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    Ixeris dentata (Thunb) Nakai Ethylacetate Extract Attenuates Sterol Regulatory Element-Binding Proteins-1c via AMP-Activated Protein Kinase Activation

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    Purpose: To investigate the molecular mechanisms underlying the role of Ixeris dentataa extract (IDE)¬†in the prevention of high glucose induced lipid accumulation in human HepG-2 hepatocytes.Methods: IDE extract was prepared by maceration in ethyl acetate. Its fractionation was carried out by¬†column chromatography. HepG-2 cells were pretreated with various concentration of IDE (0, 10, 20, 40¬†and 80 őľg/mL) and then treated with serum-free medium with normal glucose (5 mM) for 1 h, followed¬†by exposure to high glucose (30 mM D-glucose) for 24 h. Cell viability and cytotoxicity parameters were¬†measured using lactate dehydrogenase (LDH) and MTT assay while triglyceride and total cholesterol¬†levels were evaluated using respective enzymatic reagent kits. Protein expressional levels were¬†analyzed by Western blotting.Results: IDE did not influence the cell viability (up to 200 őľg/mL) and did not show any signs of¬†cytotoxicity (up to 80 őľg/mlL. IDE significantly attenuated lipid accumulation in human HepG2¬†hepatocytes when exposed to high glucose (30 mM D-glucose) in a dose-dependent manner (p < 0.05,¬†0.01 and 0.001 at 20, 40 and 80 őľg/mL concentrations, respectively). Nile red staining showed that 10,¬†20, 40 and 80 őľg/mL concentrations of IDE reduced lipid accumulation by 23.4, 34.8 (p < 0.05), 46.5 (p¬†< 0.01) and 53.2 % (p < 0.001), respectively. The increased levels of triglycerides and total cholesterol¬†were also attenuated by IDE (p < 0.001 at 80 őľg/mL). Further, IDE attenuated the expression of fatty¬†acid synthase and sterol regulatory element-binding protein-1. Adenosine monophosphate-activated¬†protein kinase was also activated by IDE treatment when exposed to high glucose level (30 mM Dglucose)¬†in human HepG2 hepatocytes.Conclusion: The findings indicate that IDE exerts hypolipidemic effect by inhibiting lipid biosynthesis,¬†mediated via AMPK signaling. This probably explains the extract‚Äôs beneficial effect in various¬†inflammation disorders.Keywords: Ixeris dentata extract, Lipogenesis, AMP-activated protein kinase, Sterol regulatory¬†element-binding protein 1 -1, Anti-obesity, HepG-2 cel

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