氯喹抑制LPS和CpG DNA诱导人外周血单个核细胞释放促炎细胞因子的研究

Study of chloroquine on pro-inflammatory cytokines released by human peripheral blood mononuclear cells induced by LPS and CpG DNA

目的 观察氯喹对脂多糖(LPS)和CpG DNA诱导人外周血单个核细胞释放促炎细胞因子的抑制作用。方法 采用密度梯度离心法从新鲜人外周血中分离单个核细胞(hPBMC)。先用LPS和CpG DNA刺激hPBMC分泌TNF-α、IL-6,再加入不同浓度氯喹进行拮抗,观察其量效关系;在不同时相点加入氯喹观察时效关系;用ELISA法检测培养上清中促炎细胞因子的含量。采用甲基四唑蓝(MTT)法检测氯喹对细胞活性的影响。结果 氯喹在50μg/ml时显著抑制LPS和CpG DNA刺激hPBMC释放TNF-α和IL-6(P<0.01)。若先给氯喹,则其拮抗细胞因子释放的作用非常显著(P<0.01),提前给予LPS或CpG DNA,再给氯喹时也可观察到显著拮抗作用(P<0.05)。氯喹浓度小于100μg/ml时对细胞活性无显著影响(P>0.05)。结论 氯喹对LPS和CpGDNA诱导人外周血hPBMC释放促炎细胞因子的抑制作用呈明显的剂量和时间依赖关系。

Objective To observe the inhibitory effect of chloroquine on pro-inflammatory cytokines released by human peripheral blood mononuclear cells (hPBMC) induced by LPS and CpG DNA. Methods Mononuclear cells were isolated from fresh human blood by density gradient centrifugation. hPBMC was stimulated by LPS or CpG DNA to release TNF-α and IL-6, then various concentrationes of chloroquin were administrated to observe its dose-dependent inhibitory effect and they were added at different time to observe its time-dependent effect. Contents of cytokines in culture supematant were detected by ELISA. Influences of various concentrations of chloroquine on viability of hPBMC were measured by MTT test. Results 50μg/ml chloroquine significantly inhibited release of TNF-α and IL-6 from hPBMC stimulated by LPS, CpG DNA(P<0.01). When chloroquine was administrated first, cytokine released by hPBMC was inhibited very markedly(P<0.01). If hPBMC was stimulated by LPS and CpG DNA prior to administration of chloroquine, release of TNF-α and IL-6 was suppressed(P<0.05). Chloroquine had no influence on the viability of hPBMC when its concentration was under 100μg/ml(P>0.05). Conclusion Chloroquine inhibits pro-inflammatory cytokines released by hPBMC in a dose-and time-dependent way.