Grider A, Bailey L B, Cousins R J
Food Science and Human Nutrition Department, University of Florida, Gainesville 32611.
Proc Natl Acad Sci U S A. 1990 Feb;87(4):1259-62. doi: 10.1073/pnas.87.4.1259.
Metallothionein concentrations in erythrocyte lysates derived from human subjects were measured by an ELISA procedure. IgG obtained from serum of sheep injected with human metallothionein 1 was used in this competitive assay. Subjects were fed a semipurified zinc-deficient diet (0.7 mg of zinc per kg of diet) for an 8-day depletion period after 3 days of acclimation. Fasting plasma zinc concentrations were reduced approximately 7%. Metallothionein in the erythrocyte lysates was significantly decreased to 59% of the initial level by the end of the depletion period. Supplementation of these depleted subjects with zinc (50 mg) did not increase erythrocyte metallothionein levels within 24 hr. Daily supplementation of control subjects with zinc (50 mg/day) increased erythrocyte metallothionein to a 7-fold maximum within 7 days. These levels were reduced by 61% within 14 days after zinc supplementation was terminated. Incubation of rat [35S]metallothionein with human erythrocyte lysate showed a time-dependent increase in 35S soluble in 20% trichloroacetic acid, indicating degradation of the labeled protein, presumably via protease activity in the lysate. It is proposed that zinc supplementation induces erythrocyte metallothionein during erythropoiesis and that low zinc intake decreases synthesis and/or accelerates degradation of the protein in reticulocytes/erythrocytes. Metallothionein levels in erythrocytes may provide a useful index upon which to assess zinc status in humans.
采用酶联免疫吸附测定(ELISA)法测量了人体受试者红细胞裂解物中的金属硫蛋白浓度。在这种竞争性测定中,使用了从注射人金属硫蛋白1的绵羊血清中获得的IgG。受试者在适应3天后,食用半纯化的缺锌饮食(每千克饮食含0.7毫克锌),为期8天的耗竭期。空腹血浆锌浓度降低了约7%。到耗竭期结束时,红细胞裂解物中的金属硫蛋白显著降至初始水平的59%。给这些缺锌受试者补充锌(50毫克)在24小时内并未提高红细胞金属硫蛋白水平。给对照受试者每日补充锌(50毫克/天),在7天内红细胞金属硫蛋白最多增加到7倍。在停止补锌后14天内,这些水平降低了61%。将大鼠[35S]金属硫蛋白与人红细胞裂解物一起孵育,结果显示,可溶于20%三氯乙酸的35S呈时间依赖性增加,表明标记蛋白发生了降解,推测是通过裂解物中的蛋白酶活性。有人提出,补锌在红细胞生成过程中诱导红细胞金属硫蛋白,而低锌摄入会降低网织红细胞/红细胞中该蛋白的合成和/或加速其降解。红细胞中的金属硫蛋白水平可能为评估人体锌状态提供一个有用的指标。