Determination of the serum metallothionein (MT)1/2 concentration in patients with Wilson's disease and Menkes disease
Determination of the serum metallothionein (MT)1/2 concentration in patients with Wilson's disease and Menkes disease
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DOI：
10.1016/j.jtemb.2014.07.013
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发表时间：
2014-01-01
影响因子：
3.5
通讯作者：
Nagamine, Takeaki
中科院分区：
文献类型：
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作者：
Nakazato, Kyoumi;Tomioka, Satoru;Nagamine, Takeaki
我们建立了一种简便、特异的酶联免疫分析方法，可同时测定人和实验动物血清中金属金属蛋白-1(MT-1)和金属蛋白酶-2(MT-2)的含量。用抗大鼠MT-2的特异性多克隆抗体建立了竞争性酶联免疫吸附试验。该抗体在人和实验动物中表现出与MT相同的交叉反应。含有乙酰化蛋氨酸的MT的NH2末端多肽是该抗体的表位。该体系对MT1/2基因敲除小鼠的肝、肾和脑组织的反应性较低，而对MT-3基因敲除小鼠的反应正常。该方法的最低检测下限为0.6 ng/ml，可从血浆中完全回收MT-1。我们调查了200例健康人血清中MT-1/2(25-75%)的正常范围，发现其范围为27-48 ng/ml，并与各种肝病患者的血清水平进行了比较。慢性丙型肝炎患者血清MT1/2水平明显低于健康对照组和其他肝病患者。在慢性肝炎患者中，MT1/I2水平逐渐升高，随着疾病的进展而发展为肝硬变和肝细胞癌。特别是，我们发现肝豆状核变性患者血浆MT1/2水平显著升高，这一水平与Long-Evans Cinnamon(LEC)大鼠(肝豆状核变性模型动物)非常相似。此外，在Menkes病患者中发现MT1/2水平显著升高。Menkes病是一种先天性铜代谢错误，如肝豆状核变性。(C)2014年爱思唯尔股份有限公司。版权所有。
We have developed an easy and specific enzyme-linked immunoassay (ELISA) for the simultaneous determination of serum metallothinein-1 (MT-1) and 2 (MT-2) in both humans and experimental animals. A competitive ELISA was established using a specific polyclonal antibody against rat MT-2. The antibody used for this ELISA had exhibited the same cross-reactivity with MT in humans and experimental animals. The NH2 terminal peptide of MT containing acetylated methionine was shown to be the epitope of this antibody. The reactivity of this ELISA system with the liver, kidney and brain in MT1/2 knock-out mice was significantly low, but was normal in an MT-3 knock-out mouse. The lowest detection limit of this ELISA was 0.6 ng/ml and the spiked MT-1 was fully recovered from the plasma.We investigated the normal range of MT1/2 (25-75%tile) in 200 healthy human serum and found it to be 27-48 ng/ml, and this was compared with the serum levels in various liver diseases. The serum MT1/2 levels in chronic hepatitis C (HCV) patients were significantly lower than healthy controls and also other liver diseases. In the chronic hepatitis cases, the MT1/I2 levels increased gradually, followed by the progression of the disease to liver cirrhosis and hepatocellular carcinoma. In particular, we found significantly elevated MT1/2 plasma levels in Wilson's disease patients, levels which were very similar to those in the Long-Evans Cinnamon (LEC) rat (model animal of Wilson's disease). Furthermore, a significantly elevated MT1/2 level was found in patients with Menkes disease, an inborn error of copper metabolism such as Wilson's disease. (C) 2014 Elsevier GmbH. All rights reserved.