Application of UPLC-Q-Exactive-MS Technology Combined With Network Pharmacology to Investigate the Mechanism of Action of AsterTataricus L. Extract in the Treatment of Drug-induced Liver Injury

Objective: The objective of this study was to speculate the Pharmacodynamic material basis and potential mechanism of Aster tataricus L. Total Flavonoids (ATF) in the treatment of DILI in C57BL/6 mice caused by Isoniazid (INH) and Rifampicin (RIF) capsules. Methods: This experiment used ultrasonic extraction to extract the ATF, and purified the ATF with AB-8 macroporous adsorbent resin. SPF-grade C57 male mice were administered INH and RIF to create DILI models, and histopathological sections were observed by HE staining; the test kit was used to determine the content of ALT and AST in serum and the content of SOD, MDA, GSH-Px, and T-AOC in liver tissue; UPLC-Q-Exactive-MS was used to analyze the ATF and the serum components of DILI mice, and network pharmacology analysis and molecular docking validation was performed. Results: The extraction rate of ATF was 31.9%, and the purification rate was 83.1%. All of the ATF administration groups could alleviate cell necrosis and inflammatory infiltration, reduce organ index, and alleviate the abnormalities of the indexes of ALT, AST, SOD, MDA, GSH-Px, and T-AOC, and the best effect was found in the low dose. 15 blood-entry ingredients were compared and analyzed using UPLC-Q-Exactive-MS. 9 prototype class components were screened out and 4 core targets were obtained, all of which possessed high binding energies. Conclusion: This experiment provides a preliminary analysis of the potential for further study of ATF in the treatment of DILI, and offers a new avenue for research into the natural products of Traditional Chinese Medicine. result: The extraction rate of ATF by ultrasonic extraction was 31.9%, and the purification rate of AB-8 macroporous adsorbent resin was 83.1%. INH combined with RIF could successfully induced DILI model, and all of the ATF administration groups could alleviate cell necrosis and inflammatory infiltration, reduce organ index, and alleviate the abnormalities of the indexes of ALT, AST, SOD, MDA, GSH-Px and T-AOC, and the best effect was found in the low dose. 15 blood-entry ingredients were compared and analyzed by UPLC-Q-Exactive-MS, including Calycosin, Quercetin, Isorhamnetin, Quercetin 3-O-glucosyl-xyloside, Quercetin 3-(6'-acetylglucoside) and so on. With 9 prototype components as target compounds, 236 intersecting targets were obtained by Wayne analysis through SwissTargetPrediction and GeneCards databases; 108 interacting gene proteins were obtained by PPI network construction; 10 each of related BP, CC and MF were screened by GO enrichment analysis; and 10 metabolic pathways for potential target compounds for DILI were screened by KEGG pathway analysis. compounds to treat 10 metabolic pathways of potential targets of DILI. The network diagram of “compound-target-metabolic pathway” was successfully constructed by Cytoscape 3.7.2 software, and the interactions among compounds, targets and metabolic pathways were strong. The molecular docking validation showed that the target compounds and the core targets screened by LAC value had high binding energy. conclusion: The ultrasonic extraction method had a relatively high extraction rate, and the AB-8 macroporous adsorbent resin was able to purify the ATF better All the groups of ATF administrations improved INH combined with RIF-induced drug-induced liver injury in mice, with the low-dose groups showing better therapeutic effects compared to the medium-dose and high-dose groups. It mainly acts on the pathways of PI3K-Akt, MAPK, EGFR, and FoxO through the core targets of IGF1R, Akt1, EGFR, and TNF to achieve the therapeutic effect of DILI. This experiment provides a preliminary analysis of the potential for further study of ATF for the treatment of DILI, and offers a new avenue for research into natural products of traditional Chinese medicine.