An Integrative Pharmacology-Based Strategy to Uncover the Mechanism of Xiong-Pi-Fang in Treating Coronary Heart Disease with Depression.

Lihong Zhang,Shaoqiang Zhang,Wuxun Du,Mingdan Zhu,Yu Zhang,Fangjun Deng,Zidong Cong,Xuefeng Xiao,Limin Pei,Jinhong Chen

doi:10.3389/fphar.2021.590602

Abstract

Objectives: This study aimed to explore the mechanism of Xiong-Pi-Fang (XPF) in the treatment of coronary heart disease (CHD) with depression by an integrative strategy combining serum pharmacochemistry, network pharmacology analysis, and experimental validation. Methods: An ultrahigh performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) method was constructed to identify compounds in rat serum after oral administration of XPF, and a component-target network was established using Cytoscape, between the targets of XPF ingredients and CHD with depression. Furthermore, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed to deduce the mechanism of XPF in treating CHD with depression. Finally, in a chronic unpredictable mild stress (CUMS)-and isoproterenol (ISO)-induced rat model, TUNEL was used to detect the apoptosis index of the myocardium and hippocampus, ELISA and western blot were used to detect the predicted hub targets, namely AngII, 5-HT, cAMP, PKA, CREB, BDNF, Bcl-2, Bax, Cyt-c, and caspase-3. Results: We identified 51 compounds in rat serum after oral administration of XPF, which mainly included phenolic acids, saponins, and flavonoids. Network pharmacology analysis revealed that XPF may regulate targets, such as ACE2, HTR1A, HTR2A, AKT1, PKIA, CREB1, BDNF, BCL2, BAX, CASP3, cAMP signaling pathway, and cell apoptosis process in the treatment of CHD with depression. ELISA analysis showed that XPF decreased Ang-II content in the circulation and central nervous system, inhibited 5-HT levels in peripheral circulation, and increased 5-HT content in the central nervous system and cAMP content in the myocardia and hippocampus. Meanwhile, western blot analysis indicated that XPF could upregulate the expression levels of PKA, CREB, and BDNF both in the myocardia and hippocampus. TUNEL staining indicated that the apoptosis index of myocardial and hippocampal cells increased in CUMS-and ISO-induced CHD in rats under depression, and XPF could increase the expression of Bcl-2, inhibit the expression of Bax, Cyt-c, and caspase-3, and rectify the injury of the hippocampus and myocardium, which exerted antidepressant and antimyocardial ischemia effects. Conclusion: Our study proposed an integrated strategy, combining serum pharmacochemistry and network pharmacology to investigate the mechanisms of XPF in treating CHD with depression. The mechanism of XPF in treating CHD with depression may be related to the activation of the cAMP signaling pathway and the inhibition of the apoptosis.

Highlights

Coronary heart disease (CHD) is a chronic and complex disease that poses a serious threat to human health (Mozaffarian et al, 2016; Blais et al, 2020)
820 targets for the 51 (Nine of them had no therapeutic targets) components were explored by using TCMSP, Swiss Target Prediction, Drugbank, and Swiss Target Prediction as shown in Supplementary Table S1, and 1817 candidate targets of CHD and depression were obtained from Drugbank, Online Mendelian Inheritance in Man (OMIM), and DisGeNet databases as shown in Supplementary Table S2
8 components, with higher degree centrality were determined to be the active ingredients of XPF, and 10 hub targets, including (ACE2, HTR1A, HTR2A, AKT1, PKIA, CREB1, BDNF, BCL2, BAX, CASP3, cAMP) were determined as hub targets of XPF in treating CHD with depression

Summary

Introduction

Coronary heart disease (CHD) is a chronic and complex disease that poses a serious threat to human health (Mozaffarian et al, 2016; Blais et al, 2020). Bupleurum scorzonerifolium Willd.) 15 g; Ligusticum wallichii (Conioselinum anthriscoides ’Chuanxiong’ and Ligusticum striatum DC.) 12 g; Rhizoma Cyperi (Cyperus rotundus L.) 12 g; Sanders (Santalum album L..) 9 g; Fructus aurantii (Citrus × aurantium L. and Citrus trifoliata L.) 9 g; Dried tangerine peel (Citrus × aurantium L.) 15 g; Pinellia ternata (Pinellia ternata (Thunb.) Makino) 15 g; Herba Menthae (Mentha haplocalyx Briq.) 10 g; Perilla Stem (Perilla frutescens (L.) Britton) 15 g; Poria Cocos (Wolfiporia extensa (Peck) Ginns) 15 g; Rhizoma Atractylodis Macrocephalae (Atractylodes Macrocephala Koidz) 15 g; Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch.) 9 g It is widely used in clinical practices, showing satisfactory therapeutic effects in relieving angina pectoris, chest pain, and depression symptoms (Hou et al, 2017). An integrated pharmacology-based strategy including computer high-throughput analytical techniques and biological experimental tools was established in this study, which provides a helpful method to reveal the potential bioactive ingredients and modern pharmacological mechanism of TCM (Park et al, 2018; Fang et al, 2019)

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Discussion

Conclusion