Wang Chong, Jin Huan, Wang Changyuan, Wu Jingjing, Meng Qiang, Zhong Ming, Sun Huijun, Wei Yuheng, Gao Ge, Kaku Taiichi, Huo Xiaokui, Liu Kexin
Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian, China.
Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning Dalian Medical University, Dalian, Liaoning, China.
Front Pharmacol. 2025 Apr 30;16:1577942. doi: 10.3389/fphar.2025.1577942. eCollection 2025.
In this study, we investigated the protective effect of JBP485 against aristolochic acid I (AAI)-induced nephrotoxicity and explored the pharmacokinetic mechanisms. The effects of JBP485 on AAI-induced cytotoxicity and nephrotoxicity were evaluated and , respectively.
To ascertain the protective effect of JBP485 against AAI-induced nephrotoxicity, we measured levels of urea nitrogen (BUN), creatinine (CRE), and indoxol sulfate in blood and urine; determined kidney weight-to-body weight ratio; and performed hematoxylin and eosin (H&E) staining. Cell viability and Western blotting assays, along with determination of malondialdehyde (MDA), superoxide dismutase (SOD), and intracellular reactive oxygen species (ROS) contents, were carried out to explore mechanisms underlying the protective effects of JBP485 against AAI-induced nephrotoxicity.
JBP485 treatment attenuated AAI-induced injuries in rat kidney while decreasing the levels of indoxyl sulfate, CRE, and BUN in plasma and increasing those of indoxyl sulfate in urine compared to that in AAI alone-treated group. The co-administration of JBP485 with AAI significantly increased the concentration and AUC of AAI in plasma, while decreasing its cumulative urinary excretion and renal clearance. Moreover, JBP485 reduced the uptake of AAI in kidney slices and human organic anion transporter 1/3 (hOAT1/3)-transfected human embryonic kidney 293 (HEK293) cells, suggesting that JBP485 ameliorated AAI-induced nephrotoxicity by reducing renal exposure to AAI via OAT inhibition. Meanwhile, JBP485 modulated the abnormal expressions of Oat1, Oat3, organic cation transporter 2 (Oct2), P-glycoprotein (P-gp), multidrug resistance-associated protein 2 (Mrp2) and multidrug and toxin extrusion proteins 1 (Mate 1) in rat kidney, suggesting that JBP485 improved tubular secretion in AAI-treated rats. Moreover, JBP485 reversed the AAI-induced changes in the expression of heme oxygenase 1 (HO-1), NAD(P) H: quinone oxidoreductase-1 (NQO1), B-cell lymphoma-2 (Bcl-2) protein expressions and Bcl-2-like protein 4 (Bax) induced by AAI in rat kidney. JBP485 increased cell viability and reduced intracellular levels of ROS in NRK-52E cells treated with AAI.
These results suggested that JBP485 protected against AAI-induced renal oxidative stress. All results indicated that JBP485 protected against AAI-induced nephrotoxicity by reducing renal exposure to AAI and alleviating oxidative stress. Our findings suggested that JBP485 has potential as a renoprotective agent for the prevention of AAI-induced nephrotoxicity.
在本研究中，我们研究了JBP485对马兜铃酸I（AAI）诱导的肾毒性的保护作用，并探讨了其药代动力学机制。分别评估了JBP485对AAI诱导的细胞毒性和肾毒性的影响。
为确定JBP485对AAI诱导的肾毒性的保护作用，我们测量了血液和尿液中的尿素氮（BUN）、肌酐（CRE）和硫酸吲哚酚水平；测定了肾重与体重比；并进行了苏木精和伊红（H&E）染色。进行细胞活力和蛋白质印迹分析，同时测定丙二醛（MDA）、超氧化物歧化酶（SOD）和细胞内活性氧（ROS）含量，以探讨JBP485对AAI诱导的肾毒性保护作用的潜在机制。
与单独使用AAI治疗的组相比，JBP485治疗减轻了AAI诱导的大鼠肾脏损伤，同时降低了血浆中硫酸吲哚酚、CRE和BUN的水平，并增加了尿液中硫酸吲哚酚的水平。JBP485与AAI联合给药显著增加了血浆中AAI的浓度和AUC，同时降低了其累积尿排泄量和肾清除率。此外，JBP485减少了肾切片和人有机阴离子转运体1/3（hOAT1/3）转染的人胚肾293（HEK293）细胞对AAI的摄取，表明JBP485通过抑制OAT减少肾脏对AAI的暴露来改善AAI诱导的肾毒性。同时，JBP485调节了大鼠肾脏中Oat1、Oat3、有机阳离子转运体2（Oct2）、P-糖蛋白（P-gp）、多药耐药相关蛋白2（Mrp2）和多药与毒素外排蛋白1（Mate 1）的异常表达，表明JBP485改善了AAI处理大鼠的肾小管分泌。此外，JBP485逆转了AAI诱导的大鼠肾脏中血红素加氧酶1（HO-1）、NAD（P）H：醌氧化还原酶-1（NQO1）、B细胞淋巴瘤-2（Bcl-2）蛋白表达以及由AAI诱导的Bcl-2样蛋白4（Bax）的变化。JBP485增加了用AAI处理的NRK-52E细胞的活力并降低了细胞内ROS水平。
这些结果表明JBP485可预防AAI诱导的肾脏氧化应激。所有结果表明，JBP485通过减少肾脏对AAI的暴露和减轻氧化应激来预防AAI诱导的肾毒性。我们的研究结果表明，JBP485有潜力作为一种肾脏保护剂来预防AAI诱导的肾毒性。
