Regulated expression of collagenases MMP-1, -8, and -13 and stromelysins MMP-3, -10, and -11 by human corneal epithelial cells

De Quan Li, Tie Yan Shang, Hyun Seung Kim, Abraham Solomon, Balakrishna L. Lokeshwar, Stephen C. Pflugfelder

Research output: Contribution to journalArticle

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Abstract

PURPOSE. This study investigated the regulated expression of collagenases (MMP-1, -8, and -13) and stromelysins (MMP-3, -10, and -11) by human corneal epithelial cells treated with IL-1β, TNF-α, and doxycycline, a medication used to treat ocular surface diseases. METHODS. Primary human corneal epithelial cell cultures were treated with IL-1β or TNF-α, with or without their corresponding inhibitors. Total RNA extracted from cells treated for 4 to 24 hours was subjected to semiquantitative RT-PCR and Northern hybridization. Conditioned media from 24-hour-treated cultures were evaluated for MMP production by ELISA and activity assays. RESULTS. Semiquantitative RT-PCR and Northern hybridization revealed that the mRNAs of MMP-1, -13, -3, -10, and -11 were dose dependently upregulated by IL-1β and TNF-α, whereas MMP-8 and -14 and tissue inhibitor of metalloproteinase (TIMP)-1 were not altered, in corneal epithelial cells. MMP ELISA and activity assays confirmed this dose-dependent increase in MMP-1, -13, -3, and -10 protein production in conditioned media by IL-1β and TNF-α. This stimulated production was inhibited by their neutralizing antibodies and by IL-1 receptor antagonist. Doxycycline suppressed stimulated MMP-1, -10, and -13 production at both the mRNA and protein levels. CONCLUSIONS. This study demonstrated that IL-1β and TNF-α upregulate collagenases (MMP-1, -13) and stromelysins (MMP-3, -10, and -11) in human corneal epithelial cells. Doxycycline suppresses stimulated MMP-1, -13, and -10 at the mRNA and protein levels, which suggests that collagenases and stromelysins may play a role in the pathogenesis of sterile corneal ulceration and other ocular surface diseases.

Original languageEnglish
Pages (from-to)2928-2936
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume44
Issue number7
DOIs
StatePublished - Jul 1 2003
Externally publishedYes

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Matrix Metalloproteinase 11
Matrix Metalloproteinases
Epithelial Cells
Interleukin-1
Doxycycline
Eye Diseases
Conditioned Culture Medium
Messenger RNA
collagenase 1
Enzyme-Linked Immunosorbent Assay
Matrix Metalloproteinase 3
Polymerase Chain Reaction
Proteins
Tissue Inhibitor of Metalloproteinase-1
Interleukin-1 Receptors
Collagenases
Neutralizing Antibodies

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Regulated expression of collagenases MMP-1, -8, and -13 and stromelysins MMP-3, -10, and -11 by human corneal epithelial cells. / Li, De Quan; Shang, Tie Yan; Kim, Hyun Seung; Solomon, Abraham; Lokeshwar, Balakrishna L.; Pflugfelder, Stephen C.

In: Investigative Ophthalmology and Visual Science, Vol. 44, No. 7, 01.07.2003, p. 2928-2936.

Research output: Contribution to journalArticle

Li, De Quan ; Shang, Tie Yan ; Kim, Hyun Seung ; Solomon, Abraham ; Lokeshwar, Balakrishna L. ; Pflugfelder, Stephen C. / Regulated expression of collagenases MMP-1, -8, and -13 and stromelysins MMP-3, -10, and -11 by human corneal epithelial cells. In: Investigative Ophthalmology and Visual Science. 2003 ; Vol. 44, No. 7. pp. 2928-2936.
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abstract = "PURPOSE. This study investigated the regulated expression of collagenases (MMP-1, -8, and -13) and stromelysins (MMP-3, -10, and -11) by human corneal epithelial cells treated with IL-1β, TNF-α, and doxycycline, a medication used to treat ocular surface diseases. METHODS. Primary human corneal epithelial cell cultures were treated with IL-1β or TNF-α, with or without their corresponding inhibitors. Total RNA extracted from cells treated for 4 to 24 hours was subjected to semiquantitative RT-PCR and Northern hybridization. Conditioned media from 24-hour-treated cultures were evaluated for MMP production by ELISA and activity assays. RESULTS. Semiquantitative RT-PCR and Northern hybridization revealed that the mRNAs of MMP-1, -13, -3, -10, and -11 were dose dependently upregulated by IL-1β and TNF-α, whereas MMP-8 and -14 and tissue inhibitor of metalloproteinase (TIMP)-1 were not altered, in corneal epithelial cells. MMP ELISA and activity assays confirmed this dose-dependent increase in MMP-1, -13, -3, and -10 protein production in conditioned media by IL-1β and TNF-α. This stimulated production was inhibited by their neutralizing antibodies and by IL-1 receptor antagonist. Doxycycline suppressed stimulated MMP-1, -10, and -13 production at both the mRNA and protein levels. CONCLUSIONS. This study demonstrated that IL-1β and TNF-α upregulate collagenases (MMP-1, -13) and stromelysins (MMP-3, -10, and -11) in human corneal epithelial cells. Doxycycline suppresses stimulated MMP-1, -13, and -10 at the mRNA and protein levels, which suggests that collagenases and stromelysins may play a role in the pathogenesis of sterile corneal ulceration and other ocular surface diseases.",
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T1 - Regulated expression of collagenases MMP-1, -8, and -13 and stromelysins MMP-3, -10, and -11 by human corneal epithelial cells

