Objective:To investigate the anti-inflammatory effects of the total flavonoids from Saussurea involucrata on lipopolysaccharides(LPS)-stimulated murine RAW264.7 macrophages and explore its underlying mechanism of acti...Objective:To investigate the anti-inflammatory effects of the total flavonoids from Saussurea involucrata on lipopolysaccharides(LPS)-stimulated murine RAW264.7 macrophages and explore its underlying mechanism of action.Methods:Total flavonoids from Saussurea involucrata were extracted using chromatographic column method.Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay.The production of nitric oxide was detected by Griess assay and the release of cytokines(IL-10 and TNF-α)and chemokines(MCP-1,MIP-1α,and CCL5/RANTES)was determined by ELISA to evaluate the anti-inflammatory activity of total flavonoids from Saussurea involucrata.Moreover,nuclear translocation of p65,c-Jun,and IRF3 was detected by immunofluorescence microscopy and Western blotting analysis was performed to determine the expression of related proteins.Results:Total flavonoids extracted from Saussurea involucrata were 751.5 mg/g and the content of rutin was 506.5 mg/g.The production of inflammatory mediators including nitric oxide,cytokines,and chemokines was effectively inhibited by total flavonoids from Saussurea involucrata.Meanwhile,total flavonoids also suppressed the nuclear translocation of p65,c-Jun,and IRF3 in LPS-stimulated RAW264.7 cells.The LPS-induced expression of iNOS and COX-2 was remarkably reduced by treatment with total flavonoids from Saussurea involucrata.Moreover,total flavonoids decreased the expression levels of p-IKKα/β,p-TBK1,p-p38,p-ERK,p-JNK,p-p65,p-c-Jun,and p-IRF3 in LPS-exposed RAW264.7 macrophages.Conclusions:Total flavonoids from Saussurea involucrata potentially inhibit the secretion of pro-inflammatory mediators,which may be related to inhibition of p65,c-Jun,and IRF3 signaling pathways in LPS-stimulated RAW264.7 cells.展开更多
基金This work was supported by the National Natural Science Foundation of China(Grant Number 81803793 and 82003957)the Young Scientist Program by Beijing University of Chinese Medicine,and the Fundamental Research Funds for the Central Universities(Grant Number 2018-JYBZZ-XJSJJ008).
文摘Objective:To investigate the anti-inflammatory effects of the total flavonoids from Saussurea involucrata on lipopolysaccharides(LPS)-stimulated murine RAW264.7 macrophages and explore its underlying mechanism of action.Methods:Total flavonoids from Saussurea involucrata were extracted using chromatographic column method.Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay.The production of nitric oxide was detected by Griess assay and the release of cytokines(IL-10 and TNF-α)and chemokines(MCP-1,MIP-1α,and CCL5/RANTES)was determined by ELISA to evaluate the anti-inflammatory activity of total flavonoids from Saussurea involucrata.Moreover,nuclear translocation of p65,c-Jun,and IRF3 was detected by immunofluorescence microscopy and Western blotting analysis was performed to determine the expression of related proteins.Results:Total flavonoids extracted from Saussurea involucrata were 751.5 mg/g and the content of rutin was 506.5 mg/g.The production of inflammatory mediators including nitric oxide,cytokines,and chemokines was effectively inhibited by total flavonoids from Saussurea involucrata.Meanwhile,total flavonoids also suppressed the nuclear translocation of p65,c-Jun,and IRF3 in LPS-stimulated RAW264.7 cells.The LPS-induced expression of iNOS and COX-2 was remarkably reduced by treatment with total flavonoids from Saussurea involucrata.Moreover,total flavonoids decreased the expression levels of p-IKKα/β,p-TBK1,p-p38,p-ERK,p-JNK,p-p65,p-c-Jun,and p-IRF3 in LPS-exposed RAW264.7 macrophages.Conclusions:Total flavonoids from Saussurea involucrata potentially inhibit the secretion of pro-inflammatory mediators,which may be related to inhibition of p65,c-Jun,and IRF3 signaling pathways in LPS-stimulated RAW264.7 cells.
