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Energy and Macronutrient Metabolism (Oral Session 10)

(OR10-01-25) The Impact of Insulinogenic and Branched-Chain Amino Acids Depletion on Glucose and Lipid Metabolism in HepG2 Cells

Monday, June 2, 2025
8:30 AM – 8:42 AM ET
Location: S230 CD
Emerging Leaders in Nutrition Science Poster Competition

    Presenting Author(s)

  • Matthew W. Gorton, BS

    Ph.D. Candidate
    Oklahoma State University
    Stillwater, Oklahoma, United States

    Disclosure(s): No relevant financial relationship(s) with ineligible companies to disclose.

    • Co-Author(s)

  • MW

    Melanine Whitmore

    Oklahoma State University
    Stillwater, Oklahoma, United States

  • GZ

    Guolong Zhang, Dr

    Oklahoma State University

  • AP

    Adel Pezeshki, Assistant Professor

    Faculty member in Animal and Food Sciences
    Oklahoma State University
    Stillwater, Oklahoma, United States

Objectives: Insulinogenic amino acids (IAA), i.e., Leu, Ile, Val, Thr, Phe, Arg, and Ala and branched-chain amino acids (BCAA), i.e., Leu, Ile, and Val are associated with impaired glucose uptake, insulin sensitivity, and increased risk of metabolic diseases. The objective of this study is to investigate the impact of IAA and BCAA depletion on hepatic glucose and lipid metabolism in HepG2 Cells.

Methods: Human hepatocellular carcinoma HepG2 cells were cultured for 48 hours in DMEM media with one of three conditions: 1) control (CON): amino acid complete media; 2) IAA depleted media (D-IAA), and 3) BCAA depleted media (D-BCAA). Following incubation with experimental treatments, cells were then challenged with or without 100 nmol/L insulin for 20 min. Glycogen and glucose assays were used to quantify the cellular glycogen content and glucose concentration in cell lysate and supernatant, respectively. The mRNA abundance of candidate genes regulating glucose and lipid metabolism were determined. All data were analyzed with SPSS® via GLM Univariate with Dunnett’s post-hoc.

Results: The concentration of cellular glycogen was reduced in both D-IAA and D-BCAA while the amount of residual glucose in the media was higher in the same groups when compared to CON. This is suggestive of an increased glycogenolysis following IAA and BCAA depletion. The mRNA abundance of phosphoenolpyruvate carboxykinase (PEPCK) decreased in D-IAA and D-BCAA with or without insulin stimulation when compared to CON suggestive of a decreased gluconeogenesis. In both insulin and noninsulin-stimulated cells, the mRNA abundance of fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and sterol regulatory element-binding protein 1 (SREBP1) was decreased while that for carnitine palmitoyltransferase 1 α (CPT1α was increased in both D-IAA and D-BCAA compared to CON. This is suggestive of a decreased lipogenesis and an increased lipolysis following IAA and BCAA depletion.

Conclusions: In conclusion, the depletion of BCAA and IAA increased glycogenolysis and lipolysis. Further, depletion of these amino acids decreased the expression of genes involved in gluconeogenesis and lipogenesis in HepG2 cells.

Funding Sources: Hatch project accession no. 1012889 from the USDA National Institute of Food and Agriculture (NIFA).