The Oxidation of Catecholamines and 6-Hydroxydopamine by Molecular Oxygen: Effect of Ascorbate

Abstract

Abstract Comparative kinetic studies on the oxidation of catecholamines (CA) (dopamine (DA), epinephrine (EP). norepinephrine (NEP)) serving as a neuromediator in the sympathetic nervous system, 3,4-dihydroxyphenylalanine (DOPA) and 6-hydroxydopamine (6-OHDA), a wellknown neurotoxic agent, were performed in the presence of ascorbate (AscH- ) in 50 mᴍ phosphate buffer, pH 7.40, at 37 °C by using a Clark electrode, EPR and the absorption spectroscopy. The oxidation of CA and DOPA alone was found to be a self-accelerating process, with quinone products (Q) acting as autocatalysts. The rate of oxygen consumption (Rox) increased with time and reached a steady-state level. A starting value of Rox increased in the order: EP < DOPA ≈ NEP ≪ DA ≪ 6-OHDA, whereas a steady-state value of Rox changed in the order: DOPA < DA < NEP ≪ EP ≪ 6-OHDA. The changes in Rox with time were found to correlate with the resistance of primary Q to the intramolecular cyclization. The effect of AscH- on CA oxidation depended dramatically on whether AscH- was added to non-oxidized or preoxidized CA. Added to non-oxidized CA and DOPA, AscH- inhibited their oxidation (but not that of 6-OHDA). For the case of DA, a pronounced lag period was observed by both a Clark electrode and spectrophotometrically. The addition of AscH- to preoxidized CA, DOPA and 6-OHDA induced an increase in Rox a steadystate concentration of the ascorbyl radical. The kinetic behaviour of the systems was determined by two major factors: 1) AscH- suppressed the formation of Q, a catalyst for CA oxidation, most likely due to the reaction of AscH- with the semiquinone formed from CA; 2) Q derived both from CA and 6-OHDA catalyzed AscH- oxidation. The elevated cytotoxicity of 6-OHDA was found to be in part caused by the condition that 6-OHDA oxidation was not inhibited by AscH- and by the high efficiency of 6-OHDA as a redox cycling agent in combination with A scH−. These observations explain the very pronounced and prolonged cytotoxicity of 6-OHDA even at low concentrations that increases at elevated concentrations of AscH- .