Mechanism of Xiaoyao San in treating non-alcoholic fatty liver disease with liver depression and spleen deficiency: based on bioinformatics, metabolomics and in vivo experiments

Abstract

Xiaoyao san (XYS) plays an important role in treatment of non-alcoholic fatty liver disease (NAFLD) with liver stagnation and spleen deficiency, but its specific mechanism is still unclear. This study aimed to investigate the material basis and mechanism by means of network pharmacology, metabolomics, systems biology and molecular docking methods. On this basis, NAFLD rat model with liver stagnation and spleen deficiency was constructed and XYS was used to intervene, and liver histopathology, biochemical detection, enzyme-linked immunosorbent assay, quantitative PCR assay and western blotting were used to further verify the mechanism. Through the above research methods, network pharmacology study showed that there were 94 targets in total for XYS in the treatment of NAFLD. Metabolomics study showed that NAFLD with liver depression and spleen deficiency had a total of 73 differential metabolites. Systems biology found that PTGS2 and PPARG were the core targets; Quercetin, kaempferol, naringenin, beta-sitosterol and stigmasterol were the core active components; AA, cAMP were the core metabolites. And molecular docking showed that the core active components can act well on the key targets. Animal experiments showed that XYS could improve liver histopathology, increase 5HT and NA, decrease INS and FBG, improve blood lipids and liver function, decrease AA, increase cAMP, down-regulate PTGS2, up-regulate PPARG, and decrease PGE2 and 15d-PGJ2. In conclusion, XYS might treat NAFLD with liver depression and spleen deficiency by down-regulating PTGS2, up-regulating PPARG, reducing AA content, increasing cAMP, improving insulin resistance, affecting glucose and lipid metabolism, inhibiting oxidative stress and inflammatory response.

Communicated by Ramaswamy H. Sarma

HIGHLIGHT

1. Network pharmacological results found that XYS might treat NAFLD through multiple targets and multiple pathways. Quercetin and kaempferol were main components of XYS in treatment of NAFLD.

2. System biology research results found that PPARG and PTGS2 were the core targets of XYS in the treatment of NAFLD.

3. Metabolomic results suggest that AA and cAMP were differential metabolites of NAFLD.

4. In vivo animal experiments showed that XYS might treat NAFLD by increasing PPARG and decreasing PTGS2, reducing AA and increasing cAMP to regulate glucose metabolism and lipid metabolism.

Keywords:

• Xiaoyao san (XYS)
• nonalcoholic fatty liver disease (NAFLD)
• network pharmacology
• metabolomics
• systems biology
• in vivo experiments

Authors’ contributions

Xiaofeng Ruan: Conceptualization ideas, Methodology, Investigation, Data collection, Animal experiment, Writing—original draft. Xiaoming Zhang: Visualization, Formal analysis, Writing—review & editing. Liming Liu: Supervision, Methodology, Project administration, Writing—review & editing. Jianjun Zhang: Supervision, Methodology, Funding acquisition, Writing—review & editing.

Disclosure statement

The authors declared there are no conflicts of interest for this study.

Additional information

Funding

This work was supported by Hubei Provincial Health and Medical Committee Chinese Medicine Scientific Research Project [Grant Number ZY2021Z012]; Hubei Medicine Top-Notch Young Talents (2019); and The Second Medical Leading Talent in Hubei Province.