Lipopolysaccharide Induces Endoplasmic Store Ca2+-Dependent Inflammatory Responses in Lung Microvessels

Abstract

The pulmonary microvasculature plays a critical role in endotoxin-induced acute lung injury. However, the relevant signaling remain unclear. Specifically the role of endothelial Ca²⁺ in the induction of endotoxin-mediated responses in lung microvessels remains undefined. Toward elucidating this, we used the isolated blood-perfused rat lung preparation. We loaded microvessels with the Ca²⁺ indicator, Fura 2 AM and then determined Ca²⁺ responses to infusions of lipopolysaccharide (LPS) into the microvessels. LPS induced a more than two-fold increase in the amplitude of cytosolic Ca²⁺ oscillations. Inhibiting inositol 1,4,5 trisphosphate receptors on endoplasmic reticulum (ER) Ca²⁺ stores with Xestospongin C (XeC), blocked the LPS-induced increase in the Ca²⁺ oscillation amplitude. However, XeC did not affect entry of external Ca²⁺ via plasma membrane Ca²⁺ channels in lung microvascular endothelial cells. This suggested that LPS augmented the oscillations via release of Ca²⁺ from ER stores. In addition, XeC also blocked LPS-mediated activation and nuclear translocation of nuclear factor-kappa B in lung microvessels. Further, inhibiting ER Ca²⁺ release blunted increases in intercellular adhesion molecule-1 expression and retention of naïve leukocytes in LPS-treated microvessels. Taken together, the data suggest that LPS-mediated Ca²⁺ release from ER stores underlies nuclear factor-kappa B activation and downstream inflammatory signaling in lung microvessels. Thus, we show for the first time a role for inositol 1,4,5 trisphosphate-mediated ER Ca²⁺ release in the induction of LPS responses in pulmonary microvascular endothelium. Mechanisms that blunt this signaling may mitigate endotoxin-induced morbidity.