Mechanism for Huanglian Jiedu Decoction-Based Therapy for MAFLD Analyzed Through Network Pharmacology and Experimental Verification

Abstract

Objective: To analyze the mechanism of Huanglian Jiedu Decoction (HLJDD) in the treatment of metabolism-associated fatty liver disease (MAFLD). Methods: The main components, targets, and pathways for treating MAFLD of HLJDD were screened through network pharmacology and molecular docking validation was done; HLJDD was used to intervene MAFLD model of rat, the levels of ALT, AST, TC, TG, GLU, HDL, and LDL were identified, HE staining was used to observe the pathological changes, lipid deposition in liver was detected by oil red O staining. MAFLD model of HepG2 (hepatocellular carcinoma cell line) was constructed by PA (palmitate-acid) incubating, and HLJDD was administered with drug-containing serum intervention, lipid droplets in HepG2 cells was observed by oil red O staining, TG and FFA of HepG2 were detected, the expressions of AMPK, mToR, and Beclin-1 were detected through Western blot. Results: Seventy components and 229 targets were obtained, and 85 targets were used to treat MAFLD, which focus on the signal passways of AMPK/mToR/PI3K-AKt/MAPK, NAFLD, autophagy-animal, insulin, etc. Molecular docking outcomes showed quercetin, kaempferol, and baicalein that were successfully docked with AMPK and mToR, and had good binding activity, compared with MAFLD group of rats, the levels of ALT, AST, TC, TG, GLU, HDL, and LDL were significantly decreased in Silybin group and each dos group of HLJDD, liver pathology and lipid deposition were significantly improved; the results in vitro experiments showed that drug-containing serum of HLJDD and Silybin could improve intracellular lipid accumulation and reduce the increase of TG and FFA levels in HepG2 cells, the therapeutic effect of HLJDD was significantly attenuated after application of AMPK inhibitor; the results of Western blot showed that HLJDD could up-regulate the protein expression of AMPK and Beclin-1,down-regulate the protein expression of mToR. Conclusion: Within process of MAFLD intervention, HLJDD could regulate AMPK-mToR signaling pathway to treat MAFLD.