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NecroX-5 对新霉素诱导斑马鱼毛细胞损伤的保护作用。

Protective role of NecroX-5 against neomycin-induced hair cell damage in zebrafish.

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, Dongguk University Ilsan Hospital, Goyang, Gyeonggi, Korea.

出版信息

Arch Toxicol. 2014 Feb;88(2):435-41. doi: 10.1007/s00204-013-1124-3. Epub 2013 Sep 13.

DOI:10.1007/s00204-013-1124-3
PMID:24030356
Abstract

NecroX-5, one of the derivatives of NecroX series compounds, is a mitochondrial reactive oxygen species and reactive nitrogen species scavenger that inhibits cell death against various kinds of oxidative stresses. The objective of the present study was to evaluate the effects of NecroX-5 on neomycin-induced ototoxicity in transgenic zebrafish (Brn3C: EGFP). Five days post-fertilization, zebrafish larvae were exposed to 125 μM neomycin and one of the following NecroX-5 concentrations for 1 h: 10, 25, 50, and 75 μM. Hair cells within the neuromasts of the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) lateral lines were analyzed using fluorescence microscopy (n = 10). The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and 2-[4-(dimethylamino) styryl]-N-ethylpyridiniumiodide (DASPEI) assay were performed for evaluation of apoptosis and mitochondrial damage. Ultrastructural changes were evaluated using scanning electron microscopy. NecroX-5 decreased neomycin-induced hair cell loss in the neuromasts (NecroX-5 50 μM: 13.4 ± 2.0 cells, 125 μM neomycin only: 8.1 ± 1.2 cells; n = 10, P < 0.05) and decreased the TUNEL reaction. The ultrastructural analysis showed that the structures of mitochondria and hair cells within the neuromasts were preserved in zebrafish exposed to 125 μM neomycin and 50 μM NecroX-5. NecroX-5 decreased apoptosis and mitochondrial damage. In conclusion, NecroX-5 attenuated neomycin-induced hair cell loss in zebrafish.

摘要

NecroX-5 是 NecroX 系列化合物的衍生物之一,是一种线粒体活性氧和活性氮物种清除剂,可抑制各种氧化应激引起的细胞死亡。本研究的目的是评估 NecroX-5 对转基因斑马鱼(Brn3C: EGFP)中新霉素诱导的耳毒性的影响。受精后 5 天,将斑马鱼幼虫暴露于 125 μM 新霉素和以下一种 NecroX-5 浓度中 1 小时:10、25、50 和 75 μM。使用荧光显微镜分析额骨(SO1 和 SO2)、耳(O1)和枕骨(OC1)侧线神经丘内的毛细胞(n = 10)。通过末端脱氧核苷酸转移酶介导的 dUTP-生物素 nick 末端标记(TUNEL)测定和 2-[4-(二甲基氨基)苯乙烯基]-N-乙基吡啶碘化物(DASPEI)测定评估细胞凋亡和线粒体损伤。使用扫描电子显微镜评估超微结构变化。NecroX-5 减少了新霉素诱导的神经丘内毛细胞丢失(NecroX-5 50 μM:13.4 ± 2.0 个细胞,125 μM 新霉素仅:8.1 ± 1.2 个细胞;n = 10,P < 0.05)并减少了 TUNEL 反应。超微结构分析表明,暴露于 125 μM 新霉素和 50 μM NecroX-5 的斑马鱼神经丘内的线粒体和毛细胞结构得到了保留。NecroX-5 减少了细胞凋亡和线粒体损伤。总之,NecroX-5 减轻了新霉素诱导的斑马鱼毛细胞丢失。

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