title: Hot water extract of adzuki (Vigna angularis) suppresses antigen-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells and passive cutaneous anaphylaxis reaction in mice.
abstract: The hot water extract of adzuki (HWEA), which is produced as a byproduct in the adzuki bean boiling process, has anti-tumor, antioxidative, and anti-diabetic activities. In this study, we fractionated HWEA to 4 fractions using stepwise gradient column chromatography with water and ethanol, and demonstrated the effects of each fraction on antigen (Ag)-stimulated degranulation in rat basophilic leukemia RBL-2H3 cells. The 40% ethanol eluate extract (EtEx.40) showed the strongest inhibition level of these fractions. To reveal the inhibitory mechanisms underlying degranulation by EtEx.40, we investigated intracellular reactive oxygen species (ROS) production, intracellular free Ca²⁺ concentration ([Ca²⁺]i), and early intracellular signaling pathways. Treatment with EtEx.40 markedly inactivated Lyn following Ag stimulation, resulting in the suppressions of intracellular elevation of [Ca²⁺]i and production of ROS. To identify the active compound in EtEx.40, we isolated 7 flavonoids from EtEx.40 and calculated their inhibition levels on Ag-stimulated degranulation. These flavonoids inhibited degranulation by about 25-60%. We further examined the in vivo effects of HWEA or EtEx.40 using a passive cutaneous anaphylaxis (PCA) reaction. Both extracts strongly suppressed the PCA reaction. These findings suggest that HWEA and/or EtEx.40 are beneficial for alleviating type I allergic symptoms.
authors.affiliation: Faculty of Agriculture, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Japan. titoh@nara.kindai.ac.jp
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Itoh
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Hori
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Atsumi
authors.forename: Kazuo
authors.initials: K
authors.lastname: Toriizuka
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakamura
authors.forename: Takeshi
authors.initials: T
authors.lastname: Maeyama
authors.forename: Masashi
authors.initials: M
authors.lastname: Ando
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Tsukamasa
authors.forename: Yoshiteru
authors.initials: Y
authors.lastname: Ida
authors.forename: Yukio
authors.initials: Y
authors.lastname: Furuichi
mesh_terms: D000818:Animals; D001491:Basophils / Q000187:drug effects*; D002118:Calcium / Q000032:analysis; D015550:Cell Degranulation / Q000187:drug effects*; D045744:Cell Line, Tumor; D007887:Fabaceae / Q000737:chemistry*; D015471:Leukemia, Basophilic, Acute / Q000473:pathology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010323:Passive Cutaneous Anaphylaxis / Q000187:drug effects*; D010936:Plant Extracts / Q000494:pharmacology*; D051381:Rats; D017382:Reactive Oxygen Species / Q000032:analysis; D015398:Signal Transduction
chemical_list: D010936:Plant Extracts; D017382:Reactive Oxygen Species; D002118:Calcium