Real-time imaging reveals endothelium-mediated leukocyte retention in LPS-treated lung microvessels
| Autor: | Geetaram Sahu, Kaushik Parthasarathi, Kathirvel Kandasamy |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Diagnostic Imaging
Lipopolysaccharides Male Pathology medicine.medical_specialty Time Factors Endothelium Intercellular Adhesion Molecule-1 Lung injury Biology Biochemistry Article Microcirculation Rats Sprague-Dawley Mice medicine Leukocytes Animals Humans L-Selectin Microvessel Lung Fluorescent Dyes Polymyxin B ICAM-1 Cell Biology Capillaries Rats Mice Inbred C57BL Perfusion medicine.anatomical_structure Microscopy Fluorescence TLR4 lipids (amino acids peptides and proteins) Endothelium Vascular Cardiology and Cardiovascular Medicine Ex vivo Signal Transduction |
| Popis: | Endotoxemia, a major feature of sepsis, is a common cause of acute lung injury and initiates rapid accumulation of leukocytes in the lung vasculature. Endothelial mechanisms that underlie this accumulation remain unclear, as current experimental models of endotoxemia are less suitable for targeted activation of the endothelium. Toward elucidating this, we used the isolated blood-perfused rat lung preparation. With a microcatheter inserted through a left atrial cannula, we cleared blood cells from a small lung region and then infused lipopolysaccharide (LPS) into microvessels. After a Ringer's wash to remove residual LPS, we infused fluorescently-labeled autologous leukocytes and imaged their transit through the treated microvessels. Image analysis revealed that leukocytes infused 90 min after LPS treatment were retained more in treated venules and capillaries than untreated vessels. Further, pretreatment with either the intercellular adhesion molecule-1 (ICAM-1) mAb or polymyxin-B blunted LPS-induced leukocyte retention in both microvessel segments. In addition, retention of leukocytes treated ex vivo with LPS in LPS-treated microvessels was higher compared to retention of untreated leukocytes. In situ immunofluorescence experiments revealed that LPS significantly increased microvessel ICAM-1 expression at 90 min post treatment. Polymyxin pretreatment inhibited this increase. Taken together, the data suggest that LPS increased leukocyte retention in both venules and capillaries and this response was mediated by the increased expression of endothelial ICAM-1. Thus, endothelial mechanisms may themselves play a major role in LPS-induced leukocyte retention in lung microvessels. Blunting the endothelial responses may mitigate endotoxin-induced morbidity. |
| Databáze: | OpenAIRE |
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