| Project/Area Number |
15591853
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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 | Osaka University |
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Principal Investigator |
KAMEI Motohiro Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助手 (40281125)
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| Co-Investigator(Kenkyū-buntansha) |
TANO Yasuo Osaka University, Graduate School of Medicine, Professor, 医学系研究科, 教授 (80093433)
OHJI Masahito Osaka University, Graduate School of Medicine, Associate Professor, 医学系研究科, 助教授 (90252650)
GOMI Fumi Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助手 (80335364)
IKUNO Yasushi Osaka University, Graduate School of Medicine, Assistant Professor, 医学系研究科, 助手 (50294096)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Diabetic macular edema / Vitreous / Proteomic analysis / Two dimensional gel electrophoresis / Mass-spectrometry |
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| Research Abstract |
To identify and analyze diabetic macular edema (DME)-related proteins in the vitreous, en masse, using two-dimensional gel (2D gel) electrophoresis and mass-spectrometry (MS). Vitreous samples (0.5ml) were collected from the pre-macular portion of 16 DME cases. The protein concentration in vitreous from patients with DME was significantly higher than in non-DME patients (1436 ± 337 vs. 967 ± 217 μg/ml, p=0.05). Samples of 15-μg were subjected to 2D gel electrophoresis. After staining with SYPRO-Ruby protein stein, spot intensities were determined using image analysis software. Clearly visible spots were excised from the gel, digested in situ with trypsin, and the proteins were identified by liquid chromatography tandem MS (LC MS/MS) sequence analysis. We identified 8 spots that were markedly higher in DME than non-DME samples. From the 8 spots, 6 proteins were identified, i.e.PEDF, Apo A-4, ApoA-1, Trip-11, PRBP, and vitamin D-binding protein. One spot was expressed only in non-DME, and it was identified Apo-H. We posit that these chemical mediators presence in the posterior vitreous may play a role in the pathogenesis of DME.
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