Therapeutic effects of Euphorbia Pekinensis and Glycyrrhiza glabra on Hepatocellular Carcinoma Ascites Partially Via Regulating the Frk-Arhgdib-Inpp5d-Avpr2-Aqp4 Signal Axis

Yanqiong Zhang,Ya Lin,Qiuyan Guo,Weiwei Tao,Yuting Li,Chen Yan,Jingao Duan,Na Lin,Xia Mao,Yuping Tang

doi:10.1038/srep41925

Abstract

To clarify unknown rationalities of herbaceous compatibility of Euphorbia Pekinensis (DJ) and Glycyrrhiza glabra (GC) acting on hepatocellular carcinoma (HCC) ascites, peritoneum transcriptomics profiling of 15 subjects, including normal control (Con), HCC ascites mouse model (Mod), DJ-alone, DJ/GC-synergy and DJ/GC-antagonism treatment groups were performed on OneArray platform, followed by differentially expressed genes (DEGs) screening. DEGs between Mod and Con groups were considered as HCC ascites-related genes, and those among different drug treatment and Mod groups were identified as DJ/GC-combination-related genes. Then, an interaction network of HCC ascites-related gene-DJ/GC combination-related gene-known therapeutic target gene for ascites was constructed. Based on nodes’ degree, closeness, betweenness and k-coreness, the Frk-Arhgdib-Inpp5d-Avpr2-Aqp4 axis with highly network topological importance was demonstrated to be a candidate target of DJ/GC combination acting on HCC ascites. Importantly, both qPCR and western blot analyses verified this regulatory effects based on HCC ascites mice in vivo and M-1 collecting duct cells in vitro. Collectively, different combination designs of DJ and GC may lead to synergistic or antagonistic effects on HCC ascites partially via regulating the Frk-Arhgdib-Inpp5d-Avpr2-Aqp4 axis, implying that global gene expression profiling combined with network analysis can offer an effective way to understand pharmacological mechanisms of traditional Chinese medicine prescriptions.

Highlights

Due to their short efficacy, limited response rates, as well as inevitable adverse effects such as intestinal obstruction, sepsis and infection[6]
We aimed to clarify the rationalities of the herbaceous compatibility of DJ and GC acting on hepatocellular carcinoma (HCC) ascites, which have not been fully elucidated
After data processing and differentially expressed genes (DEGs) screening, 2252 annotated genes were differentially expressed (>2-fold, P < 0.05), including 1418 upregulated and 834 downregulated genes in peritoneum tissues of HCC ascites mice compared to the normal mice (Supplementary Table S1)

Summary

Introduction

Due to their short efficacy, limited response rates, as well as inevitable adverse effects such as intestinal obstruction, sepsis and infection[6]. Extracts of DJ have been revealed to exhibit numerous bioactivities including anti-virus and anti-inflammatory effects[10] Another Chinese herb, Glycyrrhiza glabra (Licorice, Gancao, GC), the root of Glycyrrhiza uralensis Fisch. The mechanisms of action and scientific evidence of several herb pairs and herbal formulae have been explored based on TCM theory and practice, the available databases and computational tools in TCM network pharmacology[18,19,20,21,22,23,24,25]. We have integrated drug target prediction, network analysis and experimental validation to find that another herbal pair of Euphorbia kansui (GS) and GC may exert synergistic or antagonistic effects on HCC ascites under different combination designs, partially through regulating the Adrb1-Pik3cg-Avpr[2] signal axis[21], highlighting the efficiency of network-based approaches to the pharmacological research of herb pairs. After interaction network of DEGs was constructed by STRING, we predicted the candidate targets implicated into the combinatory effects of the herbal pair DJ and GC by calculating network topological features and performing functional enrichment analysis, followed by a series of in vivo and in vitro experimental validations (Fig. 1)

Objectives

Methods

Results