Yang Y-H, Hsieh T-J, Tsai M-L, Chen C-H, Lin H-T, Wu S-J
Dr. Shyh-Jong Wu, Ph.D. No 100, Tzyou 1 Rd, Kaohsiung Medical University, Kaohsiung City, Taiwan. Phone: +886 7 3121101 ext. 2354, Fax: +886 7 3113449, E-mail:
J Nutr Health Aging. 2014;18(2):184-90. doi: 10.1007/s12603-013-0382-3.
Oxidative stress is considered the potential risk to the development of dementia. Some medicines, vitamins, and diet supplements have been suggested to have possible benefits via the antioxidative effects to slow the decline of cognitive function in demented and non-demented individuals. However, few studies were conducted to examine their functions, especially in composite diet supplements. Hu-Yi-Neng is a composite diet supplement, including ginkgo biloba, extract of pine bark, phosphatidyl serine, docosahexaenoic acid, and folic acid, used extensively in Taiwan. Therefore, our aim is to investigate the potential protective effects of Hu-Yi-Neng on human neuron cells. MATERALS AND METHODS: H2O2-induced neuronal toxicity was characterized in SH-SY5Y human neuroblastoma cells by the decrease of cell viability using PrestoBlue™ assay and by the increase of intracellular reactive oxygen species (ROS) level using DCFH-DA (2', 7'-dichlorodihydrofluorescin diacetate) assays. HO-1 mRNA expression was detected by real-time PCR. Akt and Erk 1/2 proteins were detected by western blotting.
Pretreatment with Hu-Yi-Neng significantly reversed the decrease in cell viability induced by H2O2 in SH-SY5Y cells. Furthermore, Hu-Yi-Neng dose-dependently suppressed the elevation of intracellular reactive oxygen species (ROS) level. Hu-Yi-Neng protected SH-SY5Y cells from oxidative stress may via the increase in mRNA expression of heme oxygenase-1 (HO-1), an antioxidant enzyme. In addition, Hu-Yi-Neng inhibited H2O2-induced phosphorylation of Akt kinase but further increased the phosphorylation of Erk 1/2.
Our results suggest that Hu-Yi-Neng has protective effect against oxidative stress-induced neuron cell loss and it could be an ideal composite diet supplement for preventing neurodegenerative diseases.
氧化应激被认为是痴呆症发展的潜在风险。一些药物、维生素和膳食补充剂被认为可能通过抗氧化作用对延缓痴呆和非痴呆个体的认知功能衰退有益。然而，很少有研究对它们的功能进行检测，尤其是复合膳食补充剂。胡依能是一种复合膳食补充剂，包含银杏叶、松树皮提取物、磷脂酰丝氨酸、二十二碳六烯酸和叶酸，在台湾广泛使用。因此，我们的目的是研究胡依能对人神经元细胞的潜在保护作用。
通过使用PrestoBlue™检测法检测细胞活力的降低以及使用DCFH-DA（2',7'-二氯二氢荧光素二乙酸酯）检测法检测细胞内活性氧（ROS）水平的升高，来表征H2O2诱导的SH-SY5Y人神经母细胞瘤细胞的神经毒性。通过实时PCR检测HO-1 mRNA表达。通过蛋白质印迹法检测Akt和Erk 1/2蛋白。
用胡依能预处理可显著逆转H2O2诱导的SH-SY5Y细胞活力下降。此外，胡依能剂量依赖性地抑制细胞内活性氧（ROS）水平的升高。胡依能可能通过增加抗氧化酶血红素加氧酶-1（HO-1）的mRNA表达来保护SH-SY5Y细胞免受氧化应激。此外，胡依能抑制H2O2诱导的Akt激酶磷酸化，但进一步增加Erk 1/2的磷酸化。
我们的结果表明，胡依能对氧化应激诱导的神经元细胞损失具有保护作用，它可能是预防神经退行性疾病的理想复合膳食补充剂。
