Purpose. Lipocalins are an ubiquitous family of proteins binding lipophilic molecules, they may contribute to tear film stability; secretory lipophilins are major proteins in human tears and also bind lipids. The purpose of this work was to investigate how these proteins change in patients with dry eye due to tear increased evaporation rate. Methods. Tears were collected from 50 subjects with dry eye with BUT<10 sec and 15 normal control patients. SDS-PAGE electrophoresis utilizing pre-cast Criterion 20% Bio-Rad gels was performed on small tear volumes, gels were stained with Coomassie Blue staining solution. The densitometric volumes (with background subtracted) of the major protein bands separated in the gels (lisozyme, lactoferrin, serum albumin, tear lipocalin and lipophilin) were determined using gel-imaging software (Quantity One, Bio-Rad). Selected bands were characterized by isolation and digestion with 0.05% trypsin and peptide analysis was performed with LC ESI-Q-TOF MS/MS. Protein identification was further confirmed by peptide analysis on whole tear samples and in immunoblotting. Data output from the MS/MS was processed and searched against the NCBI non-redundant protein database using the Mascot software (Matrix Science, UK). Results were statistically evaluated by applying the independent sample t-test, the Mann-Whitney test for unpaired data (significance p< 0,05) Results. data expressed as media + Standard Deviation % Control group Dry eye patients p Lysozyme 10,69 + 2,81 10,60 + 2,58 0,920 Lactoferrin 17,41 + 3,46 8,96 + 3,86 0,000 Serum albumin 3,49 + 2,35 9,28 + 6,51 0,005 Lipocalin 10,55 + 1,01 7,35 +1,95 0,002 Lipophilin 4,14 + 1,25 3,18 + 1,36 0,010 Conclusions. Decreasing tear lipocalin and lipophilin would account for tear instability and may become a reliable marker in diagnosis and follow-up of iperevaporative dry eye. This work was supported in part through a grant from the Fondazione Cassa di Risparmio in Bologna to Prof. Emilio Campos
TEAR LIPOCALIN AND LIPOPHILIN DECREASE IN IPEREVAPORATIVE DRY EYE PATIENTS / Versura P.; Parisi D.; Coslovi C; Cioffi G; Roda A.; Piazzi M.; Campos E.C.. - STAMPA. - (2007), pp. 2-2. (Intervento presentato al convegno 5th International Conference on the Tear Film & Ocular Surface: Basic Science and Clinical Relevance tenutosi a Taormina nel Sept, 5-8).
TEAR LIPOCALIN AND LIPOPHILIN DECREASE IN IPEREVAPORATIVE DRY EYE PATIENTS.
VERSURA, PIERA;PARISI, DANIELA;COSLOVI, CHIARA;RODA, ALDO;CAMPOS, EMILIO
2007
Abstract
Purpose. Lipocalins are an ubiquitous family of proteins binding lipophilic molecules, they may contribute to tear film stability; secretory lipophilins are major proteins in human tears and also bind lipids. The purpose of this work was to investigate how these proteins change in patients with dry eye due to tear increased evaporation rate. Methods. Tears were collected from 50 subjects with dry eye with BUT<10 sec and 15 normal control patients. SDS-PAGE electrophoresis utilizing pre-cast Criterion 20% Bio-Rad gels was performed on small tear volumes, gels were stained with Coomassie Blue staining solution. The densitometric volumes (with background subtracted) of the major protein bands separated in the gels (lisozyme, lactoferrin, serum albumin, tear lipocalin and lipophilin) were determined using gel-imaging software (Quantity One, Bio-Rad). Selected bands were characterized by isolation and digestion with 0.05% trypsin and peptide analysis was performed with LC ESI-Q-TOF MS/MS. Protein identification was further confirmed by peptide analysis on whole tear samples and in immunoblotting. Data output from the MS/MS was processed and searched against the NCBI non-redundant protein database using the Mascot software (Matrix Science, UK). Results were statistically evaluated by applying the independent sample t-test, the Mann-Whitney test for unpaired data (significance p< 0,05) Results. data expressed as media + Standard Deviation % Control group Dry eye patients p Lysozyme 10,69 + 2,81 10,60 + 2,58 0,920 Lactoferrin 17,41 + 3,46 8,96 + 3,86 0,000 Serum albumin 3,49 + 2,35 9,28 + 6,51 0,005 Lipocalin 10,55 + 1,01 7,35 +1,95 0,002 Lipophilin 4,14 + 1,25 3,18 + 1,36 0,010 Conclusions. Decreasing tear lipocalin and lipophilin would account for tear instability and may become a reliable marker in diagnosis and follow-up of iperevaporative dry eye. This work was supported in part through a grant from the Fondazione Cassa di Risparmio in Bologna to Prof. Emilio CamposI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
