Yang Zhiming, Chen Xiong, Qi Guannan, Gu Jie, Liu Zheng, Zhang Xiaobo
Department of Geriatric Urology, Xiangya International Medical Center, Xiangya Hospital, Central South University, Changsha, PR China.
National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, PR China.
Kidney Dis (Basel). 2024 Nov 21;11(1):11-24. doi: 10.1159/000542471. eCollection 2025 Jan-Dec.
This study aims to explore the contribution of neutrophil extracellular traps (NETs) to kidney stones.
The microarray data from GSE73680 and bioinformatic analysis were applied to identify differentially expressed genes in patients with kidney stones. A rat model of kidney stones was established through ethylene glycol and ammonium chloride administration. The plasma was collected for examining cf-DNA, DNase I, MPO-DNA, H3Cit and NE. Superoxide dismutase, malondialdehyde, creatinine, blood urea nitrogen, and calcium were examined through biochemical analysis. MPO, H3Cit, and NE in kidney tissues were detected via immunofluorescence staining. Cell apoptosis was evaluated through TUNEL assays. HE, Periodic Acid-Schiff and Von Kossa staining were applied to determine histological structure, calcium deposits and stone formation in the kidneys. Neutrophil elastase inhibitor Sivelestat (SIVE) was administrated for NET suppression in rats.
A total of 403 differentially expressed genes including 270 upregulated and 133 downregulated genes were identified between renal papillary tissues with Randall's plaque and normal tissues. Gene ontology enrichment, KEGG pathway and protein-protein interaction network analysis of these dysregulated genes were performed. Moreover, increased NET markers including cf-DNA, DNase I, MPO-DNA, H3Cit and NE and calcium deposits were observed in patients with kidney stones. Subsequently, we established a rat model of kidney stones. We found that NET formation was significantly elevated in kidney stone rats, and renal tubular injury and apoptotic cells were enhanced as kidney stones developed. Strikingly, we found that suppression of NETs via SIVE could significantly reduce calcium deposits and apoptotic cells and alleviate tubular injury, thus improving kidney function.
NETs drive the formation of kidney stones, thus aggravating kidney injury. Our study identifies NETs as a potential diagnostic and therapeutic biomarker for nephrolithiasis.
本研究旨在探讨中性粒细胞胞外诱捕网（NETs）在肾结石形成中的作用。
应用来自GSE73680的微阵列数据和生物信息学分析，以鉴定肾结石患者中差异表达的基因。通过给予乙二醇和氯化铵建立肾结石大鼠模型。收集血浆以检测cf-DNA、脱氧核糖核酸酶I（DNase I）、髓过氧化物酶-DNA（MPO-DNA）、组蛋白H3瓜氨酸化修饰（H3Cit）和中性粒细胞弹性蛋白酶（NE）。通过生化分析检测超氧化物歧化酶、丙二醛、肌酐、血尿素氮和钙。通过免疫荧光染色检测肾组织中的MPO、H3Cit和NE。通过TUNEL法评估细胞凋亡。应用苏木精-伊红（HE）染色、过碘酸-希夫（Periodic Acid-Schiff）染色和冯科萨（Von Kossa）染色来确定肾脏的组织结构、钙沉积和结石形成。给予中性粒细胞弹性蛋白酶抑制剂西维来司他（SIVE）以抑制大鼠体内的NETs形成。
在伴有兰德尔斑的肾乳头组织与正常组织之间共鉴定出403个差异表达基因，其中270个上调基因和133个下调基因。对这些失调基因进行了基因本体富集分析、KEGG通路分析和蛋白质-蛋白质相互作用网络分析。此外，在肾结石患者中观察到包括cf-DNA、DNase I、MPO-DNA、H3Cit和NE在内的NET标志物增加以及钙沉积增加。随后，我们建立了肾结石大鼠模型。我们发现肾结石大鼠的NETs形成显著升高，并且随着肾结石的发展，肾小管损伤和凋亡细胞增加。令人惊讶的是，我们发现通过SIVE抑制NETs可显著减少钙沉积和凋亡细胞，并减轻肾小管损伤，从而改善肾功能。
NETs促进肾结石的形成，从而加重肾损伤。我们的研究将NETs鉴定为肾结石潜在的诊断和治疗生物标志物。
