R848对哮喘患儿外周血中IFN-γ、IL-33和IgE表达的影响及临床意义

The effect of R848 on the expression of IFN-γ,IL-33 and IgE in the peripheral blood of children with asthma and its clinical significance

目的:检测R848对哮喘患儿外周血中干扰素-γ(IFN-γ)、白细胞介素-33(IL-33)和IgE表达的影响及临床意义。方法:收集2018年6月—2020年5月在我院接受治疗的急性发作期哮喘患者125例,临床缓解期患者40例,分离培养外周血单个核细胞(PBMC)。ELISA实验检测PBMC分泌IFN-γ、IL-33和IgE的水平,qRT-PCR检测急性发作期哮喘患者PBMC中Toll样受体(TLR)7/8 mRNA的相对表达水平,Pearson分析急性发作期哮喘患者IFN-γ、IL-33和IgE与TLR7/8 mRNA的相对表达水平的相关性。ELISA实验检测不同浓度R848(0 mg/L、0.1 mg/L、1 mg/L、10 mg/L、100 mg/L)刺激急性发作期哮喘患者PBMC 24 h后IFN-γ、IL-33和IgE的水平。分析R84810 mg/L刺激急性发作期哮喘患者PBMC细胞6 h、12 h、24 h、36 h、48 h后IFN-γ、IL-33和IgE水平的变化。结果:急性发作期哮喘患者外周血中PBMC分泌IFN-γ的水平低于临床缓解期,分泌IL-33和IgE的水平高于临床缓解期(均P<0.05);IL-33水平与TLR7/8 mRNA相对表达水平呈负相关(r=-0.887,P<0.001;r=-0.910,P<0.001),IFN-γ水平与TLR7/8 mRNA相对表达水平呈正相关(r=0.818,P<0.001;r=0.808,P<0.001),IgE水平与TLR7/8 mRNA相对表达水平呈负相关(r=-0.850,P<0.001;r=-0.838,P<0.001)。R848刺激PBMC细胞24 h后,IL-33和IgE水平在R8481 mg/L时降低至R84810 mg/L后趋于平稳(F=56.231、42.310,均P<0.05)。IFN-γ水平在R8480.1 mg/L时逐渐升高至R84810 mg/L后趋于平稳(F=62.352,P<0.05)。R84810 mg/L刺激PBMC 6~48 h,IL-33和IgE水平在12 h时逐渐降低至36 h后趋于平稳(F=32.658、36.521,均P<0.05),IFN-γ在12 h时逐渐增加至36 h后趋于平稳(F=42.321,P<0.05)。结论:R848可抑制急性发作期患者外周血中PBMC分泌IL-33和IgE,促进IFN-γ的分泌,具有一定的抗炎作用,有望应用于哮喘的治疗。

Objective:To detect the effect of R848 on the expression of gamma interferon(IFN-γ),Interleukin-33(IL-33)and IgE in the peripheral blood of children with asthma and its clinical significance.Methods:A total of 125 patients with acute-onset asthma and 40 patients with clinical remission who received treatment in our hospital from June 2018 to May 2020 were collected,and peripheral blood mononuclear cells(PBMC)were isolated and cultured.ELISA test detected the levels of IFN-γ,IL-33 and IgE secreted by PBMC,qRT-PCR detected the relative expression level of toll-like receptor(TLR)7/8 mRNA in PBMC of acute-onset asthma patients,and Pearson detected the correlation between IFN-γ,IL-33 and IgE with the relative expression levels of TLR7/8 mRNA in patients with acute asthma.ELISA experiments were used to detect the levels of IFN-γ,IL-33 and IgE after stimulating PBMC of acute-onset asthma patients with different concentrations of R848(0 mg/L,0.1 mg/L,1 mg/L,10 mg/L,100 mg/L).The changes of IFN-γ,IL-33 and IgE levels after 6 h,12 h,24 h,36 h,48 h stimulating PBMC by R84810 mg/L in acute asthma patients were determinated.Results:The level of IFN-γsecreted by PBMC in peripheral blood of patients with acute asthma was lower than that in clinical remission,and the level of IL-33 and IgE secreted was higher than that of clinical remission(all P<0.05).The level of IL-33 was negatively correlated with the relative expression level of TLR7/8 mRNA(r=-0.887,P<0.001;r=-0.910,P<0.001),and the level of IFN-γwas positively correlated with the relative expression level of TLR7/8 mRNA(r=0.818,P<0.001;r=0.808,P<0.001),the IgE level was negatively correlated with the relative expression level of TLR7/8 mRNA(r=-0.850,P<0.001;r=-0.838,P<0.001).After R848 stimulated PBMC cells for 24 h,IL-33 and IgE gradually decreased from R8481 mg/L to R84810 mg/L and then stabilized(F=56.231,42.310,both P<0.05).The level of IFN-γgradually increased from R8480.1 mg/L to R84810 mg/L and then stabilized(F=62.352,P<0.05).R84810 mg/L stimulated PBMC for 6~48 h,IL-33 and IgE levels gradually decreased from12 h to 36 h and then stabilized(F=32.658,36.521,both P<0.05),and IFN-γgradually increased from 12 h to 36 h and then stabilized(F=42.321,P<0.05).Conclusion:R848 can inhibit the secretion of IL-33 and IgE from PBMC in the peripheral blood of patients with acute attacks,promote the secretion of IFN-γ,and has a certain anti-inflammatory effect.R848 is expected to be used in the treatment of asthma.