Network analysis and experimental verification of tanshinone ⅡA in treatment of autoimmune hepatitis

Abstract

Objective To analyze the relationship between potential targets and signaling pathways of tanshinone Ⅱ A (Tan Ⅱ A) in the treatment of autoimmune hepatitis (AIH) based on network pharmacological analysis, and verify the mechanism of Tan Ⅱ A on AIH by animal experiments. Methods The relevant gene targets of Tan ⅡA and AIH were searched and screened through multiple databases, and the intersection genes between the drug and the disease were selected. Protein-protein interaction (PPI) network was subsequently constructed with the help of String platform, and the intersection genes were enriched and analyzed by Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Then, the signaling pathway with the highest number of enriched genes was verified through animal experiments. Eighteen C57BL/6 mice were equally and randomly divided into normal, model and Tan ⅡA treatment groups. The contents of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected, and the protein and mRNA expression levels of PI3K, AKT and NF-κB were measured by Western blotting and qRT-PCR respectively. Results There were 208 effective targets screened out from Tan ⅡA, and 7 128 target genes from AIH. A total of 174 intersection gene targets were obtained. KEGG pathway analysis indicated that multiple signaling pathways were involved in the pathogenesis of AIH, among which PI3K-AKT signaling pathway played an important role in Tan ⅡA treatment of AIH. In animal experiments, the model group had significantly higher contents of ALT and AST (P < 0.05), reduced mRNA and protein levels of PI3K and AKT (P < 0.05) and elevated mRNA and protein levels of NF-κB (P < 0.05) than the control group. While, Tan ⅡA treatment resulted in decreased contents of ALT and AST (P < 0.05), increased PI3K and AKT levels (P < 0.05) and decreased NF-κB level (P < 0.05) when compared with the model mice. Conclusion Tan ⅡA contains many effective genes that may directly target AIH-related signaling pathways, among which Tan ⅡA may ameliorate liver inflammation in AIH mice by regulating PI3K-AKT signaling pathway.

Keywords

• network pharmacology
• tanshinone ⅱa
• autoimmune hepatitis
• pi3k-akt signaling pathway