Effect Of Nobiletin On Fatty Acid Metabolism In 3T3-L1 Cells

Part I: Effect of Nobiletin alone on Fatty Acid Metabolism in 3T3-L1cellsObjective: To investigate the regulatory effect of Nobiletin(NOB)on fatty acid synthesis and oxidation in mouse preadipocytes when used alone.Methods: In this study,3T3-L1 cells were treated with different concentrations of Nobiletin 12.5,50 μM(treatment group),0.1% DMSO(control group),and then the contents of intracellular acetyl-Co A carboxylase(ACC),carnitine palm pretransferase 1(CPT1),hormone-sensitive lipase(HSL),glycerol,and soft acid were determined with ELISA kit for 48 h.Real-time PCR was used to detect the expression of m RNA of fatty acid metabolism-related genes ACC,CPT1 and HSL.The expression level of factor SREBP-1c protein related to fatty acid synthesis was determined by western blotting.When treating cells with different concentrations of NOB,Akt inhibitor LY294002 was added,and all the above indicators were determined for 48 hours.Results: Treatment of 3T3-L1 cells by NOB alone promoted the synthesis of fatty acids,increased the expression of ACC enzyme,the m RNA gene ACC,and decreased the level of fatty acid oxidation,CPT1;the protein results showed increased expression of SREBP-1c factor.The results after the Akt pathway inhibition showed that there was no significant link between the gene expression of ACC and this pathway,while the expression of CPT1 and HSL was significantly reduced,and the expression of protein SREBP-1c was decreased.Conclusion:In 3T3-L1 cells,Nobiletin may promote fatty acid synthesis and oxidative decomposition by activating the Akt pathway.Part II: Effect of Nobiletin on Fatty Acid Metabolism in 3T3-L1 Cells in a cocktail systemObjective: After the introduction of cocktail system in DMEM,to investigate the regulatory effect of NOB on fatty acid synthesis and oxidation in3T3-L1 cells.Methods: In this experiment,3T3-L1 cells were treated with a cocktail of IBMX(3-isobutyl-1 methylxanthine),DEX(dexamethasone)and insulin with different concentrations of NOB for 48 hours,and acetyl-Co A carboxylase(ACC),carnitine palm pretransferase 1(CPT1),hormone-sensitive lipase(HSL),glycerol,content of soft acid;Real-time PCR was used to detect the expression of m RNA of fatty acid metabolism-related genes ACC,CPT1 and HSL.The expression level of factor SREBP-1c protein related to fatty acid synthesis was determined by western blotting.Subsequently,the AMPK inhibitor Compound C was introduced,and all the above indicators were determined in 48 hours.Results: In the cocktail culture system,NOB treatment increased the synthesis of fatty acids and the expression of ACC enzyme,reduced the expression level of synthetic gene ACC,increased the expression of fatty acid decomposition oxidation gene CPT1,and inhibited the expression of protein SREBP-1c.After using Compound C,AMPK activity was inhibited,the level of fatty acid synthesis gene ACC m RNA increased,CPT1 decreased,and the expression of protein SREBP-1c increased significantly.Conclusion: In 3T3-L1 cells,the co-treatment of the cocktail medium and NOB may promote fatty acid synthesis and breakdown through the activation of the AMPK pathway.