Effect of Repetitive Stimulation on the Contractile Response of Rabbit Urinary Bladder Subjected to in vitro Hypoxia or in vitro Ischemia Followed by Reoxygenation
Authors: Ohnishi Norio , Liu Shih-Ping , Horan Patrick , Levin Robert M. 
Published: April 2003

<b>Purpose:</b> We studied the effects of hypoxia followed by reoxygenation and an in vitro model of ischemia (hypoxia + substrate [glucose] deprivation) followed by reperfusion (reoxygenation + substrate replacement) on the contractile response of rabbit urinary bladder strips to nonrepetitive and repetitive field stimulation (FS), and correlated the results with the rate of lipid peroxidation. We view repetitive FS as a model for hyperreflexia. <b>Methods:</b> The effects of repetitive and nonrepetitive FS on the contractile responses of isolated strips of rabbit bladder to FS, carbachol, and KCl were determined in the presence of 3 different incubation media: O<sub>2</sub> + glucose (normal physiological medium); N<sub>2</sub> + glucose (in vitro hypoxia), and N<sub>2</sub> – glucose (in vitro ischemia). Then, all strips were incubated for 1 h in normal physiological medium (‘reperfusion’) followed by a final stimulation; the resultant contractile responses were correlated with the level of lipid peroxidation as determined by malonedialdehyde (MDA) concentration. <b>Results:</b> Repetitive stimulation, a model of hyperreflexia, significantly increased the rate of development of contractile dysfunction in bladder tissue strips incubated in all 3 media as compared to nonrepetitive stimulation, which caused no degradation of the contractile response in normal physiological medium. The rate of development of contractile dysfunction was significantly greater in bladder tissue strips incubated in the in vitro ischemia medium (N<sub>2</sub> – glucose) than in strips incubated in the hypoxia medium (N<sub>2</sub> + glucose); which, in turn, was significantly greater than in those incubated in the normal physiological medium (O<sub>2</sub> + glucose). Repetitive stimulation (‘hyperreflexia’) during all 3 incubation conditions resulted in increased [MDA] after reoxygenation or ‘reperfusion’. Incubation in in vitro ischemia buffer (N<sub>2</sub> – glucose) followed by 1 h reoxygenation + substrate replacement stimulated lipid peroxidation to a significantly greater extent than did incubation in hypoxia buffer (N<sub>2</sub> + glucose) followed by 1 h of reoxygenation; the level of lipid peroxidation, [MDA], paralleled the magnitude of the contractile dysfunctions present. Independent of the incubation medium, the magnitude of FS-induced contractile dysfunction after reoxygenation or ‘reperfusion’ was significantly greater than the magnitude of dysfunction in response to carbachol or KCl. <b>Conclusions:</b> The results demonstrate that the rate of contractile failure induced by in vitro ischemia is greater than that induced by in vitro hypoxia, and that the contractile response to FS is significantly more sensitive to both hypoxia and in vitro ischemia than is the contractile response to either carbachol or KCl. Repetitive stimulation (‘hyperreflexia’) increases the rate of contractile failure under all conditions tested, and the magnitude of the contractile failure may be due, in part, to the generation of free radicals and subsequent stimulation of lipid peroxidation upon reoxygenation or ‘reperfusion’.