Objective To investigate the ameliorative effects of β-hydroxybutyrate (BHB) on energy metabolism disorder in SH-SY5Y cells induced by high-concentration insulin and analyze its relationship with the SIRT2 signaling pathway. Methods An insulin resistance model was established in SH-SY5Y cells by treatment with 150 nmol/L insulin for 48 h, followed by intervention with 10 mmol/L BHB for 24 h. Cell viability was assessed using the CCK-8 assay, and glucose uptake capacity was measured with the 2-NBDG fluorescent probe. Inflammatory cytokines and oxidative stress markers were determined by ELISA. Cellular reactive oxygen species (ROS) levels and mitochondrial membrane potentials were evaluated using DCFH-DA and JC-1 fluorescent staining, respectively. ATP content, NAD⁺/NADH ratio, and the activities of mitochondrial respiratory chain complexes were measured using biochemical assay kits. Western blot was employed to detect the protein expression of phosphorylated insulin receptor substrate-1 at serine 307 [p-IRS-1 (Ser307)], sirtuin 2 (SIRT2), acetylated α-tubulin at lysine 40 [Ac-α-tubulin (Lys40)], peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and mitochondrial transcription factor A (TFAM). Results Compared with the normal control group, the model group exhibited significantly reduced cell viability and glucose uptake capacity, along with increased p-IRS-1(Ser307) expression, indicating successful construction of the insulin-resistant model. The model group also showed elevated levels of interleukin-6 (IL-6), interleukin-17 (IL-17), and tumor necrosis factor-α (TNF-α), decreased interleukin-10 (IL-10), accumulation of ROS and malondialdehyde (MDA), decreased superoxide dismutase (SOD) activity, reduced mitochondrial membrane potential, and markedly lower ATP content, NAD⁺/NADH ratio, and activities of respiratory chain complexes I, II, III, and IV. Following BHB intervention, cell viability was significantly improved, inflammatory imbalance corrected, ROS production reduced, SOD activity elevated, MDA content declined, mitochondrial membrane potential was restored, and ATP generation and respiratory chain function enhanced. Further mechanistic studies revealed that BHB significantly upregulated SIRT2 protein expression, decreased the Ac-α-tubulin (Lys40)/α-tubulin ratio, and promoted the expression of PGC-1α and TFAM. Conclusion BHB can significantly ameliorate high-concentration insulin-induced energy metabolism disorder in SH-SY5Y cells, and its mechanism is associated with the activation of SIRT2, enhancement of deacetylase function, and promotion of mitochondrial biogenesis.
Key words
β-hydroxybutyrate /
insulin resistance /
SIRT2 /
mitochondrial function /
SH-SY5Y cells
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