Chen Wei-Cheng, Lai Yi-Syuan, Lin Shih-Hang, Lu Kuan-Hung, Lin Yu-En, Panyod Suraphan, Ho Chi-Tang, Sheen Lee-Yan
Institute of Food Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
Department of Food Science, Rutgers University, New Brunswick, NJ 08901-8520, USA.
J Ethnopharmacol. 2016 Apr 22;182:190-9. doi: 10.1016/j.jep.2016.02.001. Epub 2016 Feb 16.
Gastrodia elata Blume is a highly valuable traditional Chinese medicine used in the treatment of depression. However, compounds with antidepressant effects in water extracts of G. elata Bl. (WGE) have not been identified. The aims of this study were to determine the major antidepressant compound in WGE and to evaluate the antidepressant effects of WGE and its active compounds which involved the monoaminergic system and neuronal cytoskeletal remodeling.
Gastrodin (GAS) and 4-hydroxybenzyl alcohol (HBA) in WGE, were analyzed with high-performance liquid chromatography (HPLC)-ultraviolet detection. The forced swimming test (FST) was used to induce depression-like symptoms in 9 weeks old male Sprague-Dawley rats. The open field test (OFT) was used to measure anxiety after WGE, GAS, and HBA treatments. The levels of monoamine such as serotonin (5-HT), dopamine (DA), and their metabolites 5-hydroxyindoleacetic acid (5-HIAA), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) were measured using HPLC-electrochemical detection. Western blotting was used to examine the 5-HT1A receptor and the neuronal cytoskeleton remodeling-related proteins, Slit, dihydropyrimidinase-related protein 2 (DPYSL2, also called CRMP2), Ras homologous member A (RhoA), and profilin 1 (PFN1) in vivo. Slit1 expression was evaluated in Hs683 cell line after treated with WGE (0.5mg/mL), GAS (50, 100 and 100μM), and HBA (50, 100 and 100μM).
Oral administration of WGE (500mg/kg bw), GAS (100mg/kg bw), and HBA (100mg/kg bw) exhibited the anti-depressant effect by significantly reducing the immobility time in FST, monoamine metabolism including the 5-HT to 5-HIAA in the hippocampus and DA to DOPAC and HVA ratios in the frontal cortex, amygdala, and hippocampus. In the hippocampus, the expression of the neuronal cytoskeleton remodeling-related negative regulators Slit1 and RhoA were significantly down-regulated. In addition, the positive regulators CRMP2 and PFN1 were significantly up-regulated following GAS, HBA, and WGE treatments. Moreover, WGE, GAS, and HBA were directly down-regulated Slit1 expression in Hs683 cells.
WGE, GAS, and HBA exhibited potential anti-depressant effects in rats by decreasing monoamine metabolism and modulated cytoskeleton remodeling-related protein expression in the Slit-Robo pathway. These results suggest that WGE can be used as agent for depressive prevention.
天麻是一种极具价值的传统中药，用于治疗抑郁症。然而，天麻水提取物（WGE）中具有抗抑郁作用的化合物尚未被鉴定出来。本研究的目的是确定WGE中的主要抗抑郁化合物，并评估WGE及其活性化合物的抗抑郁作用，这些作用涉及单胺能系统和神经元细胞骨架重塑。
采用高效液相色谱（HPLC）-紫外检测法分析WGE中的天麻素（GAS）和4-羟基苄醇（HBA）。采用强迫游泳试验（FST）在9周龄雄性Sprague-Dawley大鼠中诱导抑郁样症状。采用旷场试验（OFT）测量WGE、GAS和HBA处理后的焦虑水平。使用HPLC-电化学检测法测量单胺如血清素（5-HT）、多巴胺（DA）及其代谢产物5-羟基吲哚乙酸（5-HIAA）、3,4-二羟基苯乙酸（DOPAC）和高香草酸（HVA）的水平。采用蛋白质免疫印迹法检测体内5-HT1A受体以及与神经元细胞骨架重塑相关的蛋白质，即Slit、二氢嘧啶酶相关蛋白2（DPYSL2，也称为CRMP2）、Ras同源成员A（RhoA）和丝切蛋白1（PFN1）。在用WGE（0.5mg/mL）、GAS（50、100和100μM）和HBA（50、100和100μM）处理后的Hs683细胞系中评估Slit1表达。
口服WGE（500mg/kg体重）、GAS（100mg/kg体重）和HBA（100mg/kg体重）通过显著减少FST中的不动时间、单胺代谢（包括海马体中5-HT向5-HIAA的转化以及额叶皮质、杏仁核和海马体中DA向DOPAC和HVA的比率）表现出抗抑郁作用。在海马体中，与神经元细胞骨架重塑相关的负调节因子Slit1和RhoA的表达显著下调。此外，在GAS、HBA和WGE处理后，正调节因子CRMP2和PFN1显著上调。此外，WGE、GAS和HBA直接下调了Hs683细胞中Slit1的表达。
WGE、GAS和HBA通过减少单胺代谢并调节Slit-Robo途径中与细胞骨架重塑相关的蛋白表达，在大鼠中表现出潜在的抗抑郁作用。这些结果表明WGE可作为预防抑郁症的药物。
