Involvement of NADPH-oxidase enzyme in the nephroprotective effect of (-)-α-bisabolol on HK2 cells exposed to ischemia - Reoxygenation

Eur J Pharmacol. 2019 Jul 15:855:1-9. doi: 10.1016/j.ejphar.2019.04.044. Epub 2019 Apr 29.

Abstract

Acute Kidney Injury (AKI) is associated with high morbidity and mortality. Ischemia and reperfusion (I/R) are events that lead to AKI through hypoxia, reactive oxygen species (ROS) production, oxidative stress and apoptosis. We aimed to evaluate the mechanism of nephroprotection mediated by Bisabolol in human tubular kidney cells after injury by I/R in vitro. HK2 cells were exposed to I/R and treated with Bisabolol. Cell viability was accessed by MTT assay. Cells were submitted to flow cytometry to evaluate necrotic/apoptotic cells, reactive oxygen species production and mitochondrial transmembrane depolarization. TBARS and GSH were used as parameters of redox balance. Also, KIM-1 supernatant levels were measured. In order to identify an interaction between bisabolol and NOX4, molecular docking and enzymatic assays were performed. Expression of isoform NOX4 on treated cells was examined by western-blot. Finally, cells were visualized by scanning electron microscopy. Bisabolol improved cell viability and prevented cell death by apoptosis, indicated also by the decreased levels of KIM-1. It was observed a decrease on reactive oxygen species production and mitochondrial depolarization, with antioxidant regulation by increased GSH and decreased lipid peroxidation. It was also demonstrated that bisabolol treatment can inhibit NOX4. Finally, SEM images showed that bisabolol reduced I/R-induced cell damage. Bisabolol treatment protects HK2 cells against oxidative damage occasioned by I/R. This effect is related to inhibition of apoptosis, decrease on KIM-1 release, reactive oxygen species accumulation and mitochondrial dysfunction. Bisabolol inhibited NOX4 activity in the tubular cells, impairing reactive oxygen species synthesis.

Keywords: Bisabolol; Kidney ischemia/reoxygenation; NADPH oxidase.

MeSH terms

  • Apoptosis / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Cytoprotection / drug effects
  • Glutathione / metabolism
  • Hepatitis A Virus Cellular Receptor 1 / metabolism
  • Humans
  • Kidney Tubules / drug effects*
  • Kidney Tubules / metabolism
  • Kidney Tubules / pathology*
  • Monocyclic Sesquiterpenes / pharmacology*
  • NADPH Oxidases / metabolism*
  • Oxidative Stress / drug effects
  • Oxygen / metabolism*
  • Reactive Oxygen Species / metabolism
  • Reperfusion Injury / metabolism
  • Reperfusion Injury / prevention & control*
  • Thiobarbituric Acid Reactive Substances / metabolism

Substances

  • HAVCR1 protein, human
  • Hepatitis A Virus Cellular Receptor 1
  • Monocyclic Sesquiterpenes
  • Reactive Oxygen Species
  • Thiobarbituric Acid Reactive Substances
  • bisabolol
  • NADPH Oxidases
  • Glutathione
  • Oxygen