Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A

Background and Aim: Keratoconjunctivitis sicca (KCS) is a chronic inflammatory ocular disease that occurs in many dog breeds worldwide. This study aimed to investigate the tear protein pattern of healthy dogs, KCS dogs, and KCS dogs after treatment with cyclosporine A (CsA). Materials and Methods...

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Main Authors: Metita Sussadee, Rucksak Rucksaken, Phattara-Orn Havanapan, Onrapak Reamtong, Aree Thayananuphat
Format: Article
Language:English
Published: Veterinary World 2021-06-01
Series:Veterinary World
Subjects:
Online Access:http://www.veterinaryworld.org/Vol.14/June-2021/38.pdf
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author Metita Sussadee
Rucksak Rucksaken
Phattara-Orn Havanapan
Onrapak Reamtong
Aree Thayananuphat
author_facet Metita Sussadee
Rucksak Rucksaken
Phattara-Orn Havanapan
Onrapak Reamtong
Aree Thayananuphat
author_sort Metita Sussadee
collection DOAJ
description Background and Aim: Keratoconjunctivitis sicca (KCS) is a chronic inflammatory ocular disease that occurs in many dog breeds worldwide. This study aimed to investigate the tear protein pattern of healthy dogs, KCS dogs, and KCS dogs after treatment with cyclosporine A (CsA). Materials and Methods: Twenty-eight dogs of any breed were enrolled in the study. The subjects were divided into three groups: Healthy, KCS, and CsA-treated dogs. Tear samples were collected using Schirmer strips. Tear proteins extracted from the strips were analyzed using two-dimensional electrophoresis. For the first dimension, total protein from tears was separated by isoelectric focusing. The second dimension was performed using 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gel images were analyzed and the protein spots of differential expression were manually cut for protein annotation using mass spectrometry. Results: In total, 12 protein spots were excised and subjected to protein identification. Associated with KCS, six protein spots were a downregulated protein, namely, lysozyme. The other six protein spots were upregulated in KCS dogs, consisting of heat shock protein beta-1, protein S100-A12, and keratin type II cytoskeletal 1 and 5. After treatment with CsA for 45 days, the lysozyme protein was still decreasing and the inflammation protein (S100-A12) was not identified. Conclusion: Inflammatory tear proteins and proteins involved in cellular stress were present in KCS dogs and appeared to be reduced in medicated eyes. Treatment with topical CsA in the short term may not improve the activity of antibacterial proteins. Changes in the expression patterns of these four proteins might be useful for disease severity and progression assessment, as well as for exploring a novel method for dry eye management in dogs.
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spelling doaj.art-4b5cc91543124020b4611b8b9086b51d2022-12-21T20:28:26ZengVeterinary WorldVeterinary World0972-89882231-09162021-06-011461711171710.14202/vetworld.2021.1711-1717Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine AMetita Sussadee0https://orcid.org/0000-0002-1080-1528Rucksak Rucksaken1https://orcid.org/0000-0002-0833-3729Phattara-Orn Havanapan2https://orcid.org/0000-0002-1172-9193Onrapak Reamtong3https://orcid.org/0000-0002-1154-6485Aree Thayananuphat4https://orcid.org/0000-0003-3183-1491Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok, Thailand.Institute of Molecular Biosciences, Mahidol University, Salaya campus, Nakhonpathom, Thailand.Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.Background and Aim: Keratoconjunctivitis sicca (KCS) is a chronic inflammatory ocular disease that occurs in many dog breeds worldwide. This study aimed to investigate the tear protein pattern of healthy dogs, KCS dogs, and KCS dogs after treatment with cyclosporine A (CsA). Materials and Methods: Twenty-eight dogs of any breed were enrolled in the study. The subjects were divided into three groups: Healthy, KCS, and CsA-treated dogs. Tear samples were collected using Schirmer strips. Tear proteins extracted from the strips were analyzed using two-dimensional electrophoresis. For the first dimension, total protein from tears was separated by isoelectric focusing. The second dimension was performed using 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gel images were analyzed and the protein spots of differential expression were manually cut for protein annotation using mass spectrometry. Results: In total, 12 protein spots were excised and subjected to protein identification. Associated with KCS, six protein spots were a downregulated protein, namely, lysozyme. The other six protein spots were upregulated in KCS dogs, consisting of heat shock protein beta-1, protein S100-A12, and keratin type II cytoskeletal 1 and 5. After treatment with CsA for 45 days, the lysozyme protein was still decreasing and the inflammation protein (S100-A12) was not identified. Conclusion: Inflammatory tear proteins and proteins involved in cellular stress were present in KCS dogs and appeared to be reduced in medicated eyes. Treatment with topical CsA in the short term may not improve the activity of antibacterial proteins. Changes in the expression patterns of these four proteins might be useful for disease severity and progression assessment, as well as for exploring a novel method for dry eye management in dogs.http://www.veterinaryworld.org/Vol.14/June-2021/38.pdfcyclosporinedogkeratoconjunctivitis siccaproteomictear protein
spellingShingle Metita Sussadee
Rucksak Rucksaken
Phattara-Orn Havanapan
Onrapak Reamtong
Aree Thayananuphat
Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
Veterinary World
cyclosporine
dog
keratoconjunctivitis sicca
proteomic
tear protein
title Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
title_full Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
title_fullStr Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
title_full_unstemmed Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
title_short Changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine A
title_sort changes in tear protein profile in dogs with keratoconjunctivitis sicca following topical treatment using cyclosporine a
topic cyclosporine
dog
keratoconjunctivitis sicca
proteomic
tear protein
url http://www.veterinaryworld.org/Vol.14/June-2021/38.pdf
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AT rucksakrucksaken changesintearproteinprofileindogswithkeratoconjunctivitissiccafollowingtopicaltreatmentusingcyclosporinea
AT phattaraornhavanapan changesintearproteinprofileindogswithkeratoconjunctivitissiccafollowingtopicaltreatmentusingcyclosporinea
AT onrapakreamtong changesintearproteinprofileindogswithkeratoconjunctivitissiccafollowingtopicaltreatmentusingcyclosporinea
AT areethayananuphat changesintearproteinprofileindogswithkeratoconjunctivitissiccafollowingtopicaltreatmentusingcyclosporinea