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Impaired microvascular perfusion in sepsis requires activated coagulation and P-selectin-mediated platelet adhesion in capillaries

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Abstract

Purpose

Impaired microvascular perfusion in sepsis is not treated effectively because its mechanism is unknown. Since inflammatory and coagulation pathways cross-activate, we tested if stoppage of blood flow in septic capillaries is due to oxidant-dependent adhesion of platelets in these microvessels.

Methods

Sepsis was induced in wild type, eNOS−/−, iNOS−/−, and gp91phox−/− mice (n = 14–199) by injection of feces into the peritoneum. Platelet adhesion, fibrin deposition, and blood flow stoppage in capillaries of hindlimb skeletal muscle were assessed by intravital microscopy. Prophylactic treatments at the onset of sepsis were intravenous injection of platelet-depleting antibody, P-selectin blocking antibody, ascorbate, or antithrombin. Therapeutic treatments (delayed until 6 h) were injection of ascorbate or the glycoprotein IIb/IIIa inhibitor eptifibatide, or local superfusion of the muscle with NOS cofactor tetrahydrobiopterin or NO donor S-nitroso-N-acetylpenicillamine (SNAP).

Results

Sepsis at 6–7 h markedly increased the number of stopped-flow capillaries and the occurrence of platelet adhesion and fibrin deposition in these capillaries. Platelet depletion, iNOS and gp91phox deficiencies, P-selectin blockade, antithrombin, or prophylactic ascorbate prevented, whereas delayed ascorbate, eptifibatide, tetrahydrobiopterin, or SNAP reversed, septic platelet adhesion and/or flow stoppage. The reversals by ascorbate and tetrahydrobiopterin were absent in eNOS−/− mice. Platelet adhesion predicted 90% of capillary flow stoppage.

Conclusion

Impaired perfusion and/or platelet adhesion in septic capillaries requires NADPH oxidase, iNOS, P-selectin, and activated coagulation, and is inhibited by intravenous administration of ascorbate and by local superfusion of tetrahydrobiopterin and NO. Reversal of flow stoppage by ascorbate and tetrahydrobiopterin may depend on local eNOS-derived NO which dislodges platelets from the capillary wall.

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Acknowledgments

We thank HSFO for funding (NA 5941, KT and JXW), and Ms. S. Milkovich, J. MacLean, and S. Sehgal for technical help.

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Authors and Affiliations

  1. Critical Illness Research, Victoria Research Laboratories, Lawson Health Research Institute, 6th Floor, 800 Commissioners Road East, London, ON, N6C 2V5, Canada

    Dan Secor, Fuyan Li, Christopher G. Ellis & Karel Tyml

  2. Department of Physiology and Pharmacology, University of Western Ontario, London, Canada

    Dan Secor & Karel Tyml

  3. Department of Medical Biophysics, University of Western Ontario, London, Canada

    Fuyan Li, Christopher G. Ellis & Karel Tyml

  4. Department of Anesthesia and Perioperative Medicine, Program in Critical Care, University of Western Ontario, London, Canada

    Michael D. Sharpe

  5. Department of Medicine, Henderson Research Centre, McMaster University, Hamilton, ON, Canada

    Peter L. Gross

  6. Department of Exercise and Nutrition Sciences, University at Buffalo, Buffalo, NY, USA

    John X. Wilson

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  1. Dan Secor
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  2. Fuyan Li
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  3. Christopher G. Ellis
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  6. John X. Wilson
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  7. Karel Tyml
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Correspondence to Karel Tyml.

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Secor, D., Li, F., Ellis, C.G. et al. Impaired microvascular perfusion in sepsis requires activated coagulation and P-selectin-mediated platelet adhesion in capillaries. Intensive Care Med 36, 1928–1934 (2010). https://doi.org/10.1007/s00134-010-1969-3

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  • DOI: https://doi.org/10.1007/s00134-010-1969-3

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