3.4 Compound 270 ameliorates the LPS-induced inflammatory response in bone marrow derived macrophages (BMDMs)
To further clarify that whether 270 possess the direct inhibitory effect on inflammation in macrophages, we treated primary mouse BMDMs with LPS alone or combined with 270 and then detected the NF-κB and JNK signaling pathways, the secretion of inflammatory cytokines and the expression of pro-inflammatory genes. We found that the LPS-induced activation of NF-κB and JNK signaling pathways in BMDMs were blocked by the pretreatment of 270, evidenced by the reduction phosphorylation of IKK, NF-κB and JNK, and prevention degradation of IκB-α in a dose-dependent manner (Figure 4A,B). The release of TNF-α, IL-6 and MCP 1 from the BMDMs triggered by LPS was also mitigated by 270 administration (Figure 4C). Moreover, 270 remarkably down-regulated the over-expression of IL-1β, IL-6, MCP 1, Nos2 and COX2 in LPS-stimulated BMDMs (Figure 4D). In line with the inhibitory ability in RAW 264.7 macrophages. Intriguingly, the repressive effects of 270 on the LPS-induced inflammatory response were more dramatic in BMDMs than in RAW 264.7 macrophages. Our investigations revealed that NF-κB inhibitor 270 can against macrophage-mediated inflammatory response, which may have potential therapeutic effect in inflammation-related diseases.