医药导报2024，Vol.43Issue(5)：689-695,7.DOI:10.3870/j.issn.1004-0781.2024.05.003
黄酮化合物对巨噬细胞中白细胞介素介导的TLR4/MyD88/NF-κB信号转导通路的调节作用
Regulatory Effect of Flavonoid Compounds on Interleukins in Macrophage via TLR4/MyD88/NF-κB Signaling Pathway
摘要
Abstract
Objective To validate regulatory effects of two flavonoid(kumatakenin and luteolin)in macrophage con-cerning inflammatory/anti-inflammatory interleukins(IL)expression and TLR4/MyD88/NF-κB signaling.Methods Murine RAW264.7 macrophage cellular inflammation was induced by lipopolysaccharide(LPS),coupling synergistically anti-inflammato-ry treatment with different flavonoid concentrations.Expression levels of nitric oxide(NO)and IL were determined in cell culture supernatant using enzyme-linked immunosorbent assay.An effect of small-interfering RNA(siRNA)-mediated IL-6 knockdown on IL-10/IL-17 mRNA expressions was assayed by reverse transcription polymerase chain reaction(RT-PCR),and IL-6/IL-12 mR-NA regulation is performed by IL-10 knockdown.Western blotting(WB)compares a relative ratio of TLR4,p-p65,MyD88,and p-IκBα productions in macrophages blank,LPS-induced,and LPS+flavonoid-co-treated.Results IC50 values of kumatakenin and luteolin are 0.60 and 8.93 μmol·L-1,respectively,concerning the NO inhibition in LPS-induced cells.Compared to blank,LPS inducement leads to a decrease of anti-inflammatory IL-4 and IL-10,whereas the increase of inflammatory IL-6,IL-12,IL-16,IL-17,and IL-23(P＜0.01).Cellular co-treatment with LPS+kumatakenin or LPS+luteolin significantly blocks a regulatory effect of ILs in a flavonoid dose-dependent manner(P＜0.05 or P＜0.01).IL-6 and IL-12 mRNA are increased in the IL-10 siRNA,LPS+IL-10 siRNA,and LPS+IL-10 siRNA+flavonoid treatment groups,compared to the no-target siRNA negative control.IL-6 knockdown can down-regulate IL-10 and IL-17 mRNA expressions(P＜0.05 or P＜0.01).WB result shows that expression of TLR4,p-p65,MyD88,and p-IκBα is significantly increased in the LPS-induced group,while the expression level is inhibited when LPS cotreatment with flavonoid(P＜0.05 or P＜0.01).Conclusions Kumatakenin and luteolin are active against LPS-induced inflammation.They neutralize both pro-inflammatory and anti-inflammatory ILs in macrophages.IL-10 negatively regu-lates IL-6 and IL-12 expressions,while IL-6 positively regulates IL-10 and IL-17.Dual-neutralization of ILs occurs via inhibition of the TLR4/MyD88/NF-κB signaling in inflammatory macrophage.关键词
黄酮化合物/华良姜素/木犀草素/白细胞介素/核因子-κBKey words
Flavonoid compounds/Kumatakenin/Luteolin/Interleukin/Nuclear factor-κB分类
医药卫生引用本文复制引用
冯逸佳,李庆芳,董壮壮,张圆琳,乔元彪,李青山..黄酮化合物对巨噬细胞中白细胞介素介导的TLR4/MyD88/NF-κB信号转导通路的调节作用[J].医药导报,2024,43(5):689-695,7.基金项目
山西省医学科技创新团队领军项目(2020TD02) （2020TD02）
山西中医药大学博士科研启动基金资助项目(2022BK01). （2022BK01）
