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Isoespintanol, a monoterpene isolated from oxandra cf xylopioides, ameliorates the myocardial ischemia-reperfusion injury by AKT/PKCε/eNOS-dependent pathways

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Abstract

Purpose

To determine the actions of isoespintanol (Isoesp) on post-ischemic myocardial and mitochondrial alterations.

Methods

Hearts removed from Wistar rats were perfused by 20 min. After this period, the coronary flow was interrupted by half an hour and re-established during 1 h. In the treated group, Isoesp was administered at the beginning of reperfusion. To assess the participation of ε isoform of protein kinase C (PKCε), protein kinase B (PKB/Akt), and nitric oxide synthase (NOS), hearts were treated with Isoesp plus the respective inhibitors (chelerythrine, wortmannin, and N-nitro-l-arginine methyl ester). Cell death was determined by triphenyl tetrazolium chloride staining technique. Post-ischemic recovery of contractility, oxidative stress, and content of phosphorylated forms of PKCε, Akt, and eNOS were also examined. Mitochondrial state was assessed through the measurement of calcium-mediated response, calcium retention capacity, and mitochondrial potential.

Results

Isoesp limited cell death, decreased post-ischemic dysfunction and oxidative stress, improved mitochondrial state, and increased the expression of PKCε, Akt, and eNOS phosphorylated. All these beneficial effects achieved by Isoesp were annulled by the inhibitors.

Conclusion

These findings suggest that activation of Akt/eNOS and PKCε signaling pathways are involved in the development of Isoesp-induced cardiac and mitochondria tolerance to ischemia-reperfusion.

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Abbreviations

Isoesp:

Isoespintanol

NOS:

Nitric oxide synthase

l-NAME:

l-NG-Nitroarginine methyl ester

PKCε:

Protein kinase C ε

Che:

Chelerythrine

Akt:

Protein kinase B

Wort:

Wortmannin

DMSO:

Dimethyl sulfoxide

CR:

Coronary resistance

CPP:

Coronary perfusion pressure

CF:

Coronary flow

GSH:

Reduced glutathione

TBARS:

Thiobarbituric acid reactive substances

LSD:

Light scattering decrease

CRC:

Ca2+ retention capacity

ΔΨ:

Mitochondrial membrane potential

mPTP:

Mitochondrial permeability transition pore

TTC:

Triphenyltetrazolium chloride

LVDP:

Left ventricular developed pressure

+dP/dtmax :

Maximal velocity of rise of left ventricular pressure

LVEDP:

Left ventricular end diastolic pressure

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

ROS:

Radical oxygen species

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Funding

This work was supported by the Grant M-203 from the National University of La Plata, Argentina, to Dr. S. Mosca.

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Authors and Affiliations

  1. Centro de Investigaciones Cardiovasculares ¨Dr Horacio E. Cingolani¨, CCT-CONICET, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, 60 y 120, 1900, La Plata, Argentina

    Luisa F. González Arbeláez, Alejandro Ciocci Pardo, Juliana C Fantinelli & Susana M Mosca

  2. Laboratorio de Ciencias de los Alimentos, Facultad de Ciencias, Universidad Nacional de Colombia (sede Medellín), Bogotá, Colombia

    Benjamín Rojano

  3. Facultad de Ciencias Médicas, Universidad Nacional de La Plata CIC-PBA, La Plata, Argentina

    Guillermo R Schinella

Authors
  1. Luisa F. González Arbeláez
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  3. Juliana C Fantinelli
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  4. Benjamín Rojano
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  5. Guillermo R Schinella
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  6. Susana M Mosca
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Contributions

L.G.A. performed the western blots; A.C.P. performed the mitochondrial experiments; J.F. performed the isolated hearts experiments; B.J. isolated and provided the drug; G.Sch. contributed to the conception of the study; S.M. wrote the manuscript. All the authors contributed to the analysis and interpretation of data, critically reviewed, and approved the final draft.

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Correspondence to Susana M Mosca.

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González Arbeláez, L.F., Ciocci Pardo, A., Fantinelli, J.C. et al. Isoespintanol, a monoterpene isolated from oxandra cf xylopioides, ameliorates the myocardial ischemia-reperfusion injury by AKT/PKCε/eNOS-dependent pathways. Naunyn-Schmiedeberg's Arch Pharmacol 393, 629–638 (2020). https://doi.org/10.1007/s00210-019-01761-9

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