Although several estrogen receptor β (ERβ) agonists have been reported to alleviate IBD, the pivotal mechanism remains obscure. To examine the effects and mechanisms of ERβ activation on cytokine/chemokine networks in colitis mice. Dextran sulfate sodium salt (DSS) and trinitro-benzene-sulfonic acid (TNBS) were used to induce mouse colitis model. Multiple molecular biological methods were employed to evaluate the severity of mouse colitis and the level of cytokine and/or chemokine. Bioinformatics analysis, ELISA and immunofluorescence results showed that the targeted cytokines and/or chemokines associated with ERβ expression and activation is IL-1β, and the anti-colitis effect of ERβ activation was significantly attenuated by the overexpression of AAV9-IL-1β. Immunofluorescence analysis indicated that ERβ activation led to most evident downregulation of IL-1β expression in colonic macrophages as compared to monocytes and neutrophils. Given the pivotal roles of NLRP3, NLRC4, and AIM2 inflammasome activation in the production of IL-1β, we examined the influence of ERβ activation on inflammasome activity. ELISA and WB results showed that ERβ activation selectively blocked the NLRP3 inflammasome assembly-mediated IL-1β secretion. The calcium-sensing receptor (CaSR) and calcium signaling play crucial roles in the assembly of the NLRP3 inflammasome. WB and immunofluorescence results showed that ERβ activation reduced intracellular CaSR expression and calcium signaling in colonic macrophages. Combination with CaSR overexpression plasmid reversed the suppressive effect of ERβ activation on NLRP3 inflammasome assembly, and counteracting the downregulation of IL-1β secretion. Our research uncovers that the anti-colitis effect of ERβ activation is accomplished through the reduction of IL-1β levels in colonic tissue, achieved by specifically decreasing CaSR expression in macrophages to lower intracellular calcium levels and inhibit NLRP3 inflammasome assembly-mediated IL-1β production.

中文翻译：

---

雌激素受体 β 激活通过下调细胞内钙水平抑制巨噬细胞中 NLRP3 依赖性 IL-1β 的产生，从而减轻炎症性肠病


尽管据报道几种雌激素受体 β (ERβ) 激动剂可以缓解 IBD，但其关键机制仍不清楚。研究 ERβ 激活对结肠炎小鼠细胞因子/趋化因子网络的影响和机制。采用硫酸葡聚糖钠盐（DSS）和三硝基苯磺酸（TNBS）诱导小鼠结肠炎模型。采用多种分子生物学方法来评估小鼠结肠炎的严重程度以及细胞因子和/或趋化因子的水平。生物信息学分析、ELISA和免疫荧光结果表明，与ERβ表达和激活相关的靶向细胞因子和/或趋化因子是IL-1β，并且AAV9-IL-1β的过表达显着减弱ERβ激活的抗结肠炎作用。免疫荧光分析表明，与单核细胞和中性粒细胞相比，ERβ 激活导致结肠巨噬细胞中 IL-1β 表达最明显的下调。鉴于 NLRP3、NLRC4 和 AIM2 炎症小体激活在 IL-1β 产生中的关键作用，我们研究了 ERβ 激活对炎症小体活性的影响。 ELISA 和 WB 结果表明 ERβ 激活选择性阻断 NLRP3 炎性体组装介导的 IL-1β 分泌。钙敏感受体 (CaSR) 和钙信号传导在 NLRP3 炎症小体的组装中发挥着至关重要的作用。 WB 和免疫荧光结果表明 ERβ 激活降低了结肠巨噬细胞中的细胞内 CaSR 表达和钙信号传导。与CaSR过表达质粒联合逆转ERβ激活对NLRP3炎性体组装的抑制作用，并抵消IL-1β分泌的下调。 我们的研究发现，ERβ 激活的抗结肠炎作用是通过降低结肠组织中 IL-1β 水平来实现的，这是通过特异性降低巨噬细胞中 CaSR 的表达来降低细胞内钙水平并抑制 NLRP3 炎症小体组装介导的 IL-1β 产生来实现的。