Network Pharmacology and Experimental Evidence: PI3K/AKT Signaling Pathway is Involved in the Antidepressive Roles of Chaihu Shugan San

Abstract

Objective

Chaihu Shugan San (CSS) is a common antidepressant prescription in traditional Chinese medicines. However, its active ingredients and mechanisms are unknown. The aim of this study was to explore the potential active ingredients and pharmacological mechanisms of CSS for the treatment of major depressive disorder (MDD).

Methods

Active compounds in CSS were screened using the Traditional Chinese Medicine Systems Pharmacology database. Compound-related targets were retrieved using the SwissTargetPrediction database. MDD-related targets were determined using DisGeNET, Therapeutic Target Database and DrugBank databases. The common targets of active compounds in CSS and MDD were retained to construct a compound-MDD target network. Then, functional enrichment analysis and protein–protein interaction analysis were performed to identify hub targets and explore the underlying molecular mechanisms. Finally, hub-targeted genes and pathways were validated by Western blotting and immunofluorescence using chronic unpredictable mild stress (CUMS) mice with or without CSS treatment. The affinities between the active compounds in CSS and hub-targeted genes were evaluated by molecular docking.

Results

Network pharmacology analysis revealed 24 potential targets for treatment of MDD by CSS. Functional enrichment analysis showed that PI3K/AKT signaling pathway was likely to be evidently affected by CSS in the treatment of MDD. In vivo experiments showed that CSS could improve depressive-like behaviors and promote neurogenesis in CUMS mice. Furthermore, CSS could increase phosphorylated (p) PI3K/PI3K and pAKT/AKT levels and decrease the pGSK3β/GSK3β level in the hippocampus of CUMS mice. The active compounds mainly included quercetin and luteolin, which showed good docking scores targeting the PI3K protein.

Conclusion

This network pharmacological and experimental study highlights that the PI3K/AKT pathway is the potential mechanism by which CSS is involved in MDD treatment. Quercetin, luteolin, and kaempferol are probable active compounds in CSS, and these results might provide valuable guidance for further studies of MDD treatment.

Graphical Abstract

Keywords:

• Chaihu Shugan San
• major depressive disorder
• network pharmacology
• PI3K/AKT signaling pathway

Acknowledgments

This study was supported by the Beijing Natural Science Foundation (Grant No. 7204250), National Natural Science Foundation of China (Grant No. 82004109), Beijing Administration of Traditional Chinese Medicine (Grant No. JJ2018-51). We are grateful to Prof. Feng Qiu of Capital Medical University for his supports to the work of LC/MS/MS analysis. We also thank Lisa Kreiner, PhD, from Liwen Bianji, (Edanz), for editing the English text of a draft of this manuscript.

Abbreviations

ADME, absorption, distribution, metabolism, and excretion; AKT, protein kinase B; BP, biological process; BrdU, Bromodeoxyuridine; CC, cellular component; CSS, Chaihu Shugan San; CUMS, chronic unpredictable mild stress; DAVID, Database for Annotation, Visualization and Integrated Discovery; DL, drug likeness; FST, Forced swimming test; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; LC-MS/MS, Liquid chromatography-tandem mass spectrometry; MDD, major depressive disorder; MF, molecular function; OB, oral bioavailability; p, phosphorylated; PBS, phosphate-buffered saline; PI3K, phosphoinositide 3-kinase; PPI, Protein-protein interactions; SPT, Sucrose preference test; STRING, Search Tool for the Retrieval of Interacting Genes/Proteins; TCM, traditional Chinese medicines; TCMSP, Traditional Chinese Medicine System Pharmacology Database; TTD, Therapeutic Target Database; TST, tail suspension test.

Data Sharing Statement

The data that support the finding of the present study are shown in Supplementary Materials.

Ethics Approval and Informed Consent

All experiments were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Experimental Animal Ethics Committee of Beijing Friendship Hospital (Approval No.19-1006).

Author Contributions

All authors contributed to data analysis, draft, or revision, gave final approval of the version to be published, have agreed on the journal to which the article has been submitted, and agreed to be accountable for all aspects of the work.

Disclosure

The authors declare that the research has no competing interests.