Measurement of peroxiredoxin-4 serum levels in rat tissue and its use as a potential marker for hepatic disease
Measurement of peroxiredoxin-4 serum levels in rat tissue and its use as a potential marker for hepatic disease
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DOI：
10.3892/mmr.2012.935
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发表时间：
2012-08-01
影响因子：
3.4
通讯作者：
Ikeda, Yoshitaka
中科院分区：
文献类型：
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作者：
Ito, Ritsu;Takahashi, Motoko;Ikeda, Yoshitaka
过氧化物氧还蛋白（Peroxiredoxin，Prx）-4是哺乳动物Prx家族的一种分泌型内质网（Endoplasmic reticulum，ER）同工型，具有硫氧还蛋白依赖性过氧化物酶的功能。公认的是，Prx-4在过氧化氢和潜在的其他过氧化物的解毒中起作用，这些过氧化物可能在ER中蛋白质的氧化折叠和氧化应激期间产生。进行本研究是为了具体量化Prx-4的组织水平。为了实现这一点，酶联免疫吸附试验，开发了一个特定的多克隆抗体通过免疫兔与天然重组大鼠Prx-4蛋白。该试验用于检测Prx-4在0.1和10 ng/ml的范围内，并研究大鼠的组织分布。使用这种免疫测定法，我们发现，与正常大鼠相比，无症状的Long-Evans Cinnamon大鼠（威尔逊病的大鼠模型）的血清Prx-4水平显著降低。此外，用N-乙酰半胱氨酸处理大鼠肝癌细胞导致Prx-4蛋白释放到培养基中的显著增加;因此，Prx-4的分泌似乎可能与细胞内的氧化还原状态相关。这些结果表明，血清Prx-4有可能用作肝氧化应激的生物标志物。
Peroxiredoxin (Prx)-4, a secretable endoplasmic reticulum (ER)-resident isoform of the mammalian Prx family, functions as a thioredoxin-dependent peroxidase. It is acknowledged that Prx-4 plays a role in the detoxification of hydrogen peroxide, and potentially other peroxides, which may be generated during the oxidative folding of proteins and oxidative stress in the ER. The present study was undertaken in order to specifically quantify the tissue levels of Prx-4. To accomplish this, an enzyme-linked immunosorbent assay was developed using a specific polyclonal antibody produced by immunizing a rabbit with native recombinant rat Prx-4 protein. The assay was used to detect Prx-4 in the range of 0.1 and 10 ng/ml, and to investigate tissue distribution in rats. Using this immunoassay, we found that the serum levels of Prx-4 were substantially lower in asymptomatic Long-Evans Cinnamon rats, a rat model of Wilson's disease, compared to normal rats. In addition, the treatment of rat hepatoma cells with N-acetylcysteine led to a significant increase in the release of Prx-4 protein into the medium; thus, it appears likely that the secretion of Prx-4 is associated with the redox state within cells. These results suggest that serum Prx-4 has potential for use as a biomarker for hepatic oxidative stress.