Freitas Ana, Aroso Miguel, Barros António, Fernández Miriam, Conde-Sousa Eduardo, Leite Marina, Carvalho Eva Daniela, Ribeiro Cristina C, Ferreira Rita, Pêgo Ana Paula, Vitorino Rui, Gomez-Lazaro Maria
i3S-Instituto de Investigação e Inovação em Saúde, Universidade do Porto, 4200-135 Porto, Portugal.
INEB-Instituto de Engenharia Biomédica, Universidade do Porto, 4200-135 Porto, Portugal.
Antioxidants (Basel). 2021 Jul 8;10(7):1095. doi: 10.3390/antiox10071095.
Parkinson's disease's etiology is unknown, although evidence suggests the involvement of oxidative modifications of intracellular components in disease pathobiology. Despite the known involvement of the extracellular matrix in physiology and disease, the influence of oxidative stress on the matrix has been neglected. The chemical modifications that might accumulate in matrix components due to their long half-live and the low amount of extracellular antioxidants could also contribute to the disease and explain ineffective cellular therapies. The enriched striatal extracellular matrix from a mouse model of Parkinson's disease was characterized by Raman spectroscopy. We found a matrix fingerprint of increased oxalate content and oxidative modifications. To uncover the effects of these changes on brain cells, we morphologically characterized the primary microglia used to repopulate this matrix and further quantified the effects on cellular mechanical stress by an intracellular fluorescence resonance energy transfer (FRET)-mechanosensor using the U-2 OS cell line. Our data suggest changes in microglia survival and morphology, and a decrease in cytoskeletal tension in response to the modified matrix from both hemispheres of 6-hydroxydopamine (6-OHDA)-lesioned animals. Collectively, these data suggest that the extracellular matrix is modified, and underscore the need for its thorough investigation, which may reveal new ways to improve therapies or may even reveal new therapies.
帕金森病的病因尚不清楚，尽管有证据表明细胞内成分的氧化修饰参与了疾病的病理生物学过程。尽管已知细胞外基质参与生理和疾病过程，但氧化应激对基质的影响却被忽视了。由于基质成分的半衰期长且细胞外抗氧化剂含量低，可能在基质成分中积累的化学修饰也可能导致疾病，并解释细胞治疗无效的原因。通过拉曼光谱对帕金森病小鼠模型中富集的纹状体细胞外基质进行了表征。我们发现了草酸盐含量增加和氧化修饰的基质指纹图谱。为了揭示这些变化对脑细胞的影响，我们对用于重新填充该基质的原代小胶质细胞进行了形态学表征，并使用U-2 OS细胞系通过细胞内荧光共振能量转移（FRET）机械传感器进一步量化了对细胞机械应力的影响。我们的数据表明，6-羟基多巴胺（6-OHDA）损伤动物两侧半球的修饰基质会导致小胶质细胞存活和形态的改变，以及细胞骨架张力的降低。总体而言，这些数据表明细胞外基质发生了改变，并强调了对其进行深入研究的必要性，这可能会揭示改善治疗的新方法，甚至可能揭示新的治疗方法。
