Anomalin attenuates LPS-induced acute lungs injury through inhibition of AP-1 signaling

• Published in: International immunopharmacology Vol. 73; pp. 451 - 460
• Main Authors: Khan, Ashrafullah, Khan, Salman, Ali, Hassan, Shah, Kifayat Ullah, Ali, Hussain, Shehzad, Omer, Onder, Alev, Kim, Yeong Shik
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-08-2019; Elsevier BV
• Subjects: Activator protein 1; Acute Lung Injury - chemically induced; Acute Lung Injury - drug therapy; Acute Lung Injury - metabolism; Akt; AKT protein; Animals; Anomalin; Anti-oxidants; Antioxidants; Attenuation; Binding; Body temperature; Catalase; Cell activation; Coumarins - pharmacology; Coumarins - therapeutic use; Cytokines; Cytokines - metabolism; Deoxyribonucleic acid; DNA; DNA - metabolism; Extracellular signal-regulated kinase; IL-1β; Inflammation; Inhibition; Interleukin 6; Kinases; Leukocytes; Lipopolysaccharides; Lung injury; Lungs; Macrophages; Male; MAP kinase; MAPKs; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases - metabolism; Nitric Oxide - metabolism; Oxidative stress; Pain perception; Proteins; RAW 264.7 Cells; Signal transduction; Signal Transduction - drug effects; Signaling; Transcription Factor AP-1 - antagonists & inhibitors; Transcription Factor AP-1 - metabolism; Transcription factors; Tumor necrosis factor-α; Anti-oxidants; Cytokines; Anomalin; Inflammation; Akt; Lung injury; MAPKs
• ISSN: 1567-5769; 1878-1705
• DOI: 10.1016/j.intimp.2019.05.032

In the present study, the anomalin was investigated to determine the protective effects and underlying mechanism against LPS-induced acute lung injury in mice. Anomalin administration 30 min after the LPS injection, significantly attenuated the mechanical allodynia, decrease body temperature, and improved the histological changes and inhibited the infiltration of leukocytes. The anomalin treatment markedly inhibited the production of pro-inflammatory mediators such as cytokines (IL-1β, IL-6 and TNF-α) and NO in contrast to the LPS treated groups. Similarly, the anomalin also enhanced the level of anti-oxidant enzymes such as GST, GSH, Catalase and inhibited oxidative stress marker such as MDA. In order to explore the molecular mechanism the effect of anomalin was evaluated for mitogen activated protein kinases (MAPK) in LPS-stimulated RAW264.7 cells. The anomalin treatment significantly attenuated the MAPK proteins such as ERK1/2, JNK and p38 (which is downstream signaling proteins to the MAPKKKs and MAPKKs protein) in the RAW264.7 macrophages using western blot analysis. Furthermore, the western blot analysis showed that anomalin treatment significantly inhibited the activation of the Akt proteins in the RAW264.7 macrophages. The AP-1 served as downstream target for the MAPK pathways and the blocking MAPK pathways is responsible for the inhibition of the AP-1 protein. The AP-1/DNA binding was assessed in the RAW264.7 cells using EMSA. The anomalin treatment significantly restricted the AP-1/DNA binding activity and the decrease in the AP-1/DNA binding activity might be contributed due to the upstream inhibition of the MAPKs signaling.
•The anomalin treatment markedly attenuated the inflammatory mediators and induced the anti-oxidant enzymes.•The anomalin administration markedly inhibited the MAPKs and Pi3K signaling in RAW cells.•The anomalin administration significantly inhibited AP-1/DNA interaction using EMSA.