Immune Netw.  2015 Aug;15(4):206-211. 10.4110/in.2015.15.4.206.

Involvement of Protein Kinase C-delta in Vascular Permeability in Acute Lung Injury

• Ahn JJ
• Jung JP
• Park SE
• Lee M
• Kwon B
• Cho HR

• KMID: 2168048
• DOI: http://doi.org/10.4110/in.2015.15.4.206

Abstract

Pulmonary edema is a major cause of mortality due to acute lung injury (ALI). The involvement of protein kinase C-delta (PKC-delta) in ALI has been a controversial topic. Here we investigated PKC-delta function in ALI using PKC-delta knockout (KO) mice and PKC inhibitors. Our results indicated that although the ability to produce proinflammatory mediators in response to LPS injury in PKC-delta KO mice was similar to that of control mice, they showed enhanced recruitment of neutrophils to the lung and more severe pulmonary edema. PKC-delta inhibition promoted barrier dysfunction in an endothelial cell layer in vitro, and administration of a PKC-delta-specific inhibitor significantly increased steady state vascular permeability. A neutrophil transmigration assay indicated that the PKC-delta inhibition increased neutrophil transmigration through an endothelial monolayer. This suggests that PKC-delta inhibition induces structural changes in endothelial cells, allowing extravasation of proteins and neutrophils.

Keyword

Acute lung injury; Protein kinase C-delta; Vascular permeability

MeSH Terms

Acute Lung Injury*
Animals
Capillary Permeability*
Endothelial Cells
Lung
Mice
Mortality
Neutrophils
Protein Kinase C-delta*
Protein Kinases*
Pulmonary Edema
Protein Kinase C-delta
Protein Kinases

Figure

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  Figure 1 PKC-δ KO mice had no impairment in the production of proinflammatory mediators, but had severe neutrophil infiltration and perivascular edema. (A~E) BAL fluid was harvested at various time points after LPS infusion and levels of proinflammatory cytokines, TNF-α (A) and IL-6 (B), and chemokines, KC (C), MIP-2 (D) and MCP-1 (E), were measured by ELISA. n=4-10 for each group. *p<0.05 and ***p<0.001 between the indicated groups. (F) Lungs were harvested 12 h after LPS infusion and histological analysis was performed. Neutrophil infiltration and perivascular edema were more prominent in PKC-δ KO mice (right column). *perivascular cuff, V: blood vessel, Br: bronchus. Original magnification: ×40 (upper column) and ×200 (lower column). (G) Determination of myeloperoxidase activity in lung tissue. n=7-12 for each group. *p<0.05 and ***p<0.001 between the indicated groups.

• 

  Figure 2 Inhibition of PKC-δ promotes pulmonary edema and vascular permeability. (A) Total protein levels in BAL fluid at various time points after LPS infusion. n=3-14 for each group. *p<0.05, **p<0.01, and ***p<0.001 between the two groups. (B) The lung wet/dry weight ratio at 24 h after LPS infusion. n=12-13 for each group. *p<0.05. (C and D) EBD was injected 4 h after LPS infusion and lungs were harvested 30 min later. Gross observation (left column) and concentrations of EBD in extracted lung tissue (right column). n=12-13 for each group. *p<0.05. (D)δV1-1 or ΨδRACK peptide was intratracheally injected 30 min before LPS infusion. Lungs were harvested 30 min after EBD injection. n=6-8 for each group. *p<0.05 between the indicated groups.

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  Figure 3 Inhibition of PKC-δ promotes extravasation of proteins and neutrophils through an endothelial monolayer. (A) A confluent monolayer of HUVEC cells was treated with rottlerin 30 min before the addition of LPS. Four hour later, HRP was added to HUVEC cells and extravasation was allowed to proceed for 30 min. n=3 for each group. *p<0.05 and ***p<0.001 between the indicated groups. (B) PKC-δ inhibitors and LPS were treated as described in A and neutrophil transmigration was allowed for 15 h. n=3 for each group. **p<0.01 and ***p<0.001 between the indicated groups.

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