Different species of the family Clusiaceae, including Rheedia longifolia, are used in folk medicine to treat inflammatory diseases. This family is largely distributed in tropical and subtropical areas of Brazil, but their chemical and pharmacological properties have been the subject of a few studies. In previous studies, we found that the aqueous extract from R. longifolia leaves presented important anti-inflammatory and analgesic activity. We investigated the chemical profile of R. longifolia and characterized the pharmacological effect of different chemically identified fractions in pharmacological models of neurogenic and inflammatory nociception. The pharmacological tests showed that oral treatment with aqueous crude extract and fractions of methanol extract of R. longifolia leaf induced a significant antinociceptive effect using von Frey filaments. In addition, the most polar fractions presented antinociceptive activity in a neurogenic model of nociception (capsaicin model). The chromatographic analysis indicated the presence of bisflavonoids in the fractions obtained from the methanol extract. These results suggest that bisflavonoids found in methanol-extracted fractions are involved in the inhibition of inflammatory and neurogenic nociception. It is important that the R. longifolia aqueous extract treatment inhibited ulcer formation induced by indomethacin, suggesting an anti-ulcerogenic activity closely associated with its analgesic effect.