Department of Oriental Gynecology, College of Oriental Medicine, Kyung Hee University, #1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of Korea.
J Ethnopharmacol. 2011 Jan 27;133(2):934-9. doi: 10.1016/j.jep.2010.11.017. Epub 2010 Nov 11.
Dangguijakyak-san (DJS) is a multi-herbal formula that has long been widely used in traditional Oriental medicine to treat gynecologic disorders, including neurological symptoms. Recent clinical and experimental studies have reported aging and anti-neurodegenerative effects of DJS. In this study, we evaluated the neuroprotective effects of DJS on dopaminergic (DA) neurons damaged by 6-hydroxydopamine (6-OHDA).
To evaluate the protective effects of DJS, we analyzed viability in SH-SY5Y neuroblastoma cells and tyrosine hydroxylase (TH) staining in primary DA cells. To explore the possible mechanism(s) of neuroprotection, we assessed anti-oxidant activity by measuring reactive oxygen species (ROS) and glutathione (GSH) levels. To determine mitochondria-mediated apoptotic activity, we examined mitochondrial membrane potential, cytochrome c release, and caspase-3 activation.
DJS at 0.05-5 μg/mL significantly protected SH-SY5Y cells from 6-OHDA toxicity, dose-dependently, and attenuated 6-OHDA damage in primary DA cells. DJS reduced 6-OHDA-induced intracellular ROS production and GSH depletion and inhibited mitochondrial membrane instability, cytosolic cytochrome c release, and caspase-3 activation.
These results demonstrate that DJS has neuroprotective effects in DA neurons against 6-OHDA-induced toxicity through anti-oxidant and anti-mitochondrial-mediated apoptotic activities.
当归芍药散（DJS）是一种多草药配方，长期以来广泛应用于传统东方医学治疗妇科疾病，包括神经症状。最近的临床和实验研究报告了 DJS 的衰老和抗神经退行性作用。在这项研究中，我们评估了 DJS 对 6-羟多巴胺（6-OHDA）损伤的多巴胺能（DA）神经元的神经保护作用。
为了评估 DJS 的保护作用，我们分析了 SH-SY5Y 神经母细胞瘤细胞的活力和原代 DA 细胞中酪氨酸羟化酶（TH）的染色。为了探讨神经保护的可能机制，我们通过测量活性氧（ROS）和谷胱甘肽（GSH）水平来评估抗氧化活性。为了确定线粒体介导的凋亡活性，我们检查了线粒体膜电位、细胞色素 c 释放和 caspase-3 激活。
DJS 在 0.05-5μg/mL 时显著保护 SH-SY5Y 细胞免受 6-OHDA 毒性，呈剂量依赖性，并减轻了原代 DA 细胞中的 6-OHDA 损伤。DJS 减少了 6-OHDA 诱导的细胞内 ROS 产生和 GSH 耗竭，并抑制了线粒体膜不稳定、胞质细胞色素 c 释放和 caspase-3 激活。
这些结果表明，DJS 通过抗氧化和抗线粒体介导的凋亡活性对 6-OHDA 诱导的 DA 神经元毒性具有神经保护作用。
