Effect Of Naringenin On Differentiation And Lipid Metabolism Of 3T3-L1 Cells

Grapefruit peel is rich in flavonoids which range from 1%-6% and are mainly dihydroflavones,such as naringin,naringenin and hesperidin.Previous studies have shown that naringenin has good lipid-lowering,anti-inflammatory,anti-tumor and other effects.In this study,network pharmacology and 3T3-L1 cell test were used to investigate the lipidlowering effect of naringenin and its mechanism,so as to find a safer and more effective natural compound to treat obesity.In this research,we studied the molecular mechanism of naringenin in the treatment of obesity by network pharmacology,providing a new direction for the treatment of obesity.Secondly,3T3-L1 preadipocytes were treated with different concentrations of naringenin(25,50 and 100 μg/m L)for 24 h,and the cell viability was determined by MTT assay.Then,3T3-L1 preadipocytes were seeded into 6-well plates until confluent,then the growth medium was replaced with a differentiation medium(10% FBS,0.5 mmol/L 3-lsobuty-1-methylxanthine(IBMX),1 μmol/L dexamethasone(DEX),and 10 μg/m L insulin(MDI))for incubation for 48 h.Then differentiation medium was replaced with an adipocyte maintenance medium containing 10% FBS and insulin(10 μg/m L).The adipocyte maintenance medium was changed every 48 h.3T3-L1 preadipocytes were treated with or without naringenin(25,50 and 75 μg/m L)until the end of cell differentiation.After differentiation,the lipid content,TG(triglyceride)and TC(cholesterol)levels of 3T3-L1 mature adipocytes were determined.The m RNA expression levels related to fat metabolism in cells were measured by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)to study the lipid-lowering mechanism of naringenin.Finally,molecular docking technology was used to simulate the interaction mode and binding energy between naringenin and AMPK protein.KEGG pathway enrichment results showed that naringenin-mediated obesity was significantly correlated with AMPK signaling pathway.In vitro experiments showed that naringenin preconditioning inhibited the cell viability of 3T3-L1 cells in a concentrationdependent manner,and decreased the intracellular fat content,TG and TC levels.These results indicated that naringenin effectively alleviated the intracellular fat accumulation in3T3-L1 mature cells.In addition,Real-time PCR analysis showed that naringenin significantly promoted AMPK and CPT-1 m RNA expression levels in 3T3-L1 mature cells,and decreased the expression levels of ACC,FAS,SREBP-1C and HMGCR m RNA.Finally,molecular docking analysis showed that naringenin and AMPK protein were bound by hydrogen bond,van der Waals force and other interaction forces to play a lipid-lowering role.In conclusion,naringenin may alleviate obesity by activating AMPK signaling pathway,reducing the fat content of 3T3-L1 mature adipocytes and regulating the expression of adipose-related genes.