Neutrophil protein kinase Cδ as a mediator of stroke-reperfusion injury
Wen-Hai Chou, … , Donna M. Ferriero, Robert O. Messing
Wen-Hai Chou, … , Donna M. Ferriero, Robert O. Messing
Published July 1, 2004
Citation Information: J Clin Invest. 2004;114(1):49-56. https://doi.org/10.1172/JCI21655.
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Article Neuroscience

Neutrophil protein kinase Cδ as a mediator of stroke-reperfusion injury

Abstract

Thrombolysis is widely used to intervene in acute ischemic stroke, but reestablishment of circulation may paradoxically initiate a reperfusion injury. Here we describe studies with mice lacking protein kinase Cδ (PKCδ) showing that absence of this enzyme markedly reduces reperfusion injury following transient ischemia. This was associated with reduced infiltration of peripheral blood neutrophils into infarcted tissue and with impaired neutrophil adhesion, migration, respiratory burst, and degranulation in vitro. Total body irradiation followed by transplantation with bone marrow from PKCδ-null mice donors reduced infarct size and improved neurological outcome in WT mice, whereas marrow transplantation from WT donors increased infarction and worsened neurological scores in PKCδ-null mice. These results indicate an important role for neutrophil PKCδ in reperfusion injury and strongly suggest that PKCδ inhibitors could prove useful in the treatment of stroke.

Authors

Wen-Hai Chou, Doo-Sup Choi, Hong Zhang, Dezhi Mu, Tom McMahon, Viktor N. Kharazia, Clifford A. Lowell, Donna M. Ferriero, Robert O. Messing

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Figure 5

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Decreased neutrophil function in PKCδ-null mice. (A) Neutrophil adhesion...
Decreased neutrophil function in PKCδ-null mice. (A) Neutrophil adhesion stimulated by PMA (100 nM), fMLP peptide (1 & μ;M), or IL-8 (200 ng/ml). *P < 0.05 compared with WT littermates (two-tailed, unpaired t tests). (B) IL-8–stimulated neutrophil migration differed by genotype [F(1, 6) = 54.8; P = 0.0003] and treatment [F(2, 6) = 63.08; P < 0.0001] without significant interaction between these factors [F(2, 6) = 3.9; NS]. **P < 0.05 compared with PKCδ+/+ at same dose (Bonferroni test). (C) TNF-α–stimulated (20 ng/ml) superoxide anion production showed main effects of treatment [F(1, 224) = 236; P < 0.0001], genotype [F(1, 224) = 506; P < 0.0001], and time [F(6, 224) = 64.6; P < 0.0001] with an interaction between these factors [F(6, 224) = 14.7; P < 0.0001]. P < 0.05 compared with unstimulated PKCδ –/ – neutrophils, and ***P < 0.05 compared with unstimulated PKCδ+/+ and stimulated PKCδ –/ – neutrophils at the same time (Newman Keuls test). (D) fMLP-stimulated (10 & μ;M) lactoferrin release differed by genotype [F(1, 12) = 15.2; P = 0.0021] and treatment [F(1, 12) = 89.01; P < 0.0001] with an interaction between these factors [F(1, 12) = 7,24; P = 0.0197]. P < 0.05 compared with unstimulated PKCδ –/ – neutrophils, and ***P < 0.05 compared with unstimulated PKCδ+/+ and stimulated PKCδ –/ – neutrophils (Bonferroni tests). (E) PKC isozyme immunoreactivity in neutrophils. (F) PKC isozyme immunoreactivity determined by regression analysis of protein concentrations and corresponding OD values on Western blots (n = 3). Assays of neutrophil function (A–D) were performed in triplicate or quadruplicate from at least three animals of each genotype.