Ramos-Martínez Ivan, Vivanco-Rojas Oscar, Juárez-Domínguez Brenda, Hernández-Zimbrón Luis, Ochoa-de la Paz Lenin, Quiroz-Mercado Hugo, Ramírez-Hernández Eleazar, Gulias-Cañizo Rosario, Zenteno Edgar
Departamento de Bioquímica, Facultad de Medicina UNAM, Ciudad de Mexico, 04510, Mexico.
Departamento de Investigación, Asociación para Evitar la Ceguera en Mexico I.A.P. Hospital Dr. Luis Sánchez Bulnes, Mexico City, Mexico.
Clin Ophthalmol. 2021 Mar 31;15:1365-1373. doi: 10.2147/OPTH.S300242. eCollection 2021.
In order to better understand cataract development, we analyzed the glycosylation profile of human lens epithelial cells (HLECs) from anterior lens capsules of type 2 diabetes mellitus (T2DM) and non-diabetic (ND) patients undergoing routine cataract surgery.
Research Department of the Asociación para Evitar la Ceguera, Hospital "Dr. Luis Sánchez Bulnes", Mexico.
Experimental study.
Evaluation of anterior lens capsules from T2DM and ND patients undergoing phacoemulsification and free from other ocular diseases.
Hematoxylin-eosin staining revealed HLECs alterations in T2DM samples. From lectins with different sugar specificities used, concanavalin A showed significant differences, labeling homogeneously both in the cytoplasm and in cell membranes in ND capsules, while in T2DM capsules, in addition to membrane and cytoplasm labeling, there were perinuclear vesicles with high concanavalin A labeling. Two-dimensional gel electrophoresis showed that T2DM patients have a ~65-kDa spot with an isoelectric point of 5.5 with a higher density compared to ND capsules, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed 62% homology with type-1 cytokeratin. Immunohistochemistry using anti-pan cytokeratin antibody revealed co-localization with concanavalin A, and a lectin blot revealed with concanavalin A showed a band of ~65 kDa, a molecular weight that corresponds to human type 1 cytokeratin.
These results suggest that over-expression of N-glycosidically linked human type 1 cytokeratin may induce capsule disruption and affect selective permeability, allowing the entry of different molecules to the lens that facilitate cataract progression.
为了更好地理解白内障的发展，我们分析了接受常规白内障手术的2型糖尿病（T2DM）患者和非糖尿病（ND）患者前囊膜中人类晶状体上皮细胞（HLEC）的糖基化谱。
墨西哥“Dr. Luis Sánchez Bulnes”医院，预防失明协会研究部。
实验研究。
评估接受超声乳化且无其他眼部疾病的T2DM和ND患者的前囊膜。
苏木精-伊红染色显示T2DM样本中的HLEC有改变。在所使用的具有不同糖特异性的凝集素中，伴刀豆球蛋白A显示出显著差异，在ND囊膜的细胞质和细胞膜中均均匀标记，而在T2DM囊膜中，除了膜和细胞质标记外，还有高伴刀豆球蛋白A标记的核周囊泡。二维凝胶电泳显示，与ND囊膜相比，T2DM患者有一个等电点为5.5、分子量约65 kDa的斑点，密度更高，液相色谱-串联质谱（LC-MS/MS）分析显示与1型细胞角蛋白有62%的同源性。使用抗泛细胞角蛋白抗体的免疫组织化学显示与伴刀豆球蛋白A共定位，伴刀豆球蛋白A显示的凝集素印迹显示有一条约65 kDa的条带，该分子量对应于人类1型细胞角蛋白。
这些结果表明，N-糖苷连接的人类1型细胞角蛋白的过度表达可能会导致囊膜破坏并影响选择性通透性，使不同分子进入晶状体，从而促进白内障进展。
