Lertprapai Chanettee, Peerapen Paleerath, Thongboonkerd Visith
Medical Proteomics Unit, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, 6th Floor - SiMR Building, 2 Wanglang Road, Bangkoknoi, Bangkok, 10700, Thailand.
Graduate Program in Molecular Medicine, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
Cell Commun Signal. 2025 Jul 11;23(1):336. doi: 10.1186/s12964-025-02345-2.
The majority of stone formers (87.5-95.9%) exhibit mild to moderate interstitial inflammation surrounding the stone. Neutrophils and neutrophil-derived genes/proteins have been found in renal papillae, stone matrix and urine of calcium oxalate monohydrate (COM) stone formers. However, neutrophil-crystal interactions, especially responses of neutrophils to COM crystals, remained unknown.
This study addressed the effects of COM crystals on neutrophil cellular proteome and functions, including phagocytosis, activation/degranulation, neutrophil extracellular traps (NETs) formation and reactive oxygen species (ROS) production.
Label-free quantitative (LFQ) proteomics using nanoLC-ESI-Qq-TOF MS/MS with highly stringent criteria revealed that COM caused altered levels of 22 neutrophil proteins involved mainly in immune responses. Investigating neutrophil innate immune functions using flow cytometry, immunofluorescence/fluorescence imaging, ELISA and dichlorodihydrofluorescein diacetate (DCFH-DA) assay revealed that COM enhanced neutrophil phagocytic activity, NETs formation, activation/degranulation and ROS production. Moreover, secretome (a set of secretory products) from COM-treated neutrophils induced the recruitment of macrophages to phagocytose the COM-treated neutrophils.
These findings illustrate the expression and functional responses of neutrophils to COM crystals and implicate the important roles that neutrophils play in nephrolithiasis.
大多数结石形成者（87.5 - 95.9%）在结石周围表现出轻度至中度的间质炎症。在一水草酸钙（COM）结石形成者的肾乳头、结石基质和尿液中发现了中性粒细胞以及中性粒细胞衍生的基因/蛋白质。然而，中性粒细胞与晶体的相互作用，尤其是中性粒细胞对COM晶体的反应，仍然未知。
本研究探讨了COM晶体对中性粒细胞细胞蛋白质组和功能的影响，包括吞噬作用、激活/脱颗粒、中性粒细胞胞外陷阱（NETs）形成和活性氧（ROS）产生。
使用具有高度严格标准的nanoLC - ESI - Qq - TOF MS/MS进行的无标记定量（LFQ）蛋白质组学研究表明，COM导致22种主要参与免疫反应的中性粒细胞蛋白质水平发生改变。通过流式细胞术、免疫荧光/荧光成像、ELISA和二氯二氢荧光素二乙酸酯（DCFH - DA）检测来研究中性粒细胞的固有免疫功能，结果显示COM增强了中性粒细胞的吞噬活性、NETs形成、激活/脱颗粒和ROS产生。此外，COM处理的中性粒细胞分泌组（一组分泌产物）诱导巨噬细胞募集以吞噬COM处理的中性粒细胞。
这些发现阐明了中性粒细胞对COM晶体的表达和功能反应，并暗示了中性粒细胞在肾结石形成中所起的重要作用。
