AJP - Gastrointestinal and Liver Physiology, Vol 259, Issue 1 134-G139, Copyright © 1990 by American Physiological Society

ARTICLES

An in vitro model of ischemia/reperfusion-induced microvascular injury

W. Inauen, D. N. Granger, C. J. Meininger, M. E. Schelling, H. J. Granger and P. R. Kvietys
Department of Physiology and Biophysics, Louisiana State University Medical Center, Shreveport 71130.

The major objective of this study was to develop an in vitro model of ischemia/reperfusion (I/R)-induced microvascular injury. Cultured venular endothelial cells were grown to confluency, labeled with 51Cr, and exposed to different durations of anoxia (0.5, 1, 2, 3, and 4 h). 51Cr release and cell detachment (indexes of cell injury) were determined at different times after reoxygenation (1, 2, 4, 6, 8, and 18 h). Because in vivo studies have implicated neutrophils in I/R injury, in some experiments human neutrophils were added to the endothelial cells upon reoxygenation. Periods of anoxia greater than or equal to 2 h resulted in 70-80% 51Cr release and 80-95% cell detachment upon reoxygenation. Under these conditions (near maximal injury), the addition of neutrophils produced negligible effects. Periods of anoxia less than or equal to 1 h resulted in 30-40% 51Cr release and 50-60% cell detachment. Under these conditions (moderate cell injury), addition of neutrophils enhanced endothelial cell injury. Using a 30-min period of anoxia, we also assessed the effects of superoxide dismutase (SOD; 300 U/ml) and allopurinol (20 microM) on anoxia/reoxygenation (A/R)-induced injury in the presence or absence of neutrophils. In the absence of neutrophils, SOD or allopurinol did not protect against A/R-induced injury. However, in the presence of neutrophils, both SOD and allopurinol attenuated the increases in 51Cr release. The results derived using this in vitro model of I/R injury are largely consistent with published in vivo studies. Thus this in vitro model may provide further insights regarding the mechanisms involved in I/R injury.

This article has been cited by other articles:

				I. Rubio-Gayosso, S. H. Platts, and B. R. Duling Reactive oxygen species mediate modification of glycocalyx during ischemia-reperfusion injury Am J Physiol Heart Circ Physiol, June 1, 2006; 290(6): H2247 - H2256. [Abstract] [Full Text] [PDF]

				M. Sasaki, J. W. Elrod, P. Jordan, M. Itoh, T. Joh, A. Minagar, and J. S. Alexander CYP450 dietary inhibitors attenuate TNF-{alpha}-stimulated endothelial molecule expression and leukocyte adhesion Am J Physiol Cell Physiol, April 1, 2004; 286(4): C931 - C939. [Abstract] [Full Text] [PDF]

				M. Sasaki, D. Ostanin, J. W. Elrod, T. Oshima, P. Jordan, M. Itoh, T. Joh, A. Minagar, and J. S. Alexander TNF-alpha -induced endothelial cell adhesion molecule expression is cytochrome P-450 monooxygenase dependent Am J Physiol Cell Physiol, February 1, 2003; 284(2): C422 - C428. [Abstract] [Full Text] [PDF]

				N Yoshida, N Sugimoto, F Hirayama, Y Nakamura, H Ichikawa, Y Naito, and T Yoshikawa Helicobacter pylori infection potentiates aspirin induced gastric mucosal injury in Mongolian gerbils Gut, May 1, 2002; 50(5): 594 - 598. [Abstract] [Full Text] [PDF]

				A. Mizutani, K. Okajima, M. Uchiba, and T. Noguchi Activated protein C reduces ischemia/reperfusion-induced renal injury in rats by inhibiting leukocyte activation Blood, June 15, 2000; 95(12): 3781 - 3787. [Abstract] [Full Text] [PDF]

				S. Kokura, R. E. Wolf, T. Yoshikawa, D. N. Granger, and T. Y. Aw T-Lymphocyte-Derived Tumor Necrosis Factor Exacerbates Anoxia-Reoxygenation-Induced Neutrophil-Endothelial Cell Adhesion Circ. Res., February 4, 2000; 86(2): 205 - 213. [Abstract] [Full Text] [PDF]

				J. H. Park, N. Okayama, D. Gute, A. Krsmanovic, H. Battarbee, and J. S. Alexander Hypoxia/aglycemia increases endothelial permeability: role of second messengers and cytoskeleton Am J Physiol Cell Physiol, December 1, 1999; 277(6): C1066 - C1074. [Abstract] [Full Text] [PDF]

				P. Beauchamp, V. Richard, F. Tamion, F. Lallemand, J.-P. Lebreton, H. Vaudry, M. Daveau, and C. Thuillez Protective Effects of Preconditioning in Cultured Rat Endothelial Cells : Effects on Neutrophil Adhesion and Expression of ICAM-1 After Anoxia and Reoxygenation Circulation, August 3, 1999; 100(5): 541 - 546. [Abstract] [Full Text] [PDF]

				T. J. Kalogeris, F. S. Laroux, A. Cockrell, H. Ichikawa, N. Okayama, T. J. Phifer, J. S. Alexander, and M. B. Grisham Effect of selective proteasome inhibitors on TNF-induced activation of primary and transformed endothelial cells Am J Physiol Cell Physiol, April 1, 1999; 276(4): C856 - C864. [Abstract] [Full Text] [PDF]

				S. Kokura, R. E. Wolf, T. Yoshikawa, D. N. Granger, and T. Y. Aw Molecular Mechanisms of Neutrophil-Endothelial Cell Adhesion Induced by Redox Imbalance Circ. Res., March 19, 1999; 84(5): 516 - 524. [Abstract] [Full Text] [PDF]

				D. Carden, F. Xiao, C. Moak, B. H. Willis, S. Robinson-Jackson, and S. Alexander Neutrophil elastase promotes lung microvascular injury and proteolysis of endothelial cadherins Am J Physiol Heart Circ Physiol, August 1, 1998; 275(2): H385 - H392. [Abstract] [Full Text] [PDF]

				N. Okayama, H. Ichikawa, L. Coe, M. Itoh, and J. S. Alexander Exogenous NO enhances hydrogen peroxide-mediated neutrophil adherence to cultured endothelial cells Am J Physiol Lung Cell Mol Physiol, May 1, 1998; 274(5): L820 - L826. [Abstract] [Full Text] [PDF]

				H. Ichikawa, S. Flores, P. R. Kvietys, R. E. Wolf, T. Yoshikawa, D. N. Granger, and T. Y. Aw Molecular Mechanisms of Anoxia/Reoxygenation-Induced Neutrophil Adherence to Cultured Endothelial Cells Circ. Res., December 19, 1997; 81(6): 922 - 931. [Abstract] [Full Text]

				P. R. Kvietys and D. N. Granger Endothelial cell monolayers as a tool for studying microvascular pathophysiology Am J Physiol Gastrointest Liver Physiol, December 1, 1997; 273(6): G1189 - G1199. [Abstract] [Full Text] [PDF]

				L. Liao, T. Y. Aw, P. R. Kvietys, and D. N. Granger Oxidized LDL–Induced Microvascular Dysfunction : Dependence on Oxidation Procedure Arterioscler. Thromb. Vasc. Biol., December 1, 1995; 15(12): 2305 - 2311. [Abstract] [Full Text]