title: Increased expression of mitochondrial DNA-encoded genes in human renal mesangial cells in response to high glucose-induced reactive oxygen species.
abstract: Reactive oxygen species (ROS)-mediated disruption of mitochondrial respiratory function has been implicated in the complications of diabetes. The present study examined changes in the gene expression of mitochondrial DNA (mtDNA)-encoded subunits of electron transport chain complexes in response to high glucose-induced ROS overproduction in an in vitro model of diabetic nephropathy using human renal mesangial cells. Mitochondrial ROS generation was assessed by confocal microscopy and flow cytometry in the cells following culture in 5 and 25 mM glucose. The mRNA expression levels of nicotinamide adenine dinucleotide dehydrogenase 2 (ND2) of complex I, cytochrome b (CYTB) of complex III, cytochrome c oxidase (COI) of complex IV and ATPase 6 of complex V were analyzed by reverse transcription-quantitative polymerase chain reaction. The protein expression levels of ND2, CYTB, COI and ATPase 6 were analyzed by western blotting. A significant increase in mitochondrial ROS production was observed in the cells cultured in 25 mM glucose, compared with cells cultured in 5 mM glucose (P<0.05). The mRNA expression of ND2, CYTB, CO1 and ATPase 6 was significantly increased following culture in 25 mM glucose, compared with the cells cultured in 5 mM glucose (P<0.05). This increase in mRNA expression was accompanied by significant increases in protein expression following incubation in 25 mM glucose (P<0.05). The increase in mtDNA-encoded gene expression in the electron transport subunits following exposure to high glucose-induced ROS may be a compensatory response mechanism for the decline in mitochondrial function, which may be important in the development of diabetic nephropathy through enhanced ROS generation.
authors.affiliation: Department of Molecular Medicine, College of Medicine and Medical Sciences, Arabian Gulf University, Manama 26671, Bahrain.
authors.forename: Ghada
authors.initials: G
authors.lastname: Al-Kafaji
authors.affiliation: Department of Biochemistry, College of Medicine and Medical Sciences, Arabian Gulf University, Manama 26671, Bahrain.
authors.forename: Mohamed Abdalla
authors.initials: MA
authors.lastname: Sabry
authors.affiliation: Department of Molecular Medicine, College of Medicine and Medical Sciences, Arabian Gulf University, Manama 26671, Bahrain.
authors.forename: Moiz
authors.initials: M
authors.lastname: Bakhiet
mesh_terms: D004272:DNA, Mitochondrial / Q000235:genetics*; D004579:Electron Transport / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D050527:Mesangial Cells / Q000187:drug effects / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D004272:DNA, Mitochondrial; D021122:Protein Subunits; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D005947:Glucose