Zhang Zheng, Hou Lin, Li Xianghong, Ju Chuanxia, Zhang Jinyu, Li Xin, Wang Xiuli, Liu Cun, Lv Yuqiang, Wang Yuehua
Department of Biochemistry and Molecular Biology, Medical College of Qingdao University, Qingdao, Shandong Province, China.
Department of Biochemistry and Molecular Biology, Medical College of Qingdao University, Qingdao, Shandong Province, China.
Brain Res. 2016 Feb 15;1633:87-95. doi: 10.1016/j.brainres.2015.12.035. Epub 2015 Dec 29.
Animal and cell experiments showed that inositol hexaphosphate (IP6) was protective on neurons in parkinson's disease (PD) model, but the underlying mechanism of this action was not extensively elucidated. To address this question, we established 6-hydroxydopamine (6-OHDA) induced human dopaminergic cell line SH-SY5Y as PD cell model and testified the neuroprotection of IP6. Through hoechst nuclear stain method and flow cytometric analysis, apoptosis induced by 6-OHDA was blocked by IP6 pretreatment. Significant protection against reactive oxygen species (ROS) and lipid peroxidation product malondialdehyde (MDA) was observed in 6-OHDA induced cells pretreated with IP6. To further investigate the mechanism of anti-apoptotic effect of IP6, expression of mediators in mitochondrion dependent apoptotic pathway was detected. Results indicated that loss of mitochondrial membrane potential, cytochrome c releasing, upregulation of Bcl-2-associated X protein (Bax), downregulation of B-cell CLL/lymphoma 2 (Bcl-2) and caspases activation were reversed by IP6. In addition, using flow cytometric method and western blot approach, our data showed that IP6 attenuated the rise of calcium and α-synuclein aggregation in cytosol. Collectively, IP6 exerted its neuroprotection on dopaminergic cells in PD cell model and the mechanism may be associated with changes of mitochondrion mediated apoptotic pathway and α-synuclein aggregation.
动物和细胞实验表明，肌醇六磷酸（IP6）对帕金森病（PD）模型中的神经元具有保护作用，但其作用的潜在机制尚未得到广泛阐明。为了解决这个问题，我们建立了6-羟基多巴胺（6-OHDA）诱导的人多巴胺能细胞系SH-SY5Y作为PD细胞模型，并验证了IP6的神经保护作用。通过hoechst核染色法和流式细胞术分析，IP6预处理可阻断6-OHDA诱导的细胞凋亡。在用IP6预处理的6-OHDA诱导的细胞中观察到对活性氧（ROS）和脂质过氧化产物丙二醛（MDA）的显著保护作用。为了进一步研究IP6抗凋亡作用的机制，检测了线粒体依赖性凋亡途径中介质的表达。结果表明，IP6可逆转线粒体膜电位丧失、细胞色素c释放、Bcl-2相关X蛋白（Bax）上调、B细胞淋巴瘤/白血病-2（Bcl-2）下调和半胱天冬酶激活。此外，使用流式细胞术和蛋白质印迹法，我们的数据表明IP6可减轻细胞溶质中钙的升高和α-突触核蛋白的聚集。总的来说，IP6对PD细胞模型中的多巴胺能细胞发挥了神经保护作用，其机制可能与线粒体介导的凋亡途径和α-突触核蛋白聚集的变化有关。
