Department of Medical and Clinical Biochemistry, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Trieda SNP 1, 040 11 Košice, Slovakia.
Department of Medical and Clinical Biophysics, Faculty of Medicine, Pavol Jozef Šafárik University in Košice, Trieda SNP 1, 040 11 Košice, Slovakia.
Sensors (Basel). 2023 Jun 1;23(11):5251. doi: 10.3390/s23115251.
A pilot analysis of the tear fluid of patients with multiple sclerosis (MS) collected by glass microcapillary was performed using various experimental methods: liquid chromatography-mass spectrometry, Raman spectroscopy, infrared spectroscopy, and atomic-force microscopy. Infrared spectroscopy found no significant difference between the tear fluid of MS patients and the control spectra; all three significant peaks were located at around the same positions. Raman analysis showed differences between the spectra of the tear fluid of MS patients and the spectra of healthy subjects, which indicated a decrease in tryptophan and phenylalanine content and changes in the relative contributions of the secondary structures of the polypeptide chains of tear proteins. Atomic-force microscopy exhibited a surface fern-shaped dendrite morphology of the tear fluid of patients with MS, with less roughness on both oriented silicon (100) and glass substrates compared to the tear fluid of control subjects. The results of liquid chromatography-mass spectrometry showed downregulation of glycosphingolipid metabolism, sphingolipid metabolism, and lipid metabolism. Proteomic analysis identified upregulated proteins in the tear fluid of patients with MS such as cystatine, phospholipid transfer protein, transcobalamin-1, immunoglobulin lambda variable 1-47, , and ferroptosis suppressor protein 1; and downregulated proteins such as haptoglobin, prosaposin, cytoskeletal keratin type I pre-mRNA-processing factor 17, neutrophil -associated lipocalin, and . This study showed that the tear proteome in patients with MS is modified and can reflect inflammation. Tear fluid is not a commonly used biological material in clinico-biochemical laboratories. Experimental proteomics has the potential to become a promising contemporary tool for personalized medicine, and it might be applied in clinical practice by providing a detailed analysis of the tear-fluid proteomic profile of patients with MS.
采用玻璃微毛细管收集多发性硬化症 (MS) 患者的泪液，应用多种实验方法（液相色谱-质谱法、拉曼光谱法、红外光谱法和原子力显微镜）对其进行了初步分析。红外光谱法发现 MS 患者的泪液与对照光谱之间没有明显差异；三个显著峰均位于相同位置。拉曼分析显示 MS 患者泪液的光谱与健康受试者的光谱存在差异，表明色氨酸和苯丙氨酸含量降低，泪蛋白多肽链二级结构的相对贡献发生变化。原子力显微镜显示 MS 患者泪液呈现出表面蕨叶状树突形态，与健康受试者的泪液相比，在取向硅 (100) 和玻璃衬底上的粗糙度都较小。液相色谱-质谱法的结果表明糖脂代谢、鞘脂代谢和脂质代谢下调。蛋白质组学分析发现 MS 患者泪液中上调的蛋白质，如胱抑素、磷脂转移蛋白、转钴胺素-1、免疫球蛋白 lambda 可变 1-47 、 ，和铁死亡抑制蛋白 1 ；下调的蛋白质，如触珠蛋白、前蛋白原、细胞骨架角蛋白 I 前 mRNA 加工因子 17 、嗜中性粒细胞相关脂钙蛋白和 。本研究表明 MS 患者的泪液蛋白质组发生了改变，并能反映炎症。泪液不是临床生物化学实验室中常用的生物材料。实验蛋白质组学有可能成为个性化医学的一种有前途的现代工具，通过对 MS 患者泪液蛋白质组图谱进行详细分析，可能在临床实践中得到应用。
