Yang Jianing, Dai Longzhu, Chai Jingmei, Li Yunong, Wang Jiangang, Yang Yi, Zhang Yulian, Yan Guanghai, Wang Chongyang
Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases, Yanbian University, Yanji 133002, PR China; Department of Anatomy, Histology and Embryology, Yanbian University Medical College, Yanji 133002, PR China.
Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases, Yanbian University, Yanji 133002, PR China; Department of Traditional Chinese Medicine, Yanbian University Medicine College, Yanji 133002, PR China.
Phytomedicine. 2025 Aug;144:156895. doi: 10.1016/j.phymed.2025.156895. Epub 2025 Jun 3.
Although tremendous progress has been made, the challenge of preventing and treating allergic rhinitis remains on a global scale. In recent years, there has been an increasing interest in the use of natural medicines. Narirutin, as a natural medicine, plays a significant role in anti - inflammation and anti - allergy. USP15, as an important member of the deubiquitinating enzyme family, can deubiquitinate NLRP3, thereby promoting the occurrence of allergic rhinitis.
This study mainly investigated the mechanism of Narirutin inhibiting USP15 deubiquitination of NLRP3 and alleviating allergic rhinitis.
In this study, the mechanism of Narirutin regulating USP15 deubiquitination of NLRP3 in vitro was investigated by molecular docking, Western blot, RT-qPCR, immunofluorescence, co-immunoprecipitation and other experimental methods. In vivo experiments, the AR mouse models were constructed using HDM induction, the nose touching and sneezing of AR mice were recorded. The mechanism of Narirutin alleviate the symptoms of allergic rhinitis in mice was studied by flow cytometry, HE, PAS, Western blot and other experimental methods.
The results showed that Narirutin inhibited the expression of NLRP3 by downregulating USP15 in RAW264.7 macrophages. Narirutin inhibits K63-linked ubiquitination of NLRP3 by decreasing USP15 in HEK293T cells. Importantly, siUSP15 alleviates mitochondrial damage in HNEpCs. In vivo experiments, it was further confirmed that Narirutin alleviated allergic rhinitis induced by HDM. After treatment with Narirutin, the symptoms of AR in mice were alleviated.
This study confirmed that Narirutin can alleviate AR by inhibiting USP15 deubiquitination of NLRP3, providing a new direction for the treatment and prevention of AR.
尽管已取得巨大进展，但预防和治疗变应性鼻炎的挑战在全球范围内依然存在。近年来，天然药物的使用受到越来越多的关注。橙皮苷作为一种天然药物，在抗炎和抗过敏方面发挥着重要作用。USP15作为去泛素化酶家族的重要成员，可使NLRP3去泛素化，从而促进变应性鼻炎的发生。
本研究主要探讨橙皮苷抑制USP15对NLRP3的去泛素化作用及缓解变应性鼻炎的机制。
本研究通过分子对接、蛋白质免疫印迹法（Western blot）、实时荧光定量聚合酶链反应（RT-qPCR）、免疫荧光、免疫共沉淀等实验方法，研究橙皮苷在体外调节USP15对NLRP3去泛素化的机制。在体内实验中，采用屋尘螨（HDM）诱导构建变应性鼻炎小鼠模型，记录变应性鼻炎小鼠的鼻触和喷嚏情况。通过流式细胞术、苏木精-伊红染色（HE）、过碘酸-希夫染色（PAS）、蛋白质免疫印迹法等实验方法，研究橙皮苷缓解小鼠变应性鼻炎症状的机制。
结果显示，橙皮苷通过下调RAW264.7巨噬细胞中USP15的表达来抑制NLRP3的表达。橙皮苷通过降低HEK293T细胞中USP15的水平来抑制NLRP3的K63连接的泛素化。重要的是，小干扰RNA靶向USP15（siUSP15）可减轻人鼻上皮细胞（HNEpCs）中的线粒体损伤。在体内实验中，进一步证实橙皮苷可减轻HDM诱导所致的变应性鼻炎。经橙皮苷治疗后，小鼠变应性鼻炎的症状得到缓解。
本研究证实橙皮苷可通过抑制USP15对NLRP3的去泛素化作用来缓解变应性鼻炎，为变应性鼻炎的治疗和预防提供了新方向。
