Effects of adenosine pretreatment on detection of free radicals in ischemic and reperfused canine gracilis muscle flaps by use of spin-trapping electron paramagnetic resonance spectroscopy

Abstract

To determine whether adenosine pretreatment attenuates free radical production and muscle damage in ischemic and reperfused canine skeletal muscle. 9 healthy mixed-breed dogs. Dogs were anesthetized, and both gracilis muscles were isolated, leaving only the major vascular pedicle intact. Saline (0.9% NaCl) solution was injected into the artery supplying the control flap, whereas adenosine (10 mg) was injected into the contralateral artery. Ischemia was induced in both flaps for 4 hours. alpha-Phenyl-N-tert-butylnitrone was administered IV to each dog 1 hour prior to reperfusion. Following 15 minutes of reperfusion, effluent blood samples from each muscle flap were obtained and processed for spin-trapping electron paramagnetic resonance (EPR) spectroscopy. Muscle biopsy specimens were obtained for histologic evaluation, and dogs were euthanatized. EPR spectra of strong intensity were obtained from analysis of 5 of 9 paired samples. Signals identified were characteristic of oxygen- and carbon-centered free radical adducts. Signal intensity of spectra from adenosine-treated flaps was significantly less than that of control flaps; mean signal attenuation was 36% in the adenosine-treated group. Histologic evaluation of muscle flaps did not reveal significant differences between groups. Treatment of canine muscle flaps with adenosine prior to a period of ischemia reduced but did not completely attenuate free radical production after reperfusion. However, adenosine pretreatment did not affect histologic abnormalities.