Anti-inflammatory effect and mechanism of Pseurata H on inflammation model of LPS-induced RAW264.7 cells
To study the anti-inflammatory effect and mechanism of Pseurata H on peritoneal macrophages(RAW264.7 cells)inflam-mation models,RAW264.7 cells were stimulated with LPS to establish an inflammation model in vitro,and Pseurata H with the con-centrations of 10,20,and 40 μmol/L was used to interfere with this cell model.The cytotoxicity of Pseurata H in RAW264.7 cells was determined by CCK-8 colorimetric assay.Griess assay was employed to detect the content of NO in the supernatant.The release of IL-6 and TNF-α in the supernatant was assayed using ELISA.The protein expressions of COX-2,iNOS,p-NF-κB and the p-ERK were measured by Western Blot.The mRNA expressions of iNOS,TNF-α,IL-6 and IL-1β were analyzed by RT-qPCR.The results showed that Pseurata H(10,20,40 μmol/L)displayed no apparent cytotoxicity in RAW264.7 cells.The release levels of NO,iNOS,TNF-α,and IL-6 were decreased by Pseurata H(10,20,40 μmol/L).In addition,Pseurata H also inhibited the expressions of iNOS,COX-2,NF-κB,p-NF-κB,ERK,and p-ERK proteins with concentration effect.Pseurata H has a significant anti-inflammatory effect,and its mechanism of action may be through the inhibition of NF-κB/MAPK signaling pathway to exert anti-inflammatory effects.
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