Su Fengyu, Wang Ting, Lin Xiuli, Du Yang, Guo Yan, Cao Weidong, Shang Yaqiong, Zhou Anning, Huang Songming, Jia Zhanjun, Zhang Yue, Zhang Aihua, Tang Xiaomei, Chen Shuang
Department of Nephrology, Children's Hospital of Nanjing Medical University, Nanjing, China.
Jiangsu Key Laboratory of Pediatrics, Nanjing Medical University, Nanjing, China.
Kidney Dis (Basel). 2025 Apr 24;11(1):416-438. doi: 10.1159/000546033. eCollection 2025 Jan-Dec.
Purinergic signaling has been recognized as important extracellular regulator in multiple physiological and pathophysiological conditions. Adenosine triphosphate-purinergic receptor P2Y2 signaling pathway is associated with glomerular nephritis (GN), diabetic nephropathy (DN), and chronic kidney disease. Recently, there has been evidence that global knockout of P2Y2 exacerbated bilateral ischemic reperfusion-induced acute kidney injury (AKI). However, its role in cisplatin-induced AKI (CIA) remains unknown. Cisplatin is a platinum-containing antineoplastic drug widely used in variety of solid malignant tumors. Nephrotoxicity is one of the major serious side effect that limit its clinical use. In the present study, we investigated whether inhibition of P2Y2 has an effect on CIA.
We used AR-C118925 (AR-C), a selective antagonist of P2Y2, and gene transfection for interruption of the P2Y2 pathway. Mice were pretreated with AR-C (10 mg/kg/day) and then challenged with cisplatin at a dose of 20 mg/kg. Seventy-two hours after cisplatin administration, all mice developed kidney failure. Knockdown and overexpression of P2Y2 in mice and mouse proximal tubular cells (mPTCs) were employed to validate that ARC acts through P2Y2 receptor.
AR-C markedly ameliorated cisplatin-induced nephrotoxicity evidenced by improved renal function, renal morphology, and tubular injury marker expression. Further analysis of the mechanism revealed that AR-C significantly reduced kidney oxidative stress, inflammation, apoptosis, and necroptosis. Consistently, AR-C protects mPTCs from injury caused by cisplatin. To verify that AR-C acts through the P2Y2 receptor, we knocked down P2Y2 in mice or in mPTC cells. Both showed beneficial effects, while overexpression of P2Y2 promotes cisplatin-induced cell death.
Taken together, our study, for the first time revealed that P2Y2 plays an important role in CIA by regulating oxidative stress, inflammation, apoptosis, and necroptosis and its inhibitor, AR-C, is a potential drug for treating CIA.
嘌呤能信号传导已被认为是多种生理和病理生理条件下重要的细胞外调节因子。三磷酸腺苷-嘌呤能受体P2Y2信号通路与肾小球肾炎（GN）、糖尿病肾病（DN）和慢性肾脏病有关。最近，有证据表明P2Y2基因完全敲除会加重双侧缺血再灌注诱导的急性肾损伤（AKI）。然而，其在顺铂诱导的AKI（CIA）中的作用仍不清楚。顺铂是一种含铂的抗肿瘤药物，广泛用于各种实体恶性肿瘤。肾毒性是限制其临床应用的主要严重副作用之一。在本研究中，我们调查了抑制P2Y2是否对CIA有影响。
我们使用P2Y2的选择性拮抗剂AR-C118925（AR-C）以及通过基因转染来阻断P2Y2通路。小鼠先用AR-C（10毫克/千克/天）预处理，然后用20毫克/千克的顺铂进行攻击。顺铂给药72小时后，所有小鼠均出现肾衰竭。采用小鼠和小鼠近端肾小管细胞（mPTCs）中P2Y2的敲低和过表达来验证ARC是通过P2Y2受体发挥作用的。
AR-C显著改善了顺铂诱导的肾毒性，表现为肾功能改善、肾脏形态以及肾小管损伤标志物表达改善。对其机制的进一步分析表明，AR-C显著降低了肾脏氧化应激、炎症、细胞凋亡和坏死性凋亡。同样，AR-C保护mPTCs免受顺铂引起的损伤。为了验证AR-C是通过P2Y2受体发挥作用，我们在小鼠或mPTC细胞中敲低P2Y2。两者均显示出有益效果，而P2Y2过表达则促进顺铂诱导的细胞死亡。
综上所述，我们的研究首次揭示P2Y2通过调节氧化应激、炎症、细胞凋亡和坏死性凋亡在CIA中起重要作用，其抑制剂AR-C是治疗CIA的潜在药物。
