Zhu Yanrong, Guo Yilei, Guo Pengxiang, Zhang Jing, He Yue, Xia Yufeng, Wei Zhifeng, Dai Yue
Department of Pharmacology of Chinese Materia Medica, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 639 Long Mian Avenue, Nanjing 211198, China.
Department of Pharmacognosy, School of Traditional Chinese Pharmacy, China Pharmaceutical University, 639 Long Mian Avenue, Nanjing 211198, China.
J Adv Res. 2025 May;71:571-584. doi: 10.1016/j.jare.2024.06.004. Epub 2024 Jun 4.
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.
尽管已有报道称几种雌激素受体β（ERβ）激动剂可缓解炎症性肠病（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及机制。
使用葡聚糖硫酸钠（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评估小鼠结肠炎的严重程度以及细胞因子和/或趋化因子水平。
生物信息学分析、酶联免疫吸附测定（ELISA）和免疫荧光结果显示，与ERβ表达和激活相关的靶向细胞因子和/或趋化因子是白细胞介素-1β（IL-1β），腺相关病毒9-IL-1β（AAV9-IL-1β）的过表达显著减弱了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表达的下调最为明显。鉴于NLRP3、NLRC4和AIM2炎性小体激活在IL-1β产生中的关键作用，我们研究了ERβ激活对炎性小体活性的影响。ELISA和蛋白质免疫印迹（WB）结果显示，ERβ激活选择性地阻断了NLRP3炎性小体组装介导的IL-1β分泌。钙敏感受体（CaSR）和钙信号在NLRP3炎性小体的组装中起关键作用。WB和免疫荧光结果显示，ERβ激活降低了结肠巨噬细胞中细胞内CaSR的表达和钙信号。与CaSR过表达质粒联合使用可逆转ERβ激活对NLRP3炎性小体组装的抑制作用，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的表达以降低细胞内钙水平并抑制NLRP3炎性小体组装介导的IL-1β产生来实现的。
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