An analysis of high glucose and glucosamine-induced gene expression and oxidative stress in renal mesangial cells

AbstractRenal mesangial cells play an important role in the development of diabetic kidney disease. We have previously demonstrated that some of the effects of high glucose on mesangial extracellular matrix (ECM) protein expression are mediated by the hexosamine biosynthesis pathway (HBP) in which fructose-6-phosphate is converted to glucosamine-6-phosphate by the rate-limiting enzyme glutamine:fructose-6-phosphate amidotransferase (GFAT). Using Affymetrix murine expression U430 2.0 oligochips, we examined the global effects of high glucose (HG) and glucosamine (GlcN) on mRNA expression of a mouse mesangial cell line (MES-13). We sought to determine the portion of mRNA expression in MES-13 cells, which is mediated both by high glucose and glucosamine, i.e., via the HBP. Of the 34,000 genes on the chip, ∼55.7 – 60.8% genes are detected in MES-13 cells. Culturing MES-13 cells for 48 h with HG alters the expression of ∼389 genes at our preset threshold levels (at least 2-fold change) where 263 genes are up-r...