Integrated Serum Metabolomics and Network Pharmacology Approach to Reveal the Potential Mechanisms of Diwuyanggan Prescription in the Prevention of Acute Liver Injury

Objective: Acute liver injury (ALI) has become a major reason for emergency liver transplantation mortality worldwide. Diwuyanggan (DWYG), a hospital preparation of traditional chinese medicine, does a satisfactory job of treating liver disease, but its pharmacological mechanisms should be further investigated. Thus, we aimed to unravel the mechanisms of DWYG against ALI through a comprehensive approach integrating metabolomics and network pharmacology analysis in this study. Methods: An untargeted metabolomics based on ultra-high pressure liquid chromatography together with quadrupole time of flight mass spectrometry was applied to identify differential metabolites between normal mice and thioacetamide (TAA)-induced ALI mice. By utilizing network pharmacology, potential targets against ALI caused by TAA were excavated. Subsequently, the key targets were validated by real-time polymerase chain reaction assays. Results: DWYG alleviated TAA-induced ALI. Furthermore, 28 differential metabolites were identified with an association between ALI pathological processes and DWYG effects. The pathways involved in bile acid, glycerophospholipid, fatty acid, sphingolipid, tryptophan, and tyrosine metabolism. Then, we found 127 active compounds in DWYG and 26 hub genes associated with the treatment of ALI according to network pharmacology. Besides, according to Kyoto Encyclopedia of Genes and Genome analysis, sphingolipid signaling pathway was deemed as one of the main signaling pathways involved in DWYG anti-ALI, which was in agreement with the metabolomics findings. Further, a combined analysis was performed focusing on 7 key targets, including mitogen-activated protein kinase 1, mitogen-activated protein kinase 8, caspase 3, peroxisome proliferator-activated receptor gamma, albumin, carboxylesterase 1 and glucocerebrosidase, along with their related core metabolites and pathways. Confirmatory experiment further showed that DWYG could reverse the abnormal mRNA expression of these targets in TAA mice. Conclusion: This study revealed direct evidence for the preventive effect of DWYG against ALI and the related pharmacological mechanisms.