| Project/Area Number |
63570288
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内科学一般
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
HISHIDA Akira Hamamatsu University School of Medicine Internal Medicine, Assistant Professor, 医学部, 講師 (70111812)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Shuzo Hamamatsu University School of Medicine Internal Medicine, Assistant Research As, 医学部, 助手 (60195782)
YAMAMOTO Tatsuo Hamamatsu University School of Medicine Internal Medicine, Assistant Research As, 医学部, 助手 (30200819)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | glomerular basement membrane / anionic charge barrier / proteoglycan / cuprolinic blue / aminonucleoside nephrosis / ネフローゼ |
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| Research Abstract |
Despite the growing number of attempts to evaluate anionic site in glomerular basement membrane (GBM), there has not been an established method which accurately quantifies an alteration of the anionic charge of the GBM. The present work was performed to develop a method to quantify the anionic charge of the GBM and to estimate the change of the charge barrier in puromycin aminonucleoside (PAN) nephritis in rats. Cuplorinic blue which contains copper was used as a cationic probe. Left kidney was perfused with sodium acetate buffer containing cuprolinic blue. After perfusion, GBM was isolated by sieving technique and the copper content was measured with an atomic absorption spectrophotometer. Electron microscopic study of isolated GBM revealed the dye-positive fibrils on both side of the lamina densa. Isolation procedure, such as sonication did not affect the copper content of GBM. The degree of copper binding to GBM was markedly reduced in PAN nephritis rats. The degree of copper binding to GBM after PAN injection was markedly reduced at day 9 and returned to the control level at day 42, concomitantly with the recovery of albuminuria. In vito study using isolated GBM, the effects of pH on the cuprolinic blue binding was studied. At pH between 8.0 and 4.0, no significant difference was observed in the cuprolinic blue binding to GBM, but at pH between 3.0 and 4.0, the binding rate began to decrease. This pH-dependent dissociation curve of cuplrolinic blue suggested that the dye preferentially reacted with the sulfate groups in GBM. These results demonstrated that the method which we developed may be useful to quantifies an alteration of the anionic charge of GBM.
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