Protein quantification and electrophoresis in aqueous humor of pseudoexfoliation eyes

Pseudoexfoliation (PSX) eyes frequently show clinical signs of blood-aqueous barrier impairment. To analyze these alterations, the authors examined aqueous humor of human eyes with and without PSX.

After aqueous humor samples had been obtained during cataract or filtering glaucoma surgery, a modified Pierce-bicin choninic acid assay was used to quantify total aqueous protein concentration in 27 PSX eyes and 37 eyes without clinical signs of PSX (12 cataract eyes and 25 eyes with primary open-angle glaucoma). In addition, aqueous protein composition was analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis, silver staining, and laser densitometry in 27 PSX eyes and 59 eyes without PSX.

Aqueous protein concentration was significantly higher in PSX (mean 0.42 +/- 0.16 mg/ml) than in normal cataract eyes (0.22 +/- 0.08 mg/ml, P < 0.0001) and in eyes with open-angle glaucoma (0.26 +/- 0.09 mg/ml, P < 0.0001, Wilcoxon-Mann-Whitney test). Electrophoresis revealed a characteristic increase of a 12.5-kDa band in 15 of 27 PSX eyes but in only 1 of 59 eyes without PSX (P < 0.00001, chi-square test).

These results substantiate increased aqueous protein concentration and aqueous barrier impairment in PSX. The additional finding of an increased 12.5-kDa band in 56% of PSX eyes may be related to the pathogenesis of PSX in the anterior ocular segment.