Ghassemi-Barghi Nasrin, Ehsanfar Zeynab, Mohammadrezakhani Omid, Ashari Sorour, Ghiabi Shamim, Bayrami Zahra
Toxicology and Diseases Group (TDG), Pharmaceutical Sciences Research Center (PSRC), The Institute of Pharmaceutical Sciences (TIPS), Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences (TUMS), Tehran, Iran.
Vidagen Pharmed Company, Tehran, Iran.
Inflammation. 2023 Feb;46(1):342-358. doi: 10.1007/s10753-022-01737-7. Epub 2022 Sep 10.
ARA 290, an 11-amino acid linear nonhematopoietic peptide derived from the three-dimensional structure of helix B of the erythropoietin (EPO), interacts selectively with the innate repair receptor (IRR) that arbitrates tissue protection. The aim of this study was to investigate the protective effects of ARA290 against cisplatin-induced nephrotoxicity. For this purpose, HEK-293 and ACHN cells were treated with ARA290 (50-400 nM) and cisplatin (2.5 μM) in pretreatment condition. Then, cytotoxicity, genotoxicity, oxidative stress parameters (ROS, GPx, SOD, and MDA), and inflammatory markers (TNFα, IL6, and IL1β) were evaluated. Furthermore, apoptotic cell death was assessed via caspase-3 activity and tunnel assay. To determine the molecular mechanisms of the possible nephroprotective effects of ARA290, gene and protein expressions of TNFα, IL1β, IL6, Caspase-3, Bax, and Bcl2 were evaluated by real-time PCR and western blot assay, respectively. The findings indicated that ARA290 significantly reduced the DNA damage parameters of comet assay and the frequency of micronuclei induced by cisplatin. Besides, ARA290 improved cisplatin-induced oxidative stress by reducing MDA/ROS levels and enhancing antioxidant enzyme levels. In addition, reduced levels of pro-inflammatory cytokines indicated that cisplatin-induced renal inflammation was mitigated upon the treatment with ARA290. Besides, ARA290 ameliorates cisplatin-induced cell injury by antagonizing apoptosis. Furthermore, the molecular findings indicated that gene and protein levels of TNFα, IL1β, IL6, Caspase-3, and Bax were significantly decreased and gene and protein levels of Bcl2 significantly increased in the ARA290 plus cisplatin group compared with the cisplatin group. These findings revealed that ARA290 as a potent chemo-preventive agent exerted a protective effect on cisplatin-induced nephrotoxicity mostly through its anti-apoptotic, anti-inflammatory, and antioxidant potentials and also suggested that ARA290 might be a new therapeutic approach for patients with acute kidney injury.
ARA 290是一种由促红细胞生成素（EPO）螺旋B的三维结构衍生而来的含11个氨基酸的线性非造血肽，它与介导组织保护的先天修复受体（IRR）选择性相互作用。本研究的目的是探讨ARA290对顺铂诱导的肾毒性的保护作用。为此，在预处理条件下，用ARA290（50 - 400 nM）和顺铂（2.5 μM）处理人胚肾细胞293（HEK - 293）和人肾癌细胞（ACHN）。然后，评估细胞毒性、遗传毒性、氧化应激参数（活性氧（ROS）、谷胱甘肽过氧化物酶（GPx）、超氧化物歧化酶（SOD）和丙二醛（MDA））以及炎症标志物（肿瘤坏死因子α（TNFα）、白细胞介素6（IL6）和白细胞介素1β（IL1β））。此外，通过半胱天冬酶 - 3活性和隧道试验评估凋亡细胞死亡情况。为了确定ARA290可能的肾保护作用的分子机制，分别通过实时聚合酶链反应（PCR）和蛋白质免疫印迹法评估TNFα、IL1β、IL6、半胱天冬酶 - 3、Bax和Bcl2的基因和蛋白表达。研究结果表明，ARA290显著降低了彗星试验的DNA损伤参数和顺铂诱导的微核频率。此外，ARA290通过降低丙二醛/活性氧水平和提高抗氧化酶水平改善了顺铂诱导的氧化应激。此外，促炎细胞因子水平的降低表明，用ARA290治疗后顺铂诱导的肾炎症得到缓解。此外，ARA290通过拮抗凋亡减轻了顺铂诱导的细胞损伤。此外，分子研究结果表明，与顺铂组相比，在ARA290加顺铂组中，TNFα、IL1β、IL6、半胱天冬酶 - 3和Bax的基因和蛋白水平显著降低，而Bcl2的基因和蛋白水平显著升高。这些研究结果表明，ARA290作为一种有效的化学预防剂，主要通过其抗凋亡、抗炎和抗氧化潜能对顺铂诱导的肾毒性发挥保护作用，并且还表明ARA290可能是急性肾损伤患者的一种新的治疗方法。
