Study on the Inhibition of Lipopolysaccharide-Induced Cellular Inflammatory Response by Combined use of COPTIDIS RHIZOMA and MUME FRUCTUS Extracts and Its Mechanism
Objective:To investigate the effect and mechanism of combined use of COPTIDIS RHIZOMA and MUME FRUCTUS extracts on the li-popolysaccharide(LPS)-induced inflammatory response in mouse macrophage RAW264.7.Methods:LPS-induced inflammation model was established on RAW264.7 cells.In the experiment,the cells were divided into blank control group,model(LPS)group,dimethyl sulfoxide(DMSO)group and COPTIDIS RHIZOMA and MUME FRUCTUS extracts group.Cell proliferation was assessed using Cell Counting Kit-8(CCK-8).Flow cytometry was employed to measure cell apoptosis,reactive oxygen species(ROS)content,and the expressions of cell sur-face molecules CD80,CD83,and CD86.Enzyme-linked immunosorbent assay(ELISA)was used to determine the effect on inflammatory fac-tor secretion,and Western blot(WB)was used to analyze the protein expression levels of high mobility group protein B1(HMGB1),P65,and Toll-like receptor4(TLR4)in cells.Results:Compared with the blank control group,protein expressions of HMGB1 and TLR4 were up regulated in the model group(2.05±0.05,2.56±0.63,P＜0.05 or P＜0.01).The expression of cell surface molecules CD80,CD83,and CD86 were increased(19.81±1.85,3.81±0.40,and 3.23±0.40,P＜0.01),and contents of TNF-α,IL-6 and IL-1β in cell culture superna-tants were significantly increased(33.31±3.92,42.66±3.07,25.98±2.45,P＜0.01).Compared with the model group,the 343 μg/mL of COPTIDIS RHIZOMA and MUME FRUCTUS extracts down regulated the protein expression of HMGB1(1.50±0.08,P＜0.01),inhibited the expressions of cell surface molecules CD80,CD83,and CD86(8.73±2.92,1.04±0.15,1.35±0.15,P＜0.01),and significantly reduced the secretion of inflammatory factors TNF-α,IL-6 and IL-1β(19.86±3.12,29.41±8.98,16.58±0.52,P＜0.05 or P＜0.01).Conclusion:Combined use of COPTIDIS RHIZOMA and MUME FRUCTUS extracts demonstrates a potent anti-inflammatory effect in the LPS-induced in-flammation model of RAW264.7 cells.Its mechanism may be related to the inhibition of ROS/HMGB1/TLR4 signaling pathway.
COPTIDIS RHIZOMA and MUME FRUCTUS extractsMouse macrophageInflammationApoptosisROSHMGB1TLR4
NETL
NSTL
万方数据
