title: RAGE-induced cytosolic ROS promote mitochondrial superoxide generation in diabetes.
abstract: Damaged mitochondria generate an excess of superoxide, which may mediate tissue injury in diabetes. We hypothesized that in diabetic nephropathy, advanced glycation end-products (AGEs) lead to increases in cytosolic reactive oxygen species (ROS), which facilitate the production of mitochondrial superoxide. In normoglycemic conditions, exposure of primary renal cells to AGEs, transient overexpression of the receptor for AGEs (RAGE) with an adenoviral vector, and infusion of AGEs to healthy rodents each induced renal cytosolic oxidative stress, which led to mitochondrial permeability transition and deficiency of mitochondrial complex I. Because of a lack of glucose-derived NADH, which is the substrate for complex I, these changes did not lead to excess production of mitochondrial superoxide; however, when we performed these experiments in hyperglycemic conditions in vitro or in diabetic rats, we observed significant generation of mitochondrial superoxide at the level of complex I, fueled by a sustained supply of NADH. Pharmacologic inhibition of AGE-RAGE-induced mitochondrial permeability transition in vitro abrogated production of mitochondrial superoxide; we observed a similar effect in vivo after inhibiting cytosolic ROS production with apocynin or lowering AGEs with alagebrium. Furthermore, RAGE deficiency prevented diabetes-induced increases in renal mitochondrial superoxide and renal cortical apoptosis in mice. Taken together, these studies suggest that AGE-RAGE-induced cytosolic ROS production facilitates mitochondrial superoxide production in hyperglycemic environments, providing further evidence of a role for the advanced glycation pathway in the development and progression of diabetic nephropathy.
authors.affiliation: Juvenile Diabetes Research Foundation Einstein Centre for Diabetes Complications, Division of Diabetes Complications, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia. melinda.coughlan@bakeridi.edu.au
authors.forename: Melinda T
authors.initials: MT
authors.lastname: Coughlan
authors.forename: David R
authors.initials: DR
authors.lastname: Thorburn
authors.forename: Sally A
authors.initials: SA
authors.lastname: Penfold
authors.forename: Adrienne
authors.initials: A
authors.lastname: Laskowski
authors.forename: Brooke E
authors.initials: BE
authors.lastname: Harcourt
authors.forename: Karly C
authors.initials: KC
authors.lastname: Sourris
authors.forename: Adeline L Y
authors.initials: AL
authors.lastname: Tan
authors.forename: Kei
authors.initials: K
authors.lastname: Fukami
authors.forename: Vicki
authors.initials: V
authors.lastname: Thallas-Bonke
authors.forename: Peter P
authors.initials: PP
authors.lastname: Nawroth
authors.forename: Michael
authors.initials: M
authors.lastname: Brownlee
authors.forename: Angelika
authors.initials: A
authors.lastname: Bierhaus
authors.forename: Mark E
authors.initials: ME
authors.lastname: Cooper
authors.forename: Josephine M
authors.initials: JM
authors.lastname: Forbes
mesh_terms: D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000503:physiopathology*; D004579:Electron Transport; D005260:Female; D017127:Glycation End Products, Advanced / Q000502:physiology*; D007668:Kidney / Q000503:physiopathology; D007672:Kidney Cortex / Q000503:physiopathology; D008928:Mitochondria / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000502:physiology*; D013481:Superoxides / Q000378:metabolism*
chemical_list: D017127:Glycation End Products, Advanced; D017382:Reactive Oxygen Species; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D013481:Superoxides