An in vitro model of ischemia/reperfusion-induced microvascular injury.
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.
Inauen
W
W
Department of Physiology and Biophysics, Louisiana State University Medical Center, Shreveport 71130.
Granger
D N
DN
Meininger
C J
CJ
Schelling
M E
ME
Granger
H J
HJ
Kvietys
P R
PR
D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D003331:Coronary Vessels / Q000503:physiopathology*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000503:physiopathology; D006801:Humans; D000860:Hypoxia / Q000503:physiopathology; D066298:In Vitro Techniques; D007511:Ischemia / Q000503:physiopathology*; D008833:Microcirculation / Q000503:physiopathology; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology; D010196:Pancreatic Elastase / Q000097:blood; D015427:Reperfusion Injury / Q000503:physiopathology*; D013482:Superoxide Dismutase / Q000494:pharmacology; D014699:Venules / Q000503:physiopathology
D000493:Allopurinol; D013482:Superoxide Dismutase; D010196:Pancreatic Elastase