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机构地区:[1]温州医学院药学院,浙江温州325035 [2]中国医学科学院 北京协和医学院 药物研究所国家药物筛选中心,北京100050
出 处:《基础医学与临床》2013年第11期1465-1469,共5页Basic and Clinical Medicine
基 金:温州市科技局科技计划项目(S20100044)
摘 要:目的研究L-茶氨酸对叠氮钠(NaN,)诱导的人神经母细胞瘤(SH-SYSY)细胞线粒体损伤的影响其机制。方法将不同浓度的L-茶氨酸及NaN,与SH-SYSY细胞系共同孵育,用M1Tr法测定细胞存活力,以JC-1测定法和激光共聚焦显微镜来观察线粒体膜电位,Westernblot法检测细胞内caspase-3、caspase-9和细胞色素C蛋白表达水平。结果1)随着NaN,浓度的增高,细胞活性逐步降低,提前2h给予茶氨酸(0.25、0.50和1.00mmol/L)能明显降低NaN,(0.25mmol/L)孵育24h对细胞的毒性并逆转线粒体跨膜电位的下降(P〈0.05);2)茶氨酸在0.25-1.00mmol/L浓度范围内能显著逆转NaN,诱导的SH-SYSY细胞内caspase-3、caspase-9和细胞色素c蛋白表达水平的升高(P〈0.05)。结论茶氨酸可以通过稳定SH-SYSY细胞线粒体膜电位,减轻细胞线粒体的损伤。Objective To investigate the effects and its mechanism of L-theanine on mitochondrial injury induced by sodium azide in SH-SY5Y cells. Methods The SH-SY5Y cells were exposed to different concentrations of L-theanine and the sodium azide, the cell viability was measured by the MTT method, the mitochondrial membrane potential was detected by the JC-1 method and the Scanning Confocal Microscopy. The protein level of intracellular caspase-3, caspase-9 and cytochrome c was detected by the Western Blotting Method. Results 1 ) With the increase of the con- centration of sodium azide, the cell activity decreases gradually. After pretreatment by L-theanine (0. 25,0. 50, 1.00 mmol/L) for 2 hours,it significantly reduced cell toxicity and reversed decline in the mitochondrial membrane potential induced by the sodium azide (0. 25 mmol/L) for 24 hours(P 〈0. 05). 2) L-theanine(0. 25 - 1.0 mmol/L) significantly reversed the increasing of the protein expression level of the easpase-3, caspase-9 and cytoehrome c in- duced by the sodium azide in SH-SY5Y cell(P 〈0. 05). Conclusions L-Theanine can stabilize mitochondrial mem- brane potential of the SH-SY5Y cell and thus reduce the mitoehondrial damage.
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