Department of Nephrology, Children's Hospital of Nanjing Medical University, 72 Guangzhou Road, Nanjing, 210008, PR China; Nanjing Key Laboratory of Pediatrics, Children's Hospital of Nanjing Medical University, Nanjing, 210008, China; Jiangsu Key Laboratory of Pediatrics, Nanjing Medical University, Nanjing, 210029, China.
Department of Nephrology, Children's Hospital of Nanjing Medical University, 72 Guangzhou Road, Nanjing, 210008, PR China; Nanjing Key Laboratory of Pediatrics, Children's Hospital of Nanjing Medical University, Nanjing, 210008, China; Jiangsu Key Laboratory of Pediatrics, Nanjing Medical University, Nanjing, 210029, China.
Free Radic Biol Med. 2020 May 20;152:821-837. doi: 10.1016/j.freeradbiomed.2020.01.182. Epub 2020 Jan 28.
Mitochondrial dysfunction plays an important role in acute kidney injury (AKI). Thus, the agents improving the mitochondrial function could be beneficial for treating AKI. Ursodeoxycholic acid (UDCA) has been demonstrated to prevent mitochondrial dysfunction under pathology, however, its role in AKI and the underlying mechanism remain unknown. This study aimed to evaluate the effect of UDCA on cisplatin-induced AKI. In vivo, C57BL/6 J mice were treated with cisplatin (25 mg/kg) for 72 h to induce AKI through a single intraperitoneal (i.p.) injection with or without UDCA (60 mg/kg/day) administration by gavage. Renal function, mitochondrial function and oxidative stress were analyzed to evaluate kidney injury. In vitro, mouse proximal tubular cells (mPTCs) and human proximal tubule epithelial cells (HK2) were treated with cisplatin with or without UDCA treatment for 24 h. Transcriptomic RNA-seq was preformed to analyze possible targets of UDCA. Our results showed that cisplatin-induced increments of serum creatinine (Scr), blood urea nitrogen (BUN), and cystatin C were significantly reduced by UDCA along with ameliorated renal tubular injury evidenced by improved renal histology and blocked upregulation of neutrophil gelatinase associated lipocalin (NGAL) and kidney injury molecule 1 (KIM-1). Meanwhile, the apoptosis induced by cisplatin was also markedly attenuated by UDCA administration. In vitro, UDCA treatment protected against tubular cell apoptosis possibly through antagonizing mitochondrial dysfunction and oxidative stress by targeting ALDH1L2 which was screened out by an RNA-seq analysis. Knockout of ALDH1L2 by CRISPR/Cas9 greatly blunted the protective effects of UDCA in renal tubular cells. Moreover, UDCA did not diminish cisplatin's antineoplastic effect in human cancer cells. In all, our results demonstrated that UDCA protects against cisplatin-induced AKI through improving mitochondrial function through acting on the expression of ALDH1L2, suggesting a clinical potential of UDCA for the treatment of AKI.
线粒体功能障碍在急性肾损伤(AKI)中起着重要作用。因此,改善线粒体功能的药物可能有益于 AKI 的治疗。熊去氧胆酸(UDCA)已被证明可在病理条件下预防线粒体功能障碍,但其在 AKI 中的作用及其潜在机制尚不清楚。本研究旨在评估 UDCA 对顺铂诱导的 AKI 的影响。在体内,通过单次腹腔内(i.p.)注射顺铂(25mg/kg)72h,C57BL/6J 小鼠用顺铂(25mg/kg)诱导 AKI,并用灌胃给予 UDCA(60mg/kg/天)。分析肾功能、线粒体功能和氧化应激以评估肾损伤。在体外,用顺铂处理小鼠近端肾小管细胞(mPTCs)和人近端肾小管上皮细胞(HK2),并用 UDCA 处理 24h。进行转录组 RNA-seq 分析以分析 UDCA 的可能靶点。结果表明,顺铂诱导的血清肌酐(Scr)、血尿素氮(BUN)和胱抑素 C 升高,UDCA 可显著降低,同时改善的组织学和阻断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)的上调表明肾小管损伤得到改善。同时,UDCA 给药也明显减轻顺铂诱导的细胞凋亡。在体外,UDCA 治疗可能通过靶向 ALDH1L2 拮抗线粒体功能障碍和氧化应激来保护肾小管细胞免受凋亡,ALDH1L2 是通过 RNA-seq 分析筛选出来的。CRISPR/Cas9 敲除 ALDH1L2 大大减弱了 UDCA 在肾小管细胞中的保护作用。此外,UDCA 并未降低顺铂对人癌细胞的抗肿瘤作用。总之,我们的研究结果表明,UDCA 通过作用于 ALDH1L2 的表达来改善线粒体功能,从而防止顺铂诱导的 AKI,提示 UDCA 在治疗 AKI 方面具有临床潜力。