Effec to fCV159-Ca 2 /Calmoduli nBlockad eo nRedo xStatu sHepatic Ischemia-Reperfusio nInjur yi nMic eEvaluate db ya Newl yDeveloped I nViv oEP RImagin gTechnique

1,4-Dihydro-2,6-dimethyl-4-(3-nitrophenyl)-3,5-pyridine- dicarboxylic acid methyl 6-(5-phenyl-3-pyrazolyloxy)hexyl ester (CV159) exhibits selective blocking of Ca 2 / 2 overloading in living or- ganisms. The effects of this antagonist in mice with hepatic ischemia-reperfusion injury were investi- gated using electron paramagnetic resonance imaging (EPRI) and ex vivo EPR (x-band EPR) techniques. The EPRI determined that the 3-carbamoyl-2,2, 5,5-tetramethylpyrrolidine-1-oxyl half-life in CV159- treated mice was significantly shorter than that in untreated mice and was almost equal to that in the sham group. Both the cytosolic and the mitochon- drial superoxide scavenging activities in CV-treated mice were significantly higher than that in un- treated mice. Faint staining of the anti-superoxide dismutase antibody and strong staining of anti- inducible nitric oxide synthase antibody were ob- served in the liver of control group. In contrast to these findings, immunostaining of these antibodies in the liver of CV159-treated mice were reversed compared to control group. Western blotting showed that CV159 contributed to the high superoxide dis- mutase expression and low expression of inducible nitric oxide synthase. The alanine aminotransferase level in CV159-treated mice significantly decreased in comparison to that observed in the untreated mice. We conclude that CV159 retains its organ- reducing activity against radicals in hepatic reper- fusion injury, which is mediated by the inhibition of Ca 2