摘要目的 了解三氧化二砷(AS2O3)对SLE患者PBMCs Toll样受体9(TLR9)及干扰素调节因子5(IRF5) mRNA表达的影响.方法 分离15例SLE患者和15名健康人外周血PBMCs,分别与不同浓度AS2O3及环磷酰胺进行体外培养12h和24h,采用实时荧光定量PCR检测TLR9和IRF5 mRNA表达.采用t检验或方差分析进行组间差异比较.结果 与健康对照组[12 h(1.05±0.35)、24 h(0.97±0.19)]相比,SLE组体外培养[12 h(1.38±0.26)和24 h(1.28±0.35)]后TLR9 mRNA的相对表达水平显著升高,差异有统计学意义(t=2.37,P=0.03;t=2.44,P=0.02);与对照组[12 h(0.62±0.23)、24h(0.60±0.39)]相比,SLE组体外培养[12 h(0.95±0.27)和24 h(0.91±0.35)]后IRF5 mRNA的相对表达水平显著升高,差异有统计学意义(t=3.07,P=0.01;t=3.45,P＜0.01).AS2O3可以下调SLE患者PBMCs中TLR9 mRNA表达,其效应随药物浓度和作用时间的增加而增加[0.2 mg/L AS2O3组12 h (0.430±0.110)和24h(0.290±0.050),0.4 mg/LAS2O3组12 h (0.170±0.038)和24 h(0.090±0.017),0.8 mg/L AS2O3组12 h (0.023±0.011)和24 h(0.003±0.001)],与环磷酰胺[12 h(0.814±0.081)和24 h(0.755±0.139)]相比较,AS2O3对SLE患者的作用更明显,差异有统计学意义(F=165.32,P＜0.01;F=99.20,P＜0.01);环磷酰胺与AS2O3均对PBMCs中IRF5 mRNA表达无影响,各组间比较差异无统计学意义(P＞0.05).结论 SLE患者外周血PBMCs的TLR9和IRF5mRNA表达异常,AS2O3可明显抑制SLE患者TLR9 mRNA表达,可能是其治疗SLE的机制之一.

abstractsObjective To investigate the mRNA expression of toll like receptor-9 (TLR9) and interferon regulatory factors-5 (IRF5) of AS2O3 on peripheral blood mononuclear cells (PBMCs) in systemic lupus erythematosus (SLE) patients.Methods PBMCs of 15 SLE patients and 15 healthy subjects were treated with different concentrations of AS2O3 and cyclophosphamide (CTX) in vitro.Real-time quantitative polymerase chain reaction was used to amplify TLR9 and IRF5 gene before and after 12 and 24 hours drug intervention and the mRNA expressions were measured.Differences between groups were analyzed by paired t test or variance analysis.Results The mRNA expression levels of TLR9 [12 h(1.38±0.26) and 24 h (1.28±0.35)] on PBMCs in SLE patients were significantly higher than those in healthy controls [12 h(1.05±0.35) and 24 h (0.97±0.19)](t=2.37,P=0.03; t=2.44,P=0.02).The IRF5 mRNA expression levels [12 h (0.95±0.27) and 24 h (0.91 ±0.35)] in SLE patients were obviously higher than those in healthy controls [12 h (0.62 ±0.23) and 24 h (0.60±0.39)] (t =3.07,P=0.01 ; t =3.45,P＜0.01).AS2O3 could suppress the mRNA expression of TLR9 on PBMCs and the effect was gradually increasing with the increasing concentration of AS2O3 and processing time [0.2 mg/L AS2O3 group 12 h (0.430±0.110) and 24 h(0.290±0.050),0.4 mg/L AS2O3 group 12 h (0.170±0.038) and 24 h (0.090±0.017),0.8 mg/L AS2O3 group 12 h (0.023±0.011) and 24 h (0.003±0.001)].Comparing with CTX [12 h (0.814±0.081) and 24 h(0.755±0.139)],AS2O3 had a more significant strong effect on inhibiting the expression of TLR9 mRNA in SLE patients [F=165.32(12 h),P＜0.01; F=99.20 (24 h),P＜0.01].The mRNA expression of IRF5 on PBMCs was not suppressed by AS2O3 and CTX and there was no statistically significant difference between groups (P＞0.05).Conclusion There is abnormal expression of IRF5 and TLR9 mRNA in SLE patients.AS2O3 may suppress the TLR9 mRNA expression in SLE patients,which may be one mechanism of clinical effectiveness.