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Leone et al., 2007). Flavonoids from Scutellaria baicalensis (Chinese skullcap) and Acacia catechu (black catechu) have been used in many traditional medicines and pharmaceutical products for a variety of purposes, including anti-in?ammatory, antiviral, antibacterial, anticancer and cardiovascular applications (van Loon, 1997;

Chou et al., 2003;

De Clerq, 2005;

Huang et al., 2005). Flavocoxid, marketed as an FDA-regulated medical food, Limbrel?, for the clinical dietary management of the metabolic aspects of OA in the United States, is a mixed extract containing the naturally occurring ?avonoids, baica- lin and catechin, and has been shown to act as a dual inhibi- tor of COX-1 and COX-2 peroxidase enzyme activities with signi?cant inhibition of 5-LOX enzyme activity in vitro (Burnett et al., 2007;

Pillai and Burnett, unpubl. experiments). These ?ndings may help explain preliminary in vivo data in animals showing that ?avocoxid decreases, in a dose- dependent manner, edema in an AA-induced mouse ear swell- ing model, and abates the swelling and restores function in mice subjected to an intra-articular space injection of AA (Burnett et al., 2007). Flavocoxid has also been shown to be equivalent to naproxen for ef?cacy as measured by quality of life end points in at least one well-controlled human clinical trial (Levy et al., 2007). These results are, on the surface, direct inhibitory effects on proteins which process AA and thus reduce in?ammatory fatty acid metabolites. Although the relative inhibition of COX and 5-LOX AA processing by ?av- ocoxid is known, there are no data on how it affects gene expression and protein levels of in?ammatory markers from immune cells. Therefore, the aim of the present study was to investigate the effects of ?avocoxid in rat peritoneal macroph- ages stimulated by lipopolysaccharide (LPS) and its effect on the levels of gene expression and protein markers of in?ammation. Methods Animals and isolation of peritoneal macrophages All procedures complied with the standards for care and use of animal subjects as stated in the Guide for the Care and Use of Laboratory Animals (Institute of Laboratory Animal Resources, National Academy of Sciences, Bethesda, MD, USA). Peritoneal MFs were obtained from male SpragueC Dawley rats (250C275 g) by washing the abdominal cavity with RPMI 1640. The cells were centrifuged twice and resus- pended in the same medium at a concentration of

1 ?

106 . Macrophages were obtained after

2 h adhesion to plastic Petri dishes (Nunc, Roskilde, Denmark) at 37°C. The homogeneity and the viability of macrophages were greater than 98% as determined by differential staining and trypan blue exclusion. Macrophages w........