BV-2 Microglial Cells Respond to Rotenone Toxic Insult by Modifying Pregnenolone, 5α-Dihydroprogesterone and Pregnanolone Levels

Cells. 2020 Sep 13;9(9):2091. doi: 10.3390/cells9092091.


Neuroinflammation, whose distinctive sign is the activation of microglia, is supposed to play a key role in the development and progression of neurodegenerative diseases. The aim of this investigation was to determine levels of neurosteroids produced by resting and injured BV-2 microglial cells. BV-2 cells were exposed to increasing concentrations of rotenone to progressively reduce their viability by increasing reactive oxygen species (ROS) production. BV-2 cell viability was significantly reduced 24, 48 and 72 h after rotenone (50-1000 nM) exposure. Concomitantly, rotenone (50-100 nM) determined a dose-independent augmentation of ROS production. Then, BV-2 cells were exposed to a single, threshold dose of rotenone (75 nM) to evaluate the overtime release of neurosteroids. In particular, pregnenolone, pregnenolone sulfate, progesterone, 5α-dihydroprogesterone (5α-DHP), allopregnanolone, and pregnanolone, were quantified in the culture medium by liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis. BV-2 cells synthesized all the investigated neurosteroids and, after exposure to rotenone, 5αDHP and pregnanolone production was remarkably increased. In conclusion, we found that BV-2 cells not only synthesize several neurosteroids, but further increase this production following oxidative damage. Pregnanolone and 5α-DHP may play a role in modifying the progression of neuroinflammation in neurodegenerative diseases.

Keywords: BV-2 cells; ROS; microglia; neurodegeneration; neuroinflammation; neurosteroids; rotenone; steroidogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5-alpha-Dihydroprogesterone / metabolism*
  • Animals
  • Cell Line, Transformed
  • Cell Survival / drug effects
  • Chromatography, Liquid
  • Mice
  • Microglia / drug effects*
  • Microglia / metabolism*
  • Pregnanolone / metabolism*
  • Pregnenolone / metabolism*
  • Reactive Oxygen Species / metabolism
  • Rotenone / toxicity*
  • Signal Transduction / drug effects*
  • Tandem Mass Spectrometry


  • Reactive Oxygen Species
  • Rotenone
  • Pregnenolone
  • 5-alpha-Dihydroprogesterone
  • Pregnanolone