Possible involvement of enhanced arginase activity due to up-regulated arginases and decreased hydroxyarginine in accelerating intimal hyperplasia with hyperglycemia

Abstract

The present study was designed to investigate the roles of enhanced arginase activity due to up-regulated arginases and the decreased hydroxyarginine for accelerating intimal hyperplasia with hyperglycemia. Thirteen weeks after injection of alloxan or physiological saline, endothelial denudation of the carotid artery was performed to induce intimal hyperplasia. The intimal hyperplasia occurred on 4 weeks following denudation was significantly accelerated by hyperglycemia. The method to measure l-arginine, endogenous NOS inhibitors such as monomethylarginine and asymmetric dimethylarginine, and hydroxyarginine as an intermediate of NO production simultaneously was established with the aid of high-performance liquid chromatography. In hyperglycemia group, the impaired cyclic GMP production as an indicator of NO production in endothelial cells was accompanied by the enhanced arginase activity together with increased expression of arginase I and II proteins, accumulated endogenous NOS inhibitors, reduced concentration of hydroxyarginine, and decreased DDAH activity in endothelial cells. However, NOS activity per se remained unchanged in the hyperglycemia group. Authentic hydroxyarginine inhibited arginase activity in a concentration-dependent manner. The inhibition of arginase with hydroxyarginine at a reduced concentration with hyperglycemia became significantly lower than that for the control. These results suggest that the accelerated intimal hyperplasia with hyperglycemia is closely related to the impaired NO production in endothelial cells, which results from accumulation of endogenous NOS inhibitors and accelerated arginase activity together with up-regulation of arginase I and II proteins. Decreased DDAH activity would bring about the accumulation of endogenous NOS inhibitors. Furthermore, reduced concentration of hydroxyarginine with hyperglycemia possibly results in an enhanced arginase activity in vivo, implicating partly in the impairment of NO production.