Integrated network analysis and metabolomics reveal the molecular mechanism of Yinchen Sini decoction in CCl4-induced acute liver injury

Weiwei Zheng; Chao Shi; Yao Meng; Jian Peng; Yongfei Zhou; Tong Pan; Ke Ning; Qiuhong Xie; Hongyu Xiang

doi:10.3389/fphar.2023.1221046

Integrated network analysis and metabolomics reveal the molecular mechanism of Yinchen Sini decoction in CCl₄-induced acute liver injury

Front Pharmacol. 2023 Sep 25:14:1221046. doi: 10.3389/fphar.2023.1221046. eCollection 2023.

Authors

Weiwei Zheng¹, Chao Shi¹, Yao Meng¹, Jian Peng¹, Yongfei Zhou¹, Tong Pan¹, Ke Ning¹, Qiuhong Xie^{1

2

3}, Hongyu Xiang^{1

2

3}

Affiliations

¹ Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, Changchun, Jilin, China.
² National Engineering Laboratory for AIDS Vaccine, School of Life Sciences, Jilin University, Changchun, Jilin, China.
³ Institute of Changbai Mountain Resource and Health, Jilin University, Fusong, Jilin, China.

Abstract

Objective: Yinchen Sini decoction (YCSND), a traditional Chinese medicine (TCM) formula, plays a crucial role in the treatment of liver disease. However, the bioactive constituents and pharmacological mechanisms of action remain unclear. The present study aimed to reveal the molecular mechanism of YCSND in the treatment of acute liver injury (ALI) using integrated network analysis and metabolomics. Methods: Ultra-high-performance liquid chromatography coupled with Q-Exactive focus mass spectrum (UHPLC-QE-MS) was utilized to identify metabolites in YCSND, and high-performance liquid chromatography (HPLC) was applied to evaluate the quality of four botanical drugs in YCSND. Cell damage and ALI models in mice were established using CCl₄. ¹H-NMR metabolomics approach, along with histopathological observation using hematoxylin and eosin (H&E), biochemical measurements, and reverse transcription quantitative real-time PCR (RT-qPCR), was applied to evaluate the effect of YCSND on CCl₄- induced ALI. Network analysis was conducted to predict the potential targets of YCSND in ALI. Result: Our results showed that 89 metabolites in YCSND were identified using UHPLC-QE-MS. YCSND protected against ALI by reducing the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and malondialdehyde (MDA) contents and increasing those of superoxide dismutase (SOD), and glutathione (GSH) both in vivo and in vitro. The ¹H-NMRmetabolic pattern revealed that YCSND reversed CCl₄-induced metabolic abnormalities in the liver. Additionally, the Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analysis identified five pathways related to liver injury, including the PI3K-AKT, MAPK, HIF-1, apoptosis, and TNF signaling pathways. Moreover, RT-qPCR showed YCSND regulated the inflammatory response (Tlr4, Il6, Tnfα, Nfκb1, Ptgs2, and Mmp9) and apoptosis (Bcl2, Caspase3, Bax, and Mapk3), and inhibited PI3K-AKT signaling pathway (Pi3k and Akt1). Combined network analysis and metabolomics showed a link between the key targets (Tlr4, Ptgs2, and Mmp9) and vital metabolites (choline, xanthine, lactate, and 3-hydroxybutyric acid) of YCSND in ALI. Conclusion: Overall, the results contribute to the understanding of the therapeutic effects of YCSND on ALI, and indicate that the integrated network analysis and metabolomics could be a powerful strategy to reveal the pharmacological effects of TCM.

Keywords: 1 H-NMR; PI3K-AKT; Yinchen Sini decoction; acute liver injury; network analysis.

Grants and funding

This work was supported by Changchun Science and Technology Bureau under Grant 21ZGN11, Jilin Province Development and Reform Commission under Grant 2021FGWCXNLJSSZ11, and Jilin Province Science and Technology Institute under Grant 20220204067YY.