动脉粥样硬化作为多种心脑血管疾病的主要病理基础，已成为全球范围内导致死亡和残疾的主要原因之一。新的证据表明，Rho GTPase Rnd3 在心血管疾病中发挥着无可争议的作用，尽管其在动脉粥样硬化中的功能仍不清楚。在这里，我们发现 Rnd3 与主动脉内皮细胞 (EC) 焦亡之间存在显着相关性。ApoeKO 小鼠被用作动脉粥样硬化模型。使用内皮特异性转基因小鼠来破坏体内 Rnd3 的表达水平。使用液相色谱串联质谱 (LC-MS/MS)、免疫共沉淀 (Co-IP) 测定和分子对接对 Rnd3 对内皮细胞焦亡的影响进行了机制研究。来自功能获得和功能获得的证据功能丧失研究表明 Rnd3 对 EC 焦亡具有保护作用。 Rnd3 的下调使 EC 在氧化低密度脂蛋白 (oxLDL) 攻击下对细胞焦亡敏感，并加剧动脉粥样硬化，而 Rnd3 的过度表达则有效阻止了这些影响。 LC-MS/MS、Co-IP 测定和分子对接表明，Rnd3 通过与肿瘤坏死因子受体相关因子 6 (TRAF6) 的无名指结构域直接相互作用，负向调节细胞焦亡信号传导。这导致K63连接的TRAF6泛素化的抑制和K48连接的TRAF6泛素化的促进，抑制NF-κB的激活并促进TRAF6的降解。此外，TRAF6 敲低可以对抗 Rnd3 敲除引起的 EC 细胞焦亡加剧。这些发现在 EC 中建立了 Rnd3 与 TRAF6/NF-κB/NLRP3 信号通路之间的关键功能联系，表明 Rnd3 在预防细胞焦亡中的重要作用ECs。© 2023 作者。约翰·威利出版的《临床与转化医学》

As the main pathological basis for various cardiovascular and cerebrovascular diseases, atherosclerosis has become one of the leading causes of death and disability worldwide. Emerging evidence has suggested that Rho GTPase Rnd3 plays an indisputable role in cardiovascular diseases, although its function in atherosclerosis remains unclear. Here, we found a significant correlation between Rnd3 and pyroptosis of aortic endothelial cells (ECs).ApoeKO mice were utilized as a model for atherosclerosis. Endothelium-specific transgenic mice were employed to disrupt the expression level of Rnd3 in vivo. Mechanistic investigation of the impact of Rnd3 on endothelial cell pyroptosis was carried out using liquid chromatography tandem mass spectrometry (LC-MS/MS), co-immunoprecipitation (Co-IP) assays, and molecular docking.Evidence from gain-of-function and loss-of-function studies denoted a protective role for Rnd3 against ECs pyroptosis. Downregulation of Rnd3 sensitized ECs to pyroptosis under oxidized low density lipoprotein (oxLDL) challenge and exacerbated atherosclerosis, while overexpression of Rnd3 effectively prevented these effects. LC-MS/MS, Co-IP assay, and molecular docking revealed that Rnd3 negatively regulated pyroptosis signaling by direct interaction with the ring finger domain of tumor necrosis factor receptor-associated factor 6 (TRAF6). This leads to the suppression of K63-linked TRAF6 ubiquitination and the promotion of K48-linked TRAF6 ubiquitination, inhibiting the activation of NF-κB and promoting the degradation of TRAF6. Moreover, TRAF6 knockdown countered Rnd3 knockout-evoked exacerbation of EC pyroptosis in vivo and vitro.These findings establish a critical functional connection between Rnd3 and the TRAF6/NF-κB/NLRP3 signaling pathway in ECs, indicating the essential role of Rnd3 in preventing pyroptosis of ECs.© 2023 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.