Curcumin attenuates peroxynitrite-induced neurotoxicity in spiral ganglion neurons
Curcumin attenuates peroxynitrite-induced neurotoxicity in spiral ganglion neurons
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DOI：
10.1016/j.neuro.2010.09.003
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发表时间：
2011-01-01
期刊：
影响因子：
3.4
通讯作者：
Wang, Haibo
中科院分区：
文献类型：
--
作者：
Liu, Wenwen;Fan, Zhaomin;Wang, Haibo
观察姜黄素对过氧亚硝酸盐(ONOO)诱导的大鼠螺旋神经节神经元(SGN)损伤的影响。将原代培养的大鼠SGN暴露于ONOO-加或不加姜黄素预处理。用四甲基偶氮唑盐比色法测定细胞存活率。用Ho.33342和碘化丙啶(PI)双重染色及流式细胞术检测细胞凋亡。用分光光度法检测细胞内谷胱甘肽(GSH)含量、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。RT-PCR法检测Apaf-1、Caspase-9、Caspase-3、Bcl2、Bax的mRNA表达，Western印迹法检测线粒体、胞浆细胞色素C、Caspase-9、Caspase-3、Bcl-2、Bax蛋白的表达。ONOO-(100 MU M)作用于SGN后，细胞存活率显著降低，凋亡率显著升高。SOD活性和GSH水平显著降低，而丙二醛水平显著升高。姜黄素对ONOO诱导的SGN细胞损伤有保护作用，降低了细胞凋亡率，提高了SOD和GSH的水平，降低了MDA的升高。ONOO诱导SGN线粒体释放细胞色素c，进而激活Caspase-9、Caspase-3，促进细胞凋亡。同时，姜黄素可抑制细胞色素c的释放，阻断Caspase-3的激活，并改变由ONOO-引发的Bcl2家族的表达。我们的数据表明姜黄素可以通过抗氧化活性减轻ONOO对SGN的损伤，并保护线粒体免受氧化应激的影响。(C)2010 Elsevier Inc.保留所有权利。
The present study was designed to investigate the effect of curcumin on peroxynitrite (ONOO-)-induced damage in rat spiral ganglion neurons (SGNs). The primary cultured rat SGNs were exposed to ONOO- with or without curcumin pretreatment. Cell viability was measured by MTT assay. Apoptosis was determined by Ho.33342 and propidium iodide (PI) double staining and flow cytometry. The cellular glutathione (GSH) content, superoxide dismutase (SOD) activity and malonaldehyde (MDA) levels were evaluated by spectrophotometer. The mRNA expressions of Apaf-1, Caspase-9, Caspase-3, Bcl-2, and Bax were examined by RT-PCR, while, the protein expressions of mitochondrial and cytosolic cytochrome C. Caspase-9, Caspase-3, Bcl-2 and Bax proteins were determined by Western blot respectively. The cell viability was markedly reduced, while, the apoptotic rate increased significantly after exposure of ONOO- (100 mu M) to SGNs. The activity of SOD and level of GSH were notably reduced, whereas, the MDA level was significantly increased. Pretreatment with curcumin protected SGNs against ONOO--induced cell damage, declined the apoptotic rate, and improved the levels of SOD and GSH, decreased the elevation of MDA. ONOO- induced cytochrome c release from the mitochondria of SGNs and subsequently activated Caspase-9, Caspase-3 and cell apoptosis. Meanwhile, pretreatment with curcumin abrogated cytochrome c release, blocked activation of Caspase-3, and altered the expression of Bcl-2 family triggered by ONOO-. Our data indicate that curcumin can attenuate ONOO--induced damage in SGNs by the anti-oxidative activity as well as protect mitochondria from oxidative stress. (C) 2010 Elsevier Inc. All rights reserved.