Yin Shou Yu, Lee Jae Joon, Kim Yu Mi, Jin Chun Mei, Yang Yoo Jung, Kang Min Hee, Kai Masaaki, Lee Myung Koo
College of Pharmacy, and Research Center for Bioresource and Health, Chungbuk National University, San 48, Kaeshin-Dong, Heungduk-Gu, Cheongju 361-763, South Korea.
Eur J Pharmacol. 2004 Mar 19;488(1-3):71-7. doi: 10.1016/j.ejphar.2004.02.021.
(1R,9S)-beta-Hydrastine in lower concentrations of 10-50 microM inhibits dopamine biosynthesis in PC12 cells. In this study, the effects of (1R,9S)-beta-hydrastine on L-DOPA (L-3,4-dihydroxyphenylalanine)-induced cytotoxicity in PC12 cells were investigated. (1R,9S)-Hydrastine at concentrations up to 250 microM did not reduce cell viability. However, at concentrations higher than 500 microM it caused cytotoxicity in PC12 cells, as determined with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, TUNEL (terminal deoxynucleotidyltransferase dUTP nick-end labeling) method and flow cytometry. Exposure of PC12 cells to cytotoxic concentrations of (1R,9S)-beta-hydrastine (500 and 750 microM) in combination with L-DOPA (20, 50 and 100 microM) after 24 or 48 h resulted in a significant decrease in cell viability compared with the effects of (1R,9S)-beta-hydrastine or L-DOPA alone, and apoptotic cell death was observed. However, the decrease in cell viability induced by (1R,9S)-beta-hydrastine was not prevented by the antioxidant N-acetyl-L-cysteine, indicating that it is not mediated by membrane-based oxygen free radical damage. These data suggest that (1R,9S)-beta-hydrastine has a mild cytotoxic effect and at higher concentration ranges aggravates L-DOPA-induced cytotoxicity in PC12 cells.
较低浓度（10 - 50微摩尔）的(1R,9S)-β-氢化小檗碱可抑制PC12细胞中的多巴胺生物合成。在本研究中，研究了(1R,9S)-β-氢化小檗碱对PC12细胞中L-多巴（L-3,4-二羟基苯丙氨酸）诱导的细胞毒性的影响。浓度高达250微摩尔的(1R,9S)-氢化小檗碱不会降低细胞活力。然而，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、TUNEL（末端脱氧核苷酸转移酶dUTP缺口末端标记）法和流式细胞术测定，浓度高于500微摩尔时，它会在PC12细胞中引起细胞毒性。在24或48小时后，将PC12细胞暴露于细胞毒性浓度的(1R,9S)-β-氢化小檗碱（500和750微摩尔）与L-多巴（20、50和100微摩尔）组合中，与单独使用(1R,9S)-β-氢化小檗碱或L-多巴相比，细胞活力显著降低，并观察到凋亡性细胞死亡。然而，抗氧化剂N-乙酰-L-半胱氨酸并不能阻止(1R,9S)-β-氢化小檗碱诱导的细胞活力下降，这表明它不是由基于膜的氧自由基损伤介导的。这些数据表明，(1R,9S)-β-氢化小檗碱具有轻度细胞毒性作用，在较高浓度范围内会加重PC12细胞中L-多巴诱导的细胞毒性。
