HUWE1介导的TTC4泛素化与降解通过抑制PINK1/parkin依赖性线粒体自噬并促进FKBP5/NF-κB诱导的Treg/Th17失衡加重过敏性鼻炎
HUWE1-mediated ubiquitination and degradation of TTC4 aggravates allergic rhinitis via inhibiting PINK1/parkin-dependent mitophagy and promoting FKBP5/NF-κB-induced Treg/Th17 imbalance.
文献信息
| PMID | 42722119 |
|---|---|
| 原文 | 在 PubMed 查看原文 ↗ |
| 发表日期 | 2026 |
| 作者 | Yuting Huo |
| 作者单位 | Department of Otolaryngology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou, Fujian Province, 350001, China. |
| 期刊 | Archives of biochemistry and biophysics |
| SCI 分区 | Q2 |
| IF | 3.9 |
| 研究类型 | 基础研究 · 基础／转化 |
| 所属专科 | 鼻科 |
中文摘要
背景： 此前，我们证明四肽重复结构域4（TTC4）在过敏性鼻炎（AR）中保护免受炎症损伤。本研究进一步探讨HECT、UBA和WWE结构域包含蛋白1（HUWE1）是否作为E3泛素连接酶靶向TTC4参与AR发病机制。
方法： 体外用IL-13刺激HNEpCs，并用卵清蛋白致敏小鼠建立AR模型。通过CCK-8测定细胞活力。分别用DCFH-DA、MitoSOX Red和JC-1染色检测细胞内和线粒体氧化剂产生及线粒体膜电位。用流式细胞术分析Treg/Th17细胞比例。进行RT-qPCR、Western blot、免疫荧光、免疫组化和ELISA以检测mRNA和蛋白水平。通过H&E和Giemsa染色评估鼻组织病理学。共免疫沉淀和泛素化实验剖析分子机制。
结果： HUWE1在AR小鼠和IL-13刺激的HNEpCs中增加。HUWE1沉默通过抑制TTC4泛素化依赖性降解稳定TTC4。HUWE1敲低减少NLRP3炎症小体激活和线粒体氧化损伤，并在IL-13条件下增强线粒体自噬，这些作用被TTC4沉默所消除。线粒体自噬抑制剂3-MA/Mdivi-1抵消了HUWE1耗竭/TTC4过表达对IL-13诱导的HNEpCs损伤的有益作用。在AR小鼠中，HUWE1耗竭减轻鼻黏膜增厚、炎症反应和NLRP3激活，并恢复Treg/Th17平衡和失活FK506结合蛋白5（FKBP5）/NF-κB通路，这些作用在TTC4敲低后被消除。
结论： HUWE1沉默减少TTC4泛素化依赖性降解，从而增强线粒体自噬以缓解线粒体氧化损伤，并减轻FKBP5/NF-κB触发的Treg/Th17失衡，从而改善AR。本研究为AR提供了有前景的治疗靶点。
英文摘要
BACKGROUND: Previously, we proved that tetratricopeptide repeat domain 4 (TTC4) protected against inflammatory injury in allergic rhinitis (AR). This study further explored whether HECT, UBA, and WWE domain containing protein 1 (HUWE1) functions as an E3 ubiquitin ligase targeting TTC4 to participate in AR pathogenesis.
METHODS: HNEpCs were stimulated with IL-13 in vitro, and mice were sensitized with ovalbumin to establish AR model. Cell viability measured by CCK-8. Intracellular and mitochondrial oxidant productions and mitochondrial membrane potential were detected with DCFH-DA, MitoSOX Red, and JC-1 staining, respectively. The proportions of Treg/Th17 cells were analyzed with flow cytometry. RT-qPCR, Western blot, immunofluorescence, immunohistochemistry, and ELISA were conducted to examine mRNA and protein levels. Nasal histopathology was assessed by H&E and Giemsa staining. Co-immunoprecipitation and ubiquitination assay dissect the molecular mechanism.
RESULTS: HUWE1 was increased in AR mice and IL-13-stimulated HNEpCs. HUWE1 silencing stabilized TTC4 via inhibiting its ubiquitination-dependent degradation. HUWE1 knockdown diminished NLRP3 inflammasome activation and mitochondrial oxidative damage, and enhanced mitophagy under IL-13 condition, which were abolished by TTC4 silencing. Mitophagy inhibitor 3-MA/Mdivi-1 counteracted the beneficial effects of HUWE1 depletion/TTC4 overexpression on IL-13-induced HNEpCs injury. In AR mice, HUWE1 depletion relieved nasal mucosal thickening, inflammatory response and NLRP3 activation, as well as restored Treg/Th17 balance and inactivated FK506-binding protein 5 (FKBP5)/NF-κB pathway, which were abrogated upon TTC4 knockdown.
CONCLUSION: HUWE1 silencing reduced TTC4 ubiquitination-dependent degradation, which enhanced mitophagy to relieve mitochondrial oxidative damage and mitigated FKBP5/NF-κB-triggered Treg/Th17imbalance, thus improving AR. This study provided promising therapeutic targets for AR.