MECHANISMS OF HYPOCHLORITE INJURY OF TARGET-CELLS
MECHANISMS OF HYPOCHLORITE INJURY OF TARGET-CELLS
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DOI：
10.1172/jci114472
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发表时间：
1990-02-01
影响因子：
15.9
通讯作者：
COCHRANE, CG
中科院分区：
文献类型：
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作者：
SCHRAUFSTATTER, IU;BROWNE, K;COCHRANE, CG
HOCl，这是由髓过氧化物酶的作用过程中产生的刺激中性粒细胞的呼吸爆发，被用作P338 D1细胞的细胞毒性试剂。低浓度的HOC 1（1-20 μ M）引起质膜巯基氧化，确定为碘乙酰化藻红蛋白的结合减少。这些相同的低浓度的HOCl引起的各种质膜功能的干扰：他们灭活葡萄糖和氨基异丁酸的摄取，造成细胞K+的损失，并增加细胞体积。这些变化很可能是质膜SH氧化的结果，因为对氯汞苯磺酸盐（pCMBS）（一种作用于细胞表面的巯基试剂）也观察到了类似的效应。组合给予pCMBS和HOCl显示出累加效应。较高剂量的HOCl（< 50 μ M）导致-SH、甲硫氨酸和色氨酸残基的一般氧化，并形成蛋白质羰基。HOCL诱导的ATP和未降解的NAD的损失紧随细胞裂解。相比之下，NAD降解和ATP耗尽引起的H2 O2细胞死亡前几个小时。DNA链断裂的形成，H2 O2诱导的损伤的一个主要因素，没有观察到与HOCl。因此，HOCl的目标是不同的H2 O2与甘油醛-3-磷酸脱氢酶，这是由两种氧化剂失活的例外。
HOCl, which is produced by the action of myeloperoxidase during the respiratory burst of stimulated neutrophils, was used as a cytotoxic reagent in P338D1 cells. Low concentrations of HOC1 (1-20 .mu.M) caused oxidation of plasma membrane sulfhydryls determined as decreased binding of iodoacetylated phycoerythrin. These same low concentrations of HOCl caused disturbance of various plasma membrane functions: they inactivated glucose and aminoisobutyric acid uptake, caused loss of cellular K+, and an increase in cell volume. It is likely that these changes were the consequence of plasma membrane SH-oxidation, since similar effects were observed with para-chloromercuriphenylsulfonate (pCMBS), a sulfhydryl reagent acting at the cells surface. Given in combination pCMBS and HOCl showed an additive effect. Higher doses of HOCl (< 50 .mu.M) led to general oxidation of -SH, methionine and tryptophan residues, and formation of protein carbonyls. HOCL-induced loss of ATP and undegraded NAD was closely followed by cell lysis. In contrast, NAD degradation and ATP depletion caused by H2O2 preceded cell death by several hours. Formation of DNA strand breaks, a major factor of H2O2-induced injury, was not observed with HOCl. Thus targets of HOCl were distinct from those of H2O2 with the exception of glyceraldehyde-3-phosphate dehydrogenase, which was inactivated by both oxidants.