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Chae, Park, and Choi: Comparative Analysis of the Tear Protein Expression After Photorefractive Keratectomy Using Two-Dimensional Electrophoresis
Original Article

J Korean Ophthalmol Soc 2009;50(5):762-768.

Published online: 7 September 2009

DOI: https://doi.org/10.3341/jkos.2009.50.5.762

Comparative Analysis of the Tear Protein Expression After Photorefractive Keratectomy Using Two-Dimensional Electrophoresis

Jong Keun Chae, MD1, Sung Pyo Park, MD2, Tae Hoon Choi, MD3

1Department of Ophthalmology, Military Manpower Administration, Seoul, Korea

2Department of Ophthalmology, Kangdong Sacred Heart Hospital, College of Medicine, Hallym University, Seoul, Korea

3Department of Ophthalmology, Nune Eye Hospital, Seoul, Korea

Address reprint requests to Jong Keun Chae, MD Department of Ophthalmology, Military Manpower Administration #159-1, Singil 7-dong, Yeongdeungpo-gu, Seoul 150-057, Korea Tel: 82-2-820-4542, Fax: 82-2-820-4548, E-mail: sg50@lycos.co.kr

Received 21 July 2008       Accepted 8 September 2009

Copyright © 2009 Korean Ophthalmological Society

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To investigate the change in the tear protein composition of patients who underwent refractive surgery.

Methods

Tear samples were collected before photorefrative keratectomy (PRK), on the first, the second, and the third postoperative day, and then a month after the operation from 40 eyes of 20 patients. These tear samples were analyzed using two-dimensional gel electrophoresis (2-DE). Matrix-associated laser desorption/ionization time of flight (MALDI-TOF) was employed for the identification of expressed proteins. Control tear samples were collected from 40 eyes of 20 healthy volunteers who had no history of ocular surgery or pathology.

Results

On the first postoperative day, lipocalin-1 precursor, lipocalin-1, and lysozyme were up-regulated. On the second postoperative day, serum albumin precursor and serum albumin were up-regulated. The tears collected on the third postoperative day and after 1 month had similar protein expression levels to the control group. Lipocalin 1 precursor and lysozyme were up-regulated and down-regulated after reftactive surgery, respectively. However, each protein had a different molecular weight and isopotential point.

Conclusions

The tear protein composition changed uniquely in the early postoperative period, and proteins with different isopotential points were detected after PRK. We hypothesized that the healing process might influence the expression of the tear proteins.

Keywords: Matrix-associated laser desorption/ionization time of flight (MALDI-TOF), Photorefractive keratectomy (PRK), Tear proteins, Two-dimensional electrophoresis (2-DE)

