Modern Research Center for Traditional Chinese Medicine, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, No. 92, Wucheng Road, Taiyuan, 030006, Shanxi, PR China; Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province, Shanxi University, No. 92, Wucheng Road, Taiyuan, 030006, Shanxi, PR China.
Modern Research Center for Traditional Chinese Medicine, The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, No. 92, Wucheng Road, Taiyuan, 030006, Shanxi, PR China; Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050, PR China.
J Ethnopharmacol. 2023 Apr 6;305:116068. doi: 10.1016/j.jep.2022.116068. Epub 2022 Dec 24.
Bupleurum chinense DC-Paeonia lactiflora Pall (BCD-PLP) is a common clinical herb pair in traditional Chinese medicine (TCM) prescriptions commonly used to treat depression. However, its combination mechanisms with its anti-depressive effects remain highly unclear.
Here, an effective strategy has been developed to study the combination mechanisms of Bupleurum chinense DC (BCD) and Paeonia lactiflora Pall (PLP) by integrating serum pharmacochemistry analysis, metabolomics technology, and molecular docking technology.
First, the depression model rats were replicated by the chronic unpredictable mild stress (CUMS) procedure, and the difference in the chemical composition in vivo before and after the combination of BCD and PLP was analyzed by integrating background subtraction and multivariate statistical analysis techniques. Then, UPLC/HRMS-based serum metabolomics was performed to analyze the synergistic effect on metabolite regulation before and after the combination of BCD and PLP. Further, the correlation analysis between the differential exogenous chemical components and the differential endogenous metabolites before and after the combination was employed to dissect the combination mechanisms from a global perspective of combining metabolomics and serum pharmacochemistry. Finally, the molecular docking between the differential chemical components and the key metabolic enzymes was applied to verify the regulatory effect of the differential exogenous chemical components on the differential endogenous metabolites.
The serum pharmacochemistry analysis results demonstrated that the combination of BCD and PLP could significantly affect the content of 10 components in BCD (including 5 prototype components were significantly decreased and 5 metabolites were significantly increased) and 8 components in PLP (including 4 prototype components and 3 metabolites were significantly increased, 1 metabolite was significantly decreased), which indicated that the combination could enhance BCD prototype components' metabolism and the absorption of the PLP prototype components. Besides, metabolomics results indicated that the BCD-PLP herb pair group significantly reversed more metabolites (8) than BCD and PLP single herb group (5 & 4) and has a stronger regulatory effect on metabolite disorders caused by CUMS. Furthermore, the correlation analysis results suggested that saikogenin F and saikogenin G were significantly positively correlated with the endogenous metabolite itaconate, an endogenous anti-inflammatory metabolite; and benzoic acid was significantly positively correlated with D-serine, an endogenous metabolite with an antidepressant effect. Finally, the molecular docking results further confirmed that the combination of BCD and PLP could affect the activities of cis-aconitic acid decarboxylase and D-amino acid oxidase by increasing the in vivo concentration of saikogenin F and benzoic acid, which further enhances its anti-inflammatory activity and anti-depressive effect.
In this study, an effective strategy has been developed to study the combination mechanisms of BCD and PLP by integrating serum pharmacochemistry analysis, multivariate statistical analysis, metabolomics technology, and molecular docking technology. Based on this strategy, the present study indicated that the combination of BCD and PLP could affect the activities of cis-aconitic acid decarboxylase and D-amino acid oxidase by increasing the concentration of saikogenin F and benzoic acid in vivo, which further enhances its anti-depressive effect. In short, this strategy will provide a reliable method for elucidating the herb-herb compatibility mechanism of TCM.
柴胡-白芍（BCD-PLP）是中药方剂中常用的一对临床草药，常用于治疗抑郁症。然而，其与抗抑郁作用的组合机制仍不清楚。
本研究通过整合血清药化学分析、代谢组学技术和分子对接技术，开发了一种研究柴胡（BCD）和白芍（PLP）组合机制的有效策略。
首先，采用慢性不可预测轻度应激（CUMS）程序复制抑郁模型大鼠，通过背景扣除和多变量统计分析技术，分析 BCD 和 PLP 联合前后体内化学成分的差异。然后，采用 UPLC/HRMS 基于血清代谢组学的方法，分析 BCD 和 PLP 联合前后对代谢物调节的协同作用。进一步，通过相关性分析，研究 BCD 和 PLP 联合前后差异外源性化学物质与差异内源性代谢物之间的相关性，从代谢组学和血清药化学相结合的整体角度剖析组合机制。最后，应用分子对接技术研究差异化学物质与关键代谢酶之间的关系，验证差异外源性化学物质对差异内源性代谢物的调节作用。
血清药化学分析结果表明，BCD 和 PLP 的联合应用可显著影响 BCD（包括 5 个原型成分显著降低，5 个代谢产物显著增加）和 PLP（包括 4 个原型成分和 3 个代谢产物显著增加，1 个代谢产物显著降低）中的 10 个成分的含量，表明联合应用可增强 BCD 原型成分的代谢和 PLP 原型成分的吸收。此外，代谢组学结果表明，与 BCD 和 PLP 单药组（5 和 4）相比，BCD-PLP 药对组能更显著地逆转更多的代谢物（8 个），对 CUMS 引起的代谢物紊乱具有更强的调节作用。进一步的相关性分析结果表明，柴胡皂苷 F 和柴胡皂苷 G 与内源性抗炎代谢物衣康酸呈显著正相关；而苯甲酸与具有抗抑郁作用的内源性代谢物 D-丝氨酸呈显著正相关。最后，分子对接结果进一步证实，BCD 和 PLP 的联合应用可以通过增加体内 saikogenin F 和苯甲酸的浓度，影响顺乌头酸脱羧酶和 D-氨基酸氧化酶的活性，从而增强其抗炎和抗抑郁作用。
本研究通过整合血清药化学分析、多变量统计分析、代谢组学技术和分子对接技术，开发了一种研究 BCD 和 PLP 组合机制的有效策略。基于该策略，本研究表明，BCD 和 PLP 的联合应用可通过增加体内 saikogenin F 和苯甲酸的浓度，影响顺乌头酸脱羧酶和 D-氨基酸氧化酶的活性，从而增强其抗抑郁作用。总之，该策略将为阐明中药的药对配伍机制提供一种可靠的方法。
