Pediatric Urodynamic Center and Department of Urology, Institute of Clinical Medicine, The First Affiliated Hospital of Zhengzhou University, 450052, Zhengzhou, China.
Helsinki Rheumatic Diseases and Inflammation Research Group, Institute of Clinicum, University of Helsinki, Haartmaninkatu 8, 00290, Helsinki, Finland.
Pediatr Res. 2019 Dec;86(6):692-698. doi: 10.1038/s41390-019-0418-4. Epub 2019 May 13.
The treatment of nucleotide-binding domain and leucine-rich repeat containing family, pyrin domain containing 3 (NLRP3) inflammasome-mediated pediatric inflammatory diseases is challenging. Here we studied whether cyclic adenosine monophosphate (cAMP) elevator forskolin could attenuate the nigericin-induced NLRP3-inflammasome activation and interleukin-1β (IL-1β) secretion in human macrophages.
The proteins and messenger RNA (mRNA) levels of inflammasome structural proteins and proinflammatory cytokines were measured in forskolin-stimulated nigericin-activated human THP-1 macrophages and primary macrophages.
Activation of THP-1 macrophages with nigericin increased the mRNA expression of NLRP3, IL-1β, and caspase-1 (P < 0.01). Forskolin stimulation had no effect on the mRNA expression of NLRP3, caspase-1, or IL-1β in nigericin-activated cells (P > 0.05), while their protein levels were significantly decreased (P < 0.05). Forskolin-mediated increase in cytoplasmic cAMP in non-activated cells was attenuated in nigericin-activated macrophages (P < 0.05). Basal IL-1β secretion increased from 584 to 2696 pg/mL (P < 0.01) in nigericin-activated macrophages; forskolin dose-dependently reduced the nigericin-induced secretion of mature IL-1β (P < 0.01). Forskolin also inhibited the IL-1β secretion from activated human primary macrophages.
Forskolin inhibits the NLRP3 inflammasome activation and the secretion of mature IL-1β, in human macrophages. Forskolin and other cAMP elevator drugs could represent a novel approach for treatment of diseases associated with excessive inflammasome activation, like pediatric inflammatory diseases.
核苷酸结合域和富含亮氨酸重复序列家族、富含亮氨酸重复序列的pyrin 域 3（NLRP3）炎性小体介导的儿科炎症性疾病的治疗具有挑战性。在这里，我们研究了环磷酸腺苷（cAMP）提升剂福司可林是否可以减轻 Nigericin 诱导的人类巨噬细胞中 NLRP3 炎性小体的激活和白细胞介素-1β（IL-1β）的分泌。
在福司可林刺激的 Nigericin 激活的人 THP-1 巨噬细胞和原代巨噬细胞中测量炎性小体结构蛋白和前炎性细胞因子的蛋白和信使 RNA（mRNA）水平。
Nigericin 激活 THP-1 巨噬细胞增加了 NLRP3、IL-1β 和 caspase-1 的 mRNA 表达（P<0.01）。福司可林刺激对 Nigericin 激活细胞中的 NLRP3、caspase-1 或 IL-1β 的 mRNA 表达没有影响（P>0.05），而它们的蛋白水平则明显降低（P<0.05）。在 Nigericin 激活的巨噬细胞中，非激活细胞中福司可林介导的细胞质 cAMP 增加被减弱（P<0.05）。Nigericin 激活的巨噬细胞中，基础 IL-1β 的分泌从 584 增加到 2696 pg/mL（P<0.01）；福司可林剂量依赖性地减少了 Nigericin 诱导的成熟 IL-1β 的分泌（P<0.01）。福司可林还抑制了激活的人原代巨噬细胞中 IL-1β 的分泌。
福司可林抑制了人类巨噬细胞中 NLRP3 炎性小体的激活和成熟的 IL-1β 的分泌。福司可林和其他 cAMP 提升剂药物可能代表了一种治疗与过度炎性小体激活相关疾病的新方法，例如儿科炎症性疾病。
