The Active Components of Fuzheng Huayu Formula and Their Potential Mechanism of Action in Inhibiting the Hepatic Stellate Cells Viability - A Network Pharmacology and Transcriptomics Approach.

Si Chen,Xiaobo Wang,Xinrui Xing,Ling Li,Zhenyu Zhu,Langdong Chen,Yan Cao

doi:10.3389/fphar.2018.00525

Abstract

Purpose: This study aimed to identify the active components of Fuzheng Huayu (FZHY) formula and the mechanism by which they inhibit the viability of hepatic stellate cells (HSCs) by a combination of network pharmacology and transcriptomics.Methods: The active components of FZHY formula were screened out by text mining. Similarity match and molecular docking were used to predict the target proteins of these compounds. We then searched the STRING database to analyze the key enriched processes, pathways and related diseases of these target proteins. The relevant networks were constructed by Cytoscape. A network analysis method was established by integrating data from above network pharmacology with known transcriptomics analysis of quiescent HSCs-activated HSCs to identify the most possible targets of the active components in FZHY formula. A cell-based assay (LX-2 and T6 cells) and surface plasmon resonance (SPR) analysis were used to validate the most possible active component-target protein interactions (CTPIs).Results: 40 active ingredients in FZHY formula and their 79 potential target proteins were identified by network pharmacology approach. Further network analysis reduced the 79 potential target proteins to 31, which were considered more likely to be the target proteins of the active components in FZHY formula. In addition, further enrichment analysis of 31 target proteins indicated that the HIF-1, PI3K-Akt, FoxO, and chemokine signaling pathways may be the primary pathways regulated by FZHY formula in inhibiting the HSCs viability for the treatment of liver fibrosis. Of the 31 target proteins, peroxisome proliferator activator receptor gamma (PPARG) was selected for validation by experiments at the cellular and molecular level. The results demonstrated that schisandrin B, salvianolic acid A and kaempferol could directly bind to PPARG, decreasing the viability of HSCs (T6 cells and LX-2 cells) and exerting anti-fibrosis effects.Conclusion: The active ingredients of FZHY formula were successfully identified and the mechanisms by which they inhibit HSC viability determined, using network pharmacology and transcriptomics. This work is expected to benefit the clinical application of this formula.

Highlights

Liver fibrosis is a protective mechanism of organ integrity with the accumulation of extracellular matrix and collagen proteins, which occurs as a result of large-scale apoptosis and necrosis (Fagone et al, 2016)
As a combination of approaches is thought most likely to bear fruit, we hypothesized that the combination of network pharmacology and transcriptomics could increase the accuracy of target identification of network pharmacology
To improve the accuracy of target identification, we combined above network pharmacology results with known transcriptomics results, which reduced the number of possible active components of FZHY formula from 40 to 28, and the number of possible target proteins from 79 to 31

Summary

Introduction

Liver fibrosis is a protective mechanism of organ integrity with the accumulation of extracellular matrix and collagen proteins, which occurs as a result of large-scale apoptosis and necrosis (Fagone et al, 2016). It is mainly associated with chronic viral hepatitis type B or viral hepatitis type C infection, biliary diseases, alcoholic steatohepatitis, and nonalcoholic steatohepatitis (Friedman, 2008). Since the activation of hepatic stellate cells (HSCs) is the primary process of hepatic fibrosis (Schuppan and Kim, 2013), inhibiting the activation of HSCs should be an effective treatment (Liu et al, 2015). Safer and more effective treatment strategies for hepatic fibrosis are urgently needed

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