The Influence of Tear Proteins on the Film Stability of Rabbit Tear Extracts

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Abstract
This study was undertaken to gain an understanding of the significance of tear proteins in stabilizing the tear film. Either a sigma agonist, N,N-dimethyl-2-phenylethylamine HCl (AF2975), or a sigma antagonist, haloperidol, was administered to rabbit eyes in order to increase or decrease protein secretion, respectively. At 0, 10 and 60 minutes after instillation, tear proteins were extracted from Schirmer strips and measured for total protein. A portion of the extract was used for separating five major protein fractions using size-exclusion HPLC. Total protein extract or individual protein fractions were measured for surface tension by the horizontal capillary method and for in vitro break up time (in vitro BUT), a newly designed procedure. A statistically significant decrease was measured for surface tension and a concomitant increase was measured for in vitro BUT for the total protein samples at 10 and 60 minutes after instillation of AF2975 compared to the vehicle treated eye. The results for haloperidol yielded an increase in surface tension and an decrease in in vitro BUT. When the tear proteins were separated into five major fractions, only the 23 minute protein fraction was found to decrease surface tension and increase in vitro BUT following AF2975 administration. Haloperidol, a sigma antagonist, showed an exact opposite effect for the total protein and the 23 minute protein fraction.