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人皮肤中阳离子氨基酸转运体(hCATs)1和2的特性研究

Characterization of cationic amino acid transporters (hCATs) 1 and 2 in human skin.

作者信息

Jaeger Kristin, Paulsen Friedrich, Wohlrab Johannes

机构信息

Department of Anatomy and Cell Biology, Martin-Luther-University Halle-Wittenberg, Grosse Steinstrasse 52, Halle, Germany.

出版信息

Histochem Cell Biol. 2008 Mar;129(3):321-9. doi: 10.1007/s00418-007-0367-6. Epub 2008 Jan 3.

Abstract

In the present study, we characterized the distribution of human cationic amino acid transporters 1 (hCAT1) and 2 (hCAT2) in healthy skin and compared it to psoriatic skin lesions by means of immunohistochemistry. Moreover, we tested the hypothesis that L -arginine and L -ornithine influence the expression and synthesis of hCAT1 and hCAT2 in cell culture experiments by means of real-time-PCR and Western blot. Immunohistochemical comparison between healthy and psoriatic skin revealed a decreased amount of hCAT1, especially in the stratum granulosum of psoriatic skin; the distribution pattern of hCAT2 was not significantly affected in psoriatic skin. Cell culture experiments showed that supraphysiological concentrations of 15 mM L -arginine (72 h) lead to a significant increase of the hCAT1-mRNA and protein expression, whereas other concentrations had no significant influence. In contrast, L -arginine concentrations of 2 mM led to a significant increase of the hCAT2B mRNA-expression after 24 h. However, 48 and 72 h revealed no significant changes and high concentrations (15 mM L -arginine) led to a significant downregulation of the hCAT2B transporter over all time points analyzed. L -ornithine had no effect on the hCAT1 expression of mRNA and protein level. On the other hand the expression of hCAT2B was significantly up regulated at a 5-mM concentration of L -ornithine at all analyzed time points. Other concentrations had no effect. For the first time, the findings yield data about hCAT1 and hCAT2 on protein-level and suggest that L -arginine is a worthwhile object of studies, which investigated L -arginine as a possible therapeutic agent to reduce psoriatic symptoms.

摘要

在本研究中,我们通过免疫组织化学方法对健康皮肤中人类阳离子氨基酸转运体1(hCAT1)和2(hCAT2)的分布进行了表征,并将其与银屑病皮肤病变进行了比较。此外,我们在细胞培养实验中通过实时定量PCR和蛋白质印迹法检验了L-精氨酸和L-鸟氨酸影响hCAT1和hCAT2表达与合成的假设。健康皮肤与银屑病皮肤的免疫组织化学比较显示,hCAT1的数量减少,尤其是在银屑病皮肤的颗粒层;银屑病皮肤中hCAT2的分布模式未受到显著影响。细胞培养实验表明,15 mM L-精氨酸的超生理浓度(72小时)会导致hCAT1-mRNA和蛋白质表达显著增加,而其他浓度则无显著影响。相比之下,2 mM的L-精氨酸浓度在24小时后会导致hCAT2B mRNA表达显著增加。然而,在48小时和72小时时未发现显著变化,且高浓度(15 mM L-精氨酸)在所有分析时间点均导致hCAT2B转运体显著下调。L-鸟氨酸对hCAT1的mRNA和蛋白质水平表达没有影响。另一方面,在所有分析时间点,5 mM浓度的L-鸟氨酸会使hCAT2B的表达显著上调。其他浓度则无影响。这些发现首次提供了关于hCAT1和hCAT2蛋白质水平的数据,并表明L-精氨酸是一个值得研究的对象,有研究将L-精氨酸作为一种可能的治疗剂来减轻银屑病症状。

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