Investigating the Pharmacological Mechanisms of SheXiang XinTongNing Against Coronary Heart Disease Based on Network Pharmacology and Experimental Evaluation

Li-Ying Jia,Xin-Yi Lan,Gui-Yun Cao,Jia Li,Zhao-Qing Meng,Xin He,Lu Gan,Jin-Xin Li,Chun-Su Yuan,Chun-Feng Zhang,Chong-Zhi Wang

doi:10.3389/fphar.2021.698981

Abstract

SheXiang XinTongNing (XTN), which is composed of six traditional Chinese herbs, is a commercially available Chinese patent medicine that has been widely used as the treatment of coronary heart disease (CHD). Its mechanisms against coronary heart disease, however, remain largely unknown. This study aimed to investigate the pharmacological mechanisms of XTN against CHD via network pharmacology and experimental evaluation. In this study, GO enrichment and KEGG pathway enrichment were firstly performed for acquiring the potentially active constituents of XTN, the candidate targets related to coronary heart disease, the drug-components-targets network as well as the protein-protein interaction network and further predicting the mechanisms of XTN against coronary heart disease. Subsequently, a series of in vitro experiments, specifically MTT assay, flow cytometry and Real-time quantitative polymerase chain reaction analysis, and a succession of in vivo experiments, including Tunel staining and immunohistochemical staining were conducted for further verification. Results showed that Bcl-2, IGF1, CASP3 were the key candidate targets which significantly associated with multiple pathways, namely PI3K-Akt signaling pathway and MAPK signaling pathway. It indicated that the potential mechanism of XTN against CHD may be predominantly associated with cell apoptosis. The in vitro experimental results showed that XTN treatment remarkably decreased the apoptotic rate and Bax/Bcl-2 ratio of H9c2 cells. Histological results confirmed that XTN not only effectively alleviated oxidative damage caused by myocardial ischemia but inhibited cell apoptosis. Given the above, through the combined utilization of virtual screening and experimental verification, the findings suggest that XTN makes a significant contribution in protecting the heart from oxidative stress via regulating apoptosis pathways, which lays the foundations and offers an innovative idea for future research.

Highlights

Cardiovascular disease, as a leading cause of deaths worldwide, has led to 17.3 million deaths globally (Townsend et al, 2016)
We obtained 104 candidate targets connected with coronary heart disease (CHD) from Genecards platform
Our research demonstrated that XTN decreased the expression levels of Bax and caspase-3 but increased Bcl-2 expression level, suggesting XTN has the capacity of anti-apoptosis and the efficacy on anti-CHD

Summary

Introduction

Cardiovascular disease, as a leading cause of deaths worldwide, has led to 17.3 million deaths globally (Townsend et al, 2016). CHD exerted tremendous pressure to the society, and led to one third of all death cases in the adult population (Roger, 2007; Letnes et al, 2019). High cholesterol level, diabetes and obesity have been considered as major diagnostic criteria of CHD (Timmis et al, 2018). Women with an onset of type one diabetes before age 10 have a 60 times increased risk of CHD occurrence (Rawshani et al, 2018). Apart from those, it has been reported that HIV infection caused a substantially increased risk of CHD (Hsue and Waters, 2019). Regardless of the previously-mentioned treatments against CHD, the therapeutics of CHD still confront with many limitations, more precisely, the lack of comprehensive management, patients’ unwillingness in changing lifestyles and the side effects of medication especially Western medicine. (Mao et al, 2019)

Objectives

Methods

Results

Discussion

Conclusion