Oleanane triterpenoids with C-14 carboxyl group from inhibited LPS-induced macrophages activation by suppressing the NF-B signaling pathway.

BACKGROUND

Excessive inflammation poses significant risks to human physical and mental health. , a traditional Miao medicine, is renowned for its anti-inflammatory properties. However, the specific anti-inflammatory effects and mechanisms of many compounds within this plant remain unclear. This study aims to investigate the anti-inflammatory effects and mechanisms of two characteristic oleanane triterpenoids, 3-acetoxyolean-12-en-27-oic acid () and 3-acetoxyolean-12-en-27-oic acid (), isolated from , using lipopolysaccharide (LPS)-induced Macrophages.

METHODS

The anti-inflammatory effects and mechanisms of compounds and were investigated by establishing an LPS-induced inflammation model in RAW 264.7 cells and THP-1 cells. Nitric oxide (NO) levels were assessed using the Griess method. The concentrations of tumor necrosis factor-alpha (TNF-), interleukin-6 (IL-6), and interleukin-1beta (IL-1) were measured via enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was determined using western blotting and quantitative real-time PCR (qRT-PCR). Additionally, the phosphorylation level of p65 in nuclear factor-kappa B (NF-B) was assessed through western blotting. The nuclear translocation of NF-B p65 was assessed through immunofluorescence staining. Finally, the binding affinity of the compounds to NF-B p65 target was validated through molecular docking.

RESULTS

Compounds and significantly inhibited the expression of NO, TNF-, IL-6, IL-1, COX-2, and iNOS in LPS-induced Macrophages. Mechanistically, they attenuated the activation of the NF-B signaling pathway by downregulating the phosphorylation level and nuclear translocation of p65.

CONCLUSION

This study elucidates the anti-inflammatory activities and potential mechanism of the characteristic oleanane triterpenoids with C-14 carboxyl group, compounds and , in LPS-induced Macrophages by inhibiting the NF-B signaling pathway for the first time. These findings suggest that these two compounds hold promise as potential candidates for anti-inflammatory interventions in the future.