Korean J Physiol Pharmacol.  2024 Sep;28(5):479-491. 10.4196/kjpp.2024.28.5.479.

Network pharmacology prediction to discover the potential pharmacological action mechanism of Rhizoma Dioscoreae for liver regeneration

• Liu W
• Wang W
• Tian C
• Sun MZ
• Liu S
• Liu Q

• KMID: 2559140
• DOI: http://doi.org/10.4196/kjpp.2024.28.5.479

Abstract

Improving liver regeneration (LR) remains a medical issue, and there is currently a lack of safe and effective drugs for LR. Rhizoma Dioscoreae (SanYak, SY) is a traditional Chinese medicine. However, the underlying action mechanism of SY treatment for LR is yet to be fully elucidated. To explore the mechanism by which SY affects LR, we have conducted a series of methods for network pharmacological analysis, molecular docking, and in vivo experimental validation in mice. Overall, 9 compounds and 30 predicted target genes of SY were found to be associated with the therapeutic effects of LR. Compared with the model group, hematoxylin and eosin staining revealed that the mice with preoperative drug intervention possessed fewer postoperative hepatocyte bubbles and relatively regular morphology. Furthermore, the serum alanine transaminase and aspartate aminotransferase levels were reduced, immunohistochemistry revealed elevated proliferating cell nuclear antigen positivity rate, and Western blotting demonstrated that the phospho-protein kinase B (AKT)/AKT ratio was downregulated and that vascular endothelial growth factor A (VEGFA) expression levels were upregulated. This study explored dioscin, the main active ingredient of SY, and its potential therapeutic effects on LR. It repairs damaged liver following surgery and promotes liver cell proliferation. The action mechanism comprises reducing AKT phosphorylation levels and upregulating VEGFA expression levels. Thus, this study provides a new direction for further research on the mechanism of SY promoting LR.

Keyword

Diosgenin; Liver regeneration; Network pharmacology; Rhizoma Dioscoreae

Figure

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  Fig. 1 Research design flowchart. DIO, diosgenin; IR, ischemia-reperfusion; PHx, hepatectomy; AST, aspartate aminotransferase; AKT, protein kinase B; VEGFA, vascular endothelial growth factor A; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; LR, liver regeneration; SY, SanYak, Rhizoma Dioscoreae; PPI, protein–protein interaction; KEGG, Kyoto Encyclopedia of Genes and Genomes; IHC, immunohistochemistry. ****p < 0.0001.

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  Fig. 2 Venn diagram of Rhizoma Dioscoreae and disease corresponding to target at the intersection. Red represents the number of targets of the active components, and the blue represents the number of liver regeneration genes, which included 30 cross-targeted genes. LR, liver regeneration; SY, SanYak, Rhizoma Dioscoreae.

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  Fig. 3 Gene target network of the effective components of Rhizoma Dioscoreae in the “drug–component–target–disease” network. Green squares represent Rhizoma Dioscoreae, red hexagons represent active ingredients, yellow circles represent diseases, and blue diamonds represent target proteins. LR, liver regeneration; SY, SanYak, Rhizoma Dioscoreae.

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  Fig. 4 PPI network diagram of the target proteins. The size of the point represents the degree, color depth represents the compactness, and thickness of the line represents the intermediateness. Taking a degree of freedom of ≥18 as the line represents target proteins were AKT1, VEGFA, TP53, HIF1A, PTGS2, and ESR1. PPI, protein–protein interaction.

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  Fig. 5 GO and KEGG enrichment analyses of key target genes. (A) Top items of biological function are listed on the horizontal axis, including the top eight items of molecular function (MF, blue), top seven items of cell component (CC, orange), and top twenty items of biological process (BP, green). (B) Histogram of the top nine pathways based on KEGG enrichment analysis. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

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  Fig. 6 Compound–target–pathway network. The yellow circle represents the pathway, and the blue diamond represents the target point.

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  Fig. 7 Molecular docking of SY compounds and hub target proteins. (A) Binding mode of AKT and diosgenin. (B) Binding mode of AKT and diosgenin. SY, SanYak, Rhizoma Dioscoreae; AKT, protein kinase B.

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  Fig. 8 Liver tissue stained with H&E. (A) Sham group. (B) IR + 85%PHx 24 h group. (C) DIO IR + 85%PHx 24 h group (n = 4). Scale bar = 100 µm. IR, ischemia-reperfusion; PHx, hepatectomy; DIO, diosgenin.

• 

  Fig. 9 Determination of ALT and AST in serum (n = 4). (A) Significantly increased ALT levels in IR + 85%PHx 24 h group, ****p < 0.0001 (vs. Sham group); ALT levels were significantly reduced in DIO IR + 85%PHx 24 h group, ***p < 0.001 (vs. IR + 85%PHx 24 h group). (B) Significantly increased AST levels in IR + 85%PHx 24 h group, ****p < 0.0001 (vs. Sham group); AST levels were significantly reduced in the DIO IR + 85%PHx 24 h group, ****P < 0.0001 (vs. IR + 85%PHx 24 h group) (n = 4). Values are presented as mean ± SD. ALT, alanine transaminase; AST, aspartate aminotransferase; IR, ischemia-reperfusion; PHx, hepatectomy; DIO, diosgenin.

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  Fig. 10 Immunohistochemistry determines the positive rate of proliferating cell nuclear antigen (PCNA) (n = 4) (200×). (A) Sham group. (B) IR + 85%PHx 24 h group. (C) DIO IR + 85%PHx 24 h group. (D) Analysis of results: PCNA positivity increased in IR + 85%PHx 24 h group, ***p < 0.001 (vs. Sham group); PCNA positivity increased in the DIO IR + 85%PHx 24 h group, ****p < 0.0001 (vs. IR + 85%PHx 24 h group). Values are presented as mean ± SD. IR, ischemia-reperfusion; PHx, hepatectomy; DIO, diosgenin.

• 

  Fig. 11 Effect of DIO on the expression of AKT and VEGFA in IR + 85%PHx mice. (A) Western blot image showing the expression levels of p-AKT, AKT, and VEGFA in the liver tissue (n = 4). (B) Increased p-AKT/AKT expression levels in the IR + 85%PHx 24 h group, **p < 0.01 (vs. Sham group); decreased p-AKT/AKT expression levels in the DIO IR + 85%PHx 24 h group, *p < 0.05 (vs. IR + 85%PHx 24 h group). (C) No difference in VEGFA expression levels in the IR + 85%PHx 24 h group compared with the Sham group. Increased VEGFA expression levels in the DIO IR + 85%PHx 24 h group, *p < 0.05 (vs. IR + 85%PHx 24 h group). Values are presented as mean ± SD. AKT, protein kinase B; VEGFA, vascular endothelial growth factor A; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IR, ischemia-reperfusion; PHx, hepatectomy; DIO, diosgenin; ns, > 0.05.

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