目的探讨天然水蛭素对高尿酸血症肾损伤的保护作用，并基于核因子-κB（NF-κB）信号通路与M1型巨噬细胞极化阐明其机制。方法 SD大鼠随机分为7组：对照组、模型组、别嘌醇（阳性对照，30 mg·kg^-1）和水蛭素低、中、高剂量组（0.266、0.533、1.066 g·kg^-1）及水蛭素（0.533 g·kg^-1）＋NF-κB通路激活剂（A2，5 mg·kg^-1）组。除对照组外，其余各组每天上午ig氧嗪酸钾（0.75 g·kg^-1）造模，连续5周；各给药组于造模同时每天下午ig相应药物，对照组和模型组ig等体积0.9%氯化钠溶液。通过苏木精-伊红（HE）染色、酶联免疫吸附测定（ELISA）、免疫荧光、Western blotting及转录组测序技术，评估水蛭素对肾脏病理、血清炎症因子、M1型巨噬细胞极化、尿酸转运蛋白表达及肾组织转录组谱的影响。结果水蛭素可缓解高尿酸血症大鼠体质量降低（P＜0.001），减轻肾脏病理损伤（P＜0.05、0.001），减轻肾小管上皮细胞变性与间质炎症细胞浸润，减少单钠尿酸盐（MSU）晶体沉积；显著降低血清白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）及肿瘤坏死因子-α（TNF-α）水平（P＜0.01、0.001）；显著抑制肾组织CD86阳性M1型巨噬细胞的浸润（P＜0.01、0.001）；双向调节尿酸转运蛋白表达，即下调尿酸盐转运蛋白1（URAT1）、葡萄糖转运蛋白9（GLUT9），上调有机阴离子转运蛋白1（OAT1）、有机阴离子转运蛋白3（OAT3）、三磷酸腺苷结合盒转运蛋白G2（ABCG2）（P＜0.05、0.01、0.001）。联合NF-κB通路激活剂可削弱或逆转水蛭素的上述效应。转录组测序显示，水蛭素可部分逆转高尿酸血症诱导的肾脏基因表达紊乱，差异基因显著富集于白细胞介素-17（IL-17）信号通路、核苷酸结合寡聚化结构域（NOD）样受体信号通路及上皮细胞分化等过程。结论水蛭素可能通过抑制NF-κB信号通路活化，负向调控M1型巨噬细胞极化，减轻MSU晶体相关炎症反应，并恢复肾脏尿酸转运蛋白稳态，从而保护高尿酸血症肾损伤。

Objective To investigate the renoprotective effects of natural hirudin on hyperuricemia-induced renal injury, and to elucidate the underlying mechanisms with a focus on the nuclear factor-κB (NF-κB) signaling pathway and M1 macrophage polarization. Methods SD rats were randomly divided into seven groups: control group, model group, allopurinol (positive control, 30 mg·kg^-1) group, low, medium, and high-dose hirudin groups (0.266, 0.533, 1.066 g·kg^-1), and hirudin (0.533 g·kg^-1) + NF-κB pathway activator (activator 2, A2, 5 mg·kg^-1) group. Except for the control group, all other groups received model establishment by ig administration of potassium oxonate (0.75 g·kg^-1) every morning for 5 consecutive weeks. The drug administration groups received corresponding drugs by ig administration every afternoon at the same time as model establishment, while the control and model groups received ig administration of equal volumes of 0.9% sodium chloride solution. Renal histopathology, serum inflammatory cytokines, M1 macrophage polarization, urate transporter expression, and renal transcriptomic profiles were evaluated using hematoxylin–eosin (HE) staining, enzyme-linked immunosorbent assay (ELISA), immunofluorescence, Western blotting, and RNA sequencing, respectively. Results Hirudin exerted a protective effect against body weight reduction (P < 0.001) and pathological renal injury (P < 0.05, and 0.001) in hyperuricemic rats, as evidenced by reduced degeneration of renal tubular epithelial cells, infiltration of interstitial inflammatory cells, and deposition of monosodium urate (MSU) crystals. Hirudin significantly decreased the serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), and inhibited the infiltration of CD86-positive M1 macrophages in renal tissue (P < 0.01 and 0.001). In addition, hirudin exerted bidirectional regulatory effects on the expression of on urate transporters, characterized by downregulation of urate transporter 1 (URAT1) and glucose transporter 9 (GLUT9), while upregulation of organic anion transporter 1 (OAT1), organic anion transporter 3 (OAT3), and ATP-binding cassette subfamily G member 2 (ABCG2) (P < 0.05, 0.01 and 0.001). These effects of hirudin were attenuated or completely reversed by co-administration of NF-κB pathway activator. Transcriptomic analysis further demonstrated that hirudin partially restored hyperuricemia-induced dysregulation of renal gene expression, with differentially expressed genes significantly enriched in the interleukin-17 (IL-17) signaling pathway, NOD-like receptor signaling pathway, and epithelial cell differentiation processes. Conclusion Hirudin may protect against hyperuricemia-induced renal injury by inhibiting the activation of the NF-κB signaling pathway, thereby negatively regulating M1 macrophage polarization, mitigating MSU crystal-associated inflammatory responses, and restoring the homeostasis of renal urate transporters.

R965

广西自然科学基金项目面上项目（2024GXNSFAA010388）；广西自然科学基金项目面上项目（2023GXNSFAA026210）；广西中医药优秀人才研修项目（桂中医药科教发〔 2024〕 11号）；广西高校中青年教师科研基础能力提升项目（2025KY1801）