Non-canonical NF-κB contributes to endothelial pyroptosis and atherogenesis dependent on IRF-1.
Non-canonical NF-κB contributes to endothelial pyroptosis and atherogenesis dependent on IRF-1.
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DOI：
10.1016/j.trsl.2022.11.001
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发表时间：
2022-11
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影响因子：
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通讯作者：
X. Fan;Qiannan Li;Yiying Wang;Dai-Min Zhang;Jingchao Zhou;Qing Chen;Liang Sheng;A. Passerini;ChongXiu Sun
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作者：
X. Fan;Qiannan Li;Yiying Wang;Dai-Min Zhang;Jingchao Zhou;Qing Chen;Liang Sheng;A. Passerini;ChongXiu Sun
细胞炎症和死亡是导致内皮功能障碍的密切相关过程，内皮功能障碍在动脉粥样硬化中起着关键作用。NLRP3炎性小体的激活导致焦亡，即气凝胶蛋白D （GSDMD）介导的炎症细胞死亡。非典型NF-κB通路与炎症有关；然而，其在NLRP3炎症小体介导的内皮功能障碍中的作用尚未被研究。本研究探讨非典型NF-κB通路在调节内皮细胞焦亡中的作用，因为它与动脉粥样硬化的发生有关。免疫组化显示人动脉粥样硬化性动脉内皮细胞（EC）的炎性体活化。流式细胞术和Western blot分析显示，氧化低密度脂蛋白（oxLDL）激活了NLRP3炎性体，同时激活了非典型NF-κB。NF-κ b诱导激酶（NIK）是非规范通路的关键调节因子，其干扰可显著减弱oxLDL-或LPS/ atp诱导的NLRP3炎性体激活、焦亡、IL-1β和IL-18分泌。相反，过度表达niki会加重这些反应。染色质免疫沉淀显示，非典型NF-κB通路的激活通过RelB/p52结合其启动子区域-782/-770上调转录因子IRF-1。除了已知的靶基因casp1外，RNA测序进一步确定了gsdmdas IRF-1的靶基因。IRF-1而非RelB/p52与gsdmd启动子-526/-515和-11/10位点的casp1启动子相互作用，促进GSDMD的表达和casp1介导的激活。与在培养内皮中观察到的结果一致，内皮特异性缺乏NIK或IRF-1可减轻高脂肪饮食中无apoe小鼠的动脉粥样硬化。这些数据表明，非典型NF-κB途径通过依赖于IRF-1的方式激活GSDMD，参与NLRP3炎症小体介导的内皮细胞凋亡和动脉粥样硬化的发展。进一步的研究可能有助于确定动脉粥样硬化性心脏病的特异性治疗靶点。
Cell inflammation and death are closely linked processes contributing to endothelial dysfunction, which plays a critical role in atherogenesis. Activation of the NLRP3 inflammasome causes pyroptosis, the Gasdermin D (GSDMD)-mediated inflammatory cell death. The non-canonical NF-κB pathway has been implicated in inflammation; however, its role in NLRP3 inflammasome-mediated endothelial dysfunction has not been investigated. This study investigated a role for the non-canonical NF-κB pathway in regulating endothelial pyroptosis as it relates to atherogenesis. Immunohistochemistry indicated inflammasome activation in the endothelial cells (EC) of human atherosclerotic arteries. Flow cytometry and Western blot analysis revealed that oxidized low-density lipoprotein (oxLDL) activated the NLRP3 inflammasome, concomitant with the activation of non-canonical NF-κB in primary human aortic EC. Interference of NF-κB inducing kinase (NIK), the key regulator of the non-canonical pathway, significantly attenuated oxLDL- or LPS/ATP-induced NLRP3 inflammasome activation, pyroptosis, IL-1β, and IL-18 secretion. In contrast, overexpression ofNIKexacerbated these responses. Chromatin immunoprecipitation revealed that activation of the non-canonical NF-κB pathway upregulated the transcription factor IRF-1 through RelB/p52 binding to its promoter region at -782/-770. In addition to the known targetCASP1,RNA sequencing further identifiedGSDMDas a target gene of IRF-1. IRF-1 but not RelB/p52 interacted with theGSDMDpromoter at -526/-515 and theCASP1promoter at -11/10 to promote the expression andCASP1-mediated activation of GSDMD. Consistent with the observations in cultured endothelium, endothelial-specific deficiency of NIK or IRF-1 attenuated atherosclerosis in high-fat diet-fedApoe-null mice. These data demonstrate that the non-canonical NF-κB pathway contributes to NLRP3 inflammasome-mediated endothelial pyroptosis and the development of atherosclerosis through GSDMD activation in a manner dependent on IRF-1. Further investigation may facilitate the identification of specific therapeutic targets for atherosclerotic heart diseases.