Liu Hao, Mao Ping, Wang Jia, Wang Tuo, Xie Chang-Hou
Department of Neurosurgery, The First Affiliated Hospital of Xi'an Jiao Tong University, Xi'an, Shaanxi, China.
Cell Physiol Biochem. 2015;36(3):966-79. doi: 10.1159/000430271. Epub 2015 Jun 12.
Parkinson disease (PD) is a common adult-onset neurodegenerative disorder, and PD related neuronal injury is associated with oxidative stress and mitochondrial dysfunction. Allicin, the main biologically active compound derived from garlic, has been shown to exert various anti-oxidative and anti-apoptotic activities in in vitro and in vivo studies.
The present study aimed to investigate the potential protective role of allicin in an in vitro PD model induced by 6-hydroxydopamine (6-OHDA) in PC12 cells. The protective effects were measured by cell viability, decreased lactate dehydrogenase (LDH) release and flow cytometry, and the anti-oxidative activity was determined by reactive oxygen species (ROS) generation, lipid peroxidation and the endogenous antioxidant enzyme activities. Mitochondrial function in PC12 cells was detected by mitochondrial membrane potential (MMP) collapse, cytochrome c release, mitochondrial ATP synthesis, and the mitochondrial Ca(2+) buffering capacity. To investigate the potential mechanism, we also measured the expression of mitochondrial biogenesis factors, mitochondrial morphological dynamic changes, as well as detected mitochondrial dynamic proteins by western blot.
We found that allicin treatment significant increased cell viability, and decreased LDH release and apoptotic cell death after 6-OHDA exposure. Allicin also inhibited ROS generation, reduced lipid peroxidation and preserved the endogenous antioxidant enzyme activities. These protective effects were associated with suppressed mitochondrial dysfunction, as evidenced by decreased MMP collapse and cytochrome c release, preserved mitochondrial ATP synthesis, and the promotion of mitochondrial Ca(2+) buffering capacity. In addition, allicin significantly enhanced mitochondrial biogenesis and prevented fragmentation of mitochondrial network after 6-OHDA treatment. The results of western blot analysis showed that the 6-OHDA induced decrease in the expression of optic atrophy type 1 (Opa-1), increase in mitochondrial fission 1 (Fis-1) and dynamin-related protein 1 (Drp-1) were all partially revised by allicin.
In summary, our data strongly suggested that allicin treatment can exert protective effects against PD related neuronal injury through inhibiting oxidative stress and mitochondrial dysfunction with dynamic changes.
帕金森病（PD）是一种常见的成人起病的神经退行性疾病，与PD相关的神经元损伤与氧化应激和线粒体功能障碍有关。大蒜素是大蒜中的主要生物活性化合物，在体外和体内研究中已显示出具有多种抗氧化和抗凋亡活性。
本研究旨在探讨大蒜素在6-羟基多巴胺（6-OHDA）诱导的PC12细胞体外PD模型中的潜在保护作用。通过细胞活力、乳酸脱氢酶（LDH）释放减少和流式细胞术来测定保护作用，并通过活性氧（ROS）生成、脂质过氧化和内源性抗氧化酶活性来确定抗氧化活性。通过线粒体膜电位（MMP）崩溃、细胞色素c释放、线粒体ATP合成以及线粒体Ca(2+)缓冲能力来检测PC12细胞中的线粒体功能。为了研究潜在机制，我们还测量了线粒体生物发生因子的表达、线粒体形态动态变化，并通过蛋白质免疫印迹法检测线粒体动态蛋白。
我们发现，大蒜素处理显著提高了细胞活力，并减少了6-OHDA暴露后的LDH释放和凋亡细胞死亡。大蒜素还抑制了ROS生成，减少了脂质过氧化，并保留了内源性抗氧化酶活性。这些保护作用与线粒体功能障碍的抑制有关，表现为MMP崩溃和细胞色素c释放减少、线粒体ATP合成得以保留以及线粒体Ca(2+)缓冲能力增强。此外，大蒜素显著增强了线粒体生物发生，并在6-OHDA处理后防止了线粒体网络的碎片化。蛋白质免疫印迹分析结果表明，6-OHDA诱导的视神经萎缩蛋白1（Opa-1）表达降低以及线粒体分裂蛋白1（Fis-1）和动力相关蛋白1（Drp-1）表达增加均被大蒜素部分纠正。
总之，我们的数据强烈表明，大蒜素处理可通过抑制氧化应激和线粒体功能障碍及其动态变化，对与PD相关的神经元损伤发挥保护作用。
