Consequences of tributyltin chloride induced stress in Leydig cells: an ex-vivo approach

Environ Toxicol Pharmacol. 2014 Mar;37(2):850-60. doi: 10.1016/j.etap.2014.02.018. Epub 2014 Mar 3.

Abstract

Tributyltin (TBT), a member of the organotin family, is a known endocrine disruptor. It persists long in the environment and is widely used in various industrial applications. This study was planned to understand its toxic influence on Leydig cells isolated from 28 day old wistar rats. In-vitro exposure to TBT-Chloride (TBTC) (300-3000 nM) reduced cell viability (DNA fragmentation, nuclear condensation and MTT assay) and affected testosterone production. TBTC induced both apoptotic and necrotic cell death (AnnexinV/PI binding assay). Involvement of calcium (Ca(2+)), redox imbalance (ROS, GSH and TBARS) and mitochondria in TBTC toxicity was evaluated by using Ca(2+) inhibitors (BAPTA-AM, EGTA, Ruthenium Red), free radical scavengers (NAC, C-Phycocyanin) and mitochondrial permeability transition pore inhibitor (Cyclosporine A). Protein expression analysis of phosphorylated MAPKinases (ERK1/2, JNK1/2, & p38), steroidogenic proteins (3β-HSD, StAR & TSPO) and apoptotic proteins (Bax, Bcl2) illustrates the cytotoxic and anti-steroidogenic activity of TBTC.

Keywords: Leydig; Oxidative stress; Steroidogenesis; Testosterone; Tributyltin chloride.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Hydroxysteroid Dehydrogenases / metabolism
  • Animals
  • Apoptosis / drug effects
  • Calcium / metabolism
  • Carrier Proteins / metabolism
  • Cell Survival / drug effects
  • Cells, Cultured
  • Endocrine Disruptors / toxicity*
  • Environmental Pollutants / toxicity*
  • Glutathione / metabolism
  • Leydig Cells / drug effects*
  • Leydig Cells / metabolism
  • Lipid Peroxidation / drug effects
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Mitogen-Activated Protein Kinases / metabolism
  • NF-E2-Related Factor 2 / metabolism
  • Oxidative Stress
  • Phosphoproteins / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism
  • Receptors, GABA-A / metabolism
  • Testosterone / metabolism
  • Trialkyltin Compounds / toxicity*
  • bcl-2-Associated X Protein / metabolism

Substances

  • Bax protein, rat
  • Carrier Proteins
  • Endocrine Disruptors
  • Environmental Pollutants
  • NF-E2-Related Factor 2
  • Nfe2l2 protein, rat
  • Phosphoproteins
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • Receptors, GABA-A
  • Trialkyltin Compounds
  • bcl-2-Associated X Protein
  • steroidogenic acute regulatory protein
  • Tspo protein, rat
  • Testosterone
  • tributyltin
  • 3-Hydroxysteroid Dehydrogenases
  • Mitogen-Activated Protein Kinases
  • Glutathione
  • Calcium