Different action of IBMX, isoproterenol and rutin on orthovanadate-induced nitric oxide release in mouse macrophage cells

Abstract

The effects of cAMP-elevating compounds IBMX (3-isobutyl-1-methyl­xanthine) and isoproterenol, and that of rutin (an effective superoxide scavenger) were studied on orthovanadate- (a putative protein-phosphotyrosine phosphatase inhibitor) induced nitric oxide (NO) production in J774A.1 mouse macrophage cells. As we previously reported (Koncz and Horváth, 2000), rutin and sodium orthovanadate act synergistically to induce production of high amount of NO in J774A.1 cells. IBMX, an agent that can elevate cAMP level in the cells, can reduce the production of both the LPS- and rutin + orthovanadate-induced NO in macrophages. In contrast, isoproterenol, a non-selective ß-adrenergic receptor agonist, that reduced the LPS-induced NO production in macrophage cells, was unable to reduce the rutin + orthovanadate-induced NO production without negatively affecting cell viability. Moreover, isoproterenol dramatically enhanced the orthovanadate-induced NO synthesis in J774A.1 cells. Our previous study clarified that rutin and orthovanadate, in a specific concentration ratio of both, were able to produce hydrogen peroxide (H2O2). Using 2',7'-dichlorofluorescein-diacetate as a marker for H2O2, isoproterenol alone induced its oxidation but the rutin plus orthovanadate-induced H2O2production was reduced by isoproterenol. These observations have revealed that, in some cases, H2O2and superoxide (O2-) scavengers can act in a reverse mode on macrophage cells depending on the presence or absence of orthovanadate.