Interleukin-17A Induces IL-1β Secretion From RPE Cells Via the NLRP3 Inflammasome
Interleukin-17A Induces IL-1β Secretion From RPE Cells Via the NLRP3 Inflammasome
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IL-17 A通过NLRP 3炎症体诱导RPE细胞分泌IL-1β
DOI：
10.1167/iovs.15-17578
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发表时间：
2016-02-01
影响因子：
4.4
通讯作者：
Wu, Jihong
中科院分区：
文献类型：
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作者：
Zhang, Shujie;Yu, Ning;Wu, Jihong
目的.已经提出炎性小体活化和IL-1 β产生在年龄相关性黄斑变性（AMD）中具有重要作用。越来越多的证据表明白细胞介素-17A（IL-17 A）参与AMD的发病机制。我们研究了IL-17 A对原代人RPE细胞中炎性小体活化和IL-1 β产生的影响。分离并培养原代人RPE细胞用于以下实验。通过RT-PCR、流式细胞术和免疫荧光分析IL-17受体A（IL-17 RA）、IL-17受体C（IL-17 RC）和ACT 1的表达模式。将IL-17 A加入细胞培养物中，并使用实时RT-PCR、ELISA、Western印迹、流式细胞术和小干扰RNA进行细胞因子表达、信号传导途径和炎性小体机制研究。视网膜色素上皮细胞组成型表达IL-17 RA、IL-17 RC和ACT 1。IL-17 A上调pro-IL-1 β、IL-8、CCL 2和CCL 20的mRNA水平以及IL-1 β的蛋白水平。IL-17 A诱导RPE细胞中Akt、Erk 1/2、p38 MAPK和NF-κ B p65的磷酸化。阻断NF-κ B可减弱IL-17 A诱导的IL-1 β前体mRNA表达。IL-17 A增强caspase-1和NLRP 3 mRNA表达。抑制半胱天冬酶-1活性和沉默NLRP 3降低IL-1 β分泌，证实NLRP 3是转录后水平上的IL-17 A应答性炎性体。IL-17 A激活NLRP 3并随后分泌IL-1 β的机制与活性氧的产生有关。我们的研究结果表明，IL-17 A通过NLRP 3炎性体激活从RPE细胞中触发关键炎症介质IL-1 β，从而具有治疗AMD的潜力。
PURPOSE. Inflammasome activation and IL-1 beta production have been proposed to have an important role in age-related macular degeneration (AMD). Growing evidence is emerging for involvement of interleukin-17A (IL-17A) in AMD pathogenesis. We investigated the effects of IL-17A on the activation of inflammasome and production of IL-1 beta in primary human RPE cells.METHODS. Primary human RPE cells were isolated and cultured for the following experiments. Expression patterns of IL-17 receptor A (IL-17RA), IL-17 receptor C (IL-17RC), and ACT1 were analyzed by RT-PCR, flow cytometry, and immunofluorescence. IL-17A was added to the cell cultures, and cytokine expression, signaling pathways, and inflammasome machinery were investigated using real-time RT-PCR, ELISA, Western blot, flow cytometry, and small interfering RNA.RESULTS. Retinal pigment epithelial cells constitutively expressed IL-17RA, IL-17RC, and ACT1. IL-17A upregulated the mRNA levels of pro-IL-1 beta, IL-8, CCL2, and CCL20, as well as the protein level of IL-1 beta. IL-17A induced the phosphorylation of Akt, Erk1/2, p38 MAPK, and NF-kappa B p65 in RPE cells. Blocking NF-kappa B attenuated IL-17A-induced expression of pro-IL-1 beta mRNA. IL-17A enhanced pro-caspase-1 and NLRP3 mRNA expression. Inhibiting caspase-1 activity and silencing NLRP3 decreased IL-1 beta secretion, confirming NLRP3 as the IL-17A-responsive inflammasome on the posttranscriptional level. The mechanism of IL-17A-triggered NLRP3 activation and subsequent IL-1 beta secretion was found to involve the generation of reactive oxygen species.CONCLUSIONS. Our results suggest that IL-17A triggers a key inflammatory mediator, IL-1 beta, from RPE cells, via NLRP3 inflammasome activation, holding therapeutic potential for AMD.