Neuroprotective effect of clavulanic acid on trimethyltin (TMT)-induced cytotoxicity in PC12 cells

Drug Chem Toxicol. 2019 Mar;42(2):187-193. doi: 10.1080/01480545.2018.1468772. Epub 2018 May 15.

Abstract

Trimethyltin (TMT) is a short-chain trialkyltin with various applications in industry. In addition, it is a known neurotoxin, producing significant and selective neurodegeneration in the limbic system of both human and animals. Recently, effect of clavulanic acid (CA) in nervous system has been mentioned. Therefore, in this study, the role of CA in TMT-induced toxicity in PC12 cells was evaluated. For this study, PC12 cells were cultured and exposed to different concentrations of CA for 24 h. Then, TMT (20 µM) was added to cells. After that, MTT test was performed to assay cytotoxicity. Reactive oxygen species production (ROS) was determined using 2,7-dichlorofluorescein diacetate (DCFH-DA) method. Additionally, the levels of Bax, Bcl-2, caspase-3, CERB and p-CREB proteins were evaluated using Western blot analysis. The exposure of PC12 cells to TMT reduced cell viability, increased intracellular ROS production, elevated Bax/Bcl-2 ratio and enhanced the expression of caspase-3 (Pro and cleaved forms) protein. Pretreatment of cells with CA before TMT, significantly reduced ROS generation, diminished upregulation of proapoptotic Bax protein and attenuated caspase-3 protein expression. In conclusion, CA exhibited significant neuroprotective effects against neurotoxicity of TMT mainly throughout reduction of ROS production and regulation of proteins, which are involved in apoptosis pathway.

Keywords: Trimethyltin; apoptosis; clavulanic acid; cytotoxicity.

MeSH terms

  • Animals
  • Blotting, Western
  • Caspase 3 / metabolism
  • Cell Survival / drug effects
  • Clavulanic Acid / pharmacology*
  • Dose-Response Relationship, Drug
  • Neuroprotective Agents / pharmacology*
  • Neurotoxins / antagonists & inhibitors
  • Neurotoxins / pharmacology*
  • PC12 Cells / drug effects*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Rats
  • Reactive Oxygen Species / metabolism
  • Trimethyltin Compounds / antagonists & inhibitors
  • Trimethyltin Compounds / pharmacology*
  • bcl-2-Associated X Protein / metabolism

Substances

  • Bax protein, rat
  • Neuroprotective Agents
  • Neurotoxins
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • Trimethyltin Compounds
  • bcl-2-Associated X Protein
  • trimethyltin
  • Clavulanic Acid
  • Caspase 3