Department of Otolaryngology-Head and Neck Surgery, Provincial Hospital affiliated to Shandong University, Jinan, PR China.
Neurotoxicology. 2011 Jan;32(1):150-7. doi: 10.1016/j.neuro.2010.09.003. Epub 2010 Sep 24.
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(-) (100μM) 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.
本研究旨在探讨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。原代培养的大鼠 SGN 暴露于 ONOO(-)或姜黄素预处理。通过 MTT 测定法测量细胞活力。通过 Ho.33342 和碘化丙啶（PI）双重染色和流式细胞术测定细胞凋亡。通过分光光度计评估细胞内谷胱甘肽（GSH）含量、超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平。通过 RT-PCR 检测 Apaf-1、Caspase-9、Caspase-3、Bcl-2 和 Bax 的 mRNA 表达，同时通过 Western blot 分别检测线粒体和细胞质细胞色素 c、Caspase-9、Caspase-3、Bcl-2 和 Bax 蛋白的蛋白表达。暴露于 ONOO(-)（100μM）后，SGN 的细胞活力明显降低，而凋亡率显著增加。SOD 活性和 GSH 水平明显降低，而 MDA 水平显著升高。姜黄素预处理可保护 SGN 免受 ONOO(-)诱导的细胞损伤，降低凋亡率，并改善 SOD 和 GSH 水平，降低 MDA 升高。ONOO(-)诱导 SGN 线粒体细胞色素 c 释放，并随后激活 Caspase-9、Caspase-3 和细胞凋亡。同时，姜黄素预处理可阻断细胞色素 c 释放，阻断 Caspase-3 的激活，并改变 ONOO(-)触发的 Bcl-2 家族的表达。我们的数据表明，姜黄素通过抗氧化活性减轻 ONOO(-)诱导的 SGN 损伤，并保护线粒体免受氧化应激。
