Doi Kent, Suzuki Yoshifumi, Nakao Akihide, Fujita Toshiro, Noiri Eisei
Department of Nephrology and Endocrinology, University of Tokyo, Tokyo, Japan.
Kidney Int. 2004 May;65(5):1714-23. doi: 10.1111/j.1523-1755.2004.00567.x.
Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) is a potent scavenger of free radicals and has the antioxidant ability to inhibit lipid peroxidation. Its protective effect on brain ischemia has been shown. This study aimed to elucidate its possible therapeutic effects on renal oxidative stress in a rat ischemia/reperfusion model.
Ischemic acute renal failure was induced by 45-minute clamp of the left renal artery followed by administration of edaravone. Renal function and pathologic findings were examined. The generation of free radicals was observed by the fluorogenic probe, dichlorodihydrofluorescein (CM-H(2)DCFDA), and lipid peroxidation was observed by immunochemical staining and Western analysis of 4-hydroxy-2-hexenal (HHE). The ability of edaravone to reduce in vitro oxidative stress was investigated using cultured human renal tubular cells (HKC-8) and the cell viability assay with Alamar blue.
Edaravone attenuated renal function and pathologic findings significantly. The protective effect of edaravone showed the dose-response was at the same level as that of dimethylthiourea. In addition, edaravone significantly reduced the generation of free radicals in the tubular cells indicated by CM-H(2)DCFDA. Immunochemical staining and Western analysis of HHE showed significant suppression of lipid peroxidation in edaravone-treated kidney. In HKC-8 cells loaded with CM-H(2)DCFDA, a marked elevation of fluorescence was observed after exposure to hydrogen peroxide (0.5 mmol/L, for 1 hour), which was reduced by edaravone. The cell viability assay with Alamar blue also showed the protective effect of edaravone.
Edaravone ameliorates renal ischemia/reperfusion injury by scavenging free radicals produced in renal tubular cells and inhibiting lipid peroxidation.
依达拉奉（3 - 甲基 - 1 - 苯基 - 2 - 吡唑啉 - 5 - 酮）是一种强效自由基清除剂，具有抑制脂质过氧化的抗氧化能力。其对脑缺血的保护作用已得到证实。本研究旨在阐明其在大鼠缺血/再灌注模型中对肾脏氧化应激可能的治疗作用。
通过夹闭左肾动脉45分钟诱导缺血性急性肾衰竭，随后给予依达拉奉。检测肾功能和病理结果。用荧光探针二氯二氢荧光素（CM - H₂DCFDA）观察自由基的产生，通过免疫化学染色和对4 - 羟基 - 2 - 己烯醛（HHE）的蛋白质免疫印迹分析观察脂质过氧化。使用培养的人肾小管细胞（HKC - 8）和阿拉玛蓝细胞活力测定法研究依达拉奉降低体外氧化应激的能力。
依达拉奉显著改善肾功能和病理结果。依达拉奉的保护作用呈剂量依赖性，与二甲基硫脲处于同一水平。此外，依达拉奉显著减少了CM - H₂DCFDA所示肾小管细胞中自由基的产生。对HHE的免疫化学染色和蛋白质免疫印迹分析显示依达拉奉处理的肾脏中脂质过氧化得到显著抑制。在加载CM - H₂DCFDA的HKC - 8细胞中，暴露于过氧化氢（0.5 mmol/L，1小时）后观察到荧光显著升高，而依达拉奉可使其降低。阿拉玛蓝细胞活力测定法也显示了依达拉奉的保护作用。
依达拉奉通过清除肾小管细胞中产生的自由基并抑制脂质过氧化来改善肾脏缺血/再灌注损伤。
