Gardenia fruit has been traditionally used as a folk medicine for centuries in Asian countries. Extraction with ethanol was used to obtain an extract (GFE) that contains two known constituents, geniposide and genipin, which were subsequently evaluated for anti-inflammatory activity. GFE, genipin, and geniposide showed acute anti-inflammatory activities in carrageenan-induced rat paw edema. In a dose-dependent manner, GFE also inhibited vascular permeability induced by acetic acid. Both genipin and geniposide inhibited production of exudate and nitric oxide (NO) in the rat air pouch edema model. However, genipin possessed stronger anti-inflammatory activity than geniposide, as demonstrated by the results with carrageenan-induced rat paw edema, carrageenan-induced air pouch formation, and measurement of NO content in the exudates. GFE caused a dose-dependent inhibition of acetic acid-induced abdominal writhing in mice. Collectively, genipin, rather than geniposide, is the major anti-inflammatory component of gardenia fruit.