Hydroxychloroquine inhibits proinflammatory signalling pathways by targeting endosomal NADPH oxidase
Hydroxychloroquine inhibits proinflammatory signalling pathways by targeting endosomal NADPH oxidase
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DOI：
10.1136/annrheumdis-2016-210012
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发表时间：
2017-05-01
影响因子：
27.4
通讯作者：
Lackner, Karl J.
中科院分区：
文献类型：
--
作者：
Mueller-Calleja, Nadine;Manukyan, Davit;Lackner, Karl J.
目的羟基氯喹(HCQ)几十年来一直被用于治疗风湿性疾病，如系统性红斑狼疮(SLE)、类风湿性关节炎(RA)或抗磷脂综合征(APS)。我们推测HCQ可能针对参与细胞因子和抗磷脂抗体(APL)信号转导的NADPH氧化酶(NOX)。方法体外实验用肿瘤坏死因子α(TNFα)、白介素1β(IL-1β)或人单抗APL刺激单核细胞，用流式细胞仪检测NOX的活性。已知由这些刺激诱导的基因的表达通过定量逆转录聚合酶链式反应进行定量。用激光共聚焦扫描显微镜进行活细胞成像。结果HCQ能有效抑制或完全阻断TNFa、IL-1β和APL诱导的人单核细胞和MonoMac1细胞产生NOX。因此，这些刺激物对下游基因的诱导减少或取消。HCQ的这种作用不是通过直接干扰激动剂而是通过抑制NOX2的催化亚单位(Gp91Phox)移位到内体而实现的。在体内，HCQ对APL诱导的小鼠血栓形成和NOX2介导的血栓形成具有保护作用。结论本研究描述了一种新的作用机制，即干扰内体NOX2的组装。由于内体NOX2参与许多炎症和血栓前信号通路，HCQ的这种活性可能解释了它在包括APS在内的风湿性疾病中的许多有益作用。
Objectives Hydroxychloroquine (HCQ) has been used for decades to treat patients with rheumatic diseases, for example, systemic lupus erythematosus (SLE), rheumatoid arthritis or the antiphospholipid syndrome (APS). We hypothesise that HCQ might target endosomal NADPH oxidase (NOX), which is involved in the signal transduction of cytokines as well as antiphospholipid antibodies (aPL).Methods For in vitro experiments, monocytic cells were stimulated with tumour necrosis factor a (TNF alpha), interleukin-1 beta (IL-1 beta) or a human monoclonal aPL and the activity of NOX was determined by flow cytometry. The expression of genes known to be induced by these stimuli was quantified by quantitative reverse transcription PCR. Live cell imaging was performed by confocal laser scanning microscopy. Finally, the effects of HCQ on NOX-induced signal transduction were analysed in an in vivo model of venous thrombosis.Results HCQ strongly reduces or completely prevents the induction of endosomal NOX by TNFa, IL-1 beta and aPL in human monocytes and MonoMac1 cells. As a consequence, induction of downstream genes by these stimuli is reduced or abrogated. This effect of HCQ is not mediated by direct interference with the agonists but by inhibiting the translocation of the catalytic subunit of NOX2 (gp91phox) into the endosome. In vivo, HCQ protects mice from aPL-induced and NOX2-mediated thrombus formation.Conclusions We describe here a novel mechanism of action of HCQ, that is, interference with the assembly of endosomal NOX2. Since endosomal NOX2 is involved in many inflammatory and prothrombotic signalling pathways, this activity of HCQ might explain many of its beneficial effects in rheumatic diseases including the APS.