References

1. Trokel SL, Srinivasan R, Braren B. Excimer laser surgery of the cornea. Am J Ophthalmol. 1983; 96:710–5.
crossref
2. Shyn KH, Yoon SC. Refractive survey 2005 in Korea. J Korean Ophthalmol Soc. 2008; 49:570–6.
3. Murphy PJ, Corbett MC, O'Brart DP, et al. Loss and recovery of corneal sensitivity following photorefracitve keratectomy for myopia. J Refract Surg. 1999; 15:38–45.
4. Perez-Santonja JJ, Sakla HF, Cardona C, et al. Corneal sensitivity after photorefractive keratectomy and laser in situ keratomileusis for low myopia. Am J Ophthalmol. 1999; 127:497–504.
5. Kanellapoulos AJ, Pallikaris IG, Donnenfeld ED, et al. Comparison of corneal sensation following photorefractive keratectomy and laser in situ keratomileusis. J Cataract Refract Surg. 1997; 23:34–8.
6. Campos M, Hertzog L, Garbus JJ, McDonnell PJ. Corneal sensitivity afterphotorefractive keratectimy. Am J Ophthalmol. 1992; 114:51–4.
7. Kauffmann T, Bodanowitz S, Hesse L, Kroll P. Corneal reinner-vation after photorefractive keratectomy and laser in situ keratomileusis: an in vivo study with a confocal videomicroscope. Ger J Ophthalmol. 1996; 5:508–12.
8. Tervo K, Latvala T, Tervo T. Recovery of corneal innervation following photorefractive keratoabration. Arch Ophthalmol. 1994; 112:1466–70.
9. Tervo T, Virtanen T, Honkanen N, et al. Tear fluid plasmin activity after excimer laser photorefractive keratectomy. Invest Ophthalmol Vis Sci. 1994; 35:3045–50.
10. Vesaluoma MH, Tervo TT. Tenascin and cytokines in tear fluid after photorefractive keratectomy. J Refract Surg. 1998; 14:447–54.
crossref
11. Mertaniemi P, Ylatuoa S, Partanen P, Tervo T. Increased release of immunoreactive calcitonin gene-related peptide (CGRP) in tears after excimer laser keratectomy. Exp Eye Res. 1995; 60:659–65.
12. Virtanen T, Ylatupa S, Mertaniemi P, et al. Tear fluid fibronectin levels after photorefractive keratectomy. J Refract Surg. 1995; 11:106–12.
13. Bilgihan A, Bilgihan K, Toklu Y, et al. Ascobic acid levels in human tear after photorefractive keratectomy, transepithelial photorefractive keratectomy, and laser in situ keratomileusis. J Cataract Refract Surg. 2001; 27:585–8.
14. Song WK, Lee HK, Kim HC, et al. Nerve growth factor concentration and implications in photorefractive keratectomy versus laser in situ keratomileusis. J Korean Ophthalmol Soc. 2006; 47:1349–57.
15. Csutak A, Silver DM, Tozser J, et al. Plasminogen activator inhibitor in human tear after laser refractive surgery. J Cataract Refract Surg. 2008; 34:897–901.
16. Fust A, Veres A, Kiszel P, et al. Changes in tear protein pattern after photorefractive keratectomy. Eur J Ophthalmol. 2003; 13:525–31.
17. Joo H, Han J. Proteomics. 1st ed.Seoul: Panmun;2004. p. 38–87.
18. Grus FH, Augustin AJ. Protein analysis methods in diagnosis of sicca syndrome. Ophthalmologe. 2000; 97:54–61.
19. Grus FH, Augustin AJ. Analysis of tear protein patterns by a neural network as diagnostical tool for the detection of dry eyes. Electrophoresis. 1999; 20:875–80.
20. Grus FH, Sabuncruo P, Augustin AJ. Quantitative analysis of tear protein profile for soft contact lenses-a clinical study. Klin Monatsbl Augenheilkd. 2001; 218:239–42.
21. Herber S, Grus FH, Sabuncuo P, Augustin AJ. Two-dimensional analysis of tear fluid patterns of diabetes patients. Electrophoresis. 2001; 22:1838–44.
22. Janssen PT, van Bijsterveld OP. Origin and biosynthesis of human tear fluid proteins. Invest Ophthalmol Vis Sci. 1983; 24:623–30.

Figure 1.
Representative 2-DE gel map of control and PRK patient tear proteins. (A) The 2-DE gel map of control subject. (B) The 2-DE gel map of preoperative subject.
jkos-50-762f1.tif
Figure 2.
Representative 2-DE gel map of PRK patient tear proteins. (A) The 2-DE gel map of the first post-operative day subject. (B) The 2-DE gel map of the second postoperative day subject.
jkos-50-762f2.tif
Figure 3.
Representative 2-DE gel map of PRK patient tear proteins. (A) The 2-DE gel map of the third post-operative day subject. (B) The 2-DE gel map of the subject obtained after a month postoperatively
jkos-50-762f3.tif
Table 1.
Demographic features of PRK and Control
PRK Control p-value
Age Mean± SD 25.35±4.23 26.65±2.60 0.25
Range 19∼30 19∼31
Sex (male/female) 4/16 6/14 0.48
Table 2.
Identification of the down-regulated proteins in PRK patients
Spot No. Protein Mw* (KDa) pI† Accession No.
1 Lipocalin 1 precursor 20 4.8 gi|4504963
2 Neutrophil lactoferrin 97 8.4 gi|34412
3 Lysozyme 31 9.2 gi|1421350
4 Lysozyme 15 9.2 gi|15825835
5 Neutrophil lactoferrin 97 4.1 gi|186818
6 MGC27165 Protein 18 4.8 gi|21619010
7 Zn-alpha2-glycoprotein 15 4.5 gi|38026
8 Lipocalin 1 precursor 59 5.9 gi|4504963
9 Cystain S precusor; Cystain 4 45 4.5 gi|55667085
10 Lacrimal praline-rich protein 31 4.4 gi|3318722
11 Lacritin precursor 26 4.7 gi|15187164

* Molecular weight

† Isopotential point.

Table 3.
Identification of the up-regulated proteins in PRK patients
Spot No. Protein Mw* (KDa) pI† Accession No.
12 Lipocalin 1 precursor; Lipocalin 1 20 4.8 gi|4504963
13 Lipocalin 1 precursor; Lipocalin 1 20 5.6 gi|4504963
14 Lipocalin 1 precursor; Lipocalin 1 20 6.7 gi|4504963
15 Lysozyme 15 5.1 gi|12084401
16 Lysozyme 15 6.8 gi|6729881
17 Lysozyme 15 8.1 gi|4930014
18 Human serum albumin 69 4.3 gi|31615330
19 Human serum albumin 69 5.8 gi|3212456
20 Human serum albumin precursor 69 6.9 gi|6013427

* Molecular weight

† Isopotential point.

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