Abstract:

To explore the therapeutic effects and mechanism of water extract of Orychophragmus violaceus (L.) O.E.Schulz seeds (OVSWE) on the treatment of nonalcoholic steatohepatitis (NASH).The China National Knowledge Infrastructure (CNKI) was used to retrieve and obtain the water-soluble active ingredients of OVSWE.This study used diverse disease databases to obtain genes information related to NASH.And this study extracted the intersection targets of OVSWE and NASH,further visualized the network of "drugs-potential active ingredients-potential targets".This study imported intersection targets into the STRING database to construct a protein-protein interaction network;performed GO function and KEGG pathway enrichment analysis and visualization,and then used Autodock software to construct molecular docking models.Finally,the targets and pathways predicted by network pharmacology were experimentally validated through animal experiments.The analysis of network pharmacology showed that the core targets of OVSWE was Akt serine/threonine kinase 1(AKT1) and phosphoinositide-3-kinase regulatory subunit 1(PIK3R1),etc;the core components were schizophylline A,schizophyllin B and schizophyllin C,the treatment of OVSWE is closely related to insulin resistance (IR) and lipid metabolism signal pathway.Molecular docking results suggested that the core ingredient in OVSWE have strong binding ability to target AKT1.The NASH rat model was induced with high-fat and high-sugar diet combined with CCl4.Hematoxylin-eosin staining(HE) and Sirius red staining showed that OVSWE could alleviate liver lipid accumulation and damage.Biochemical tests showed that OVSWE could significantly reduce the content of triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in liver tissue (P< 0.05 or P< 0.01).Serum glucose (GLU),free fatty acid (FFA),fasting insulin content and IR index were also significantly decreased (P< 0.05 or P< 0.01).Not targeting lipid omics analysis showed that OVSWE could regulate a variety of lipid differentials.The enrichment of their pathways showed that the disorders of lipid metabolism mainly focused on the sphigolipid metabolism and glycerol phospholipid metabolism in NASH rats.Western blot showed that OVSWE could significantly increase insulin receptor(InsR) and P-Akt/AKT protein expression levels in liver tissue (P< 0.05 or P< 0.01).These findings suggested that the therapeutic mechanism of the OVSWE on NASH rats induced by the high-fat and high-sugar diet combined with CCl4 might be related to its activation of the InsR/PI3K/AKT signaling pathway,improvement of IR,regulation of lipid metabolism and reduction of inflammatory response.

Key words:

nonalcoholic steatohepatitis, Orychophragmus violaceus (L.) O.E.Schulz seeds, network pharmacology, Lipid metabolism, insulin resistance