| Project/Area Number |
11558068
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
環境影響評価(含放射線生物学)
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| Research Institution | Tohoku University |
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Principal Investigator |
KIKUCHI Hideaki INSTITUTE OF DEVELOPMENT, AGING AND CANCER, TOHOKU UNIVERSITY, ASSOCIATE PROFESSOR, 加齢医学研究所, 助教授 (60006111)
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| Co-Investigator(Kenkyū-buntansha) |
BABA Tuneo DAICEL CHEMICAL INDUSTRIES, LTD., DIVISION ON TOXICOLOGY AND PHARMACOLOGY, RESEARCH DIRECTOR, 生理活性研究所, 主任研究員
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥13,800,000 (Direct Cost: ¥13,800,000)
Fiscal Year 2001: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2000: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1999: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Dioxin / T cell / Immunotoxicity / Apoptosis / カスパーゼ3 |
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| Research Abstract |
Our aim of this project is a development of assay system for immunotoxicity by dioxin compounds. We tried to find a method for a large number of samples, high sensitivity and low background for the detection of T cell apoptosis. We tested caspase 3 assay system as a possible candidate method. In the process of apoptosis by TCDD, caspase 3 was activated in L-MAT cells. Therefore, we utilized fluorescence dye conjugated to peptide of PARP as a substrate to improve the sensitivity of apoptosis assay. The apoptosis of L-MAT cells by TCDD can be detected as low as 10^5 cells by this method. Using fluorometric-measuring equipment with 96 well culture dish, we developed a mass-screening system.
Furthermore, we analyzed the pathway of apoptotic signal to caspase 3. A specific inhibitor of Protein kinase C-theta (PKC-θ) blocked the apoptosis of L-MAT cells by TCDD. 1,2-Diacylglycerol (DAG) is known to activate PKC-θ, therefore, DAG producing enzyme, phospholipase Cγ(PLCγ), was blocked by a specific inhibitor, U73122. The blockade of PLCγ suppressed the apoptosis of L-MAT cells by TCDD. We concluded that TCDD activates PLCγ by some mechanism.
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