Ischemic preconditioning ameliorates microcirculatory disturbance through downregulation of TNF-alpha production in a rat cremaster muscle model.
Ischemia-reperfusion (I/R) injury is a complex process involving the generation and release of inflammatory cytokines, and the accumulation and infiltration of neutrophils and macrophages, which disturbs the microcirculatory hemodynamics. Nonetheless, ischemic preconditioning (IPC) is known to produce immediate tolerance to subsequent prolonged I/R insults, although its underlying mechanism largely remains unknown. Our study investigated the role of the IkappaB-alpha-NF-kappaB-TNF-alpha (tumor necrosis factor-alpha) pathway in IPC's ability to ameliorate I/R-induced microcirculatory disturbances in rat cremaster muscle flaps. Male Sprague-Dawley rats were randomized (n = 8 per group) into 3 groups: a sham-operated control group, an I/R group (4 h of pudic epigastric artery ischemia followed by 2 h of reperfusion), and an IPC+I/R group (3 cycles of 10 min of ischemia followed by 10 min reperfusion before I/R). Intravital microscopy was used to observe leukocyte/endothelial cell interactions and quantify functional capillaries in cremaster muscles. I/R markedly increased the number of rolling, adhering, and migrating leukocytes. It was also observed that I/R significantly increased TNF-alpha expression in these injured tissues. On the other hand, IPC prevented I/R-induced increases in leukocyte rolling, adhesion, and transmigration. Moreover, TNF-alpha protein production and its mRNA expression were downregulated in the IPC group. Finally, I/R-induced IkappaB-alpha phosphorylation and NF-kappaB (p65) nuclear translocation were both suppressed by IPC. These results indicated that IPC attenuated NF-kappaB activation and subsequently reduced TNF-alpha expression, which resulted in the amelioration of microcirculatory disturbances in I/R-injured cremaster muscles.
Hung
Li-Man
LM
Department of Life Science, College of Medicine, Chang Gung University, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
Wei
William
W
Hsueh
Yi-Jen
YJ
Chu
Wing-Keung
WK
Wei
Fu-Chan
FC
D021581:Active Transport, Cell Nucleus; D000818:Animals; D015153:Blotting, Western; D002196:Capillaries; D002465:Cell Movement; D015536:Down-Regulation; D020672:I-kappa B Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D007249:Inflammation; D007511:Ischemia; D019194:Ischemic Preconditioning; D007962:Leukocytes / Q000378:metabolism; D008264:Macrophages; D008833:Microcirculation; D018715:Microscopy, Video; D009132:Muscles / Q000378:metabolism / Q000473:pathology*; D009206:Myocardium / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606206:Nfkbia protein, rat; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha