Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin
Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin
复制标题
DOI：
10.1016/j.lfs.2003.09.074
复制
发表时间：
2004-04-30
期刊：
影响因子：
6.1
通讯作者：
Liu, GQ
中科院分区：
文献类型：
--
作者：
Hong, H;Liu, GQ
本研究探讨了抗氧化剂灯盏乙素对H2 O2暴露于PC 12细胞产生的细胞毒性的保护作用。这通过测定MTT（3，（4，5-二甲基噻唑-2-基）2，5-二苯基-溴化四唑）还原和乳酸脱氢酶（LDH）释放来完成。以DCFH和Fura 2-AM为探针，用荧光酶标仪检测细胞内活性氧（ROS）和钙离子（Ca 2+）的含量。使用硫代巴比妥酸反应物质（TBARS）定量脂质过氧化。线粒体膜电位（NIMP）的保留罗丹明123（Rh 123），一个特定的荧光阳离子染料，很容易被活跃的线粒体，这取决于他们的跨膜电位进行评估。流式细胞仪检测细胞DNA含量和凋亡率。维生素E，一种有效的抗氧化剂，被用作比较剂。用灯盏乙素预孵育PC 12细胞可防止H2 O2诱导的细胞毒性。灯盏乙素能显著降低细胞内ROS、Ca ~（2+）和H_2O_2引起的脂质过氧化产物的积累。与H2 O2孵育的细胞引起MMP的显着下降，这是显着抑制灯盏乙素。流式细胞仪检测显示H2 O2处理后的PC 12细胞发生凋亡。在不同浓度的灯盏乙素存在下，这种H2 O2诱导的细胞凋亡的百分比降低。与抗氧化剂维生素E相比，灯盏乙素具有更高的效力。本研究结果表明，灯盏乙素减弱H2 O2诱导的细胞毒性，细胞内ROS和Ca 2+的积累，脂质过氧化，MMP和DNA的损失，这可能是其神经保护作用的细胞机制。(C)2004爱思唯尔公司All rights reserved.
The present study investigated the protective actions of the antioxidant scutellarin against the cytotoxicity produced by exposure to H2O2 in PC12 cells. This was done by assaying for MTT (3,(4,5-dimethylthiazole-2-yl) 2,5-diphenyl-tetrazolium bromide) reduction and lactate dehydrogenase (LDH) release. Reactive oxygen species (ROS) and Ca2+ in cells were evaluated by fluorescent microplate reader using DCFH and Fura 2-AM, respectively, as probes. Lipid peroxidation was quantified using thiobarbituric acid-reactive substances (TBARS). Mitochondrial membrane potential (NIMP) was assessed by the retention of rhodamine123 (Rh123), a specific fluorescent cationic dye that is readily sequestered by active mitochondria, depending on their transmembrane potential. The DNA content and percentage of apoptosis were monitored with flow cytometry. Vitamin E, a potent antioxidant, was employed as a comparative agent. Preincubation of PC12 cells with scutellarin prevented cytotoxicity induced by H2O2. Intracellular accumulation of ROS, Ca2+ and products of lipid peroxidation, resulting from H2O2 were significantly reduced by scutellarin., Incubation of cells with H2O2 caused a marked decrease in MMP, which was significantly inhibited by scutellarin. PC12 cells treated with H2O2 underwent apoptotic death as determined by flow cytometric assay. The percentage of this H2O2-induced apoptosis in the cells was decreased in the presence of different concentrations Of scutellarin. Scutellarin exhibited significantly higher potency compared to the antioxidant vitamin E. The present findings showed that scutellarin attenuated H2O2- induced cytotoxicity, intracellular accumulation of ROS and Ca2+, lipid peroxidation, and loss of MMP and DNA, which may represent the cellular mechanisms for its neuroprotective action. (C) 2004 Elsevier Inc. All rights reserved.