Effects of neohesperidin on the TLR4/NLRP3 signaling pathway in LPS-induced macrophage inflammatory model
Objective To investigate the effect of neohesperidin(NHP)on the Toll-like receptor 4(TLR4)/NOD-like receptor family pyrin domain containing 3(NLRP3)signaling pathway in lipopolysaccharide(LPS)-induced macrophage inflammation model.Methods RAW264.7 cells were divided into five groups:Control group,model group,experimental-L group,experimental-H group and MCC950 group.The control group was cultured normally,while the other groups were stimulated with 100 ng·mL-1 of LPS to induce the macrophage inflammation model.The experimental-L,-H groups were treated with 100 and 400 μmol·L-1 of NHP,respectively,while the MCC950 group was treated with 30 µmol·L-1 of the NLRP3 inhibitor MCC950.All interventions lasted for 24 hours.After the intervention,quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression levels of interleukin-1β(IL-1β),TLR4 and NLRP3 in RAW264.7 cells.Western blotting was used to detect the protein expression levels of TLR4 and NLRP3 in RAW264.7 cells.Results The mRNA expression levels of IL-1β in the control,model,experimental-L,experimental-H and MCC950 groups were 1.00±0.08,6.45±1.19,3.87±0.55,1.96±0.32 and 3.26±0.16,respectively;the mRNA expression levels of TLR4 were 1.00±0.13,2.69±0.35,1.92±0.22,1.32±0.23 and 3.38±0.33,respectively;the mRNA expression levels of NLRP3 were 1.00±0.14,1.28±0.19,0.83±0.02,0.87±0.15 and 0.95±0.25,respectively;the protein expression levels of TLR4 were 0.63±0.05,0.86±0.04,0.68±0.08,0.64±0.08 and 0.71±0.08,respectively;the protein expression levels of NLRP3 were 0.44±0.02,0.66±0.03,0.56±0.07,0.52±0.05 and 0.54±0.07,respectively.The differences between the model group and the control group were statistically significant(all P＜0.05);the differences between the experimental-H group and the model group were statistically significant(all P＜0.05).Conclusion NHP can improve macrophage inflammation,and its mechanism is related to inhibition of TLR4/NLRP3 signaling pathway.
neohesperidinlipopolysaccharidemacrophageToll-like receptor 4NOD-like receptor family pyrin domain containing 3
万方数据
维普
