The fruits of Piper cubeba L. are used traditionally to treat respiratory disorders in Indonesia. Inorder to determine the compounds responsible for this activity, the fruits were extracted with n-hexane followed by ethanol to give n-hexane and ethanol extracts. Based on tracheospasmolyticassay on these two extracts, the n-hexane extract was more active to inhibit trachea contractionthan that of ethanol extract. Upon bioassay guided isolation of the n-hexane extract, a tracheospasmolyticactive compound was isolated and identified as dihydrocubebin [(3,4),(3’,4’)-bis-methylenedioxy-9,9’dihydroxylignan] (1). Compound 1 was tested further for its ability to inhibit histamine releasedfrom mast cells, using rat basophilic leukemia (RBL-2H3) cell line and rat peritoneal mast cells(RPMCs) as models; and DNP24-BSA, thapsigargin, ionomycin, compound 48/80 and PMA wereused as inducers for histamine released from mast cell. The test result showed that 1 inhibitedhistamine release from RBL-2H3 cells induced by DNP24-BSA, thapsigargin and ionomycin. Inaddition, 1 suppressed histamine release from RPMC induced by either thapsigargin or ionomycin.However, 1 did not inhibit histamine release from RPMC induced by either compound 48/80 orcombination PMA-sub optimum dose of ionomycin. Therefore, it was concluded that the inhibitory effectsof 1 on the histamine released from mast cells may involve mechanisms related to intracellular Ca2+signaling events or downstream processes of intracellular Ca2+ signaling in mast cells.