Postgraduate Program in Pharmaceutical Sciences, Federal University of Ceará, Fortaleza, CE, Brazil.
Department of Clinical and Toxicological Analysis, Federal University of Ceará, Fortaleza, CE, Brazil.
Life Sci. 2022 Feb 15;291:120271. doi: 10.1016/j.lfs.2021.120271. Epub 2021 Dec 30.
Amphotericin B (AmB), used for systemic fungal infections, has a limited clinical application because of its high nephrotoxicity. Natural antioxidant and anti-inflammatory substances have been widely studied for protection against drug-induced nephrotoxicity. α-Bisabolol (BIS) has demonstrated a nephroprotective effect on both in vitro and in vivo models.
The aim of this work was to evaluate the effect of BIS against AmB-induced nephrotoxicity in vitro.
LLC-MK2 cells were pre- and post-treated with non-toxic BIS concentrations and/or AmB IC50 (13.97 μM). Cell viability was assessed by MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)] assay. Flow cytometry analyses were used to assess cell death mechanism, production of reactive oxidative stress (ROS) and mitochondrial transmembrane potential. Kidney Injury Molecule-1 (KIM-1) levels were measured via ELISA.
The present work showed that BIS pretreatment (125; 62.5 and 31.25 μM) increased cell viability when compared to the group treated only with AmB IC50. AmB treatment induced both necrosis (7-AAD-labeled cells) and late apoptosis (AnxV-labeled). BIS was able to prevent the occurrence of these events. These effects were associated with a decrease of ROS accumulation, improving transmembrane mitochondrial potential and protecting against tubular cell damage, highlighted by the inhibition of KIM-1 release after BIS treatment.
BIS presented a potential effect on model of renal cytotoxicity induced by AmB, bringing perspectives for the research of new nephroprotective agents.
两性霉素 B（AmB）用于治疗系统性真菌感染，但由于其肾毒性较高，临床应用受到限制。天然抗氧化剂和抗炎物质已被广泛研究用于预防药物引起的肾毒性。α- 姜烯醇（BIS）已在体外和体内模型中显示出对肾脏的保护作用。
本研究旨在评估 BIS 对 AmB 诱导的体外肾毒性的作用。
LLC-MK2 细胞用无毒 BIS 浓度和/或 AmB IC50（13.97 μM）进行预和后处理。通过 MTT [（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物）] 测定评估细胞活力。通过流式细胞术分析评估细胞死亡机制、活性氧（ROS）的产生和线粒体跨膜电位。通过 ELISA 测量肾损伤分子 1（KIM-1）的水平。
本研究表明，与仅用 AmB IC50 处理的组相比，BIS 预处理（125；62.5 和 31.25 μM）可提高细胞活力。AmB 处理诱导坏死（7-AAD 标记细胞）和晚期凋亡（AnxV 标记）。BIS 能够预防这些事件的发生。这些作用与 ROS 积累的减少、线粒体跨膜潜能的改善以及对肾小管细胞损伤的保护有关，这一点在 BIS 处理后 KIM-1 释放受到抑制时得到了强调。
BIS 对 AmB 诱导的肾毒性模型具有潜在作用，为研究新的肾保护剂带来了前景。
