Suppression of Microglia-Mediated Neuroninflammation by Small Molecule NF-kB Inhibitors

Brian Astry, Rainer Grant, Katherine Bradley, Kathryn Brumbaugh, Jim Rivard, Makenna Carlson-Habegger, Andrew Hudacek, and Greta Wegner


It has been shown that long-term production of proinflammatory cytokines by activated microglia plays a role in neurodegenerative disorders like Parkinson’s disease and multiple sclerosis. Here we investigated if inhibition of the
NF-κB signaling pathway, an important factor in the inflammatory response, could alter the chemokine and cytokine profile of the BV-2 mouse microglia cell line. Using the Toll-like Receptor 4 (TLR4) agonist lipopolysaccharide (LPS), we stimulated BV-2 cells and subsequently treated them with two different small molecule inhibitors, MLN4924 (a NEDD8 E1 Activating Enzyme inhibitor) and Celastrol (a NF-κB inhibitor). Cell supernatants and lysates were measured for 26 mouse cytokines/chemokines/growth factors using a R&D Systems Mouse Magnetic Luminex® Assay. LPS stimulation increased multiple proinflammatory cytokines, including IL-1α/IL-1F1, IL-1β/IL-1F2, IL-6, IL-27, and TNF-α, and the chemokines CCL4/MIP-1β, CCL5/RANTES, CXCL1/KC and CXCL10/IP-10/CRG-2. Treatment with either MLN4924 or Celastrol resulted in significant decreases in the levels of the proinflammatory cytokines and chemokines. To determine the effects of the inhibitors on the NF-κB pathways, cell lysates of activated cells were analyzed for inhibition of the canonical (IκBα degradation) pathway. The data suggest that MLN4924 and Celastrol inhibits the degradation of IκBα. These results indicate that the small molecule inhibitors, MLN4924 and Celastrol, may be used to suppress microglia, NF-κB-mediated neuroinflammation.

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