Ischemia-reperfusion injury in rats affects hydraulic conductivity in two phases that are temporally and mechanistically separate.
Ischemia-reperfusion (IR) injury is a major insult to postcapillary venules. We hypothesized that IR increases postcapillary venular hydraulic conductivity and that IR-mediated changes in hydraulic conductivity result from temporally and mechanistically separate processes. A microcannulation technique was used to determine hydraulic conductivity (Lp) in rat mesenteric postcapillary venules serially throughout ischemia (45 min) and reperfusion (5 h) induced by superior mesenteric artery occlusion and release. Mesenteric IR resulted in a biphasic increase in Lp. White blood cell (WBC) adhesion slowly increased with maximal adhesion corresponding to the second peak (P < 0.005). After IR, tissue was harvested for RT-PCR analysis of ICAM-1, E-selectin, and P-selectin mRNA. Intercellular adhesion molecule-1 (ICAM-1) mRNA in the gut showed the most significant upregulation. Quantitative real-time PCR revealed that ICAM-1 mRNA was upregulated 60-fold in the gut. An ICAM-1 antibody was therefore used to determine the effect of WBC adhesion on Lp during IR. ICAM-1 inhibition attenuated Lp during the first peak and completely blocked the second peak (P < 0.005). When rats were fed a tungsten diet to inhibit xanthine oxidase and then underwent IR, Lp was dramatically attenuated during the first peak and mildly decreased the second peak (P < 0.005). Inhibition of xanthine oxidase by oxypurinol decreased Lp during IR by over 60% (P < 0.002). Tempol, a superoxide dismutase mimetic, decreased Lp during IR by over 30% (P < 0.01). We conclude that IR induces a biphasic increase in postcapillary hydraulic conductivity. Reactive oxygen species impact both the first transient peak and the sustained second peak. However, the second peak is also dependent on WBC-endothelial cell adhesion. These serial measurements of postcapillary hydraulic conductivity may lead the way for optimal timing of pharmaceutical therapies in IR injury.
Victorino
Gregory P
GP
Department of Surgery, University of California San Franciso-East Bay, Alameda County Medical Center, Oakland, CA 94602, USA. victorinog@surgery.ucsf.edu
Ramirez
René M
RM
Chong
Terry J
TJ
Curran
Brian
B
Sadjadi
Javid
J
D000818:Animals; D000906:Antibodies; D002448:Cell Adhesion / Q000187:drug effects; D006224:Cricetinae; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004195:Disease Models, Animal; D019040:E-Selectin / Q000235:genetics / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008643:Mesentery / Q000098:blood supply*; D008647:Mesocricetus; D010117:Oxypurinol; D019007:P-Selectin / Q000235:genetics / Q000378:metabolism; D010539:Permeability; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015427:Reperfusion Injury / Q000378:metabolism*; D013113:Spin Labels; D013997:Time Factors; D017973:Tungsten Compounds / Q000494:pharmacology; D014699:Venules / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism
D000906:Antibodies; D003497:Cyclic N-Oxides; D019040:E-Selectin; D004791:Enzyme Inhibitors; D019007:P-Selectin; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D013113:Spin Labels; D017973:Tungsten Compounds; D018799:Intercellular Adhesion Molecule-1; C025399:sodium tungstate(VI); D014969:Xanthine Oxidase; D010117:Oxypurinol; C001803:tempol
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