Zhu Yuan-gui, Chen Xiao-chun, Chen Zhi-zhe, Zeng Yu-qi, Shi Guang-bin, Su Yan-hua, Peng Xu
Fujian Institute of Geriatrics, Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
Acta Pharmacol Sin. 2004 Dec;25(12):1606-12.
To investigate the effect of curcumin on tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in rat cortical neurons and to explore the possible mechanism.
Primary cultured rat cortical neurons were performed in vitro and cell viability was measured by MTT assay. DNA fragmentation was used to evaluate cell apoptosis. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (Deltapsim) was determined by flow cytometric assay. Cellular glutathione (GSH) content was measured by spectrophotometer. Bcl-2 family proteins, cytochrome c, cleaved caspase-3, and poly (ADP-ribose) polymerase (PARP) were detected by Western blot.
Exposure of tBHP 100 micromol/L to neurons for 60 min resulted in DYm loss and cytochrome c release from mitochondria and subsequent activation of caspase-3 and PARP cleavation, and cell apoptosis. After removal of tBHP and then further treatment with curcumin (2.5-20 micromol/L) for 18 h, curcumin abrogated Deltapsim loss and cytochrome c release, blocked activation of caspase 3, and altered the expression of Bcl-2 family. Further curcumin treatment also prevented cellular GSH and decreased intracellular ROS generation markedly. Curcumin eventually attenuated tBHP-induced apoptosis in cortical neurons.
Curcumin may attenuate oxidative damages in cortical neurons by reducing intracellular production of ROS and protecting mitochondria from oxidative damage.
研究姜黄素对叔丁基过氧化氢（t-BHP）诱导的大鼠皮质神经元氧化损伤的影响，并探讨其可能机制。
体外培养原代大鼠皮质神经元，采用MTT法检测细胞活力。用DNA片段化评估细胞凋亡。通过流式细胞术检测细胞内活性氧（ROS）和线粒体膜电位（Δψm）。用分光光度计测量细胞内谷胱甘肽（GSH）含量。通过蛋白质免疫印迹法检测Bcl-2家族蛋白、细胞色素c、裂解的半胱天冬酶-3和聚（ADP-核糖）聚合酶（PARP）。
100 μmol/L tBHP作用于神经元60分钟导致线粒体膜电位丧失、细胞色素c从线粒体释放，随后半胱天冬酶-3激活和PARP裂解，以及细胞凋亡。去除tBHP后，再用姜黄素（2.5 - 20 μmol/L）进一步处理18小时，姜黄素消除了线粒体膜电位丧失和细胞色素c释放，阻断了半胱天冬酶3的激活，并改变了Bcl-2家族的表达。进一步的姜黄素处理还可预防细胞内GSH的减少，并显著降低细胞内ROS的生成。姜黄素最终减轻了tBHP诱导的皮质神经元凋亡。
姜黄素可能通过减少细胞内ROS的产生和保护线粒体免受氧化损伤来减轻皮质神经元的氧化损伤。
