Lan Fang, Long Chunli, Huang Huimin, Xie Yongxiang, Shi Wei
Guangxi University of Traditional Chinese Medicine - The First Affiliated Hospital - Department of Nephrology - Nanning (Guangxi) - China.
Guangxi University of Traditional Chinese Medicine - School of Nursing - Nanning (Guangxi) - China.
Acta Cir Bras. 2025 Apr 28;40:e403325. doi: 10.1590/acb403325. eCollection 2025.
To reveal the role and underlying mechanism of hirudin in renal fibrosis.
The unilateral ureteral obstruction (UUO) rat model and ferroptosis activator RSL3-induced human kidney proximal tubular epithelial cells (HK-2) were established. Hematoxylin-eosin staining, commercial kits, and immunohistochemistry were used to assess the effect of hirudin on renal function and renal fibrosis. Cell counting kit-8 assay was employed to test cell viability. Ferroptosis indicator levels were detected using commercial kits. The protein levels were examined by Western blot. The STAT3 activator colivelin was introduced to verify the role of the STAT3/NLRP3 signaling pathway in ferroptosis.
Hirudin alleviated renal injury and improved renal fibrosis in UUO rats. The cell viability of RSL3-treated HK-2 cells was increased after hirudin treatment. In the model group, GPX4, SLC7A11, and glutathione expression decreased, while malondialdehyde and iron content levels increased, indicating that ferroptosis was activated. Besides, p-STAT3 and NLRP3 protein levels were also upregulated. However, hirudin treatment reversed these changes. When the STAT3 activator colivelin was added, the effect of hirudin was altered.
Hirudin improved renal fibrosis by inhibiting ferroptosis via the STAT3/NLRP3 signaling pathway.
揭示水蛭素在肾纤维化中的作用及潜在机制。
建立单侧输尿管梗阻（UUO）大鼠模型和铁死亡激活剂RSL3诱导的人肾近端小管上皮细胞（HK-2）模型。采用苏木精-伊红染色、商用试剂盒和免疫组织化学方法评估水蛭素对肾功能和肾纤维化的影响。使用细胞计数试剂盒-8法检测细胞活力。采用商用试剂盒检测铁死亡指标水平。通过蛋白质印迹法检测蛋白质水平。引入STAT3激活剂colivelin以验证STAT3/NLRP3信号通路在铁死亡中的作用。
水蛭素减轻了UUO大鼠的肾损伤并改善了肾纤维化。水蛭素处理后，RSL3处理的HK-2细胞的细胞活力增加。在模型组中，GPX4、SLC7A11和谷胱甘肽表达降低，而丙二醛和铁含量水平升高，表明铁死亡被激活。此外，p-STAT3和NLRP3蛋白水平也上调。然而，水蛭素处理逆转了这些变化。当加入STAT3激活剂colivelin时，水蛭素的作用发生了改变。
水蛭素通过STAT3/NLRP3信号通路抑制铁死亡，从而改善肾纤维化。
