The role of osteopontin in modulating macrophage phagocytosis of calcium oxalate crystals.

IF 2 2区 医学 Q2 UROLOGY & NEPHROLOGY
Tatsuya Hattori, Kazumi Taguchi, Ryosuke Chaya, Shuzo Hamamoto, Atsushi Okada, Takahiro Yasui
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Abstract

In inflammation, osteopontin (OPN) acts as both a stone matrix component for calcium oxalate (CaOx) crystal formation and an inflammatory mediator. While previous studies have demonstrated the individual roles of OPN and macrophages (Mφ) in renal CaOx stone formation during inflammation, their interaction remains poorly understood. This study aimed to elucidate the role of OPN in modulating Mφ function during crystal formation, using an ex vivo model. Bone marrow-derived macrophages (BMDM) were isolated from eight-week-old male C57BL/6J wild-type and OPN knockout mice. BMDMs from OPN-positive (BMDMOPN+) and OPN-negative (BMDMOPN-) mice were co-cultured with fluorescently labeled CaOx monohydrate (COM) crystals for phagocytosis assays and analyzed using the IN Cell Analyzer 6000. We further performed real-time quantitative reverse transcription PCR and RNA sequencing to identify gene expression profiles and clarify the role of OPN in Mφ function. The assay analysis demonstrated that phagocytosis rates were significantly higher in BMDMOPN- than in BMDMOPN+. Inflammatory markers, such as IL-6, TNF, CD44, were upregulated following COM exposure, and IL-6 expression was significantly lower in BMDMOPN- than in BMDMOPN+. RNA sequencing revealed that BMDMOPN- exhibited a less pro-inflammatory and more anti-inflammatory phenotype (Csf2low, Irf5low, Itgaxlow, Csf1high, Cd163high), resembling M2-like Mφs. Further functional analysis indicated that OPN knockdown in Mφs increased the S100 family and CREB signaling, which enhanced the M2-like phenotype shift and phagosome formation. In conclusion, OPN plays a critical role in enhancing pro-inflammatory Mφ function, potentially limiting COM phagocytosis. Modulating OPN expression in circulating Mφs may represent a therapeutic approach for kidney stone disease.

骨桥蛋白在调节草酸钙晶体巨噬细胞吞噬中的作用。
在炎症中,骨桥蛋白(OPN)既是草酸钙(CaOx)晶体形成的石基质成分,也是炎症介质。虽然先前的研究已经证明了OPN和巨噬细胞(Mφ)在炎症期间肾CaOx结石形成中的个体作用,但它们之间的相互作用仍然知之甚少。本研究旨在通过体外模型阐明OPN在晶体形成过程中调节Mφ功能的作用。从8周龄雄性C57BL/6J野生型和OPN敲除小鼠中分离骨髓源性巨噬细胞(BMDM)。将来自opn阳性(BMDMOPN+)和opn阴性(BMDMOPN-)小鼠的BMDMs与荧光标记的氧化钙(COM)晶体共培养进行吞噬实验,并使用IN细胞分析仪6000进行分析。我们进一步通过实时定量反转录PCR和RNA测序来鉴定基因表达谱,并阐明OPN在Mφ功能中的作用。实验分析表明,BMDMOPN-的吞噬率明显高于BMDMOPN+。炎症标志物,如IL-6、TNF、CD44在COM暴露后上调,IL-6在BMDMOPN-中的表达明显低于BMDMOPN+。RNA测序结果显示,BMDMOPN-具有较低的促炎和较强的抗炎表型(Csf2low, Irf5low, Itgaxlow, Csf1high, Cd163high),类似于m2样的m - φs。进一步的功能分析表明,m - φs中OPN的敲低增加了S100家族和CREB信号,从而增强了m2样表型转移和吞噬体的形成。综上所述,OPN在增强促炎Mφ功能中起关键作用,可能限制COM吞噬。调节循环Mφs中OPN的表达可能是肾结石疾病的一种治疗方法。
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来源期刊
Urolithiasis
Urolithiasis UROLOGY & NEPHROLOGY-
CiteScore
4.50
自引率
6.50%
发文量
74
期刊介绍: Official Journal of the International Urolithiasis Society The journal aims to publish original articles in the fields of clinical and experimental investigation only within the sphere of urolithiasis and its related areas of research. The journal covers all aspects of urolithiasis research including the diagnosis, epidemiology, pathogenesis, genetics, clinical biochemistry, open and non-invasive surgical intervention, nephrological investigation, chemistry and prophylaxis of the disorder. The Editor welcomes contributions on topics of interest to urologists, nephrologists, radiologists, clinical biochemists, epidemiologists, nutritionists, basic scientists and nurses working in that field. Contributions may be submitted as full-length articles or as rapid communications in the form of Letters to the Editor. Articles should be original and should contain important new findings from carefully conducted studies designed to produce statistically significant data. Please note that we no longer publish articles classified as Case Reports. Editorials and review articles may be published by invitation from the Editorial Board. All submissions are peer-reviewed. Through an electronic system for the submission and review of manuscripts, the Editor and Associate Editors aim to make publication accessible as quickly as possible to a large number of readers throughout the world.
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