In this study, we aimed to study the role of
growth factor receptor-bound protein 2 (Grb2) in
palmitic acid-induced steatosis and other "
fatty liver" symptoms in vitro. HepG2 cells, with or without stably suppressed Grb2 expression, were incubated with
palmitic acid for 24 h to induce typical clinical "
fatty liver" features, including steatosis, impaired
glucose metabolism, oxidative stress, and apoptosis. MTT and
Oil Red O assays were applied to test cell viability and fat deposition, respectively.
Glucose uptake assay was used to evaluate the
glucose utilization of cells. Quantitative polymerase chain reaction and Western blot were used to measure expressional changes of key markers of
insulin signaling,
lipid/
glucose metabolism, oxidative stress, and apoptosis. After 24-h
palmitic acid induction, increased fat accumulation, reduced
glucose uptake, impaired
insulin signaling, enhanced oxidative stress, and increased apoptosis were observed in HepG2 cells. Suppression of Grb2 in HepG2 significantly reduced fat accumulation, improved
glucose metabolism, ameliorated oxidative stress, and restored the activity of
insulin receptor substrate-1/Akt and MEK/ERK pathways. In addition, Grb2 deficiency attenuated hepatic apoptosis shown by reduced activation of
caspase-3 and fluorescent staining. Modulation of Bcl-2 and Bak1 also contributed to reduced apoptosis. In conclusion, suppression of Grb2 expression in HepG2 cells improved hepatic steatosis,
glucose metabolism, oxidative stress, and apoptosis induced by
palmitic acid incubation partly though modulating the
insulin signaling pathway.