Chen Jiajun, Li Tian, Qin Xuemei, Du Guanhua, Zhou Yuzhi
Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, China.
The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, China.
Front Pharmacol. 2022 Jun 30;13:900459. doi: 10.3389/fphar.2022.900459. eCollection 2022.
DC (Chaihu) Pall (Baishao) is among the most accepted herb pairs in many classic antidepressant prescriptions. Our previous study has shown that the Chaihu-Baishao herb pair (CBHP) had a better antidepressant effect than Chaihu or Baishao. Nevertheless, the synergistic antidepressant mechanism of this herb pair was not clearly understood. This study aimed to investigate the compatibility mechanism of Chaihu and Baishao for treating depression through a strategy of non-targeted metabolomics combined with targeted quantitative analysis and molecular biology techniques. First, the compatibility effects of CBHP were assessed by the chronic unpredictable mild stress (CUMS) rat model. Next, cortex metabolomics based on ultra-high-performance liquid chromatography combined with quadrupole orbitrap mass spectrometry (UPLC-Q-Orbitrap/MS) was used to discover the metabolic pathway that was synergistically regulated by CBHP. Based on the results of metabolomics analysis, metabolites were quantitatively validated by UPLC-MS/MS combined with the MRM mode in the crucial metabolic pathway. In addition, the signaling pathway associated with this metabolic pathway was detected by molecular biology techniques to further identify the biological meaning of the crucial metabolite on the synergistic antidepressant effect of CBHP. The antidepressant effect of CBHP was significantly better than that of Chaihu or Baishao single administrated in the behavioral test. According to cortex metabolomics, a total of 21 differential metabolites were screened out, and purine metabolism was selected as the crucial metabolic pathway by the enrichment analysis of differential metabolites. Subsequently, purine metabolism was confirmed as disorder in the CUMS group by targeted quantitative analysis, CBHP regulated more purine metabolites (six) than individual administration (two and two). The results showed that purine metabolism was modulated by CBHP through synergistically decreasing xanthine levels and inhibiting the conversion of xanthine dehydrogenase (XDH) to xanthine oxidase (XOD). Finally, the synergistic regulation effect of CBHP on xanthine synthesis was found to be related to inhibition of malondialdehyde (MDA) production, Nod-like receptor protein 3 (NLRP3) inflammasome expression, and interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)- secretion. The present study demonstrated that the regulation of purine metabolism, the suppression of oxidative stress, and inflammatory responses in the cortex were involved in the synergistic antidepressant effect of CBHP.
柴胡-白芍药对是众多经典抗抑郁方剂中最常用的药对之一。我们之前的研究表明，柴胡-白芍药对（CBHP）的抗抑郁效果优于柴胡或白芍单味药。然而，该药对协同抗抑郁的机制尚不清楚。本研究旨在通过非靶向代谢组学结合靶向定量分析及分子生物学技术，探讨柴胡与白芍配伍治疗抑郁症的机制。首先，采用慢性不可预测温和应激（CUMS）大鼠模型评估CBHP的配伍效应。其次，基于超高效液相色谱结合四极杆轨道阱质谱（UPLC-Q-Orbitrap/MS）的皮质代谢组学技术，发现CBHP协同调控的代谢通路。基于代谢组学分析结果，在关键代谢通路中，采用UPLC-MS/MS结合MRM模式对代谢产物进行定量验证。此外，运用分子生物学技术检测与该代谢通路相关的信号通路，进一步明确关键代谢产物对CBHP协同抗抑郁作用的生物学意义。行为学实验结果显示，CBHP的抗抑郁效果显著优于柴胡或白芍单味药。通过皮质代谢组学分析，共筛选出21种差异代谢产物，经差异代谢产物富集分析，选择嘌呤代谢作为关键代谢通路。随后，通过靶向定量分析证实CUMS组嘌呤代谢紊乱，CBHP调控的嘌呤代谢产物（6种）多于单味药组（2种和2种）。结果表明，CBHP通过协同降低黄嘌呤水平及抑制黄嘌呤脱氢酶（XDH）向黄嘌呤氧化酶（XOD）的转化来调节嘌呤代谢。最后，发现CBHP对黄嘌呤合成的协同调节作用与抑制丙二醛（MDA）生成、Nod样受体蛋白3（NLRP3）炎性小体表达及白细胞介素（IL）-1、IL-6和肿瘤坏死因子（TNF）分泌有关。本研究表明，CBHP协同抗抑郁作用涉及对皮质嘌呤代谢的调节、氧化应激及炎症反应的抑制。
