In Vitro Antioxidant and Anti-Inflammatory Activities of Protocatechualdehyde Isolated from Phellinus gilvus

Abstract

Summary Although various biological activities of Phellinus gilvus (PG) have been reported, the active compounds responsible for these effects are not known. Here, we evaluated the activity of various solvent extracts of PG, and found the ethyl acetate extract (Fd) to be the most active fraction, showing a strong DPPH free radical scavenging activity, and inhibitory effects on LPS-induced nitric oxide (NO) production and COX-2 mRNA expression in RAW264.7 macrophages. Six major compounds were identified from the ethyl acetate extract of PG, and protocatechualdehyde (PCA) was supposed to be the major phenolic compound of PG responsible for its DPPH free radical scavenging activity and its inhibitory effects on LPS-induced NO production in RAW264.7 cells. Further in vitro and in vivo experiments are currently underway to confirm this observation and to investigate the detailed molecular mechanisms involved in the process as well as the biological activities of other fractions of Fd. Key Words Phellinus gilvus , protocatechualdehyde, DPPH, NO, RAW264.7 cellsPhellinus gilvus (PG) is a medicinal mushroom belonging to Hymenochaetaceae basidiomycetes , and has advantages over many Phellinus species due to its short growth period (3 mo), making it cheaper to produce ( 1-3 ). Previous studies in our laboratory and by other investigators have established various biological activities of PG. Among the reported benefits of PG are inhibition of pulmonary inflammation, prevention of intraperitoneal adhesion under infectious circumstances, promotion of dermal wound healing, anti-platelet aggregation and antitumor activities ( 1-4 ). In a previous study with various solvent extracts of PG, we have demonstrated marked DPPH free radical scavenging and xanthine oxidase (XO)-inhibitory activities of the ethyl acetate extract of PG, along with the enhancement of cellular immunity by hot water extracts of PG and inhibition of thrombin-induced platelet aggregation by chloroform-, methanol-and butanol extracts of PG ( 3 ). In the current investigation, we determined the major components of the ethyl acetate extract of PG responsible for its biological activities and further compared the magnitude of the antioxidant/anti-inflammatory activities of components with the various fractional extracts of PG.The fruiting body of PG was provided by Gyeongbuk Agriculture Technology Administration (Daegu, Korea). The preparation of PG fractional extracts was reported earlier ( 3 ). Briefly, fruiting bodies of the mushroom were cut into small pieces and dried at 40-50˚C for 48 h. The pieces were homogenized, extracted with water (1 : 25) at 100˚C for 10 h, and the aqueous phase (Fa) concentrated at 80˚C in a rotary evaporator (Büchi Rotavapor R-114, Switzerland) and the final volume reduced to 1/10 of the initial volume. The Fa was then mixed with 95% ethanol (1 : 3, v/v) and stored at 4˚C overnight. The solution was then centrifuged at 15,000 ϫ g for 30 min, the precipitate dialyzed (1 : 100,000) in water, filtered, centrifuged, re-dissolved and ly...

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