| 研究課題/領域番号 |
19K07187
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| 研究機関 | 東北大学 |
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研究代表者 |
李 宣和 東北大学, 薬学研究科, 准教授 (60519776)
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| 研究分担者 |
大江 知行 東北大学, 薬学研究科, 教授 (10203712)
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| 研究期間 (年度) |
2019-04-01 – 2022-03-31
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| キーワード | Pyridoxamine / Dopamine / Parkinson`s disease / Oxidative stress |
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| 研究実績の概要 |
Confirmation of the formation of PL-DA adduct in the intracellular-like conditions:
1. Dopamine (DA) was reacted with pyridoxamine (PM) in the presence of tyrosinase. Reaction time- and tyrosinase concentration-dependent decrease in DA and increase in pyridoxal (PL) and PL-DA adduct were observed. This result indicates that PM can scavenge DA quinone (DAQ) that is produced by enzymatic oxidation of DA.
2. DA was reacted with PM in the presence of glutathione (GSH). DAQ was formed via autooxidation and reacted with GSH as well as PM to produce GSH-DA and PL-DA adducts. The level of PL-DA adduct was increased in a PM dose-dependent manner, but not GSH-DA adduct. The same results were obtained when the reaction was carried out in the presence of GSH and tyrosinase.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
As planned, PL-DA adduct obtained from the reaction of DA with PM was characterized by MS/MS, UV, and NMR analyses. The mechanism for the adduct formation was proposed based on the reaction of DA analogs and PM. PM was shown to scavenge DAQ in the presence of tyrosinase and/or GSH, indicating that PM could inhibit DA-induced neurotoxicity by directly scavenging DAQ.
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| 今後の研究の推進方策 |
To investigate the inhibition effect of PM:
1. On the modification of α-synuclein (Syn) by DA: DA will be reacted with Syn to characterize the modification. Changes in levels of DA-modified Syn and PL-DA adduct will be monitored by adding PM.
2. On DA-induced aggregation of Syn: DA-induced aggregation of Syn will be detected by Thioflavin T fluorescence assay. Changes of aggregation status will be monitored by adding PM
3. On DA-induced oligomerization of Syn and cytotoxicity in human dopaminergic neuroblastoma cell line SH-SY5Y: DA-induced oligomerization of Syn will be detected by SDS page or Western blotting. Cytotoxicity will be evaluated by MTT assay. Changes in oligomerization/aggregation/cytotoxicity will be monitored by adding PM.
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