The perfusion for 15 min of isolated rat hearts with 100 microM t-butylhydroperoxide leads to a 75% diminuition of the tissue GSH/GSSG ratio. 2. After t-butylhydroperoxide infusion, the isoproterenol-stimulated heart ODC was strongly inhibited. The addition of 2 mM DTT in the assaying buffer removed the ODC inactivation. 3. The inhibited ODC had an eluition profile similar to active ODC when chromatographed on a Sephacryl S-200 column; moreover, the ODC activity recovered after a thiol affinity chromatography as unbound fraction, was two times increased in the t-butylhydroperoxide perfused hearts in comparison to control. 4. The hearts perfused with 1 mM acetylcysteine after 15 min of perfusion with t-butylhydroperoxide recovered almost completely the initial ODC activity.
摘要
用100微摩尔叔丁基过氧化氢对离体大鼠心脏灌注15分钟,会导致组织谷胱甘肽/氧化型谷胱甘肽比值降低75%。2. 输注叔丁基过氧化氢后,异丙肾上腺素刺激的心脏鸟氨酸脱羧酶(ODC)受到强烈抑制。在测定缓冲液中加入2毫摩尔二硫苏糖醇(DTT)可消除ODC的失活。3. 当在Sephacryl S - 200柱上进行色谱分析时,受抑制的ODC具有与活性ODC相似的洗脱图谱;此外,经硫醇亲和色谱作为未结合部分回收后,叔丁基过氧化氢灌注心脏中的ODC活性与对照相比增加了两倍。4. 在叔丁基过氧化氢灌注15分钟后,用1毫摩尔乙酰半胱氨酸灌注的心脏几乎完全恢复了初始ODC活性。