Liu Yanqi, Xin Yu, Yuan Mengyao, Liu Yuhan, Song Yuchen, Shen Lifeng, Xiao Yu, Wang Xinran, Wang Dawei, Liu Linqiong, Liu Yuxi, Luo Yinghao, Huang Pengfei, Zhang Qianqian, Zhang Weiting, Li Hongxu, Zhou Yuxin, Wang Xibo, Yu Kaijiang, Wang Changsong
Department of Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, 150001, China; Heilongjiang Provincial Key Laboratory of Critical Care Medicine, Harbin, 150001, China; Central Laboratory of the First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China.
Department of Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, 150001, China; Department of Critical Care Medicine, Harbin Medical University Cancer Hospital, No. 150 Haping Rd, Nangang District, Harbin, 150081, China; Heilongjiang Provincial Key Laboratory of Critical Care Medicine, Harbin, 150001, China; Central Laboratory of the First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China.
Arch Biochem Biophys. 2025 Mar;765:110318. doi: 10.1016/j.abb.2025.110318. Epub 2025 Jan 23.
Ischemia-reperfusion injury (IRI) often results in renal impairment. While the presence of neutrophil extracellular traps (NETs) is consistently observed, their specific impact on IRI is not yet defined. Sivelestat sodium, an inhibitor of neutrophil elastase which is crucial for NET formation, may offer a therapeutic approach to renal IRI, warranting further research.
A mouse model was established for early-stage renal IRI, confirmed by injury markers and histological assessments. The involvement of NETs in renal I/R was demonstrated using immunofluorescence and Western blot. Renal function and pathology were further evaluated through a comprehensive set of methods, including Periodic Acid-Schiff staining (PAS) and Terminal Deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) staining, enzyme-linked immunosorbent assay (ELISA), Real time Glomerular Filtration Rate (RT-GFR) monitoring, Polymerase Chain Reaction (PCR), biochemical analysis, and additional Western blot and immunofluorescence assays.
We firstly quantified NET expression in renal IRI mice, noting a peak at 24 h. Subsequently, sivelestat sodium treatment was administered, resulting in decreased MPO, CitH3, and attenuated tubular damage. Moreover, it resulted in a decrease in serum levels of creatinine, blood urea nitrogen (BUN), as well as neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1). Additionally, it lowered the abundance of renal tissue inflammatory markers interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), and mitigated the levels of oxidative stress indicators malondialdehyde (MDA) and 4 Hydroxynonenal (4HNE), accompanied by a decline in renal cell apoptosis and an enhancement of GFR in renal I/R mice.
Sivelestat sodium ameliorates renal IRI by downregulating neutrophil NETs, reducing inflammation, oxidative stress, and apoptosis, thereby enhancing renal function.
缺血再灌注损伤（IRI）常导致肾功能损害。虽然一直观察到中性粒细胞胞外陷阱（NETs）的存在，但其对IRI的具体影响尚未明确。西维来司他钠是一种对NET形成至关重要的中性粒细胞弹性蛋白酶抑制剂，可能为肾IRI提供一种治疗方法，值得进一步研究。
建立早期肾IRI小鼠模型，通过损伤标志物和组织学评估进行确认。使用免疫荧光和蛋白质印迹法证明NETs参与肾缺血/再灌注。通过一系列综合方法进一步评估肾功能和病理，包括高碘酸-希夫染色（PAS）和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色、酶联免疫吸附测定（ELISA）、实时肾小球滤过率（RT-GFR）监测、聚合酶链反应（PCR）、生化分析以及额外的蛋白质印迹和免疫荧光测定。
我们首先对肾IRI小鼠中的NET表达进行了定量，发现在24小时达到峰值。随后给予西维来司他钠治疗，导致髓过氧化物酶（MPO）、瓜氨酸化组蛋白H3（CitH3）减少，肾小管损伤减轻。此外，它还导致血清肌酐、血尿素氮（BUN）以及中性粒细胞明胶酶相关脂质运载蛋白（NGAL）和肾损伤分子-1（KIM-1）水平降低。此外，它降低了肾组织炎症标志物白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的丰度，并减轻了氧化应激指标丙二醛（MDA）和4-羟基壬烯醛（4HNE）的水平，同时肾缺血/再灌注小鼠的肾细胞凋亡减少，肾小球滤过率提高。
西维来司他钠通过下调中性粒细胞NETs、减轻炎症、氧化应激和细胞凋亡来改善肾IRI，从而增强肾功能。
