Key Laboratory of Modern Preparation of Traditional Chinese Medicines, Ministry of Education, Jiangxi University of Chinese Medicine, 330004, Nanchang, China.
School of Pharmacy, Jiangxi University of Chinese Medicine, 330004, Nanchang, China.
J Ethnopharmacol. 2022 Nov 15;298:115662. doi: 10.1016/j.jep.2022.115662. Epub 2022 Aug 27.
Gui-Zhi-Jia-Ge-Gen decoction (GJGD) is a classical Chinese medicine prescription that has been widely used in clinical practice for centuries. In recent times, TCM has received considerable attention for its potential efficacy in treating a wind-cold type of common cold. However, the effect of the Gui-Zhi-Jia-Ge-Gen decoction on the wind-cold type of common cold is still not fully understood, which presents challenges for both quality control, research and development. Furthermore, the identification of potential pharmacodynamic ingredients (PPIs) is important for developing quality control procedures for industrial and large-scale production.
The aim of this study was to investigate the potential curative effect of Gui-Zhi-Jia-Ge-Gen decoction on wind-type of common cold using multidimensional qualitative analysis that combined water-decoction spectrums, in vivo plasma spectrums, and molecular docking to identify key constituents of GJGD.
Water-based GJGDs were formulated according to the clinical usage documented in ancient medical texts. Ultra-high-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF-MS) was combined with computer-aided modeling screening to identify GJGD PPIs in rats following oral administration. Molecular docking experiments were carried out to predict the binding affinity of the PPIs to tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin-1β (IL-1β). Finally, the active ingredients of GJGD were further validated through pharmacodynamic experiments by assessing their efficacy in treating a wind-cold type of common cold in rats.
A total of 61 compounds were identified in the GJGD, 8 of which were detected in rat blood samples, providing stronger evidence for PPIs. Molecular docking also confirmed that these 8 compounds had a better affinity for TNF-α, IL-6, and IL-1β. In animal studies, various doses of the GJGD groups and the positive control groups caused significant elevations (P < 0.05) in the levels of white blood cell count and lymphocyte ratio and caused a significant decrease (P < 0.05) in the monocyte ratio and neutrophilic granulocyte ratio compared to the model group. Organ indexes of the GJGD treated groups were higher than the model group (P < 0.05). Significant neutrophil infiltration, hemorrhage, compensatory vacuole, and interstitium proliferation were observed in the lung tissue of the model group. However, the lung tissues of the various dose groups that received GJGD showed a near normal appearance, except for slight thickening, interstitium proliferation, and compensatory vacuole in some areas. The GJGD was found to be effective against a cold-wind type of common cold, which is in accordance with molecular docking studies suggesting that GJGD may be effective against a cold-wind type of common cold. Finally, based on multidimensional analysis, 8 potential compounds in GJGD were identified as PPIs (puerarin, 3'-hydroxy puerarin, 3'- methoxy puerarin, daidzin, cinnamic acid, paeoniflorin, liquiritin, and glycyrrhizic acid).
The present study combined water decoction spectral analysis, molecular docking, and in vivo blood plasma spectrum analysis to develop a multidimensional qualitative approach for the development of GJGD and to assess its effectiveness in a wind type of common cold in Sprague Dawley rats. Meanwhile, 8 compounds in the GJGD were identified as PPIs in this study, which may be useful in developing quality standards for complex TCM prescriptions.
桂枝加葛根汤（GJGD）是一种经典的中药方剂，已在临床实践中广泛应用了数个世纪。最近，中医因其在治疗风寒型普通感冒方面的潜在疗效而受到了相当多的关注。然而，桂枝加葛根汤治疗风寒型普通感冒的效果仍未被充分了解，这为质量控制、研究和开发带来了挑战。此外，鉴定潜在药效成分（PPIs）对于开发工业和大规模生产的质量控制程序非常重要。
本研究旨在采用多维定性分析方法，结合水提谱、体内血浆谱和分子对接，鉴定桂枝加葛根汤治疗风寒型普通感冒的潜在疗效，以识别 GJGD 的关键成分。
根据古代医书记载的临床使用情况，配制了水基桂枝加葛根汤。采用超高效液相色谱-四极杆飞行时间质谱联用（UHPLC-Q-TOF-MS）结合计算机辅助建模筛选，鉴定大鼠口服后桂枝加葛根汤的药效成分。分子对接实验用于预测药效成分与肿瘤坏死因子-α（TNF-α）、白细胞介素 6（IL-6）和白细胞介素-1β（IL-1β）的结合亲和力。最后，通过评估桂枝加葛根汤治疗大鼠风寒型普通感冒的疗效，进一步通过药效学实验验证了 GJGD 的活性成分。
在 GJGD 中总共鉴定出 61 种化合物，其中 8 种在大鼠血样中被检测到，为药效成分提供了更强的证据。分子对接也证实，这 8 种化合物与 TNF-α、IL-6 和 IL-1β具有更好的亲和力。在动物研究中，与模型组相比，各剂量 GJGD 组和阳性对照组的白细胞计数和淋巴细胞比值显著升高（P<0.05），单核细胞比值和中性粒细胞比值显著降低（P<0.05）。GJGD 治疗组的器官指数高于模型组（P<0.05）。模型组肺组织可见明显的中性粒细胞浸润、出血、代偿性空泡和间质增生。然而，接受 GJGD 各剂量治疗的肺组织除了在某些区域出现轻微的增厚、间质增生和代偿性空泡外，外观基本正常。研究结果表明，GJGD 对风寒型普通感冒有效，这与分子对接研究表明 GJGD 可能对风寒型普通感冒有效一致。最后，基于多维分析，鉴定出 GJGD 中的 8 种潜在药效成分（葛根素、3'-羟基葛根素、3'-甲氧基葛根素、大豆苷元、桂皮酸、芍药苷、甘草苷和甘草酸）。
本研究结合水提谱分析、分子对接和体内血浆谱分析，为桂枝加葛根汤的开发建立了多维定性方法，并评估了其在 Sprague Dawley 大鼠风寒型普通感冒中的疗效。同时，本研究在桂枝加葛根汤中鉴定出 8 种化合物为药效成分，这可能有助于开发复杂中药方剂的质量标准。
