Department of Retina and Vitreous Services, Aravind Eye Hospital; Department of Proteomics, Aravind Medical Research Foundation, Madurai, Tamil Nadu, India.
Department of Proteomics, Aravind Medical Research Foundation, Madurai, Tamil Nadu, India.
Indian J Ophthalmol. 2023 Aug;71(8):3069-3079. doi: 10.4103/IJO.IJO_276_23.
To explore the vitreous humor proteome from type 2 diabetes subjects with proliferative diabetic retinopathy (PDR) in the Indian population.
We performed mass spectrometry-based label-free quantitative analysis of vitreous proteome of PDR (n = 13) and idiopathic macular hole (IMH; control) subjects (n = 14). Nine samples of PDR and 10 samples of IMH were pooled as case and control, respectively, and compared. Four samples each of PDR and IMH were analyzed individually without pooling to validate the results of the pooled analysis. Comparative quantification was performed using Scaffold software which calculated the fold changes of differential expression. Bioinformatics analysis was performed using DAVID and STRING software.
We identified 469 proteins in PDR and 517 proteins in IMH vitreous, with an overlap of 172 proteins. Also, 297 unique proteins were identified in PDR and 345 in IMH. In PDR vitreous, 37 proteins were upregulated (P < 0.05) and 19 proteins were downregulated compared to IMH. Protein distribution analysis clearly demonstrated a separation of protein expression in PDR and IMH. Significantly upregulated proteins included fibrinogen gamma chain, fibrinogen beta chain, and carbonic anhydrase 1 and downregulated proteins included alpha-1-antitrypsin, retinol-binding protein 3, neuroserpin, cystatin C, carboxypeptidase E and cathepsin-D.
Diabetic retinopathy pathogenesis involves proteins which belong to inflammation, visual transduction, and extracellular matrix pathways. Validation-based experiments using enzyme-linked immunosorbent assay (ELISA) or western blotting are needed to establish cause and effect relationships of these proteins to the disease state, to develop them as biomarkers or drug molecules.
探索印度 2 型糖尿病增殖性糖尿病视网膜病变（PDR）患者的玻璃体蛋白质组。
我们对 PDR（n=13）和特发性黄斑裂孔（IMH；对照）患者的玻璃体蛋白质组进行了基于质谱的无标记定量分析。9 份 PDR 和 10 份 IMH 样本分别进行了混合，作为病例和对照，分别进行了比较。没有混合的情况下，每个 PDR 和 IMH 样本各分析了 4 个，以验证混合分析的结果。使用 Scaffold 软件进行比较定量分析，该软件计算了差异表达的倍数变化。使用 DAVID 和 STRING 软件进行生物信息学分析。
我们在 PDR 和 IMH 玻璃体中分别鉴定出 469 种和 517 种蛋白质，有 172 种蛋白质重叠。此外，在 PDR 玻璃体中鉴定出 297 种独特蛋白质，在 IMH 玻璃体中鉴定出 345 种独特蛋白质。在 PDR 玻璃体中，与 IMH 相比，有 37 种蛋白质上调（P<0.05），19 种蛋白质下调。蛋白质分布分析清楚地表明了 PDR 和 IMH 中蛋白质表达的分离。显著上调的蛋白质包括纤维蛋白原γ链、纤维蛋白原β链和碳酸酐酶 1，下调的蛋白质包括α-1-抗胰蛋白酶、视黄醇结合蛋白 3、神经丝氨酸蛋白酶、胱抑素 C、羧肽酶 E 和组织蛋白酶-D。
糖尿病视网膜病变的发病机制涉及属于炎症、视觉转导和细胞外基质途径的蛋白质。需要使用酶联免疫吸附测定（ELISA）或 Western blot 进行基于验证的实验，以确定这些蛋白质与疾病状态的因果关系，将其开发为生物标志物或药物分子。
