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In vivo monitoring of .OH generation on jejunal ischemic injury by dialysis technique.

We applied an in vivo microdialysis technique to examine free radical generation following ischemic injury in the rat jejunum. For this purpose, we used a microdialysis probe holding system. The salicylate reacts with hydroxyl free radical (.OH) and generates 2,3-dihydroxybenzoic acid (DHBA), which can be measured electrochemically in picomole amounts by high-performance liquid chromatography (HPLC). Therefore, the former was used for trapping the .OH formation. Reperfusion of the jejunum produced progressive elevations in the levels of 2,3-DHBA achieving the maximums at 20 min, after which they decreased progressively to the baseline. These elevations were significant only after 10 min- and 20 min-ischemia. However, this change was not significant in the case of 30 min-ischemia. Ischemia-reperfusion of rat intestine caused morphological changes. These results suggest that .OH production is significantly reduced after 30 min-ischemia providing thus the possibility for more severe intestinal damage. In conclusion our dialysis probe can be useful for the measurement of .OH generation following ischemia-reperfusion injury.