Antiparasitic effect of (-)-α-bisabolol against Trypanosoma cruzi Y strain forms

Diagn Microbiol Infect Dis. 2019 Nov;95(3):114860. doi: 10.1016/j.diagmicrobio.2019.06.012. Epub 2019 Jun 28.

Abstract

Chagas disease is caused by Trypanosoma cruzi and affects about 7 million people worldwide. Benznidazole and nifurtimox have low efficacy and high toxicity. The present study was designed to identify the trypanocidal effect of (-)-α-Bisabolol (BIS) and investigate its mechanism. Epimastigotes and trypomastigotes were cultured with BIS and the viable cells were counted. BIS antiamastigote effect was evaluated using infected LLC-MK2 cells. MTT assay was performed to evaluate BIS cytotoxicity. Growth recovery was assessed to evaluate BIS effect after short times of exposure. BIS mechanism was investigated through flow cytometry, with 7-AAD and Annexin V-PE. DCFH-DA, rhodamine 123 (Rho123) and acridine orange (AO). Finally, enzymatic and computational assays were performed to identify BIS interaction with T. cruzi GAPDH (tcGAPDH). BIS showed an inhibitory effect on epimastigotes after all tested periods, as well on trypomastigotes. It caused cytotoxicity on LLC-MK2 cells at higher concentrations, with selectivity index (SeI) = 26.5. After treatment, infected cells showed a decrease in infected cells, the number of amastigotes per infected cell and the survival index (SuI). Growth recovery demonstrated that BIS effect causes rapid death of T. cruzi. Flow cytometry showed that BIS biological effect is associated with apoptosis induction, increase in cytoplasmic ROS and mitochondrial transmembrane potential, while reservosome swelling was observed at a late stage. Also, BIS action mechanism may be associated to tcGAPDH inhibition. Altogether, the results demonstrate that BIS causes cell death in Trypanosoma cruzi Y strain forms, with the involvement of apoptosis and oxidative stress and enzymatic inhibition.

Keywords: Apoptosis; Bisabolol; Trypanosoma cruzi.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Chagas Disease / drug therapy
  • Chagas Disease / parasitology
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / antagonists & inhibitors
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism
  • Inhibitory Concentration 50
  • Macaca mulatta
  • Molecular Docking Simulation
  • Molecular Structure
  • Monocyclic Sesquiterpenes / chemistry
  • Monocyclic Sesquiterpenes / pharmacology*
  • Oxidative Stress / drug effects
  • Trypanocidal Agents / pharmacology*
  • Trypanosoma cruzi / drug effects*
  • Trypanosoma cruzi / physiology

Substances

  • Monocyclic Sesquiterpenes
  • Trypanocidal Agents
  • bisabolol
  • Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)