Department of Food and Nutrition, College of Natural Science, Chosun University, 301, Pilmun-daero, Dong-gu, Gwangju 501-759, Korea.
Nutr Res Pract. 2013 Aug;7(4):249-55. doi: 10.4162/nrp.2013.7.4.249. Epub 2013 Aug 7.
In this study, the protective effects of EGCG on L-3,4-dihydroxyphenylalanine (L-DOPA)-induced oxidative cell death in catecholaminergic PC12 cells, the in vitro model of Parkinson's disease, were investigated. Treatment with L-DOPA at concentrations higher than 150 µM caused cytotoxicity in PC12 cells, as determined using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry detection. The apoptotic ratio was similar in cells treated with 100 µM EGCG plus 150 µM L-DOPA (5.02%) and the control (0.96%) (P > 0.05), and was lower than that of cells treated with L-DOPA only (32.24%, P < 0.05). The generation level of ROS (% of control) in cells treated with EGCG plus L-DOPA was lower than that in cells treated with L-DOPA only (123.90% vs 272.32%, P < 0.05). The optical density in production of TBARS in cells treated with L-DOPA only was higher than that in the control (0.27 ± 0.05 vs 0.08 ± 0.04, P < 0.05), and in cells treated with EGCG only (0.14 ± 0.02, P < 0.05), and EGCG plus L-DOPA (0.13 ± 0.02, P < 0.05). The intracellular level of GSH in cells treated with EGCG plus L-DOPA was higher than that in cells treated with L-DOPA only (233.25 ± 16.44 vs 119.23 ± 10.25, P < 0.05). These results suggest that EGCG protects against L-DOPA-induced oxidative apoptosis in PC12 cells, and might be a potent neuroprotective agent.
在这项研究中,我们研究了 EGCG 对 L-3,4-二羟基苯丙氨酸(L-DOPA)诱导的儿茶酚胺能 PC12 细胞(帕金森病的体外模型)氧化细胞死亡的保护作用。用浓度高于 150µM 的 L-DOPA 处理 PC12 细胞会导致细胞毒性,这可以通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定和流式细胞术检测来确定。用 100µM EGCG 和 150µM L-DOPA 处理的细胞的凋亡比例与对照(0.96%)相似(5.02%)(P>0.05),低于单独用 L-DOPA 处理的细胞(32.24%,P<0.05)。用 EGCG 和 L-DOPA 处理的细胞的 ROS 生成水平(对照的%)低于单独用 L-DOPA 处理的细胞(123.90%对 272.32%,P<0.05)。单独用 L-DOPA 处理的细胞中 TBARS 生成的光密度高于对照(0.27±0.05 对 0.08±0.04,P<0.05)和单独用 EGCG 处理的细胞(0.14±0.02,P<0.05),以及 EGCG 和 L-DOPA 处理的细胞(0.13±0.02,P<0.05)。用 EGCG 和 L-DOPA 处理的细胞内 GSH 水平高于单独用 L-DOPA 处理的细胞(233.25±16.44 对 119.23±10.25,P<0.05)。这些结果表明,EGCG 可防止 L-DOPA 诱导的 PC12 细胞氧化凋亡,可能是一种有效的神经保护剂。