AU - Li, De Quan

AU - Shang, Tie Yan

AU - Kim, Hyun Seung

AU - Solomon, Abraham

AU - Lokeshwar, Balakrishna L.

AU - Pflugfelder, Stephen C.

PY - 2003/7/1

Y1 - 2003/7/1

N2 - PURPOSE. This study investigated the regulated expression of collagenases (MMP-1, -8, and -13) and stromelysins (MMP-3, -10, and -11) by human corneal epithelial cells treated with IL-1β, TNF-α, and doxycycline, a medication used to treat ocular surface diseases. METHODS. Primary human corneal epithelial cell cultures were treated with IL-1β or TNF-α, with or without their corresponding inhibitors. Total RNA extracted from cells treated for 4 to 24 hours was subjected to semiquantitative RT-PCR and Northern hybridization. Conditioned media from 24-hour-treated cultures were evaluated for MMP production by ELISA and activity assays. RESULTS. Semiquantitative RT-PCR and Northern hybridization revealed that the mRNAs of MMP-1, -13, -3, -10, and -11 were dose dependently upregulated by IL-1β and TNF-α, whereas MMP-8 and -14 and tissue inhibitor of metalloproteinase (TIMP)-1 were not altered, in corneal epithelial cells. MMP ELISA and activity assays confirmed this dose-dependent increase in MMP-1, -13, -3, and -10 protein production in conditioned media by IL-1β and TNF-α. This stimulated production was inhibited by their neutralizing antibodies and by IL-1 receptor antagonist. Doxycycline suppressed stimulated MMP-1, -10, and -13 production at both the mRNA and protein levels. CONCLUSIONS. This study demonstrated that IL-1β and TNF-α upregulate collagenases (MMP-1, -13) and stromelysins (MMP-3, -10, and -11) in human corneal epithelial cells. Doxycycline suppresses stimulated MMP-1, -13, and -10 at the mRNA and protein levels, which suggests that collagenases and stromelysins may play a role in the pathogenesis of sterile corneal ulceration and other ocular surface diseases.

AB - PURPOSE. This study investigated the regulated expression of collagenases (MMP-1, -8, and -13) and stromelysins (MMP-3, -10, and -11) by human corneal epithelial cells treated with IL-1β, TNF-α, and doxycycline, a medication used to treat ocular surface diseases. METHODS. Primary human corneal epithelial cell cultures were treated with IL-1β or TNF-α, with or without their corresponding inhibitors. Total RNA extracted from cells treated for 4 to 24 hours was subjected to semiquantitative RT-PCR and Northern hybridization. Conditioned media from 24-hour-treated cultures were evaluated for MMP production by ELISA and activity assays. RESULTS. Semiquantitative RT-PCR and Northern hybridization revealed that the mRNAs of MMP-1, -13, -3, -10, and -11 were dose dependently upregulated by IL-1β and TNF-α, whereas MMP-8 and -14 and tissue inhibitor of metalloproteinase (TIMP)-1 were not altered, in corneal epithelial cells. MMP ELISA and activity assays confirmed this dose-dependent increase in MMP-1, -13, -3, and -10 protein production in conditioned media by IL-1β and TNF-α. This stimulated production was inhibited by their neutralizing antibodies and by IL-1 receptor antagonist. Doxycycline suppressed stimulated MMP-1, -10, and -13 production at both the mRNA and protein levels. CONCLUSIONS. This study demonstrated that IL-1β and TNF-α upregulate collagenases (MMP-1, -13) and stromelysins (MMP-3, -10, and -11) in human corneal epithelial cells. Doxycycline suppresses stimulated MMP-1, -13, and -10 at the mRNA and protein levels, which suggests that collagenases and stromelysins may play a role in the pathogenesis of sterile corneal ulceration and other ocular surface diseases.

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