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Mitochondria-targeted antioxidant SkQR1 reduces TNF-induced endothelial permeability in vitro

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Abstract

Prolonged or excessive increase in the circulatory level of proinflammatory tumor necrosis factor (TNF) leads to abnormal activation and subsequent damage to endothelium. TNF at high concentrations causes apoptosis of endothelial cells. Previously, using mitochondria-targeted antioxidants of SkQ family, we have shown that apoptosis of endothelial cells is dependent on the production of reactive oxygen species (ROS) in mitochondria (mito-ROS). Now we have found that TNF at low concentrations does not cause cell death but activates caspase-3 and caspase-dependent increase in endothelial permeability in vitro. This effect is probably due to the cleavage of β-catenin–an adherent junction protein localized in the cytoplasm. We have also shown that extracellular matrix metalloprotease 9 (MMP9) VE-cadherin shedding plays a major role in the TNF-induced endothelial permeability. The mechanisms of the caspase-3 and MMP9 activation are probably not related to each other since caspase inhibition did not affect VE-cadherin cleavage and MMP9 inhibition had no effect on the caspase-3 activation. Mitochondria-targeted antioxidant SkQR1 inhibited TNF-induced increase in endothelial permeability. SkQR1 also inhibited caspase-3 activation, β-catenin cleavage, and MMP9-dependent VE-cadherin shedding. The data suggest that mito-ROS are involved in the increase in endothelial permeability due to the activation of both caspase-dependent cleavage of intracellular proteins and of MMP9-dependent cleavage of the transmembrane cell-to-cell contact proteins.

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Abbreviations

Ac-DEVD-MCA:

L-acetyl-Asp-Glu-Val-Aspα-(4-methylcoumaryl-7-amide

DEVD:

N-carbobenzyloxyAsp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethyl ketone

MMP:

matrix metalloprotease

NAC:

N-acetyl-D-cysteine

PARP:

poly(ADP-ribose) polymerase

ROS:

reactive oxygen species

SkQ:

conjugates of plastoquinone and penetrating cations

SkQ1:

plastoquinonyl-10(6′-decyltriphenyl)phosphonium

SkQR1:

10-(6′-plastoquinonyl)decylrhodamine 19

TNF:

tumor necrosis factor

ZVAD:

N-benzyloxycarbonyl-ValAla-Asp-trifluoromethyl

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Authors and Affiliations

  1. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    I. I. Galkin, O. Yu. Pletjushkina, R. A. Zinovkin, V. V. Zakharova, B. V. Chernyak & E. N. Popova

  2. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    R. A. Zinovkin

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  1. I. I. Galkin
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  2. O. Yu. Pletjushkina
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  3. R. A. Zinovkin
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  4. V. V. Zakharova
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  5. B. V. Chernyak
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  6. E. N. Popova
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Correspondence to I. I. Galkin.

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Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM16-170, September 12, 2016.

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Galkin, I.I., Pletjushkina, O.Y., Zinovkin, R.A. et al. Mitochondria-targeted antioxidant SkQR1 reduces TNF-induced endothelial permeability in vitro . Biochemistry Moscow 81, 1188–1197 (2016). https://doi.org/10.1134/S0006297916100163

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  • DOI: https://doi.org/10.1134/S0006297916100163

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