Quantitative Assessment of the Effect of Nitric Oxide Synthase Inhibition on Tumor Vascular Activity Using Dynamic Contrast-Enhanced Computed Tomography

Purpose: The purpose of this study was to develop a method to quantitatively assess the effect of nitric oxide synthase (NOS) inhibition on tumor vascular activity using dynamic contrast-enhanced computed tomography (DCE-CT) and to investigate its usefulness using animal experiments. Mate-rials and Methods: The DCE-CT studies were performed in anesthetized Fisher rats bearing tumors using a 4-row multi-slice CT. The scanning started 4 s before a bolus injection of iodinated contrast agent (CA) (150 mgI/kg) from the tail vein using an automatic injector and lasted 60 s at 1-s in-tervals. The contrast enhancement (CE) images were generated by subtracting the CT images before and after the administration of CA. First, the DCE-CT studies were performed before and 15, 30, and 45 min after administration of N-nitro-L-arginine (L-NNA) (1, 3, and 10 mg/kg) or vehicle, and the relative CE values were calculated by normalizing the CE image at each time point by that obtained from the first DCE-CT study. Second, we investigated the case when L-arginine (L-ARG) (200 mg/kg) and L-NNA (1, 3, and 10 mg/kg) were administered after the first and second DCE-CT studies, respectively. Third, we investigated the case when L-NNA (1, 3, and 10 mg/kg) and L-ARG (200 mg/kg) were administered after the first and second DCE-CT studies, respectively. Finally, we investigated the case when L-NNA (1, 3, and 10 mg/kg) and L-ARG (200 mg/kg) were administered simultaneously after the first DCE-CT study. Results: The relative CE value significantly decreased after L-NNA administration in a dose-dependent manner (p-values = 0.0074 and <0.0001 for 0 vs. 3 mg/kg and 0 vs. 10 mg/kg, respectively, at 15 min, 0.0003 and <0.0001 for 0 vs. 3 mg/kg and 0 vs. 10 mg/kg, respectively, at 30 min, and 0.0367 and 0.0004 for 0 vs. 3 mg/kg and 0 vs. 10 mg/kg, respectively, at 45 min). When L-ARG was administered prior to the administration of 1 mg/kg L-NNA, the relative CE value at 45 min was significantly higher than that at 15 min. When L-ARG was administered after L-NNA administration, there was no significant difference between the relative CE values at 15 min and 45 min. These results suggest that when using L-NNA in combination with L-ARG, their effect on tumor vascular activity differs depending on the order of their administration. When L-NNA and L-ARG were administered simultaneously, there was a tendency for the relative CE value to be higher than that when only L-NNA was administered, at all injected doses of L-NNA. Conclusion: Our method using DCE-CT is useful for monitoring the effect of NOS inhibition on tumor vascular activity and for determining the optimal injected dose and timing of NOS inhibitors for anticancer therapy.