Reactive oxygen species are involved in insulin-dependent regulation of autophagy in primary rat podocytes

Int J Biochem Cell Biol. 2016 Jun:75:23-33. doi: 10.1016/j.biocel.2016.03.015. Epub 2016 Mar 26.

Abstract

Autophagy is an intracellular defense mechanism responsible for the turnover of damaged or non-functional cellular constituents. This process provides cells with energy and essential compounds under unfavorable environmental conditions-such as oxidative stress and hyperglycemia, which are both observed in diabetes. The most common diabetes complication is diabetic nephropathy (DN), which can lead to renal failure. This condition often includes impaired podocyte function. Here we investigated autophagic activity in rat podocytes cultured with a high insulin concentration (300nM). Autophagy was activated after 60min of insulin stimulation. Moreover, this effect was abolished following pharmacological (apocynin) or genetic (siRNA) inhibition of NAD(P)H oxidase activity, indicating that insulin-dependent autophagy stimulation involved reactive oxygen species (ROS). We also observed a continuous and time-dependent increase of podocyte albumin permeability in response to insulin, and this process was slightly improved by autophagy inhibition following short-term insulin exposure. Our results suggest that insulin may be a factor affecting the development of diabetic nephropathy.

Keywords: Autophagy; Filtration barrier permeability; Insulin; Podocyte; Reactive oxygen species.

MeSH terms

  • Acetophenones / pharmacology
  • Albumins / metabolism
  • Animals
  • Autophagy / drug effects*
  • Autophagy-Related Protein 12 / metabolism
  • Autophagy-Related Protein 5 / metabolism
  • Gene Expression Regulation / drug effects
  • Gene Knockdown Techniques
  • Insulin / pharmacology*
  • Microtubule-Associated Proteins / metabolism
  • NADPH Oxidase 4
  • NADPH Oxidases / deficiency
  • NADPH Oxidases / genetics
  • Permeability / drug effects
  • Podocytes / cytology*
  • Podocytes / drug effects
  • Podocytes / metabolism*
  • Rats
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism*

Substances

  • Acetophenones
  • Albumins
  • Autophagy-Related Protein 12
  • Autophagy-Related Protein 5
  • Insulin
  • LC3 protein, rat
  • Microtubule-Associated Proteins
  • Reactive Oxygen Species
  • acetovanillone
  • NADPH Oxidase 4
  • NADPH Oxidases
  • Nox4 protein, rat