| Project/Area Number |
07670862
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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 |
Pediatrics
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| Research Institution | Osaka University |
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Principal Investigator |
SHIMA Masaaki Osaka University, Faculty of Medicine, assistant professor, 医学部, 助手 (10252660)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Takehisa Osaka University, Faculty of Medicine, assistant professor, 医学部, 助手 (30231674)
YAMAOKA Kanji Osaka University, Faculty of Medicine, assistant professor, 医学部, 助手 (60144490)
松本 小百合 大阪大学, 医学部, 助手
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | chronic glomerulonephritis / matrix metalloproteinase / matrix / マトリックス / matrix metallapnoteinrre / 腎炎 |
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| Research Abstract |
Purpose
In this reseach, we have examined a role of matrix metalloproteinase for the development of pediatric glomerulonephritis.
Method
Glomerolus are microdissected from the kidney tissue in patients with glomerulonephritis, and cultured in 96 wells for 48 hours under serum-unsupplimented condition. The supernatant was obtainted and stored before the anaysis of matrix metalloproteinase. Samples were diluted into an equal volume of sample buffer, loaded without boiling and separated at 4ﾟC.After the electrophoresis, the gels were washed twice for 15 min.in 25% Triton X-100. The gels were incubated in a solution of 50 mM Tris-HCL containing 5 mM CaCl_2 and 1 mM ZnCl_2 for 48hours at 37ﾟC to allow for the enzymatic digestion of gelatin substrate.
Results
1.MMP-9,92 Kd protein, was found in the urine supecimens.
2.In the supernatant of the glomerular culture, MMP-9 activity was confirmed.
3.In patients with focal GN,MMP-9 was not observed, but was found in urine samples of the patients with diffuse proliferative GN.
4.The urinary MMP-9 level was correlated with the activity of the disease.
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