Figure 5
KNG deficiency has profound blood-brain barrier stabilizing and antiedematous effects in ischemic stroke. (A left) Representative corresponding coronal brain sections from a wild-type (WT) mouse, a Kng−/− mouse, and a Kng−/− mouse reconstituted with bradykinin (BK) on day 1 after tMCAO after injection of the vascular tracer Evans blue. Vascular leakage was significantly decreased in the absence of KNG after stroke as confirmed by the concentration of Evans blue detectable in the brain parenchyma (right). BK reconstitution restored edema formation in Kng−/− mice (N = 7-8 per group). (B) Edema formation as reflected by the brain water content in the ischemic hemispheres of WT mice and Kng−/− mice on day 1 after tMCAO (N = 6-9 per group). (C) Relative gene expression of endothelin-1 (Edn-1) in the cortices and basal ganglia of WT mice and Kng−/− mice 24 hours after tMCAO or sham operation (N = 4 per group). (D) Expression of occludin on day 1 after tMCAO in the hemispheres of WT mice and Kng−/− mice. Immunohistochemistry suggests that occludin is predominately located in the gaps between vascular endothelial cells (indicated by the marker CD31). Occludin expression was markedly reduced in WT mice but preserved in mice lacking KNG. Hoechst staining (blue) depicts cell nuclei. One representative panel per group of 3 independent experiments is shown. Scale bar represents 50 μm. (E top) Occludin expression in the cortex or basal ganglia (BG) of WT mice or Kng−/− mice on day 1 after tMCAO or sham operation as determined by immunoblot. One representative immunoblot of each group is shown. (Bottom) Densitometric quantification of occludin immunoreactivity in the mouse groups indicated above (N = 4 per group). The i indicates ipsilateral (ischemic) hemisphere; c, contralateral (healthy) hemisphere. (A,B) ***P < .001, *P = .0148, unpaired Student t test. (C) ***P < .001, **P < .01, *P < .05, ##P < .01, #P < .05, 1-way ANOVA followed by Bonferroni multiple comparison test compared with sham-operated mice (* symbol) or WT (# symbol) mice. (E) *P < .05, 2-way ANOVA, followed by Bonferroni multiple comparison test, group comparisons as indicated in the figure.

KNG deficiency has profound blood-brain barrier stabilizing and antiedematous effects in ischemic stroke. (A left) Representative corresponding coronal brain sections from a wild-type (WT) mouse, a Kng−/− mouse, and a Kng−/− mouse reconstituted with bradykinin (BK) on day 1 after tMCAO after injection of the vascular tracer Evans blue. Vascular leakage was significantly decreased in the absence of KNG after stroke as confirmed by the concentration of Evans blue detectable in the brain parenchyma (right). BK reconstitution restored edema formation in Kng−/− mice (N = 7-8 per group). (B) Edema formation as reflected by the brain water content in the ischemic hemispheres of WT mice and Kng−/− mice on day 1 after tMCAO (N = 6-9 per group). (C) Relative gene expression of endothelin-1 (Edn-1) in the cortices and basal ganglia of WT mice and Kng−/− mice 24 hours after tMCAO or sham operation (N = 4 per group). (D) Expression of occludin on day 1 after tMCAO in the hemispheres of WT mice and Kng−/− mice. Immunohistochemistry suggests that occludin is predominately located in the gaps between vascular endothelial cells (indicated by the marker CD31). Occludin expression was markedly reduced in WT mice but preserved in mice lacking KNG. Hoechst staining (blue) depicts cell nuclei. One representative panel per group of 3 independent experiments is shown. Scale bar represents 50 μm. (E top) Occludin expression in the cortex or basal ganglia (BG) of WT mice or Kng−/− mice on day 1 after tMCAO or sham operation as determined by immunoblot. One representative immunoblot of each group is shown. (Bottom) Densitometric quantification of occludin immunoreactivity in the mouse groups indicated above (N = 4 per group). The i indicates ipsilateral (ischemic) hemisphere; c, contralateral (healthy) hemisphere. (A,B) ***P < .001, *P = .0148, unpaired Student t test. (C) ***P < .001, **P < .01, *P < .05, ##P < .01, #P < .05, 1-way ANOVA followed by Bonferroni multiple comparison test compared with sham-operated mice (* symbol) or WT (# symbol) mice. (E) *P < .05, 2-way ANOVA, followed by Bonferroni multiple comparison test, group comparisons as indicated in the figure.

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