| Budget Amount *help |
¥4,020,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
1. The scavenging activity of polyphenol for superoxide anion radical was evaluated by electron spin resonance (ESR) to clarify the mechanism of enhancing effect polyphenol radicals on wound heading. Polyphenol chemical compounds quenched superoxide anion radical in a dose-dependent manner. Polyphenol compounds were reduced to polyphenol radicals by superoxide anion radical form the enzymatic reaction using hypoxanthine and xanthine oxidase. In addition, polyphenol compounds were reduced polyphenol radicals in water DMSO solution on alkaline state.
2. Although the evaluation of superoxide anion radical scavenging activity of lipophilic antioxidant substances have been an object of study for a long time, there is little agreement as to conduct buffer solution for the lipophilic antioxidant substances in enzymatic reaction. lipid-soluble Sikonin (from herbal medicine: Kampo medicine) or catechin (from green tea) have scavenging activity for superoxide anion radical activity in DMSO-water
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solution, although it need evaluation method in lipid-solution for the activity. UV irradiation cause formation of free radicals in lipid solution, which has been inhibited by a -tocopherol (VE). The lipid radicals (L) from unsaturated fatty acid irradiated UV were detected by using spin trapping reagent with N-tett-buthyl- a -phenylnitrone (PBN) in a dose-dependent manner. The reaction between L and PBN was inhibited by VE, indicating a competitive reaction between DMPO and VE for L The method suggested that lipid radical were generation system by UV irradiation and PBN-adduct produced by the competitive reaction between PBN and VE for L. It was suggested that this PBN UV method was effective for active grade of alipid-soluble antioxidant
3. Isolation of skin from heirless mice with spin probe (4-hydroxy TENPO or Carbamoyl-PRXYL) were set in tissue-type quartz cell before epidermal side UV irradiation (256nm/365nm). The decay rate of ESR signals in process of UV irradiation were more rapid than that except UV irradiation. Therefore, in the case of homogenate of liver-tissue administrated spin probe, they were exposed to UV light, and just after others, the ESR signals were decayed quickly. The height of signals was nothing after administration of potassium ferricyanide solution on the sample for the re-oxidation. It was suggested that UV irradiation (1.11〜2.4mW/cm_2) was jack up REDOX reaction or another metabolism in the tissue.
4. The inflamed skin of mice after UV irradiation (24hr) was isolated and set on tissue-cell with two spin trapping agents (DMPO and 2- (5, 5-Dimethy1-2-oxo-2-5-[1, 3, 2]-dioxaphosphinan-2-y1)-2-methyl-3, 4-dihydro-2H-pyrrole 1-oxide: CYPMPO). In the case of using DMPO, ESR signals were showed that DMPO-adduct constituted by isotopic hexa-signals. These spectrum patterns were similar to DMPO-OOH or DMPO-OOL in water/DMSO solution. In another case of CYPMPO, ESR spectrum showed that involved signal patterns were composed of a number of CYPMPO adducts. Furthermore, there is ascorbyl radical is observed by the case. We come now to the point at which it is necessary to deal more carefully with check of the generation of free radical from skin by UV irrasiation. Less
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