| Project/Area Number |
16K09654
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Kidney internal medicine
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| Research Institution | Kitasato University |
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Principal Investigator |
Nooguchi Hiroshi 北里大学, 北里大学メディカルセンター, 医員(医師) (30218341)
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| Co-Investigator(Kenkyū-buntansha) |
泉 裕一郎 熊本大学, 大学院生命科学研究部(医), 助教 (20736243)
安岡 有紀子 北里大学, 医学部, 講師 (50348504)
河原 克雅 北里大学, 医学部, 名誉教授 (70134525)
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| Project Period (FY) |
2016-04-01 – 2020-03-31
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| Project Status |
Completed (Fiscal Year 2019)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | エリスロポエチン / 低酸素 / 貧血 / ウェスタンブロット / 脱糖鎖 / 糖タンパク / 腎臓 / アルドステロン / バゾプレシン / HIF2α / RAS系 / 腎尿細管 / 低酸素誘導因子 |
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| Outline of Final Research Achievements |
We investigated the detection of erythropoietin protein by Western blot analysis.Epo protein was detected at 34-43 kDa, which was shifted to 22 kDa by degycosylation. Using this new method, we found thaterythropoietin production by the kidney is increased by 600-fold in response to severe hypoxia. Tke Epo production by the liver was not stimulated by severe hypoxia.
We furthere succeeded in detecting Epo in urine anf blood by Western blotting. Urinary Epo was detected by direct application of urine sample to SDS-PAGE. In contrast, Epo band was not seen by direct application of blood to SDS-PAGE. Incubation of blood sample with deglycosykation buffer without PNGase showed Epo band at 34-43 kDa. The band at 34-43 kDa shifted to 22 kDa by deglycosulation by PNGase.
Using our new Western blotting, the detection of EPo protein in tissue, blood and urine has become possible.
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| Academic Significance and Societal Importance of the Research Achievements |
エリスロポエチンの検出は、ドーピングも行われており、私たちの方法を用いることで、簡単に血液や尿中のエリスロポエチンの検出ができるようになり、種々のスポーツ競技の公正な実施に役立てることが可能であり、社会的な意義は極めて高いと思われる。
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