木犀草素对哮喘患儿外周血单核细胞TNF-α和IL-6分泌的影响及机制研究

Effects of Luteolin on secretion of TNF-α and IL-6 in peripheral blood mononuclear cells of asthmatic children and its mechanism of action

目的:本研究旨在分析木犀草素对哮喘患儿外周血单核细胞(PBMCs)促炎性因子肿瘤坏死因子α(TNF-α)、IL-6分泌的影响及其可能的作用机制。方法:采集、分离24例缓解期过敏性哮喘患儿PBMCs,分别采用MTT法检测不同浓度(0.1、0.5、2 μmol/L)木犀草素对其活性的影响,酶联免疫吸附测定法(ELISA)检测培养48 h后各组PBMCs培养上清液中TNF-α、IL-6的浓度,免疫印迹法检测PBMCs 细胞中Toll样受体4(TLR4)、核因子κB(NF-κB)和髓样分化因子88 (MyD88)蛋白表达水平的变化。结果:不同浓度的木犀草素对PBMCs细胞活性无明显影响,不同浓度木犀草素刺激PBMCs细胞后,PBMCs上清液中TNF-α、IL-6的浓度明显降低,其抑制作用呈线性相关;此外,TLR4、NF-κB和MyD88蛋白表达水平均较空白对照组明显降低( P <0.05)。结论:木犀草素可抑制哮喘患儿PBMCs促炎性细胞因子TNF-α、IL-6的分泌,其可能通过作用于TLR4/NF-κB信号通路发挥抗炎效应,对哮喘具有潜在的治疗作用。

Objective: To investigate the effect of Luteolin on the secretion of TNF-α and IL-6 in peripheral blood mononuclear cells of asthmatic children and the underlying mechanisms. Methods: PBMCs were collected and separated from 24 children with allergic asthma in remission stage.Cell viability was examined by MTT assay at different concentrations (0.1,0.5,2 μmol/L) of Luteolin pretreated PBMCs cells.The effect of Luteolin on the secretion of TNF-α and IL-6 in the supernatant of PBMCs cultured after 48 h was evaluated by ELISA.Western blot was used to detect the expression of TLR4,NF-κB and MyD88 levels in PBMCs cells. Results: Cell viability had not significantly changes stimulated by different concentrations of Luteolin,and a significantly inhibition role showed in the release of TNF-α and IL-6 from PBMCs cells.However,the results showed that the expression levels of TLR4,NF-κB and MyD88 were decreased( P <0.05). Conclusion: Luteolin suppresses the secretion of pro-inflammatory cytokines TNF-α and IL-6 in PBMCs of asthmatic children via acting the TLR4/NF-κB signaling pathway to exert its anti-inflammatory effect and has a potential therapeutic effect on asthma.