Radical scavenger edaravone developed for clinical use ameliorates ischemia/reperfusion injury in rat kidney
Radical scavenger edaravone developed for clinical use ameliorates ischemia/reperfusion injury in rat kidney
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DOI：
10.1111/j.1523-1755.2004.00567.x
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发表时间：
2004-05-01
影响因子：
19.6
通讯作者：
Noiri, E
中科院分区：
文献类型：
--
作者：
Doi, K;Suzuki, Y;Noiri, E
背景资料。依达拉奉(3-甲基-1-苯基-2-吡唑啉-5-酮)是一种有效的自由基清除剂，具有抗氧化能力，抑制脂质过氧化。其对脑缺血的保护作用已被证明。本研究旨在阐明其对大鼠肾缺血/再灌流模型肾脏氧化应激的可能治疗作用。左肾动脉夹闭45分钟后给予依达拉奉造成急性缺血性肾功能衰竭。检查肾功能和病理结果。用荧光探针二氯二氢荧光素(CM-H，DCFDA)观察自由基的产生，免疫化学染色和4-羟基-2-己烯醛(HHE)的Western分析观察脂质过氧化反应。用体外培养的人肾小管上皮细胞(HKC-8)观察依达拉奉对体外氧化应激的抑制作用，并用Alamar蓝比色法检测细胞活力。依达拉奉显著减轻肾功能和病理改变。依达拉奉的保护作用与二甲基硫脲的剂量效应相当。此外，依达拉奉显著减少CM-H、DCFDA所示的肾小管上皮细胞内自由基的产生。免疫化学染色和Western分析显示依达拉奉治疗组大鼠肾脏脂质过氧化反应受到明显抑制。在负载CM-H(2)DCFDA的HKC-8细胞中，H_2O_2(0.5 mmol/L，作用1h)可显著增强HKC-8细胞的荧光强度，而依达拉奉可使其荧光强度降低。Alamar蓝检测细胞活力也显示依达拉奉对细胞的保护作用。依达拉奉通过清除肾小管细胞产生的自由基和抑制脂质过氧化来减轻肾缺血/再灌注损伤。
Background. 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.Methods. 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,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.Results. 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,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.Conclusion. Edaravone ameliorates renal ischemia/ reperfusion injury by scavenging free radicals produced in renal tubular cells and inhibiting lipid peroxidation.