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  • Oxidative stress damage in the protozoan parasite Trypanosoma cruzi is inhibited by Cyclosporin A.

Oxidative stress damage in the protozoan parasite Trypanosoma cruzi is inhibited by Cyclosporin A.

Parasitology (2015-04-01)
Patricia L Bustos, Alina E Perrone, Natalia Milduberger, Miriam Postan, Jacqueline Bua
ABSTRACT

Cyclosporin A (CsA) specifically inhibits the mitochondrial permeability transition pore (mPTP). Opening of the mPTP, which is triggered by high levels of matrix [Ca2+] and/or oxidative stress, leads to mitochondrial dysfunction and thus to cell death by either apoptosis or necrosis. In the present study, we analysed the response of Trypanosoma cruzi epimastigote parasites to oxidative stress with 5 mm H2O2, by studying several features related to programmed cell death and the effects of pre-incubation with 1 μ m of CsA. We evaluated TcPARP cleavage, DNA integrity, cytochrome c translocation, Annexin V/propidium iodide staining, reactive oxygen species production. CsA prevented parasite oxidative stress damage as it significantly inhibited DNA degradation, cytochrome c translocation to cytosol and TcPARP cleavage. The calcein-AM/CoCl2 assay, used as a selective indicator of mPTP opening in mammals, was also performed in T. cruzi parasites. H2O2 treatment decreased calcein fluorescence, but this decline was partially inhibited by pre-incubation with CsA. Our results encourage further studies to investigate if there is a mPTP-like pore and a mitochondrial cyclophilin involved in this protozoan parasite.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MOPS, ≥99.5% (titration)
Sigma-Aldrich
MOPS, BioPerformance Certified, suitable for cell culture, ≥99.5% (titration)
Sigma-Aldrich
Cyclosporin A, 98.5-101.5% (on dried basis)
Sigma-Aldrich
Fluorescein diacetate, used as cell viability stain
Sigma-Aldrich
Calcein-AM, BioReagent, suitable for fluorescence, ≥95.0% (HPLC)
Sigma-Aldrich
Calcein AM solution, 4 mM in DMSO, ≥90% (HPLC), solution
Sigma-Aldrich
Cyclosporin A, from Tolypocladium inflatum, ≥95% (HPLC), solid
Sigma-Aldrich
Calcein-AM, Small Package (20 X 50 μg ), ≥95.0% (HPLC)
Sigma-Aldrich
Cyclosporin A, BioReagent, from Tolypocladium inflatum, for molecular biology, ≥95%
Sigma-Aldrich
MOPS, BioXtra, ≥99.5% (titration)
SAFC
MOPS
Sigma-Aldrich
MOPS, anhydrous, free-flowing, Redi-Dri, ≥99.5%