CAJ | 학술논문

目的：初步探讨羟氯喹对Toll样受体-9（TLR-9）介导的人外周血单个核细胞（PB-MC）中IFN-α表达水平的影响。方法提取健康志愿者的PBMC。分别将PBMC与0、0.375、0.75、1.5、3.0、6.0μg/ml TLR-9激动剂ODN 2216共同培养12 h，以及与1.5μg/ml ODN 2216共同培养0、6、12、24、48 h，从细胞水平模拟SLE高表达IFN-α。然后用1.5μg/ml ODN 2216与PBMC共同培养6 h，加入羟氯喹1.25μg/ml （实验组）及等量的培养基（阳性对照组），在未加入ODN 2216的PBMC内加入等量的培养基（空白对照组），继续培养12 h后用实时荧光定量PCR法检测并比较各组PBMC中IFN-αmRNA表达水平。结果 PBMC与1.5μg/ml ODN 2216共同培养6 h时，IFN-αmRNA表达水平最高（P＜0.01）。阳性对照组PBMC中IFN-αmRNA表达水平明显高于空白对照组（P＜0.01）。与阳性对照组相比，实验组PBMC中IFN-αmRNA表达水平明显降低（P＜0.05）。结论羟氯喹可明显抑制经TLR-9介导的IFN-α高表达。
목적：초보탐토간록규대Toll양수체-9（TLR-9）개도적인외주혈단개핵세포（PB-MC）중IFN-α표체수평적영향。방법제취건강지원자적PBMC。분별장PBMC여0、0.375、0.75、1.5、3.0、6.0μg/ml TLR-9격동제ODN 2216공동배양12 h，이급여1.5μg/ml ODN 2216공동배양0、6、12、24、48 h，종세포수평모의SLE고표체IFN-α。연후용1.5μg/ml ODN 2216여PBMC공동배양6 h，가입간록규1.25μg/ml （실험조）급등량적배양기（양성대조조），재미가입ODN 2216적PBMC내가입등량적배양기（공백대조조），계속배양12 h후용실시형광정량PCR법검측병비교각조PBMC중IFN-αmRNA표체수평。결과 PBMC여1.5μg/ml ODN 2216공동배양6 h시，IFN-αmRNA표체수평최고（P＜0.01）。양성대조조PBMC중IFN-αmRNA표체수평명현고우공백대조조（P＜0.01）。여양성대조조상비，실험조PBMC중IFN-αmRNA표체수평명현강저（P＜0.05）。결론간록규가명현억제경TLR-9개도적IFN-α고표체。
Objectives To preliminarily investigate the effect of hydroxychloroquine on interferon-α(IFN-α)expression mediated by Toll like receptor-9 (TLR-9 )in human peripheral blood mononuclear cells (PBMCs). Methods PBMCs were freshly isolated from healthy donors. PBMCs were cultured with 0, 0.375,0.75,1.5,3.0,6.0 μg/ml TLR-9 agonist (ODN2216)for 12 h,or with 1.25 μg/ml ODN2216 for 0,6,12,24,48 h to mimic high expression of IFN-αin patients with SLE at the cell level. Then PBMCs were cultured with 1.5 μg/ml ODN 2216 for 6 h,and then treated with 1.25 μg/ml hydroxychloroquine (experi-ment group)or the same volume of medium (positive control group),and cultured for 12 h. PBMCs without any treatment were used as control group. Afterward,the expressions of IFN-αamong three groups were detec-ted by real-time quantitative PCR. Results When PBMCs were treated with 1.5 μg/ml ODN 2216 for 6 h, IFN-αmRNA expression reached the highest level (P<0.01). The expression of IFN-αmRNA in PBMC in positive control group was significantly higher than that of the control group(P<0.01). Compared with the positive control group,the mRNA expression of IFN-αin experiment group was significantly lower (P<0.05).Conclusions High expression of IFN-αmediated by TLR-9 may be inhibited by Hydroxychloroquine.