Regulation of 1-Cys peroxiredoxin expression in the process of stromal wound healing after photorefractive keratectomy

Hungwon Tchah, Myoung Joon Kim, Tae-im Kim, Hyun Jeung Choi, Jae Yong Kim, Mi Jung Kim, Jhang Ho Pak

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

PURPOSE. To investigate 1-cys peroxiredoxin (1-cysPrx) expression during the corneal wound-healing process after PRK and the effect of growth factors on 1-cysPrx expression in cultured bovine keratocytes (BKs). METHODS. Rat corneas were excised at 4 hours, 12 hours, 1 day, 3 days, and 7 days after PRK. Expression of 1-cysPrx in the corneas was examined by immunohistochemical, Northern blot, and immunoblot analyses. Keratocytes were isolated from bovine corneas and subcultured to study the effects of TGF-β1, keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and H2O2 on 1-cysPrx expression at different concentrations and time intervals. Generation and proliferation of intracellular reactive oxygen species (ROS) in cultured BKs stimulated by these growth factors were measured by the DCF (2′,7′-dichlorofluorescein) assay, the CCK-8 assay, and immunoblot analysis with a polyclonal proliferating cell nuclear antigen (PCNA) antibody, respectively. RESULTS. Intense staining of 1-cysPrx was observed in the epithelia and the anterior stromas of wounded corneas 4 hours after PRK and had extended to the entire stroma by day 3. By day 7, the expression almost returned to nonsurgical control level in epithelia, although notable expression was still detectable in the stroma. Concomitant augmentation of 1-cysPrx mRNA and protein was seen in the corneas at 12 hours to 7 days. Growth factor treatment in cultured BKs resulted in 1-cysPrx induction in a dose- and time-dependent manner. Growth factor-stimulated cells showed strong DCF fluorescence and increased proliferation during a 24-hour incubation, during which an upregulation of 1-cysPrx occurred. CONCLUSIONS. These observations provide new information for the regulation of 1-cysPrx expression during the corneal wound-healing process.

Original languageEnglish
Pages (from-to)2396-2403
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume46
Issue number7
DOIs
Publication statusPublished - 2005 Dec 1

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Photorefractive Keratectomy
Peroxiredoxins
Wound Healing
Cornea
Intercellular Signaling Peptides and Proteins
Epithelium
Fibroblast Growth Factor 7
Sincalide
Hepatocyte Growth Factor
Platelet-Derived Growth Factor
Proliferating Cell Nuclear Antigen
Northern Blotting
Reactive Oxygen Species
Up-Regulation
Fluorescence
Staining and Labeling
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Tchah, Hungwon ; Kim, Myoung Joon ; Kim, Tae-im ; Choi, Hyun Jeung ; Kim, Jae Yong ; Kim, Mi Jung ; Pak, Jhang Ho. / Regulation of 1-Cys peroxiredoxin expression in the process of stromal wound healing after photorefractive keratectomy. In: Investigative Ophthalmology and Visual Science. 2005 ; Vol. 46, No. 7. pp. 2396-2403.
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title = "Regulation of 1-Cys peroxiredoxin expression in the process of stromal wound healing after photorefractive keratectomy",
abstract = "PURPOSE. To investigate 1-cys peroxiredoxin (1-cysPrx) expression during the corneal wound-healing process after PRK and the effect of growth factors on 1-cysPrx expression in cultured bovine keratocytes (BKs). METHODS. Rat corneas were excised at 4 hours, 12 hours, 1 day, 3 days, and 7 days after PRK. Expression of 1-cysPrx in the corneas was examined by immunohistochemical, Northern blot, and immunoblot analyses. Keratocytes were isolated from bovine corneas and subcultured to study the effects of TGF-β1, keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and H2O2 on 1-cysPrx expression at different concentrations and time intervals. Generation and proliferation of intracellular reactive oxygen species (ROS) in cultured BKs stimulated by these growth factors were measured by the DCF (2′,7′-dichlorofluorescein) assay, the CCK-8 assay, and immunoblot analysis with a polyclonal proliferating cell nuclear antigen (PCNA) antibody, respectively. RESULTS. Intense staining of 1-cysPrx was observed in the epithelia and the anterior stromas of wounded corneas 4 hours after PRK and had extended to the entire stroma by day 3. By day 7, the expression almost returned to nonsurgical control level in epithelia, although notable expression was still detectable in the stroma. Concomitant augmentation of 1-cysPrx mRNA and protein was seen in the corneas at 12 hours to 7 days. Growth factor treatment in cultured BKs resulted in 1-cysPrx induction in a dose- and time-dependent manner. Growth factor-stimulated cells showed strong DCF fluorescence and increased proliferation during a 24-hour incubation, during which an upregulation of 1-cysPrx occurred. CONCLUSIONS. These observations provide new information for the regulation of 1-cysPrx expression during the corneal wound-healing process.",
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Regulation of 1-Cys peroxiredoxin expression in the process of stromal wound healing after photorefractive keratectomy. / Tchah, Hungwon; Kim, Myoung Joon; Kim, Tae-im; Choi, Hyun Jeung; Kim, Jae Yong; Kim, Mi Jung; Pak, Jhang Ho.

