Laboratory of Micronutrient, Institute of Nutrition and Food Technology (INTA), University of Chile, Santiago, Chile.
Biometals. 2012 Apr;25(2):383-91. doi: 10.1007/s10534-011-9511-9. Epub 2011 Nov 29.
Assessment of proteins in blood and other tissues has failed to identify markers of early copper effects on health. Studies in animal models show that chaperone of SOD (CCS) respond to changes of copper status. Evidence about other copper chaperones (COXIV, ATOX) is not clear. The aim of this study was to assess by means of an in vitro challenge the mRNA relative abundance of ccs, sod1, coxIV, mtIIa and atox in peripheral mononuclear cells (PMNCs) obtained from healthy individuals, acutely and chronically supplemented with small-to-moderate amounts of copper. Healthy participants received 8 mg Cu/d (supplemented group, SG) or placebo, (placebo group, PG) for 2 months. Biochemical indicators were assessed at basal (T0) and after 2 (T2) and 60 days (T60). At these times PMNCs were obtained, challenged with 1, 5 or 20 μM Cu-histidine for 20 h and the mRNA relative abundance of the selected genes assessed by real time PCR. The results showed that at T0, intracellular copper was not different between experimental and control groups. This increased at T2 and T60 when the copper in the media increased (two-way ANOVA, P < 0.001). In PG, CCS mRNA transcripts showed no significant changes (two-way ANOVA) at T2 and T60. In SG, CCS changed by treatment, time and interaction (two-way ANOVA, all P < 0.001). SOD, ATOX and COXIV expressions changed in both PG and SG showing various patterns of response, requiring further study. MTII responded as expected. We conclude that using healthy individuals as a human model, CCS but not SOD, ATOX or COXIV responded consistently to controlled changes of copper availability in an in vitro copper challenge.
评估血液和其他组织中的蛋白质未能确定早期铜对健康影响的标志物。动物模型研究表明，SOD 的伴侣蛋白（CCS）对铜状态变化有反应。关于其他铜伴侣蛋白（COXIV、ATOX）的证据尚不清楚。本研究旨在通过体外挑战评估来自健康个体的外周单核细胞（PMNC）中 ccs、sod1、coxIV、mtIIa 和 atox 的 mRNA 相对丰度，这些个体急性和慢性补充少量至中等量的铜。健康参与者每天接受 8 毫克 Cu（补充组，SG）或安慰剂（安慰剂组，PG）治疗 2 个月。在基线（T0）和 2 天后（T2）和 60 天后（T60）评估生化指标。此时从 PMNC 中获得，用 1、5 或 20 μM Cu-组氨酸刺激 20 小时，并通过实时 PCR 评估所选基因的 mRNA 相对丰度。结果表明，在 T0 时，实验组和对照组之间细胞内铜没有差异。当培养基中的铜增加时，这种情况在 T2 和 T60 时增加（双因素方差分析，P < 0.001）。在 PG 中，CCS mRNA 转录本在 T2 和 T60 时没有明显变化（双因素方差分析）。在 SG 中，CCS 因处理、时间和相互作用而变化（双因素方差分析，所有 P < 0.001）。SOD、ATOX 和 COXIV 的表达在 PG 和 SG 中均发生变化，表现出不同的反应模式，需要进一步研究。MTII 反应如预期。我们得出结论，使用健康个体作为人体模型，CCS 而不是 SOD、ATOX 或 COXIV 对体外铜挑战中铜可用性的受控变化始终有反应。
