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P2X7R antagonism after subfailure overstretch injury of blood vessels reverses vasomotor dysfunction and prevents apoptosis

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Abstract

Human saphenous vein (HSV) is harvested and prepared prior to implantation as an arterial bypass graft. Injury and the response to injury from surgical harvest and preparation trigger cascades of molecular events and contribute to graft remodeling and intimal hyperplasia. Apoptosis is an early response after implantation that contributes the development of neointimal lesions. Here, we showed that surgical harvest and preparation of HSV leads to vasomotor dysfunction, increased apoptosis and downregulation of the phosphorylation of the anti-apoptotic protein, Niban. A model of subfailure overstretch injury in rat aorta (RA) was used to demonstrate impaired vasomotor function, increased extracellular ATP (eATP) release, and increased apoptosis following pathological vascular injury. The subfailure overstretch injury was associated with activation of p38 MAPK stress pathway and decreases in the phosphorylation of the anti-apoptotic protein Niban. Treatment of RA after overstretch injury with antagonists to purinergic P2X7 receptor (P2X7R) antagonists or P2X7R/pannexin (PanX1) complex, but not PanX1 alone, restored vasomotor function. Inhibitors to P2X7R and PanX1 reduced stretch-induced eATP release. P2X7R/PanX1 antagonism led to decrease in p38 MAPK phosphorylation, restoration of Niban phosphorylation and increases in the phosphorylation of the anti-apoptotic protein Akt in RA and reduced TNFα-stimulated caspase 3/7 activity in cultured rat vascular smooth muscle cells. In conclusion, inhibition of P2X7R after overstretch injury restored vasomotor function and inhibited apoptosis. Treatment with P2X7R/PanX1 complex inhibitors after harvest and preparation injury of blood vessels used for bypass conduits may prevent the subsequent response to injury that lead to apoptosis and represents a novel therapeutic approach to prevent graft failure.

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Acknowledgments

We would like to thank the Vanderbilt cardiac surgical teams for providing human specimens for this study.

Funding

This work was supported by NIH grants R01HL70715-09 (to CB) R01HL105731-01 (to JC), a VA merit award I01BX002036 (to CB), and in part by CTSA award No. UL1TR000445 from the National Center for Advancing Translational Sciences (to JC). Its contents are solely the responsibility of the authors and do not necessarily represent official views of the National Center for Advancing Translational Sciences or the National Institutes of Health.

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Authors and Affiliations

  1. Department of Surgery, Vanderbilt University Medical Center, Nashville, TN, USA

    Weifeng Luo, Daniel Feldman, Reid McCallister, Colleen Brophy & Joyce Cheung-Flynn

  2. VA Tennessee Valley Healthcare System, Nashville, TN, USA

    Colleen Brophy

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  1. Weifeng Luo
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  2. Daniel Feldman
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  3. Reid McCallister
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  4. Colleen Brophy
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Correspondence to Joyce Cheung-Flynn.

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Conflict of interest

Drs. Brophy and Cheung-Flynn are inventors on U.S. Patent 8691556 and disclose that they have financial relationships with VasoPrep Surgical, Inc. (Morristown, NJ).

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Luo, W., Feldman, D., McCallister, R. et al. P2X7R antagonism after subfailure overstretch injury of blood vessels reverses vasomotor dysfunction and prevents apoptosis. Purinergic Signalling 13, 579–590 (2017). https://doi.org/10.1007/s11302-017-9585-0

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  • DOI: https://doi.org/10.1007/s11302-017-9585-0

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