The First Hospital, Hunan University of Chinese Medicine, Changsha, Hunan Province, China.
Hunan University of Chinese Medicine, Changsha, Hunan Province, China.
J Ethnopharmacol. 2024 Jan 30;319(Pt 1):117090. doi: 10.1016/j.jep.2023.117090. Epub 2023 Aug 26.
Anxiety disorders leads to a decline in quality of life and increased risk of morbidity and mortality. The Baihe Dihuang decoction (BDD) is a classic Chinese medical formula that has been widely used to treat anxiety disorders for thousands of years in China. However, the pharmacodynamic material that is responsible for the antianxiety of BDD remains unclear.
To screen the main ingredients of anti-anxiety in BDD based on the establishment of spectrum-effect relationship and verified experiment.
The UPLC-Q-TOF/MS technique was utilized to establish fingerprints of various fractions of BDD and identify the main compounds. The anti-anxiety effects of BDD were comprehensively evaluated through multiple assessments, including the open field test, elevated plus maze test, and neurotransmitters tests. Then, the spectrum-effect relationship was established through Pearson correlation analysis, gray correlation analysis, orthogonal partial least squares regression analysis. The spectrum-effect relationship results were confirmed through various measures on an anxiety condition cell model, induced by a corticosterone and lipopolysaccharide intervention. These measures included assessing neuronal cell viability, morphology, apoptosis, synaptic damage, and the expression of neurotransmitters and inflammatory factors.
In the UPLC-Q-TOF-MS fingerprint, 46 common peaks were identified. The pharmacological results indicated that different fractions of BDD have strong effects on improving anxiety-like behavior and regulating neurotransmitters. Among them, butanol fraction has the highest comprehensive evaluation score of anti-anxiety efficacy, which is main active fraction of BDD for anti-anxiety. The analysis of the spectrum-effect relationship revealed that the 46 peaks exhibited varying degrees of correlation with the anti-anxiety efficacy indicators of BDD. Among them, 14 components have a high correlation with the anti-anxiety efficacy indicators, which may be the potential anti-anxiety efficacy components of BDD. The in vitro activity verification of active components verified our prediction, regaloside A, B, C, D, H, acteoside, and isoacteoside improved neuronal cell viability, cell morphology, apoptosis, and synaptic damage. Additionally, regaloside A, B, C, D, H and acteoside regulated the neurotransmitter levels, while regaloside A, B, C, D, acteoside and isoacteoside inhibited the levels of inflammatory cytokines.
The butanol fraction was found to be the main active fraction of BDD, and 14 compounds were the major anxiolytic active components. The results of verifying the major active components were consistent with the predicted results of the spectrum-effect analysis. The developed spectrum-effect analysis in this study demonstrates high accuracy and reliability for screening active components in TCMs.
焦虑症会导致生活质量下降，增加发病率和死亡率的风险。百合地黄汤（BDD）是一种经典的中药方剂，在中国已经被广泛用于治疗焦虑症数千年。然而，负责 BDD 抗焦虑作用的药效物质仍然不清楚。
基于谱效关系的建立和验证实验，筛选 BDD 中的主要抗焦虑成分。
采用 UPLC-Q-TOF/MS 技术建立 BDD 各馏分的指纹图谱并鉴定主要化合物。通过旷场试验、高架十字迷宫试验和神经递质试验等综合评价 BDD 的抗焦虑作用。然后，通过皮尔逊相关分析、灰色关联分析、正交偏最小二乘回归分析建立谱效关系。通过皮质酮和脂多糖干预诱导的焦虑状态细胞模型，采用评估神经元细胞活力、形态、凋亡、突触损伤以及神经递质和炎症因子表达等多种措施，对谱效关系结果进行验证。
在 UPLC-Q-TOF-MS 指纹图谱中，共鉴定出 46 个共有峰。药理结果表明，BDD 的不同馏分对改善焦虑样行为和调节神经递质具有很强的作用。其中，正丁醇馏分具有最高的抗焦虑综合评价评分，是 BDD 抗焦虑的主要活性馏分。谱效关系分析表明，46 个峰与 BDD 的抗焦虑疗效指标呈不同程度的相关性。其中，有 14 个成分与抗焦虑疗效指标具有高度相关性，可能是 BDD 的潜在抗焦虑功效成分。活性成分的体外活性验证验证了我们的预测， regaloside A、B、C、D、H、acteoside 和 isoacteoside 提高了神经元细胞活力、细胞形态、凋亡和突触损伤。此外，regaloside A、B、C、D、H 和 acteoside 调节神经递质水平，而 regaloside A、B、C、D、acteoside 和 isoacteoside 抑制炎症细胞因子水平。
正丁醇馏分被发现是 BDD 的主要活性馏分，14 种化合物是主要的抗焦虑活性成分。主要活性成分的验证结果与谱效分析的预测结果一致。本研究中建立的谱效分析方法对筛选中药中的活性成分具有较高的准确性和可靠性。
