Impact of evodiamine on IL-1β-induced chondrocyte inflammatory injury by regulating the Nrf2/HO-1/NF-κB signaling pathway
Objective To investigate the impact of evodiamine(Evo)on interleukin-1 beta(IL-1(3)-induced chondrocyte inflammatory injury and its regulation on the nuclear factor erythrocyte 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/nuclear factor kappa B(NF-κB)signaling pathway.Methods Human chondrocyte cells line(HCCs)were grouped into the control group,model group,low-dose group,high-dose group,and high-dose Evo+Nrf2 inhibitor ML385 group.In addition to HCCs treated with blank control in the control group,those in the remaining groups were induced with 50 ng/mL IL-1β.Treatment of 10 µmol/L Evo,30 µmol/L Evo,and 30-min induction of 2 µmol/L ML385 followed by 30 µmol/L Evo treatment was conducted in the latter three groups,respectively.Cell viability and apoptosis were detected by cell counting kit-8(CCK-8)assay and flow cytometry,respectively.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of interleukin-6(IL-6),IL-18,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),superoxide dismutase(SOD)in the supernatant of HCCs cells.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was applied to detect the mRNA levels of inflammatory mediators cyclooxygenase 2(COX-2)and inducible nitric oxide synthase(iNOS)in HCCs cells.Western blot was applied to detect the protein expressions of Nrf2/HO-1/NF-κB pathway-related proteins in HCCs cells.Results Compared with those of the control group,HCCs in the model group had significantly higher apoptotic rate,supernatant levels of IL-6,IL-18,TNF-α and MDA,mRNA levels of COX-2 and iNOS,protein levels of p-IκB-α,cytoplasmic Nrf2 and nuclear p65 NF-κB,but lower cell viability,SOD activity,and protein levels of HO-1,IκB-α,nuclear Nrf2 and cytoplasmic p65 NF-κB(P＜0.05).Compared with those of the model group,HCCs in the low-dose and high-dose Evo groups had significantly lower apoptotic rate,supernatant levels of IL-6,IL-18,TNF-α and MDA,mRNA levels of COX-2 and iNOS,protein levels of p-IκB-α,cytoplasmic Nrf2 and nuclear p65 NF-κB,but higher cell viability,SOD activity,and protein levels of HO-1,IκB-α,nuclear Nrf2 and cytoplasmic p65 NF-κB(P＜0.05).The above indicators were significantly different between the low-dose and high-dose Evo groups(P＜0.05).ML385 treatment significantly reversed the regulatory effect of high-dose Evo on the above indicators(P＜0.05).Conclusion Evo may improve IL-1β-induced chondrocyte inflammatory injury by activating Nrf2 and HO-1 and inhibiting the downstream transcription factor NF-κB.
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