myo-Inositol Oxygenase Overexpression Accentuates Generation of Reactive Oxygen Species and Exacerbates Cellular Injury following High Glucose Ambience: A NEW MECHANISM RELEVANT TO THE PATHOGENESIS OF DIABETIC NEPHROPATHY

J Biol Chem. 2016 Mar 11;291(11):5688-5707. doi: 10.1074/jbc.M115.669952. Epub 2016 Jan 20.

Abstract

Diabetic nephropathy (DN) is characterized by perturbations in metabolic/cellular signaling pathways with generation of reactive oxygen species (ROS). The ROS are regarded as a common denominator of various pathways, and they inflict injury on renal glomerular cells. Recent studies indicate that tubular pathobiology also plays a role in the progression of DN. However, the mechanism(s) for how high (25 mm) glucose (HG) ambience induces tubular damage remains enigmatic. myo-Inositol oxygenase (MIOX) is a tubular enzyme that catabolizes myo-inositol to d-glucuronate via the glucuronate-xylulose (G-X) pathway. In this study, we demonstrated that G-X pathway enzymes are expressed in the kidney, and MIOX expression/bioactivity was up-regulated under HG ambience in LLC-PK1 cells, a tubular cell line. We further investigated whether MIOX overexpression leads to accentuation of tubulo-interstitial injury, as gauged by some of the parameters relevant to the progression of DN. Under HG ambience, MIOX overexpression accentuated redox imbalance, perturbed NAD(+)/NADH ratios, increased ROS generation, depleted reduced glutathione, reduced GSH/GSSG ratio, and enhanced adaptive changes in the profile of the antioxidant defense system. These changes were also accompanied by mitochondrial dysfunctions, DNA damage and induction of apoptosis, accentuated activity of profibrogenic cytokine, and expression of fibronectin, the latter two being the major hallmarks of DN. These perturbations were largely blocked by various ROS inhibitors (Mito Q, diphenyleneiodonium chloride, and N-acetylcysteine) and MIOX/NOX4 siRNA. In conclusion, this study highlights a novel mechanism where MIOX under HG ambience exacerbates renal injury during the progression of diabetic nephropathy following the generation of excessive ROS via an unexplored G-X pathway.

Keywords: diabetes; diabetic nephropathy; extracellular matrix; kidney; myo-inositol oxygenase (MIOX); reactive oxygen species (ROS).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Animals
  • Diabetic Nephropathies / genetics
  • Diabetic Nephropathies / metabolism
  • Diabetic Nephropathies / pathology*
  • Glucose / metabolism*
  • Hydrogen Peroxide / metabolism
  • Inositol Oxygenase / analysis
  • Inositol Oxygenase / genetics
  • Inositol Oxygenase / metabolism*
  • Kidney / metabolism
  • Kidney / pathology*
  • LLC-PK1 Cells
  • Male
  • Mice
  • Mitochondria / genetics
  • Mitochondria / metabolism
  • Mitochondria / pathology
  • NAD / metabolism
  • Oxidative Stress
  • Reactive Oxygen Species / metabolism*
  • Swine
  • Up-Regulation

Substances

  • Reactive Oxygen Species
  • NAD
  • Hydrogen Peroxide
  • Inositol Oxygenase
  • Glucose