In: Investigative Ophthalmology and Visual Science, Vol. 46, No. 7, 01.12.2005, p. 2396-2403.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Regulation of 1-Cys peroxiredoxin expression in the process of stromal wound healing after photorefractive keratectomy

AU - Tchah, Hungwon

AU - Kim, Myoung Joon

AU - Kim, Tae-im

AU - Choi, Hyun Jeung

AU - Kim, Jae Yong

AU - Kim, Mi Jung

AU - Pak, Jhang Ho

PY - 2005/12/1

Y1 - 2005/12/1

N2 - PURPOSE. To investigate 1-cys peroxiredoxin (1-cysPrx) expression during the corneal wound-healing process after PRK and the effect of growth factors on 1-cysPrx expression in cultured bovine keratocytes (BKs). METHODS. Rat corneas were excised at 4 hours, 12 hours, 1 day, 3 days, and 7 days after PRK. Expression of 1-cysPrx in the corneas was examined by immunohistochemical, Northern blot, and immunoblot analyses. Keratocytes were isolated from bovine corneas and subcultured to study the effects of TGF-β1, keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and H2O2 on 1-cysPrx expression at different concentrations and time intervals. Generation and proliferation of intracellular reactive oxygen species (ROS) in cultured BKs stimulated by these growth factors were measured by the DCF (2′,7′-dichlorofluorescein) assay, the CCK-8 assay, and immunoblot analysis with a polyclonal proliferating cell nuclear antigen (PCNA) antibody, respectively. RESULTS. Intense staining of 1-cysPrx was observed in the epithelia and the anterior stromas of wounded corneas 4 hours after PRK and had extended to the entire stroma by day 3. By day 7, the expression almost returned to nonsurgical control level in epithelia, although notable expression was still detectable in the stroma. Concomitant augmentation of 1-cysPrx mRNA and protein was seen in the corneas at 12 hours to 7 days. Growth factor treatment in cultured BKs resulted in 1-cysPrx induction in a dose- and time-dependent manner. Growth factor-stimulated cells showed strong DCF fluorescence and increased proliferation during a 24-hour incubation, during which an upregulation of 1-cysPrx occurred. CONCLUSIONS. These observations provide new information for the regulation of 1-cysPrx expression during the corneal wound-healing process.

AB - PURPOSE. To investigate 1-cys peroxiredoxin (1-cysPrx) expression during the corneal wound-healing process after PRK and the effect of growth factors on 1-cysPrx expression in cultured bovine keratocytes (BKs). METHODS. Rat corneas were excised at 4 hours, 12 hours, 1 day, 3 days, and 7 days after PRK. Expression of 1-cysPrx in the corneas was examined by immunohistochemical, Northern blot, and immunoblot analyses. Keratocytes were isolated from bovine corneas and subcultured to study the effects of TGF-β1, keratinocyte growth factor (KGF), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGF), and H2O2 on 1-cysPrx expression at different concentrations and time intervals. Generation and proliferation of intracellular reactive oxygen species (ROS) in cultured BKs stimulated by these growth factors were measured by the DCF (2′,7′-dichlorofluorescein) assay, the CCK-8 assay, and immunoblot analysis with a polyclonal proliferating cell nuclear antigen (PCNA) antibody, respectively. RESULTS. Intense staining of 1-cysPrx was observed in the epithelia and the anterior stromas of wounded corneas 4 hours after PRK and had extended to the entire stroma by day 3. By day 7, the expression almost returned to nonsurgical control level in epithelia, although notable expression was still detectable in the stroma. Concomitant augmentation of 1-cysPrx mRNA and protein was seen in the corneas at 12 hours to 7 days. Growth factor treatment in cultured BKs resulted in 1-cysPrx induction in a dose- and time-dependent manner. Growth factor-stimulated cells showed strong DCF fluorescence and increased proliferation during a 24-hour incubation, during which an upregulation of 1-cysPrx occurred. CONCLUSIONS. These observations provide new information for the regulation of 1-cysPrx expression during the corneal wound-healing process.

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U2 - 10.1167/iovs.05-0107

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