1999 Fiscal Year Final Research Report Summary
Role of elastase and matrix metalloproteinases in pseudomonal keratitis
| Project/Area Number |
10671647
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | Kumamoto University School of Medicine |
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Principal Investigator |
MATSUMOTO Koki Kumamoto University Ophthalmology Associate Professor, 医学部, 助教授 (70173896)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATA Akira Kumamoto University Ophthalmology, 医学部・附属病院, 助手 (60295144)
MURATA Yasuhiro Kumamoto University Ophthalmology, 医学部, 助手 (80229997)
MIYAGAWA Shin-ichi Kumamoto University Ophthalmology, 医学部, 助手 (10260738)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Pseudomonas aeruginosa / Corneal ulcer / elastase / Keratocyte / MMP-2 / MMP-9 / protease / neutrophil |
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| Research Abstract |
Major pathogenic factors that contribute to ulceration in pseudomonal keratitis could be proteases. We investigated the role of pseudomonal elastase and MMPs in pseudomonal keratitis. Pseudomonal keratitis was produced in animals by injecting live bacteria into the corneas and proteases produced in the corneas were analyzed with gelatin and casein zymography. Several MMPs and caseinases with molecular weight of 24-27 kDa were detected. The MMPs included MMP-2 (gelatinase A), MMP-9 (gelatinase B), activated forms of these MMPs and gelatinase with molecular weight of >200 kDa. However, pseudomonal elastase was not detected. In order to clarify the origin of these proteases, culture media conditioned by corneal stromal cells with or without pseudomonal elastase were analyzed with zymography. MMP-2 was detected if the corneal cells were cultured without pseudomonal elastase. However, MMP-2, MMP-9 and activated forms of these MMPs were detected when pseudomonal elastase was added to the medium. Neutrophil extracts contained MMP-9, gelatinase with molecular weight of >200 kDa and caseinase with molecular weight of 24-27 kDa. Immunohistochemistry (ABC method) showed the presence of pseudomonal elastase in the corneas. From these results it is indicated that proteases that contribute to ulceration could be derived from bacteria, corneal cells and neutrophil, and the MMPs can be activated by pseudomonal elastase.
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