Research On Tripterygium Wilfordii Seeds Extract Affect The Anti-inflammatory Activity Of Macrophages In Vitro Through Nfκb Signaling Pathway

Objective : To explore the effect of Triplaris gardneriana seeds extract on the anti-inflammatory activity of macrophages in vitro by regulating NFκB signal.Methods : The normal cultured cells were used as the control group;the cells were treated with 100ng/ml LPS for 6 hours as the model group;the low,medium,and high-dose experimental groups were treated with 15,30,60 μmol/L Triplaris gardneriana seeds extract.Quantitative reverse transcription PCR(q RT-PCR)method was used to detect m RNA expression level of inducible nitric oxidesynthase(i NOS),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),enzyme-linked immunosorbent assay kit was used to detect secretion levels of TNF-α,IL-1β,interleukin-6(IL-6),Western blotting method was used to detect NF-κB p65 protein expression level,flow cytometry was used to detect cellular reactive oxygen species(ROS)content.Results : The relative expression levels of i NOS m RNA in control group,model group,low,medium and high dose experimental groups were(1.00 ± 0.05),(8.57 ± 0.59),(6.86 ± 0.62),(4.73 ± 0.69)and(3.66 ± 0.19),respectively;the expression levels of IL-1β were(1.00 ± 0.11),(294.62 ± 8.15),(228.79 ± 6.09),(182.13 ± 4.18)and(146.71 ±6.63),respectively The expression of TNF-α was(17.59 ± 6.97),(931.19 ± 32.44),(641.10 ± 16.22),(534.38 ± 11.64)and(389.26 ± 15.79)respectively;the secretion of TNF-α was(17.59 ± 6.97),(931.19 ± 32.44),(641.10 ± 16.22),(534.38 ± 11.64)and(389.26 ± 15.79)respectively;the secretion of IL-1 β was(12.96 ± 5.88),(1637.25 ±32.07)respectively)The levels of IL-6 secretion were(6.31 ± 3.59),(398.71 ± 14.02),(310.36 ± 7.74),(251.00 ± 14.46)and(206.74 ± 10.09),respectively;the relative levels of NF-κ B phosphorylation were(0.411),(1.399),(1.041),(0.423)and(0.500),respectively;the production rate of reactive oxygen species(ROS)was(0.411),(1.399),(1.041),(0.423)and(0.500),respectively(0.23±0.00)%,(1.00±0.02)%,(0.84±0.01)%,(0.73±0.01)%,(0.57±0.01)%。 Compared with the control group,the m RNA expression levels of i NOS,TNF-α and IL-1 β,the secretion levels of cytokines TNF-α,IL-1 β and IL-6,the protein levels of phosphorylated NF-κB and the production levels of intracellular reactive oxygen species in the low,medium and high dose experimental groups were significantly different(P < 0.05).Conclusion : Tripterygium wilfordii seeds extract enhances the anti-inflammatory activity of macrophages in vitro through the NFκB signaling pathway.