A novel synthetic compound MCAP suppresses LPS-induced murine microglial activation in vitro via inhibiting NF-kB and p38 MAPK pathways

Abstract

Aim: To investigate the anti-neuroinflammatory activity of a novel synthetic compound, 7-methylchroman-2-carboxylic acid N-(2-trifluoromethyl) phenylamide (MCAP) against LPS-induced microglial activation in vitro. Methods: Primary mouse microglia and BV2 microglia cells were exposed to LPS (50 or 100 ng/mL). The expression of iNOS and COX-2, proinflammatory cytokines, NF-κB and p38 MAPK signaling molecules were analyzed by RT-PCR, Western blot and ELISA. The morphological changes of microglia and nuclear translocation of NF-ĸB were visualized using phase contrast and fluorescence microscopy, respectively. Results: Pretreatment with MCAP (0.1, 1, 10 μmol/L) dose-dependently inhibited LPS-induced expression of iNOS and COX-2 in BV2 microglia cells. Similar results were obtained in primary microglia pretreated with MCAP (0.1, 0.5 μmol/L). MCAP dose-dependently abated LPS-induced release of TNF-α, IL-6 and IL-1β, and mitigated LPS-induced activation of NF-κB by reducing the phosphorylation of IκBα in BV2 microglia cells. Moreover, MCAP attenuated LPS-induced phosphorylation of p38 MAPK, whereas SB203580, a p38 MAPK inhibitor, significantly potentiated MCAP-caused inhibition on the expression of MEF-2 (a transcription factor downstream of p38 MAPK). Conclusion: MCAP exerts anti-inflammatory effects in murine microglia in vitro by inhibiting the p38 MAPK and NF-κB signaling pathways and proinflammatory responses. MCAP may be developed as a novel agent for treating diseases involving activated microglial cells.

Interference with Protease-activated Receptor 1 Alleviates Neuronal Cell Death Induced by Lipopolysaccharide-Stimulated Microglial Cells through the PI3K/Akt Pathway

“…This phenomenon led us to hypothesize that SCH may interact with proteins in the NF-κB pathway. Studies confirm that NF-κB can be activated by a series of stimuli42, and that activated NF-κB can be translocated from the cytoplasm to the nucleus where it interacts with κB elements in the promoter region of a variety of inflammatory response genes, and activates their transcription43. In spite of previous reports consistent with our results, the molecular mechanism underlying neuroprotective activity of SCH still remains to be elucidated.…”

Chlorogenic Acid Attenuates Dextran Sodium Sulfate-Induced Ulcerative Colitis in Mice through MAPK/ERK/JNK Pathway

“…p38 is an important protein in ulcerative colitis, which can be activated by a variety of cytokines, such as hormones, IL-1, and TNF- α . p38 can make TATA binding protein phosphorylation in the downstream nuclear NF- κ B complex, regulate the transcription activity of NF- κ B, and then regulate the inflammatory response [66]. After ERK is activated, it can regulate the downstream targets NF- κ B, Bcl-2, and so on, thus affecting the inflammatory response and cell apoptosis [67].…”

Scoparone Inhibits LPS-Simulated Inflammatory Response by Suppressing IRF3 and ERK in BV-2 Microglial Cells

“…The BV2 microglia cells were acquired as described previously [35,36]. The BV-2 microglial cells were cultured in DMEM supplemented with 5% FBS and 1% of 100 units/mL of penicillin/streptomycin at 37 °C in a humidified 5%-CO 2 incubator.…”