Fig 1.
Hyperglycemic HepG2 cells exhibit lipid accumulation.
A. H&E histology reveals successive passaging of HepG2 cells in hyperglycemic conditions show progressive phenotypic changes as compared with normoglycemic HepG2 cells. B. Brightfield microscopy also reveal these phenotypic changes within hyperglycemic HepG2 cells as compared with normoglycemic cells. C. Representative cultured cells examined at the ultrastructural level by electron microscopy demonstrate accumulations of numerous lipid vacuoles (red arrows) and water influx (clear cytoplasmic inclusion) in hyperglycemic HepG2 cells in contrast to cells grown in normoglycemic culture media.
Fig 2.
Nile red staining for triglycerides is elevated in hyperglycemic HepG2 cells.
A. Fluorescent imaging reveals increased Nile red staining in hyperglycemic HepG2 cells as compared with normoglycemic cells. B. Brightfield microscopy with fluorescent overlay reveals the accumulation of lipid within hyperglycemic HepG2 cells as compared with normoglycemic cells. C. Quantification of Nile red fluorescence reveals elevated levels in hyperglycemic HepG2 cells as compared with normoglycemic cells. Data presented indicate the mean ± SEM (n = 3 wells per group). *p<0.05 vs. normoglycemic group.
Fig 3.
qPCR analysis shows trends in hyperglycemic HepG2 cells consistent with NAFLD.
A. CEACAM1 gene expression is depressed in hyperglycemic HepG2 cells as compared with normoglycemic cells. B. CD36 gene expression is elevated in hyperglycemic HepG2 cells as compared with normoglycemic cells. C. ALT and ALP gene expression is elevated in hyperglycemic HepG2 cells as compared with normoglycemic cells. Data presented indicate the mean ± SEM (n = 3 per group). **p<0.01 vs. normoglycemic group.
Table 1.
Hepatotoxicity gene array reveals regulation of key genes found in NAFLD.
Fig 4.
qPCR analysis shows trends in hyperglycemic HepG2 cells consistent with insulin resistance.
A. PCK1 and G6PC gene expression is elevated in hyperglycemic HepG2 cells compared with normoglycemic HepG2 cells. B. GLUT2 and C. PON1 gene expression is depressed in hyperglycemic HepG2 cells compared with normoglycemic HepG2 cells. Data presented indicate the mean ± SEM (n = 3 per group). *p<0.05, ***p<0.001, ****p<0.0001 vs. normoglycemic group.
Fig 5.
MTT and LDH assay for HepG2 cells reveal no significant differences between cell conditions.
A. MTT assay demonstrated no differences in mitochondrial function between hyperglycemic and normoglycemic HepG2 cells. B. LDH assay demonstrated no differences in cytotoxicity between hyperglycemic and normoglycemic HepG2 cells.
Fig 6.
Oil Red O staining is elevated in livers of Leprdb/J mice.
Livers from Leprdb/J mice revealed increased Oil Red O staining as compared with livers from WT C57 mice.
Fig 7.
qPCR analysis shows trends in Leprdb/J mice consistent with NAFLD.
A. CEACAM1 gene expression is depressed in Leprdb/J mouse livers as compared with WT C57 mouse livers B. CD36 gene expression is elevated in Leprdb/J mouse livers as compared with WT C57 mouse livers C. ALT and ALP gene expression is elevated in Leprdb/J mouse livers as compared with WT C57 mouse livers. Data presented indicate the mean ± SEM (n = 5–6 per group). **p<0.01, ***p<0.001, and ****p<0.0001 vs. WT C57 group.