Non-Steroidal Anti-Inflammatory Drugs Alter Nuclear Factor- kβ in A Class Specific Manner

Abstract

The capacity of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) to affect the NF-κβ dependent pathway was examined. The NSAIDs were co-cultured with RAW 264.7 cells transfected with a plasmid encoding Secreted Embryonic Alkaline Phosphatase (SEAP) under the control of NF-κβ. SEAP production was initiated by stimulating the cells with E. coli derived Lipopolysaccharide (LPS). Diclofenac, indomethacin, celecoxib, tolfenamic acid, flufenamic acid, and piroxicam inhibited SEAP production. Ketoprofen, phenylbutazone, aspirin, and acetaminophen did not affect SEAP production. TNFα stimulation of HEK293 cells transfected with a SEAP reporter gene under the control of NF-κβ was used to determine whether the NSAIDs were working directly on the NF-κβ pathway or on TLR4 signaling. TNFα stimulated HEK293 cells showed that diclofenac and flunixin inhibited SEAP production, but aspirin did not. Meloxicam and diclofenac both inhibited the amount of p65 released after LPS stimulation of the RAW 264.7 cells while flunixin did not affect p65 release. Flunixin had no effect on an NF-κβ gel shift assay. These results demonstrate that NSAIDs in four of the six NSAID families can inhibit NFκβ pathway, albeit via different points in the pathway.