Bioinformatics Reveals Mechanism of Schisandrin B in Inhibiting Ferroptosis to Ameliorate Methionine and Choline Deficiency-induced Fatty Liver Disease in Mice
Objective:Nonalcoholic fatty liver disease(NAFLD)is a metabolic stress liver injury.Ferroptosis is involved in the occurrence and development of NAFLD.Exploring the efficacy and mechanism of schisandrin B in treating NAFLD facilitates the development of strategies for the prevention and treatment of NAFLD.Methods:The molecular structure of schisandrin B was obtained by searching against PubChem,and the related targets were predicted by SwissTargetPrediction.The active ingredients and their targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the high-throughput experiment-and reference-guide database of traditional Chinese medicine(HERB).GeneCards and FerrDb were searched for the targets of NAFLD and ferroptosis.The common targets were taken as the core targets,and the protein-protein interaction network of the core targets was established.DAVID was used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.Finally,molecular docking was performed between schisandrin B and core targets,and the binding energy was calculated.C57BL/6 mice were fed with a methionine and choline-deficiency(MCD)diet for the modeling of NAFLD.Mice were randomized into normal,model,positive drug(essentiale),and low-and high-dose schisandrin B groups.The body mass and liver index of mice were measured after drug administration.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the serum and those of total cholesterol(TC),triglyceride(TG),malondialdehyde(MDA),glutathione(GSH),and Fe2+in the liver homogenate were measured by biochemical assay kits.The pathological changes of the liver tissue were observed by hematoxylin-eosin(HE)and red oil O staining.Enzyme-linked immunosorbent assay was employed to determine the levels of interleukin(IL)-6,IL-1β,tumor necrosis factor(TNF)-α,and 4-hydroxynonenal(4-HNE)in the serum.Western blotting and real-time PCR were employed to determine the protein and mRNA levels,respectively,of solute carrier family 7 member 11(SLC7A11),solute carrier family 3 member 2(SLC3A2),glutathione peroxidase 4(GPX4),transferrin,and ferritin heavy chain(FTH)in the liver tissue.Results:A total of 2 370,2 547,and 1 451 targets of schisandrin B,NAFLD,and ferroptosis were obtained,in which 90 common targets were shared by the three.Enrichment analyses predicted 505 GO terms and 92 KEGG pathways.Molecular docking suggested that schizandrin B had strong binding affinity with the key targets of ferropstosis(SLC7A11 and SLC3A2).Animal experiments showed that schizandrin B significantly decreased the liver index,lowered the levels of ALT,AST,TC,TG,IL-6,IL-1β,and TNF-α,alleviated hepatocyte ballooning and inflammatory cell infiltration,and reduced lipid accumulation in the liver of NAFLD mice.In addition,schisandrin B significantly lowered the levels of MDA,4-HNE,and Fe2+,elevated the level of GSH,up-regulated the protein and mRNA levels of SLC7A11,SLC3A2,and GPX4,and down-regulated the protein and mRNA levels of transferrin in the liver tissue.Conclusion:Schisandrin B can alleviate NAFLD by inhibiting ferroptosis in hepatocytes.
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