2005 Fiscal Year Final Research Report Summary
CYR61 Expression and Function in rapidly progressive glomerulonephritis
| Project/Area Number |
16590792
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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 |
Kidney internal medicine
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| Research Institution | Kumamoto University |
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Principal Investigator |
KOHDA Yukimasa Kumamoto University, Nephrology, Faculty, Research Associate, 医学部附属病院, 助手 (20332885)
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| Project Period (FY) |
2004 – 2005
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| Keywords | RPGN / CYR61 / glomerulonephritis |
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| Research Abstract |
To certify the expression of CYR61 protein, 174 human renal biopsy samples were immuno-stained with anti-CYR61 antibody. 19 of 174 cases were rapidly progressive glomerulonephritis. 6 (30%) of 19 rapidly progressive glomerulonephritis showed positive reaction with anti-CYR61 Antibody. 4 cases showed positive reaction in cellular Crescents. 2 cases showed positive reaction in tubles. 25 (16.1%) of 155 chronic glomerulonephritis samples showed positive reaction with anti-CYR61 antibody. No reaction was seen in glomeruli of chronic glomerulonephritis samples. 25 positive samples showed the reaction in tubles. Serum creatinine was 7.2mg/dl in the patients of "crescent positive stained RPGN", and 4.3mg/dl in "crescent negative stained RPGN". There were no differences between both groups. In chronic Glomerular samples, CYR61 positive group had more severe interstitial damages compare to CYR61 negative group. But there were any clinical differences in both groups.
Because of cross reactivity o
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f antibodies, CYR61 expression was checked by another method, RT-PCR. mRNA was extracted from frozen renal biopsy samples. Small amount of renal biopsy samples were quickly frozen at the time of renal biopsy for immuno-fluorescent staining. Several frozen tissue were cut in 5μm thickness and quickly fixed with 70% ethanol to remove OCT compound. After 5 minutes, tissue sections were quickly air dried and RNA was extracted by 4M GTC solution. RT reaction was performed and PCR was performed with this cDNA. By regular 35cycle PCR, CYR61 mRNA products were seen. By real time PCR, there was no relation between the clinical future and CYR expression. Several reason of this negative result were the quality of mRNA, number of samples, especially less number of rapidly progressive glomerulonephritis.
To determine the urinaly expression of CYR61, sensitivity of Western Blot was not enough to see CYR61. Sample collection and concentration method should be changed.
The Expression of CYR61 was seen in severe glomerulonephritis, rapidly progressive glomerulonephritis. Especially the reaction was seen in crescents in rapidly progressive glomerulonephritis samples. So CYR61 will perform some reaction in human rapidly progressive glomerulonephritis. Less
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Research Products
(8 results)
