OBJECTIVE To investigate the efficacy and mechanism of the active components from Meconopsis integrifolia(Maxim.) Franch in improving nonalcoholic fatty liver disease(NAFLD) using network pharmacology and cell experiments.

METHODS An free fatty acid(FFA)-induced HepG2 steatosis cell model was employed to screen the active components of Meconopsis integrifolia for NAFLD improvement via triglyceride(triglyceride, TG) detection and oil red O staining. Ultra-high performance liquid chromatography-quadrupole-exactive tandem mass spectrometry(UPLC-Q-Exactive-MS/MS) was used to identify the components of the active fractions. Network pharmacology was applied to collect the overlapping targets between the active components of Meconopsis integrifolia and NAFLD, construct a “drug-component-target-disease” network and a protein-protein interaction(PPI) network, and screen the core components and targets. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analyses were performed to obtain key signaling pathways. Using the FFA-induced HepG2 steatosis cell model, an ELISA kit was used to detect the effect of the active components on high mobility group box 1(HMGB1). Western blotting was used to determine the protein expression levels of nuclear factor kappa B p65 subunit(NF-κB p65), phosphorylated NF-κB p65(p-NF-κB p65), Toll-like receptor 4(TLR4), HMGB1, B-cell lymphoma 2(Bcl-2), and tumor protein P53(TP53). Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression levels of the gene encoding NF-κB p65, interleukin-1β(IL-1β), and tumor necrosis factor-α(TNF-α).

RESULTS Cell experiment results showed that all polar fractions of Meconopsis integrifolia exerted a certain improvement effect on FFA-induced elevation of TG content and lipid accumulation, among which the 70% ethanol elution fraction exhibited the best efficacy. Network pharmacology results revealed 88 active components and 161 overlapping targets, including 21 core targets. The lipid metabolism and atherosclerosis pathways were enriched with more core targets, and the HMGB1/TLR4/NF-κB signaling pathway was selected for verification. ELISA results indicated that compared with the model group, the HMGB1 level in cells of the administration group was significantly decreased. Western Blot results showed that the phosphorylation level of the NF-κB p65/NF-κB p65 ratio protein, as well as the protein levels of TLR4, HMGB1, and TP53, were significantly reduced, while the Bcl-2 protein level was significantly increased. RT-qPCR results demonstrated that the mRNA expression levels of NF-κB, IL-1β, and TNF-α in cells of the administration group were significantly decreased.

CONCLUSION The study indicates that the active components of Meconopsis integrifolia can alleviate FFA-induced inflammatory response and cell apoptosis by regulating the HMGB1/TLR4/NF-κB signaling pathway, thereby improving NAFLD.