부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.mvr.2005.11.003 Byline: Jennifer L. Shelton, Lefeng Wang, Gediminas Cepinskas, Martin Sandig, Richard Inculet, David G. McCormack, Sanjay Mehta Keywords: Sepsis; Edema; Acute lung injury; Endothelial cell; Neutrophil Abstract: Human pulmonary microvascular endothelial cell (HPMVEC) injury is central to the pathophysiology of human lung injury. However, septic HPMVEC barrier dysfunction and the contribution of neutrophils have not been directly addressed in vitro. Instead, human EC responses are often extrapolated from studies of human umbilical vein EC (HUVEC). We hypothesized that HUVEC was not a good model for investigating HPMVEC barrier function under septic conditions. HPMVEC was isolated from lung tissue resected from lung cancer patients using magnetic bead-bound anti-PECAM-1 antibody. In confluent monolayers in 3-[mu]m cell-culture inserts, we assessed trans-EC Evans-Blue (EB)-conjugated albumin leak under basal, unstimulated conditions and following stimulation with either lipopolysaccharide or a mixture of equal concentrations of TNF-[alpha], IL-1[beta] and IFN-[gamma] (cytomix). Basal EB-albumin leak was significantly lower across HPMVEC than HUVEC (0.64 [+ or -] 0.06% vs. 1.13 [+ or -] 0.10%, respectively, P < 0.001). Lipopolysaccharide and cytomix increased leak across both HPMVEC and HUVEC in a dose-dependent manner, with a similar increase relative to basal leak in both cell types. The presence of neutrophils markedly and dose-dependently enhanced cytomix-induced EB-albumin leak across HPMVEC (P < 0.01), but had no effect on EB-albumin leak across HUVEC. Both cytomix and lipopolysaccharide-induced albumin leak was not associated with a loss of cell viability. In conclusion, HPMVEC barrier dysfunction under septic conditions is dramatically enhanced by neutrophil presence, and HUVEC is not a suitable model for studying HPMVEC septic barrier responses. The direct study of HPMVEC septic responses will lead to a better understanding of human lung injury. Author Affiliation: Centre for Critical Illness Research, Lawson Health Research Institute, Division of Respirology, Departments of Medicine, Physiology and Pharmacology, London Health Sciences Center, University of Western Ontario, South Street Campus, Colborne Building Room 216, 375 South Street, London, Ontario, Canada N6A 4G5 Article History: Received 11 March 2005; Revised 2 September 2005; Accepted 1 November 2005