Selective peroxynitrite scavenging activity of 3-methyl-1,2-cyclopentanedione from coffee extract.
It has been known that reactive oxygen and nitrogen species such as nitric oxide (NO), superoxide radical (*O2-) and their byproduct peroxynitrite (ONOO-) induce cellular and tissue injury, ultimately resulting in several human diseases. In this study, we examined scavenging effects of 3-methyl-1,2-cyclopentanedione (MCP) from coffee extract on the reactivity of those toxic molecules. MCP significantly inhibited both the oxidation of 2,7-dichlorodihydrofluorescein diacetate (H2DCFDA) by reactive oxygen species (ROS) (mainly *O2-) from kidney homogenate (41% at 100 microM) and the generation of fluorescent 4,5-diaminofluorescein (DAF-2) by NO from sodium nitroprusside (IC50 (concn producing 50% inhibition), 63.8 microM). More potently, however, MCP suppressed the oxidation of dihydrorhodamine 123 (DHR 123) to fluorescent rhodamine 123 mediated by authentic ONOO- with an IC50 value of 3.3 microM. The neutralizing effect of the reactivity of ONOO- by MCP was due to electron donation, not nitration of the compound. Additionally, MCP also decreased ONOO- formation of nitrotyrosine adducts of glutathione (GSH) reductase, and consequently protected the enzyme activity of GSH reductase against decreasing by ONOO-, indicating that MCP may prevent ONOO- -induced damage of GSH reductase. Furthermore, MCP only weakly suppressed NO production, which is one of the upstream sources of ONOO- in-vivo, suggesting that NO production may be not a pharmacological target for MCP. Taken together, our results suggest that MCP may be regarded as a selective regulator of ONOO- -mediated diseases via direct scavenging activity of ONOO-.
Kim
Ae Ra
AR
College of Pharmacy, Pusan National University, Busan 609-735, Korea.
Zou
Yani
Y
Kim
Hyung Suk
HS
Choi
Jae Sue
JS
Chang
Gregory Youngnam
GY
Kim
You Jung
YJ
Chung
Hae Young
HY
D015153:Blotting, Western; D002478:Cells, Cultured; D003069:Coffee / Q000737:chemistry*; D003517:Cyclopentanes / Q000302:isolation & purification / Q000494:pharmacology*; D016166:Free Radical Scavengers / Q000302:isolation & purification / Q000494:pharmacology*; D005980:Glutathione Reductase / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects; D009566:Nitrates / Q000737:chemistry; D009569:Nitric Oxide / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000737:chemistry*; D011506:Proteins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
D003069:Coffee; D003517:Cyclopentanes; D016166:Free Radical Scavengers; D008070:Lipopolysaccharides; D009566:Nitrates; D011506:Proteins; D017382:Reactive Oxygen Species; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; C471838:3-methyl-1,2-cyclopentanedione; D005980:Glutathione Reductase