ABSTRACT
ObjectiveTo investigate the mechanism of Xiaozheng Huoluo prescription (XZHLF) in the prevention and treatment of liver fibrosis based on network pharmacology. MethodsTCMSP database, Chemistry Database, ETCM database, chemical source network database, ChemSrc database, and PubChem database were searched and a literature review was performed to collect the chemical components of each traditional Chinese medicine of XZHLF, Swiss ADME database was used to screen out the active components of each traditional Chinese medicine of XZHLF, and Swiss Target Prediction database was used to predict the targets of these active components; GeneCards and OMIN databases were used to collect the disease targets of liver fibrosis, and Venn diagrams were used to obtain the potential targets of XZHLF in the prevention and treatment of liver fibrosis. Cytoscape 3.7.1 software was used to establish a “drug-active component” network for XZHLF and an “active component-potential target” network for XZHLF in the prevention and treatment of liver fibrosis. Metascape database was used to perform GO and KEGG enrichment analysis of potential targets, and bubble charts were plotted for the top 20 pathways with the highest number of enriched genes. The MAPK signaling pathway among the top 20 KEGG pathways was analyzed to plot an “active component-potential target-pathway” network. Healthy Sprague-Dawley rats were randomly divided into blank control group (K group), model group (M group), colchicine-positive control group (Y group), high-dose XZHLF group (G group), middle-dose XZHLF group (Z Group), and low-dose XZHLF group (D group), and a rat model of liver fibrosis was established by CCl4; the drug was administered simultaneously for 8 weeks. Western blot was used to measure the protein expression of ERK5, p-ERK5, MEK5, and MEKK3 in liver tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsA total of 110 active components were screened out from XZHLF and they acted on 923 targets, which were mapped to 6823 disease targets of liver fibrosis to obtain 647 potential targets. XZHLF might act on multiple protein targets such as EGFR, AKT1, IKBKB, MAPK8, and PDGFRB through the pathways including the MAPK signaling pathway and the biological processes including the regulation of MAPK cascade, thereby playing a role in the prevention and treatment of liver fibrosis. The M group had significant increases in the protein levels of ERK5, p-ERK5, MEK5, and MEKK3 in liver tissue compared with the K, Y, G, Z, and D groups (all P ＜0.05), and the K group had significant reductions in the protein levels of ERK5, p-ERK5, MEK5, and MEKK3 in liver tissue compared with the Y, G, Z, and D groups (all P ＜0.05). ConclusionBased on the method of network pharmacology, it is predicted that XZHLF may prevent and treat liver fibrosis via the MAPK signaling pathway, and it is verified through experiments that XZHLF prevents and treats liver fibrosis via the ERK5 pathway in the MAPK signaling pathway family. The high-dose XZHLF group shows the most obvious anti-liver fibrosis effect.
ABSTRACT
Objective:To study the inhibitory effect of salidroside on the proliferation of fibroblast-like synoviocytes with rheumatoid arthritis in human(HFLS-RA) induced by tomor necrossi factor-α(TNF-α),and to clarify the molecular mechanism of its control effect on rheumatoid arthritis(RA).Methods:The HFLS-RA were cultured in vitro,then treated with TNF-α and different concentrations of salidroside.The cells were divided into normal control group(0 μg·L-1TNF-α),model control group(10.0 μg·L-1TNF-α)and 12.5,25.0,50.0,and 100.0 μmol·L-1 salidroside groups(10.0 μg·L-1TNF-α+salidroside).The proliferation activity was detected by MTT mehthod;the expression levels of β-catenin,matrix metalloproteinase-7(MMP-7),and Cyclin-D1 in supernatant of the cells were detected by ELISA method;the expression level of β-catenin protein in cells was detected by Western blotting method.Results:Compared with normal control group,the proliferation activity of the HFLS-RA in model control group was significantly increased (P0.05),and the proliferation activities of the HFLS-RA in 50.0 and 100.0 μmol·L-1 salidroside groups were significantly decreased (P0.05);the expression levels of β-catenin,MMP-7,and Cyclin-D1 in the supernatant of the cells in 50.0 and 100.0 μmol·L-1 salidroside groups were decreased(P0.05);the expression levels of β-catenin protein in the cells in 50.0 and 100.0 μmol·L-1 salidroside groups were lower than that in model control group(P<0.05).Conclusion:Salidroside could inhibit the proliferation of HFLS-RA,and its control effect might be related to the regulation of Wnt/β-catenin single pathway.