State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, China.
CAS Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, The Chinese Academy of Sciences, Wuhan, China.
Phytochem Anal. 2021 Sep;32(5):767-779. doi: 10.1002/pca.3022. Epub 2020 Dec 17.
The roots of Polygonum multiflorum (PM) serve as a classical traditional Chinese medicine (TCM), which has multiple biological activities. However, many cases of hepatotoxicity in PM have been reported in recent years. Processing PM with black beans decoction is one of the typical processing methods to reduce the hepatotoxicity of PM since ancient times.
To find potential effective constituents, as well as the optimal variety and origin of black beans for the processing of PM.
Based on ultrahigh-performance liquid chromatography Q-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS) analysis, we measured the contents of the two potential toxic compounds (emodin-8-O-glucoside and torachrysone-O-hexose) in raw PM (R-PM), PM processed with big black beans (B-PM) and PM processed with small black beans (S-PM). The flow cytometry method analysed the effects of different processed products of PM on apoptosis of L02 cells in different drug concentration. Proton nuclear magnetic resonance ( H-NMR) and UHPLC-Q-Orbitrap-MS together with multivariate statistical analysis were used to systematically analyse the different components between small black beans (Small-BB) and big black beans (Big-BB) from 30 different habitats.
The toxicity was ranked from small to large: S-PM < B-PM < R-PM. Processing PM with black beans could significantly decrease the apoptosis rate of L02 cells, especially when the drug concentration is 80 μg/mL. Besides, we find five differential compounds (α-arabinose, α-galactose, proline, isomer of daidzein and isomer of genistein) may be potential active ingredients. In terms of the black beans collected from 30 producing areas, we find that Small-BB from Weifang in Shandong province was optimum to processing PM, followed by Shangqiu in Henan province, Jilin and Liaoning province.
The ingredients that affect the processing of PM may be attributed to α-arabinose, α-galactose, proline, isomer of daidzein and isomer of genistein in black beans. When the drug concentration is higher, the effect of reducing the hepatotoxicity of PM is better. Besides, Small-BB was more effective than Big-BB for reducing the toxicity of PM, especially Small-BB from Weifang in Shandong, Shangqiu in Henan province and northeast China.
何首乌的根是一种经典的中药，具有多种生物活性。然而，近年来何首乌引起肝毒性的病例屡有报道。自古以来，用黑豆汁炮制何首乌是降低何首乌肝毒性的典型炮制方法之一。
寻找潜在的有效成分，以及用于炮制何首乌的最佳黑豆品种和产地。
基于超高效液相色谱-轨道阱质谱（UHPLC-Q-Orbitrap-MS）分析，我们测量了生何首乌（R-PM）、用大黑黑豆炮制的何首乌（B-PM）和用小黑黑豆炮制的何首乌（S-PM）中两种潜在有毒化合物（大黄素-8-O-葡萄糖苷和虎杖酮-O-己糖）的含量。采用流式细胞术分析不同炮制何首乌对不同药物浓度下 L02 细胞凋亡的影响。质子核磁共振（1 H-NMR）和 UHPLC-Q-Orbitrap-MS 与多元统计分析相结合，系统分析了来自 30 个不同产地的小黑豆（Small-BB）和大黑豆（Big-BB）之间的不同成分。
毒性从低到高依次为：S-PM＜B-PM＜R-PM。用黑豆炮制何首乌能显著降低 L02 细胞的凋亡率，特别是在药物浓度为 80μg/ml 时。此外，我们发现 5 种差异化合物（α-阿拉伯糖、α-半乳糖、脯氨酸、大豆苷元异构体和染料木素异构体）可能是潜在的活性成分。就来自 30 个产地的黑豆而言，我们发现来自山东潍坊的 Small-BB 是炮制何首乌的最佳选择，其次是河南商丘、吉林和辽宁。
影响何首乌炮制的成分可能归因于黑豆中的α-阿拉伯糖、α-半乳糖、脯氨酸、大豆苷元异构体和染料木素异构体。当药物浓度较高时，降低何首乌肝毒性的效果更好。此外，小黑豆比大黑豆更能降低何首乌的毒性，尤其是来自山东潍坊、河南商丘和中国东北的小黑豆。
