Integrated Pharmaco-Bioinformatics Approaches and Experimental Verification To Explore the Effect of Britanin on Nonalcoholic Fatty Liver Disease

Abstract

Nonalcoholic fatty liver disease (NAFLD) is a prevalent global liver disorder, posing substantial health risks. Britanin, a bioactive sesquiterpene lactone extracted from Inula japonica, has demonstrated antidiabetic, hypolipidemic, and hepatoprotective attributes. Nonetheless, the precise impact of Britanin on NAFLD and the intricate biological mechanisms underpinning this interaction remain unexplored. We integrated computer-aided methods to unearth shared biological targets and signaling pathways associated with both Britanin and NAFLD. A network was constructed by compiling putative targets associated with Britanin and NAFLD, followed by a stringent screening of key targets and mechanisms through protein-protein interaction analysis along with GO and KEGG pathway enrichment analyses. Molecular docking was integrated as an evaluation tool, culminating in the identification of HO-1 as the pivotal therapeutic target, showcasing a satisfactory binding affinity. The primary mechanism was ascribed to biological processes and pathways linked to oxidative stress, as evidenced by the outcomes of enrichment analyses. Of these, the AMPK/SREBP1c pathway assumed centrality in this mechanism. Furthermore, in vivo experiments substantiated that Britanin effectively curtailed NAFLD development by ameliorating liver injury, modulating hyperlipidemia and hepatic lipid accumulation, and alleviating oxidative stress and apoptosis. In summary, this study demonstrates the potential of Britanin as a promising therapeutic drug against NAFLD.

Figure 1

Target selection for NAFLD. (A) Distribution of standardized samples from the GSE63067 data set. (B) Heatmap of DEGs in GSE63067. (C) Volcano plot illustrating DEGs in GSE63067. (D) Gene set enrichment analysis (GSEA) of the upregulated gene set in GSE63067. (E) Gene set enrichment analysis (GSEA) of the downregulated gene set in GSE63067.

Figure 2

Core gene network. (A) Venn diagram depicting potential core targets of Britanin in NAFLD treatment. (B) Distribution of core targets based on degree centrality. (C) Distribution of core targets based on closeness centrality. (D) Distribution of core targets based on betweenness centrality.

Figure 3

Enrichment analysis. (A) Bar chart depicting GO functional analysis results. (B) Chord diagram illustrating enriched GO terms. (C) Chord diagram illustrating enriched KEGG pathways. (D) Schematic representation of the endocrine resistance signaling pathway.

Figure 4

Relative gene expression and ROC model construction. (A) Relative expression levels of core genes in GSE63067 data set. (B) ROC model for core DEGs in GSE63067 samples.

Figure 5

Molecular docking models. (A1) Macroscopic 3D molecular docking model of LRRK2-Britanin. (A2) Microscopic 3D molecular docking model of LRRK2-Britanin. (A3) 2D molecular docking model of LRRK2-Britanin. (B1) Macroscopic 3D molecular docking model of CXCR2-Britanin. (B2) Microscopic 3D molecular docking model of CXCR2-Britanin. (B3) 2D molecular docking model of CXCR2-Britanin. (C1) Macroscopic 3D molecular docking model of DPP4-Britanin. (C2) Microscopic 3D molecular docking model of DPP4-Britanin. (C3) 2D molecular docking model of DPP4-Britanin. (D1) Macroscopic 3D molecular docking model of HMOX1-Britanin. (D2) Microscopic 3D molecular docking model of HMOX1-Britanin. (D3) 2D molecular docking model of HMOX1-Britanin. (E1) Macroscopic 3D molecular docking model of MAPK8-Britanin. (E2) Microscopic 3D molecular docking model of MAPK8-Britanin. (E3) 2D molecular docking model of MAPK8-Britanin. (F1) Macroscopic 3D molecular docking model of MAPK14-Britanin. (F2) Microscopic 3D molecular docking model of MAPK14-Britanin. (F3) 2D molecular docking model of MAPK14-Britanin.

Figure 6

Effects of Britanin on hepatic injury in a high-fat diet-induced NAFLD mouse model. (A) Body weight (n = 8). (B) Liver weight (n = 8). (C) Liver index (n = 8). (D) Serum AST activity (n = 8). (E) Serum ALT activity (n = 8). (F) Liver H&E staining. Typical images were chosen from each experimental group. Data are shown as mean ± SD. *P < 0.05 vs HFD.

Figure 7

Effects of Britanin on lipid accumulation in a high-fat diet-induced NAFLD mouse model. (A) Serum TC levels (n = 8). (B) Serum TG levels (n = 8). (C) Serum LDL-c levels (n = 8). (D) Serum HDL-c levels (n = 8). (E) Liver Oil Red O staining. Typical images were chosen from each experimental group. (F) Hepatic protein expression of p-AMPK and AMPK (n = 8). (G) Hepatic protein expression of SREBP-1c (n = 8). Data are shown as mean ± SD. *P < 0.05 vs HFD.

Figure 8

Effects of Britanin on oxidative stress and apoptosis in a high-fat diet-induced NAFLD mouse model. (A) Serum MDA activity (n = 8). (B) Serum SOD activity (n = 8). (C) Hepatic protein expression of HO-1 (n = 8). (D) Liver immunohistochemistry staining. (E) Hepatic protein expression of Caspase3, Bax, and Bcl2. Data are shown as mean ± SD. *P < 0.05 vs HFD.