The role of Nitric oxide synthase andCytochrome P450 enzyme on the inhibitory effect of sevoflumae on vascular relaxation

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<Isometric Force tension Recordings>After obtaining the various results as control, we checked both the Ach-induced relaxation of the rings pretreated with chlorzoxasone, a specific substrate of CYP450-2E1 and that of the rings from the rats to which CYP450-2E1 was induced by feeding with the water containing ethanol. From these results, the interaction between sevoflurane and CYP450-2E1 is suggested to be included in the mechanism underlying sevoflurane-induced inhibition of endothelium dependent vascular relaxation.<The measurement of nitric oxide synthase activity by Radioimmunoassay>The activity of nitric oxide synthase was tested with the method of measuring the conversion of radiolabelled L-arginine to L-citrulline. Sevoflurane did not affect the conversion L-arginine to L-citrulline which suggests that sevoflurane has no effect on nitric oxide synthase activity.<The confirmation of CYP450-2E1 induction with RT-PCR>The rings from the rats treatment with ethanol drinking were assured to overexpress CYP450-2E1 mRNA with the method of competitive RT-PCR.<The nitric oxide assay with chemoluminescence>After loading DAF FM-DA, nitroc oxide sensitive luminescence into the cultured bovine aortic endothelial cells, we tested the effect of sevoflurane on the intensity induced by bradykinin with or without the treatment of CYP450-2E1 induction. Under the treatment of CYP450-2E1 induction, the sevoflurane-induced inhibition of intensity was abolished.From these findings, it is suggested that CYP450-2E1 is one of the mediators producing Ach-induced endotherium dependent vascular relaxation, and sevoflurane expresses the inhibitory effect on the relaxation th* interference to the activity of CYP450-2E1 inside endothelial cells.