【脑老化.2019】硫辛酸和维生素D3联合作用对星形胶质细胞的作用，预防氧化应激和铁积累诱导导致的脑老化

文献DOI：10.1155/2019/2843121

文献PMID：30944691

文献原文链接：http://doi.org.sci-hub.tw/10.1155/2019/2843121

Role of Combined Lipoic Acid and Vitamin D3 on Astrocytes as a Way to Prevent Brain Ageing by Induced Oxidative Stress and Iron Accumulation

硫辛酸和维生素D3联合作用对星形胶质细胞的作用，预防氧化应激和铁积累诱导导致的脑老化

【Abstract】

Brain ageing is a complex multifactorial process characterized by gradual and continuous loss of neuronal functions. It is hypothesized that at the basis of brain ageing as well as age-related diseases, there is an impairment of the antioxidant defense system leading to an increase of oxidative stress. In this study, two different biological aspects involved in brain ageing and neurodegeneration have been investigated: oxidative stress and iron accumulation damage. In primary mouse astrocytes, the stimulation with 50 μM lipoic acid (LA) and 100 nM vitamin D (vitD) was first investigated in a time-course study to determine the dosages to be used in combination and then in a permeability test using an in vitro blood-brain barrier. In a second set of experiments, the role of oxidative stress was investigated pretreating astrocytes with 200 μM H2O2 for 30 min. The ability of vitD and LA alone and combined together to prevent or repair the damage caused by oxidative stress was investigated after 24h of stimulation by the MTT test, mitochondrial membrane potential measurement, and Western blot analysis. To induce neurodegeneration, cells were pretreated with 300 μM catalytic iron for 6 days and then treated with vitD and LA alone and combined for additional 6 days to investigate the protection exerted by combination, analyzing viability, ROS production, iron concentration, and activation of intracellular pathways. In our study, the combination of LA and vitD showed beneficial effects on viability of astrocytes, since the substances are able to cross the brain barrier. In addition, combined LA and vitD attenuated the H2O2-induced apoptosis through the mitochondrial-mediated pathway. The combination was also able to counteract the adverse conditions caused by iron, preventing its accumulation. All these data support the hypothesis of the synergistic and cooperative activity exerted by LA and vitD in astrocytes indicating a possible new strategy to slow down ageing.

摘要翻译：

脑老化是一个复杂的多因素过程，其特征是神经元功能的逐渐和持续丧失。据推测，在脑老化以及与年龄相关的疾病的基础上，抗氧化防御系统受损导致氧化应激增加。在这项研究中，已经研究了涉及脑老化和神经变性的两个不同的生物学方面：氧化应激和铁积累损伤。在原代小鼠星形胶质细胞中，首先在时间过程研究中研究用50μM硫辛酸（LA）和100nM维生素D（vitD）刺激以确定组合使用的剂量，然后在体外做血脑屏障的渗透性测试。在第二组实验中，研究了氧化应激的作用，用200μMH2O2预处理星形胶质细胞30分钟。通过MTT试验，线粒体膜电位测量和蛋白质印迹分析，在刺激24小时后研究了单独的vitD和LA以及组合在一起以预防或修复由氧化应激引起的损伤的能力。为了诱导神经变性，将细胞用300μM催化铁预处理6天，然后单独用vitD和LA处理，再合并用6天，研究组合的保护作用，分析细胞活力、ROS产生、铁浓度和细胞内信号活化途径。在我们的研究中，LA和vitD的组合显示出对星形胶质细胞活力的有益影响，因为这些物质能够穿过脑屏障。此外，联合LA和vitD通过线粒体介导的途径减弱H2O2诱导的细胞凋亡。该组合还能够抵消由铁引起的不利条件，防止其积累。所有这些数据都支持LA和vitD在星形胶质细胞中发挥的协同和合作活动的假设，这表明可能有一种减缓衰老的新策略。