Department of Alternative Medicine, Gwangju University, Gwangju, Republic of Korea.
J Pharm Pharmacol. 2011 Oct;63(10):1358-67. doi: 10.1111/j.2042-7158.2011.01342.x. Epub 2011 Aug 30.
The present study investigated the neuroprotective effects of Rhus verniciflua Stokes (RVS) leaf extract on rotenone-induced apoptosis in human dopaminergic cells, SH-SY5Y.
Cells were pretreated with RVS extract for 1 h then treated with vehicle or rotenone for 24 h. Cell viability, cell cytotoxicity, cell morphology and nuclear morphology were examined by MTT assay, lactate dehydrogenase release assay, phase contrast microscopy and staining with Hoechast 33342, respectively. Reactive oxygen species were measured by 2'7'-dichlorofluorescein diacetate and fragmented DNA was observed by TUNEL assay. Mitochondrial membrane potential was determined by Rhodamine 123. Pro-apoptotic and anti-apoptotic proteins and tyrosine hydroxylase were analysed by Western blotting.
Results showed that RVS suppressed rotenone-induced reactive oxygen species generation, cellular injury and apoptotic cell death. RVS also prevented rotenone-mediated changes in Bax/Bcl-2 levels, mitochondrial membrane potential dissipation and Caspase 3 activation. Moreover, RVS pretreatment increased the tyrosine hydroxylase levels in SH-SY5Y cells.
These findings demonstrate that RVS protects SH-SY5Y cells against rotenone-induced injury and suggest that RVS may have potential therapeutic value for neurodegenerative disease associated with oxidative stress.
本研究旨在探讨漆树（Rhus verniciflua Stokes）叶提取物对鱼藤酮诱导的人多巴胺能细胞（SH-SY5Y）凋亡的神经保护作用。
用漆树叶提取物预处理细胞 1 h，然后用载体或鱼藤酮处理 24 h。通过 MTT 测定法、乳酸脱氢酶释放测定法、相差显微镜和 Hoechast 33342 染色分别检测细胞活力、细胞细胞毒性、细胞形态和核形态。通过 2'7'-二氯荧光素二乙酸酯测量活性氧，通过 TUNEL 测定法观察断裂的 DNA。通过 Rhodamine 123 测定线粒体膜电位。通过 Western blot 分析促凋亡和抗凋亡蛋白以及酪氨酸羟化酶。
结果表明，漆树提取物抑制了鱼藤酮诱导的活性氧生成、细胞损伤和凋亡细胞死亡。漆树提取物还防止了鱼藤酮介导的 Bax/Bcl-2 水平变化、线粒体膜电位耗散和 Caspase 3 激活。此外，漆树提取物预处理增加了 SH-SY5Y 细胞中的酪氨酸羟化酶水平。
这些发现表明漆树提取物对鱼藤酮诱导的损伤具有保护作用，并表明漆树提取物可能对与氧化应激相关的神经退行性疾病具有潜在的治疗价值。
