histopathological changes and inflammation. Importantly, these protective effects were due to substantial macrophage phenotypic changes observed in LPS-stimulated macrophages treated with Aza+TSA as compared with untreated LPS-induced macrophages or LPS-stimulated macrophages treated with either drug alone. Further, we observed significantly lower levels of pro-inflammatory molecules and higher levels of anti-inflammatory molecules in LPS-induced macrophages treated with Aza+TSA than in LPS-induced macrophages treated with either drug alone. The protection was ascribed to dual effects by an inhibition of MAPK-HuR-TNF and activation of STAT3-Bcl2 pathways. Combinatorial treatment with Aza+TSA reduces inflammation and promotes an anti-inflammatory M2 macrophage phenotype in ALI, and has a therapeutic potential for patients with sepsis.

Topics: Acute Lung Injury; Animals; Azacitidine; Decitabine; Drug Combinations; Endotoxemia; Epigenesis, Genetic; Histone Deacetylases; Humans; Hydroxamic Acids; Inflammation; Lipopolysaccharides; Macrophages; Methyltransferases; Mice; Sepsis; Signal Transduction

Impairment of tissue fluid homeostasis and migration of inflammatory cells across the vascular endothelial barrier are crucial factors in the pathogenesis of acute lung injury (ALI). The goal for treatment of ALI is to target pathways that lead to profound dysregulation of the lung endothelial barrier. Although studies have shown that chemical epigenetic modifiers can limit lung inflammation in experimental ALI models, studies to date have not examined efficacy of a combination of DNA methyl transferase inhibitor 5-Aza 2-deoxycytidine and histone deacetylase inhibitor trichostatin A (herein referred to as Aza+TSA) after endotoxemia-induced mouse lung injury. We tested the hypothesis that treatment with Aza+TSA after lipopolysaccharide induction of ALI through epigenetic modification of lung endothelial cells prevents inflammatory lung injury. Combinatorial treatment with Aza+TSA mitigated the increased endothelial permeability response after lipopolysaccharide challenge. In addition, we observed reduced lung inflammation and lung injury. Aza+TSA also significantly reduced mortality in the ALI model. The protection was ascribed to inhibition of the eNOS-Cav1-MLC2 signaling pathway and enhanced acetylation of histone markers on the vascular endothelial-cadherin promoter. In summary, these data show for the first time the efficacy of combinatorial Aza+TSA therapy in preventing ALI in lipopolysaccharide-induced endotoxemia and raise the possibility of an essential role of DNA methyl transferase and histone deacetylase in the mechanism of ALI.

Topics: Acetylation; Acute Lung Injury; Animals; Azacitidine; Blotting, Western; Capillary Permeability; Cell Proliferation; Cell Survival; Chromatin Immunoprecipitation; Decitabine; Disease Models, Animal; Drug Therapy, Combination; Endothelial Cells; Endotoxemia; Enzyme Inhibitors; Flow Cytometry; Fluorescent Antibody Technique; Hydroxamic Acids; In Situ Nick-End Labeling; Inflammation; Lung; Male; Methylation; Mice; Mice, Inbred C57BL; Real-Time Polymerase Chain Reaction

The histone deacetylase (HDAC) inhibitors have emerged as the useful reagents that epigenetically modulate the expression of various genes. In the present study, the effects of HDAC inhibitors on the expression of inflammation-related genes and lung injury during sepsis were investigated.. Mice were pretreated with two structurally unrelated HDAC inhibitors, Trichostatin A (TSA) and sodium butyrate (SB). Thirty minutes later, mice underwent cecal ligation and puncture (CLP)-induced sepsis. Lung injury and the expression of inflammation-related molecules were determined. In addition, survival was assessed post-CLP.. Our results indicated that administration of TSA or SB alleviated sepsis-induced lung injury. This was accompanied by reduced neutrophil infiltration, decreased intercellular adhesion molecule-1 (ICAM-1) and E-selectin expression in lung tissue, and lower interleukin-6 (IL-6) level in plasma. In addition, treatment with HDAC inhibitors significantly prolonged the survival time of CLP mice.. These data indicated that the HDAC inhibitors, based on modulating the key enzymes linked to acetylation modification, effectively attenuate intrapulmonary inflammatory response, thus significantly alleviating lung injury during sepsis.

Topics: Acetylation; Acute Lung Injury; Animals; Butyrates; Cecum; E-Selectin; Histone Deacetylase Inhibitors; Hydroxamic Acids; Intercellular Adhesion Molecule-1; Interleukin-6; Ligation; Lung; Male; Mice; Mice, Inbred C57BL; Punctures; Sepsis