Anti-inflammatory effect of corynoline isolated from Corydalis bungeana Turcz

To isolate corynoline from Corydalis bungeana Turcz. and study its anti-inflammatory mechanism via TLRs/NF-kappa B signal pathway. Corynoline was extracted by 80% ethanol and purified by silicagel column chromatography. The structure and purity of corynoline was determined by UPLC, MS, H-1 NMR and C-13 NMR. In the course of experiment, the cytotoxicity of corynoline was evaluated by MTT assay. And the inflammation model was established by RAW264.7 macrophages induced by lipopolysaccharide(LPS), which was intervened by corynoline. The expression levels of TLR4, TLR2 and nuclear transcription factor-kappa B(NF-kappa B) signaling pathways related proteinsin RAW264. 7 macrophages were detected by Western blot. Furthermore, the expression of NF-kappa B p65 mRNA and nuclear p65 were determined by the real-time fluorescence quantitative PCR(RT-qPCR) and Western blot. Results showed that 5-40 mu mol/L corynoline reduced the expression level of TLR4 and TLR2, and inhibited the phosphorylation level of I kappa B alpha and the phosphorylation and nuclear translocation of p65 at gene and protein levelin a dose-dependent manner in LPS-induced RAW264. 7 cells. This study indicated the protective effect of corynoline on LPS-induced RAW264. 7 macrophages may be related with the inhibition of TLRs/NF-kappa B inflammatory signaling pathway.