Shimada Y, Yokoyama K, Goto H, Sekiya N, Mantani N, Tahara E, Hikiami H, Terasawa K
Department of Japanese Oriental Medicine, Faculty of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, Toyama 930-0194, Japan.
Phytomedicine. 2004 Jul;11(5):404-10. doi: 10.1016/j.phymed.2003.04.002.
Keishi-bukuryo-gan (Gui-Zhi-Fu-Ling-Wan) (KBG) is a traditional Chinese/Japanese medical (Kampo) formulation that has been administered to patients with "Oketsu" (blood stagnation) syndrome. In the process of neuronal cell death induced by brain ischemia, excessive generation of nitric oxide (NO) free radicals is implicated in the neurotoxicity. In the present study, we examined the protective effects of KBG and its constituent medicinal plants against NO donors, sodium nitroprusside (SNP) and 2,2'-(hydroxynitrosohydrazino)bis-ethanamine (NOC18)-induced neuronal death in cultured rat cerebellar granule cells (CGCs). MTT assay showed cell viability to be significantly increased by the addition of KBG extract (KBGE) (100 microg/ml), Cinnamomi Cortex extract (CCE) (3, 10 and 30 microg/ml), Paeoniae Radix extract (PRE) (100 microg/ml) and Moutan Cortex extract (MCE) (10 and 30 microg/ml) compared with exposure to SNP (30 microM, 24 h) only. Also, cell viability was significantly increased by the addition of KBGE (100 and 300 microg/ml), CCE (30 and 100 microg/ml), PRE (100 and 300 microg/ml) and MCE (30 and 100 microg/ml) compared with exposure to NOC 18 (100 microM, 48 h) only. Persicae Semen extract and Hoelen extract did not protect against NO donor-induced neuronal death. These results suggest that KBG has protective effect against NO-mediated neuronal death in cultured CGCs and that it is derived from Cinnamomi Cortex, Paeoniae Radix and Moutan Cortex.
桂枝茯苓丸（Keishi-bukuryo-gan，KBG）是一种传统的中医/日本汉方制剂，用于治疗“瘀血”（血液停滞）综合征患者。在脑缺血诱导的神经元细胞死亡过程中，过量产生的一氧化氮（NO）自由基与神经毒性有关。在本研究中，我们研究了桂枝茯苓丸及其组成药用植物对NO供体硝普钠（SNP）和2,2'-（羟基亚硝基肼基）双乙胺（NOC18）诱导的培养大鼠小脑颗粒细胞（CGCs）神经元死亡的保护作用。MTT分析表明，与仅暴露于SNP（30μM，24小时）相比，添加桂枝茯苓丸提取物（KBGE）（100μg/ml）、肉桂皮提取物（CCE）（3、10和30μg/ml）、芍药根提取物（PRE）（100μg/ml）和牡丹皮提取物（MCE）（10和30μg/ml）可显著提高细胞活力。此外，与仅暴露于NOC 18（100μM，48小时）相比，添加KBGE（100和300μg/ml）、CCE（30和100μg/ml）、PRE（100和300μg/ml）和MCE（30和100μg/ml）也可显著提高细胞活力。桃仁提取物和茯苓提取物对NO供体诱导的神经元死亡没有保护作用。这些结果表明，桂枝茯苓丸对培养的CGCs中NO介导的神经元死亡具有保护作用，且这种作用源自肉桂皮、芍药根和牡丹皮。
