Bupleurum marginatum Wall.ex DC in Liver Fibrosis: Pharmacological Evaluation, Differential Proteomics, and Network Pharmacology

Abstract

Liver fibrosis is a common pathological feature of many chronic liver diseases. Bupleurum marginatum Wall.ex DC (ZYCH) is a promising therapeutic for liver fibrosis. In this study, 25 compounds were isolated from ZYCH, and the effects of ZYCH on DMN-induced liver fibrosis in rats were evaluated. The optimal effect group (H-ZYCH group) was selected for further proteomic analysis, and 282 proteins were altered in comparison to the DMN model group (FC > 1.2 or < 0.83, p < 0.05). Based on GO annotation analysis, clusters of drug metabolism, oxidative stress, biomolecular synthesis and metabolism, positive regulation of cell growth, extracellular matrix deposition, and focal adhesion were significantly regulated. Then networks of the altered proteins and compounds was generated by Cytoscape. Importantly, triterpenoid saponins and lignans had possessed high libdock scores, numerous targets, important network positions, and strong inhibitory activity. These findings may suggest that triterpenoid saponins and lignans are important active compounds of ZYCH in liver fibrosis and targeted by proteins involved in liver fibrosis. The combination of network pharmacology with proteomic analysis may provide a forceful tool for exploring the effect mechanism of TCM and identifying bioactive ingredients and their targets.

Keywords: Bupleurum marginatum Wall.ex DC; bioactive ingredients; differential proteomics; liver fibrosis; network pharmacology.

Figure 1

Body weight increase, liver index and biochemical assays results after ZYCH treatment in DMN-induced liver fibrosis. (A,B) body weight increase and liver index; (C-G) serum ALT, AST, AKP, Alb, TBIL level; (H,I) the production of Hyp and TNF-α in liver tissues. #p < 0.05, ##p < 0.01 vs. control group, ^*p < 0.05, ^**p < 0.01 vs. model group.

Figure 2

Effects of ZYCH on hepatic histopathological changes of DMN-induced liver injury rats. Hepar sections by hematoxylin-eosin (HE) staining at a magniication of ×100. (A). control group, (B). model group, (C). SC group, (D). L-ZYCH group, (E). M-ZYCH group, (F). H-ZYCH group, and (G). HR-ZYCH group.

Figure 3

Effects of ZYCH on α-SMA IHC changes of DMN-induced liver injury rats. Original magnification: 100×. (A). control group, (B). model group, (C). SC group, (D). L-ZYCH group, (E). M-ZYCH group, (F). H-ZYCH group, and (G). HR-ZYCH group.

Figure 4

Volcano plot analysis of differentially expressed proteins between H-ZYCH and DMN-model groups. Volcano plot of ratios and p-value represented the protein abundance changes in the comparison between H-ZYCH and DMN-model groups. Proteins with p < 0.05 and above/below 1.2-fold changes are identified as proteins with significant changes. Down-regulated with p < 0.01 (dark blue dots), down-regulated with 0.01 ≤ p < 0.05 (light blue dots), up-regulated with p < 0.01 (red dots), and up-regulated with 0.01 ≤ p < 0.05 (yellow dots).

Figure 5

GO classification of up-regulated altered proteins and down-regulated altered proteins between H-ZYCH and DMN-model groups. (A). GO classification of up-regulated proteins; (B). GO classification of down-regulated proteins.

Figure 6

Compound–target networks related to ZYCH effects on DMN-induced liver fibrosis. (A) C-T network of triterpenoid saponins and targets. (B) C-T network of lignans and targets. Red ellipses are up-regulated proteins, and green ellipses are down-regulated proteins in proteomics. Blue ellipses are compounds.