The Medicinal Plant Pair Bupleurum chinense- Scutellaria baicalensis - Metabolomics and Metallomics Analysis in a Model for Alcoholic Liver Injury

Abstract

Traditional Chinese Medicine (TCM), a complex natural herbal medicine system, has increasingly attracted attention from all over the world. Most research has illustrated the mechanism of TCM based on the active components or single herbs. It was fruitful and effective but far from satisfactory as it failed to gain insights into the interactivity and combined effects of TCM. In this work, we used Bupleurum chinense (B. chinense DC, a species in the genus Bupleurum, family Apiaceae) and Scutellaria baicalensis (S. baicalensis Georgi, a species in the genus Scutellaria, family Lamiaceae), an herbal pair in TCM, to illustrate the combined effect. We compared the diverse effects between the B. chinense-S. baicalensis herbal pair and its compositions in an animal model of Alcoholic Liver Injury to highlight the advantages of the formula. Biochemical and histological indicators revealed that the effect of B. chinense-S. baicalensis was better than its individual parts. Furthermore, metabolite profiling of the serum, liver tissue, and feces were conducted to reveal that the herbal pair largely presented its effects through enhanced tissue penetration to maintain liver-located intervention with less global and symbiotic disturbance. Furthermore, we analyzed the distribution of the metal elements in extracts of the serum and liver tissue and found that the herbal pair significantly regulated the distribution of endogenous selenium in liver tissue. As selenium plays an important role in the anti-oxidative and hepatoprotective effects, it may be the reason for combined effects in BS formula. This research could open new perspectives for exploring the material basis of combined effects in natural herbal medicine.

Keywords: Bupleurum chinense-Scutellaria baicalensis; alcoholic liver injury; herbal pair; metabolomics; metallomics.

FIGURE 1

Biochemical and histological analysis. (A) Biochemical analysis of major biomarkers for liver functions in blood of rates. The biochemical indicators of Bupleurum chinense-Scutellaria baicalensis and their composition were compared to the normal control group; (B) liver tissue histopathology (H&E staining) of every group; (C) the quantized results of liver tissue histopathology. The results were based on the area of intercellular space and lipid droplets to present liver cell swelling and lipid droplets accumulation. N: Normal control; M: Alcoholic Liver Injury Modeling; ALI: Alcoholic Liver Injury; BS: decoctions of B. chinense-S. baicalensis; BU: decoctions of B. chinense; SC: decoctions of S. baicalensis; SA: water solution of Saikosaponin; BA: water solution of Baicalin; MI: the mixture water solution of Baicalin and Saikosaponin; Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Glucose (GLU), Cholesterol (CHOL), Triglyceride (TG).

FIGURE 2

Metabolite and pathway analysis of drug-induced perturbations. (A) Numbers of drug-intervened metabolites in ALI; (B) numbers of drug-intervened pathways in ALI. The summarization of drug-intervened global, local and symbiotic pathways is presented in the center. BS: decoctions of B. chinense-S. baicalensis; SC: decoctions of S. baicalensis; BU: decoctions of B. chinense; BA: water solution of baicalin; SA: water solution of saikosaponin; MI: the mixture of baicalin and saikosaponin.

FIGURE 3

B. chinense-S. baicalensis herbal pair altered the distribution of metal contents in the serum and liver tissue. (A) Comparison of metal contents between average of BU and SC (white column) and BS (black column). (B) Comparison of serum metal contents between average of BU-treated and SC-treated rats (white column) and BS-treated rats (black column). (C) Comparison of hepatic metal contents between average of BU-treated and SC-treated rats (white column) and BS-treated rats (black column). BU: decoctions of B. chinense; SC: decoctions of S. baicalensis; BS: decoctions of Bupleurum chinense-S. baicalensis.

FIGURE 4

Schematic diagram of the combined effects of the B. chinense-S. baicalensis herbal pair in Alcoholic Liver Injury. Alcoholic Liver Injury was used to research the characteristics of B. chinense- S. baicalensis, which is controlled by B. chinense and S. baicalensis, and their main active ingredients baicalin and saikosaponin. The biochemical indicators and histology indicated that B. chinense-S. baicalensis presented the best efficacy, which could be further illustrated by global, local, and symbiotic metabolic analysis. Finally, metallomic analysis was employed to uncover the distribution changes of selenium which, unlike other metal elements, was significantly increased by B. chinense-S. baicalensis in liver tissue to demonstrate multiple benefits such as anti-oxidative stress and hepatoprotective effect, to relieve drug pressure.