Li Xinxin, Li Jianchao, Zhou Liangliang, Li Yingbin, Wu Xufeng, Chen Jianjun
Department of Emergency Intensive Care Medicine and Emergency Medicine, Yancheng First Hospital, Affiliated Hospital of Nanjing University Medical School, No. 166, Yulong West Road, Tinghu District, Yancheng, 224000, Jiangsu, China.
Clin Exp Nephrol. 2025 Apr 30. doi: 10.1007/s10157-025-02690-z.
Sepsis, a systemic inflammatory disease, often leads to severe complications, including septic acute kidney injury (AKI). Despite its prevalence, effective treatments for septic AKI remain elusive. MicroRNAs (miRNAs) are critical regulators of gene expression and play essential roles in sepsis pathogenesis. This study investigates the function and regulatory pathway of miR-760 in septic AKI.
Septic AKI models were developed using lipopolysaccharide (LPS)-treated mice and HK-2 renal tubular epithelial cells. Renal pathology was assessed via H&E staining, while renal function was assessed through the measurement of blood urea nitrogen (BUN) and creatinine levels. Apoptosis, inflammation, and oxidative stress markers were analyzed using functional assays, ELISA, and flow cytometry. Bioinformatics analysis, RT-qPCR, and dual luciferase reporter assays were used to identify and validate miR-760's target, enkurin domain containing 1 (ENKD1).
Database analysis and experimental validation revealed markedly decreased level of miR-760-3p levels in LPS-induced septic AKI mice model. MiR-760-3p overexpression in vivo ameliorated renal damage, improved kidney function, and reduced tubular apoptosis, inflammation, and oxidative stress. In vitro, miR-760 mimics enhanced cell proliferation while inhibiting apoptosis, inflammation, and oxidative stress in LPS-stimulated HK-2 cells. Mechanistically, ENKD1 was identified as a direct target of miR-760. ENKD1 overexpression negated the protective effects of miR-760, exacerbating renal injury, apoptosis, and inflammatory responses.
This study demonstrates that miR-760 alleviates septic AKI by targeting ENKD1, reducing apoptosis, inflammation, and oxidative stress. The miR-760/ENKD1 axis offers a promising therapeutic avenue to address septic AKI.
脓毒症是一种全身性炎症性疾病，常导致严重并发症，包括脓毒症急性肾损伤（AKI）。尽管其发病率很高，但针对脓毒症AKI的有效治疗方法仍然难以捉摸。微小RNA（miRNA）是基因表达的关键调节因子，在脓毒症发病机制中起重要作用。本研究调查了miR-760在脓毒症AKI中的功能和调控途径。
使用脂多糖（LPS）处理的小鼠和HK-2肾小管上皮细胞建立脓毒症AKI模型。通过苏木精-伊红（H&E）染色评估肾脏病理学，通过测量血尿素氮（BUN）和肌酐水平评估肾功能。使用功能测定、酶联免疫吸附测定（ELISA）和流式细胞术分析细胞凋亡、炎症和氧化应激标志物。采用生物信息学分析、逆转录定量聚合酶链反应（RT-qPCR）和双荧光素酶报告基因测定法来鉴定和验证miR-760的靶标，即含enkurin结构域1（ENKD1）。
数据库分析和实验验证显示，在LPS诱导的脓毒症AKI小鼠模型中，miR-760-3p水平显著降低。体内过表达miR-760-3p可减轻肾脏损伤，改善肾功能，并减少肾小管细胞凋亡、炎症和氧化应激。在体外，miR-760模拟物可增强LPS刺激的HK-2细胞的增殖，同时抑制细胞凋亡、炎症和氧化应激。机制上，ENKD1被鉴定为miR-760的直接靶标。ENKD1过表达消除了miR-760的保护作用，加重了肾损伤、细胞凋亡和炎症反应。
本研究表明，miR-760通过靶向ENKD1减轻脓毒症AKI，减少细胞凋亡、炎症和氧化应激。miR-760/ENKD1轴为治疗脓毒症AKI提供了一条有前景的治疗途径。
