Reperfusion following ischemia leads to neutrophil recruitment into injured tissue. Selectins and β2-integrins regulate neutrophil interaction with the endothelium during neutrophil rolling and firm adhesion. Excessive neutrophil infiltration into tissue is thought to contribute to ischemia-reperfusion injury damage. Hydrogen sulfide mitigates the damage caused by ischemia-reperfusion injury. This study's objective was to determine the effect of hydrogen sulfide on neutrophil adhesion receptor expression.
Human neutrophils were either left untreated or incubated in 20 μM hydrogen sulfide and/or 50 μg/ml pharmacologic ADAM-17 inhibitor TAPI-0; activated by interleukin-8, fMLP, or TNF-α; and labeled against P-selectin glycoprotein ligand-1, leukocyte function associated antigen-1, Mac-1 α, L-selectin, and β2-integrin epitopes CBRM1/5 or KIM127 for flow cytometry. Cohorts of three C57BL/6 mice received an intravenous dose of saline vehicle or 20 μM hydrogen sulfide with or without 50 μg/ml TAPI-0 before unilateral tourniquet-induced hind-limb ischemia for 3 hours followed by 3 hours of reperfusion. Bilateral gastrocnemius muscles were processed for histology before neutrophil infiltration quantification.
Hydrogen sulfide treatment significantly increased L-selectin shedding from human neutrophils following activation by fMLP and interleukin-8 in an ADAM-17–dependent manner. Mice treated with hydrogen sulfide to raise bloodstream concentration by 20 μM before ischemia or reperfusion showed a significant reduction in neutrophil recruitment into skeletal muscle tissue following tourniquet-induced hind-limb ischemia-reperfusion injury.
Hydrogen sulfide administration results in the down-regulation of L-selectin expression in activated human neutrophils. This leads to a reduction in neutrophil extravasation and tissue infiltration and may partially account for the protective effects of hydrogen sulfide seen in the setting of ischemia-reperfusion injury.
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Ithaca, New York, and Rochester, N.Y.
From the Department of Biomedical Engineering, Cornell University; the Division of Plastic Surgery, Laboratory of Bioregenerative Medicine and Surgery, Weill Cornell Medical College; and the Department of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester.
Received for publication May 3, 2012; accepted August 31, 2012.
Presented at the Biomedical Engineering Society Annual Meeting, in Hartford, Connecticut, October 11 through 16, 2011; the 56th Annual Meeting of the Plastic Surgery Research Council, in Louisville, Kentucky, April 27 through 30, 2011; the 6th Annual Academic Surgical Congress, in Huntington Beach, California, February 1 through 3, 2011; and the 97th Annual American College of Surgeons Clinical Congress, in San Francisco, California, October 23 through 27, 2011.
Disclosure: The authors have no financial conflicts of interest to disclose.
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This work was supported by THE PLASTIC SURGERY FOUNDATION.
Michael R. King, Ph.D.; 526 Campus Road, 205 Weill Hall, Cornell University, Ithaca, N.Y. 14853, email@example.com