Measuring Lipid peroxidation in the matrix using ESR spin labeling
| Project/Area Number |
08455413
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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 | 一般 |
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| Research Field |
有機工業化学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
TAKAHASHI Mareyuki The Univ. of Tokyo, Research Center for Advanced Science and Techology, Assistant Lecturer, 先端科学技術研究センター, 助手 (60280955)
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| Co-Investigator(Kenkyū-buntansha) |
NOGUCHI Noriko The Univ. of Tokyo, Research Center for Advanced Science and Technology, Assista, 先端科学技術研究センター, 助手 (40198578)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | election spin resonance / spin label technique / lipid peroxidation / liposumes / cultured cells / plasma membrane / 細胞膜 / ESRスピンラベル法 / 酸化劣化 / 不均一系 / 脂質二分子膜 |
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| Research Abstract |
The novel method for measuring lopid peroxidation in the heterogeneous micro environments such as lopid bilayrs has been studied. This method might be useful for studying the degradation of newly synthesized materials such as functional polymers and biomembranes and also be useful for evaluating the activity of antioxidans which are designed to reside within the matrix.
Followings are the results obtained. First, we evaluated the degree of lipid peroxidation taking place in the artificial lipid bilayrs (liposome) prepared with phosphatidylcholine, using ESR spin label technique with 5,10 and 16NS as spin probes. When azo compound AAPH was used as a free radical generator, ESR signal intensity of spin probes were decreased with time-and dose-dependently. By comparing with the formation of lipid hydroperoxides this method was proved to be useful to evaluate lipid peroxidation of the artificial membranes quantiatively. We next tried to apply this method to biological materials. Human leukemia cell line THP-1 or U937 was labeled with spin probes 5NS and 16NS by incubating cells with thin film formed on the inner surface of a round-bottom flask. The various kinds of reactive oxygen species were applied. In the case of AAPH,an extracellular peroxyl radical generator, the signal intensity of 5NS was diminished faster than that of 16NS,suggesting that the amount of free radicals near the surface was greater than deep inside the membranes. On the other hand, hydrogen peroxide, one of the most common reactive oxygen species, did not reduce the signal intensities of these two spin probes significantly, suggesting that hydrogen peroxide itself has little interaction with the plasma membrane of the cell. Thus this method was shown to be useful to evaluate lipid peroxidation in the microenvironment.
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Report
(3 results)
Research Products
(15 results)
