Iizumi Kyoichi, Kawasaki Hiroaki, Shigenaga Ayako, Tominaga Mitsutoshi, Otsu Ayaka, Kamo Atsuko, Kamata Yayoi, Takamori Kenji, Yamakura Fumiyuki
Juntendo University Faculty of Health and Sports Science, 1-1 Hirakagakuendai, Inzai, Chiba 270-1695, Japan.
Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Tomioka, Urayasu, Chiba 279-0021, Japan.
J Clin Biochem Nutr. 2018 Nov;63(3):197-204. doi: 10.3164/jcbn.18-53. Epub 2018 Sep 15.
To reduce the incidence and severity of atopic dermatitis, detection and treatment at an early stage are urgently required, but no effective biomarker has been reported. In this study, we attempted to detect a candidate biomarker of early stage atopic dermatitis by focusing on the levels of nitrated residues in the plasma proteins of atopic dermatitis model mice (NC/Nga mice). We found that the immunoglobulin (Ig) light chain was more highly nitrated in the plasma of the animal model than that of control mice. Western blot analysis showed a statistically significant difference between the 6-nitrotryptophan content of the Ig light chain in the NC/Nga mice before onset of atopic dermatitis symptoms and that of the control mice. LC-ESI-MS/MS analysis demonstrated that these light chains contained nitrotryptophan (Trp56) and nitrotyrosine (Tyr66). Immunofluorescence staining revealed a significant increase in manganese superoxide dismutase and inducible nitric oxide synthase production in the skin lesions of the NC/Nga mice. Furthermore, we found protein-bound 6-nitrotryptophan and 3-nitrotyrosine only in the lesioned skin, where their signals partially overlapped with the IgG signal. Our findings suggest that the 6-nitrotryptophan content of Ig light chains could be a new biomarker for detecting early stage atopic dermatitis.
为降低特应性皮炎的发病率和严重程度,迫切需要在早期进行检测和治疗,但尚未有有效的生物标志物被报道。在本研究中,我们试图通过关注特应性皮炎模型小鼠(NC/Nga小鼠)血浆蛋白中硝化残基的水平,来检测早期特应性皮炎的候选生物标志物。我们发现,动物模型血浆中的免疫球蛋白(Ig)轻链硝化程度高于对照小鼠。蛋白质印迹分析表明,在NC/Nga小鼠出现特应性皮炎症状之前,其Ig轻链的6-硝基色氨酸含量与对照小鼠相比存在统计学显著差异。液相色谱-电喷雾串联质谱分析表明,这些轻链含有硝基色氨酸(Trp56)和硝基酪氨酸(Tyr66)。免疫荧光染色显示,NC/Nga小鼠皮肤病变中锰超氧化物歧化酶和诱导型一氧化氮合酶的产生显著增加。此外,我们仅在病变皮肤中发现了与蛋白质结合的6-硝基色氨酸和3-硝基酪氨酸,它们的信号部分与IgG信号重叠。我们的研究结果表明,Ig轻链的6-硝基色氨酸含量可能是检测早期特应性皮炎的一种新生物标志物。