LeDay Angela M, Awe Sunday O, Kulkarni Kaustubh, Harris Lydia C, Opere Catherine, Dash Alekha, Ohia Sunny E
Department of Pharmacy Sciences, School of Pharmacy and Health Professions, Creighton University Medical Center, Omaha, Nebraska, USA.
Neurochem Res. 2004 Apr;29(4):811-8. doi: 10.1023/b:nere.0000018855.77589.5d.
In the present study, we investigated the effect of naturally occurring and synthetic peroxides on K+-depolarization-evoked release of [3H]D-aspartate from bovine isolated retinae. Furthermore, effect of peroxides on endogenous glutamate concentrations were measured by HPLC in bovine neural retinae and vitreous humor of eyes treated with hydrogen peroxide (H2O2) ex vivo. Both naturally occurring H2O2 (1-100 microM) and synthetic (cumene hydroperoxide, cuOOH; 1-100 microM) peroxides caused a concentration-dependent inhibition of K+-evoked [3H]D-aspartate release without affecting basal tritium efflux. The antioxidant, trolox (2 mM) prevented the inhibition of evoked [3H]D-aspartate overflow elicited by both H2O2 (30 microM) and cuOOH (10 microM). Inhibition of catalase by 3-amino-triazole (3- AT 100 mM) enhanced an inhibitory effect of a low concentration of H2O2 (1 microM) but antagonized the effect of H2O2 (30 microM) on K+-induced [3H]D-aspartate release. In ex vivo experiments, exogenously applied H2O2 (1-100 microM) also caused a concentration-related decrease in glutamate levels in the bovine retina. We conclude that peroxides can inhibit K+-evoked release of [3H]D-aspartate and also decrease endogenous glutamate concentrations in the bovine retina.
在本研究中,我们调查了天然存在的和合成的过氧化物对牛离体视网膜中钾离子去极化诱发的[3H]D-天冬氨酸释放的影响。此外,通过高效液相色谱法(HPLC)测量了过氧化物对过氧化氢(H2O2)离体处理的牛神经视网膜和眼玻璃体液中内源性谷氨酸浓度的影响。天然存在的H2O2(1-100微摩尔)和合成过氧化物(氢过氧化异丙苯,cuOOH;1-100微摩尔)均引起钾离子诱发的[3H]D-天冬氨酸释放呈浓度依赖性抑制,而不影响基础氚外流。抗氧化剂生育三烯酚(2毫摩尔)可防止H2O2(30微摩尔)和cuOOH(10微摩尔)引起的诱发[3H]D-天冬氨酸溢出的抑制。用3-氨基-1,2,4-三唑(3-AT,100毫摩尔)抑制过氧化氢酶可增强低浓度H2O2(1微摩尔)的抑制作用,但拮抗H2O2(30微摩尔)对钾离子诱导的[3H]D-天冬氨酸释放的作用。在离体实验中,外源性应用H2O2(1-100微摩尔)也导致牛视网膜中谷氨酸水平呈浓度相关下降。我们得出结论,过氧化物可抑制钾离子诱发的[3H]D-天冬氨酸释放,并降低牛视网膜中的内源性谷氨酸浓度。