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

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.

Integrating Pharmacokinetics Study, Network Analysis, and Experimental Validation to Uncover the Mechanism of Qiliqiangxin Capsule Against Chronic Heart Failure

“…Recently, integrated strategies based on bioinformatics, system biology, and high-throughput analytical techniques have emerged as a holistic and efficient tool to solve this issue (Ning et al, 2018; Xing et al, 2018; Huang et al, 2019). Network pharmacology gives us helpful tools to screen potential bioactive ingredients and understand the molecular mechanisms underlying the therapeutic effects by constructing component–target and target–disease networks (Li and Zhang, 2013; Yuan et al, 2017).…”

Histological Outcome of Fuzheng Huayu plus Entecavir Combination Therapy in Chronic Hepatitis B Patients with Significant Liver Fibrosis

“…The content of major ingredients was measured on-site during the manufacturing processes, and as a quality inspection standard; as such, the procedures can be fully reproducible. 7 The fact that the active components of FZHY formula and their potential mechanism in inhibiting hepatic stellate cells' viability have been identified using network pharmacology and transcriptomics 24 has provided the evidence underlying its known antifibrotic action.…”

Active Compounds Derived from Fuzheng Huayu Formula Protect Hepatic Parenchymal Cells from Apoptosis Based on Network Pharmacology and Transcriptomic Analysis

“…Advanced liver cirrhosis is still incurable and can lead to liver failure and death [20]. Although the molecular mechanisms of FZHY in HPC protection and its antifibrotic effects are not fully clear, many reports have shown its antifibrotic effects without adverse effects [9,21,22]. Thus, evaluating the antifibrotic and HPC-protecting effects of FZHY and clarifying its mechanisms of action is important.…”

“…Major compounds and targets were selected using topological parameters and a structure herb-compound-target-signaling pathway network of FZHY, which resulted in the selection of 6 compounds, 39 potential targets and 12 pathways. Previous studies have reported that the selected major compounds, such as kaempferol and tanshinone IIA, inhibit the proliferation of HSCs [21], luteolin has hepatoprotective activity through the regulation of SREBP-1c phosphorylation and AMP-activated protein kinase [27], beta-sitosterol prevents hepatotoxicity [28], and hederagenin can attenuate non-alcoholic hepatic steatosis [29].…”