机构地区:[1]江苏大学附属医院风湿科,镇江212001 [2]江苏大学附属医院检验科,镇江212001
出 处:《中华风湿病学杂志》2013年第9期616-619,共4页Chinese Journal of Rheumatology
基 金:江苏大学临床医学科技发展基金(JLY20080008)
摘 要:目的探讨类风湿关节炎(RA)患者外周血CD4^+T细胞胞内白细胞介素(IL)-9、IL-17、干扰素-吖的表达及意义。方法取RA患者活动期25例、稳定期25例和健康人20名外周血单个核细胞(PBMC),免疫磁珠阴选获得CD4^+T细胞,通过流式细胞术(FCM)检测0CD4^+T细胞内IL-9、IL-17、干扰素叫水平;应用实时荧光定量反转录聚合酶链反应(RT-PCR)检测IL-9、IL-17、孤核受体(RORγt)、干扰素-γ mRNA水平。采用方差分析法进行组间比较,采用Pearson相关分析法进行相关性分析。结果免疫磁珠阴选CD4^±T细胞纯度达90%以上。RA患者病情活动组IL-9、IL-17、干扰素-1的水平显著高于病情稳定组,分别为(1.62±0.23)%与(1.15±0.24)%(P〈0.01)、(1.47±0.20)%与(1.04±0.26)%(P〈0.01)、(8.1±0.6)%与(6.9±1.0)%(P〈0.01);RA组显著高于健康对照组(P〈0.01);RA病情活动组IL-9、IL-17、RORγt、干扰素-γ mRNA水平高于病情稳定组,分别为(3.0±0.6)与(1.8±0.4)(P〈0.01)、(4.2±0.9)与(2.3±0.7)(P〈0.01)、(4.1±0.7)与(2.9±0.3)(P〈0.01)、(4.0±0.8)与(2.3±0.6)(P〈0.01),RA组IL-17、RORγt、干扰素.1mRNA表达又显著高于健康对照组(P〈0.01);RA患者胞内IL-9水平与胞内IL-17(r=0.632,P=0.001)、干扰素-γ(r=0.515,P=0.008)、DAS28(r=0.519,P=0.009)、外周血ESR水平(r=0.857,P=0.038)呈正相关,而与C反应蛋白水平(r=0.38,P=0.61)无明显相关性。结论RA患者体内IL-9、IL-17和干扰素-叫表达增高,可能参与RA炎症。Objective To detect the levels of IL-9, IL-17, and IFN-γ in CD4^+ T cells from patients with rheumatoid arthritis (RA). Methods The peripheral blood mononuelear cells (PBMCs) from patients with RA and,healthy controls were obtained, then the CD4^+ T lymphocytes were tested by immunomagnetic beads. The protein levels of IL-9, IFN-γ and IL-17 were measured by flow eytometry (FCM). The mRNA levels of IL-9, IL-17, RORγt and IFN-γ were also detected by qRT-PCR. Data were analyzed by comparison between groups using variance analysis, and Pearson's correlation analysis was used for linear correlation analysis. Results The isolation of untouched human CD4^+ T cells from PBMC was effective and its purity was over 90%. The protein levels of IL-9, IL-17, IFN-γ were higher in patients with active RA as compared with patients with inactive RA (P〈0.01) Which were (1.62±0.23)% vs (1.15±0.24)%(P〈0.01), (1.47±0.20)% vs (1.04±0.26)%(P〈0.01) and (8.1±0.6)% vs (6.9±1.0)%(P〈0.01) respectively, so did the patients with RA when compared with healthy controls (P〈0.01). The mRNA levels of IL-9, IL-17, RORγt and IFN-γ were higher in patients with active RA as compared with inactive RA patients (P〈0.01), which were (3.0±0.6) vs (1.8±0.4) (P〈0.01) (4.2±0.9) vs (2.3±0.7) (P〈0.01), (4.1±0.7) vs (2.9±0.3) (P〈0.01) and (4.0±0.8) vs (2.3±0.6) (P〈0;01) respectively, so did the patients with RA when compared with healthy controls (P〈0.01). Intracelluar IL-9 levels were positively correlated with IL-17 (r=0.632, P=0.001), IFN-γ (r=0.515, P= 0.008), DAS28 (r=0.519, P=0.009) and ESR (r=0.857,P=0.038) but had no correlation with CRP (r=0.38, P=0.61). Conclusion The levels of IL-17, IL-9, IFN-γ are higher in the PBMCs of RA patients, and these cytokines may participate in the pathogenesis of RA.
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