Abstract
Cataracts are one of the most common eye diseases that can cause blindness. Discovering susceptibility factors in the proteome that contribute to cataract development would be helpful in gaining new insights in the molecular mechanisms of the cataract process. We used label-free nanoflow ultra-high-performance liquid chromatography–tandem mass spectrometry to compare aqueous humor protein expressions in cataract patients with different cataract risk factors such as diabetes mellitus (DM) and smoking and in controls (with cataract) without risk exposure. Eight patients with diabetes and who smoked (with double risk factors), five patients with diabetes and five patients who smoked (both with a single risk factor), and nine aged-matched cataract controls patients (non-risk exposure) were enrolled. In total, 136 aqueous humor proteins were identified, of which only alpha-2-Heremans–Schmid (HS)-glycoprotein was considered to be significantly risk-associated because it was differentially expressed in these three groups and exhibited increased expression with increasing risk factors. Significant changes in the aqueous humor level of alpha-2-HS-glycoprotein between DM and control samples and between smoking and control samples were confirmed using ELISA. The alpha-2-HS-glycoprotein, called fetuin-a, could be a potential aqueous biomarker associated with DM and smoking, which were cataract risk factors.
Highlights
In developed countries, cataracts are one of the most common causes of blindness [1].They are classified by cause as age-related cataracts, pediatric cataracts, and cataracts secondary to other causes
An alpha-2-Heremans–Schmid (HS)-glycoprotein Enzyme-Linked Immunosorbent Assay (ELISA) assay was performed to measure concentrations of aqueous humor (AH) samples among the single-risk group, double-risk group, and the age-matched cataract controls with a Human Alpha-2-HS-glycoprotein ELISA Kit (EH310RB, ThermoFisher Scientific), as per the manufacturer’s protocol
A subgroup analysis was performed to confirm that Diabetes mellitus (DM) and smoking risk factors significantly influenced the ELISA concentration compared to the cataract control group (Figure 6)
Summary
Cataracts are one of the most common causes of blindness [1]. Used isobaric tags for the relative and absolute quantitation (iTRAQ) methodology to compare AH protein profiles among high myopia, glaucoma, and vitrectomy surgery patients, and controls They identified multiple candidate protein biomarkers associated with cataract development in each group. To examine the protein compositions of aqueous solutions obtained from human cataract eyes of patients who had a single risk factor of either DM or cigarette smoking, those who had double risk factors of DM and cigarette smoking, and aged-matched cataract controls (with neither risk factor) This sensitive proteomics approach could help examine the underlying pathophysiology of cataract formation using relatively scarce amounts of aqueous samples, thereby favoring the methodological approach for this investigation. This study may reveal valuable insights into the molecular changes in the AH in the course of cataract pathogenesis
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