title: Gangliosides inhibit bee venom melittin cytotoxicity but not phospholipase A(2)-induced degranulation in mast cells.
abstract: Sting accident by honeybee causes severe pain, inflammation and allergic reaction through IgE-mediated anaphylaxis. In addition to this hypersensitivity, an anaphylactoid reaction occurs by toxic effects even in a non-allergic person via cytolysis followed by similar clinical manifestations. Auto-injectable epinephrine might be effective for bee stings, but cannot inhibit mast cell lysis and degranulation by venom toxins. We used connective tissue type canine mast cell line (CM-MC) for finding an effective measure that might inhibit bee venom toxicity. We evaluated degranulation and cytotoxicity by measurement of β-hexosaminidase release and MTT assay. Melittin and crude bee venom induced the degranulation and cytotoxicity, which were strongly inhibited by mono-sialoganglioside (G(M1)), di-sialoganglioside (G(D1a)) and tri-sialoganglioside (G(T1b)). In contrast, honeybee venom-derived phospholipase A(2) induced the net degranulation directly without cytotoxicity, which was not inhibited by G(M1), G(D1a) and G(T1b). For analysis of distribution of Gα(q) and Gα(i) protein by western blotting, lipid rafts were isolated by using discontinuous sucrose gradient centrifuge. Melittin disrupted the localization of Gα(q) and Gα(i) at lipid raft, but gangliosides stabilized the rafts. As a result from this cell-based study, bee venom-induced anaphylactoid reaction can be explained with melittin cytotoxicity and phospholipase A(2)-induced degranulation. Taken together, gangliosides inhibit the effect of melittin such as degranulation, cytotoxicity and lipid raft disruption but not phospholipase A(2)-induced degranulation in mast cells. Our study shows a potential of gangliosides as a therapeutic tool for anaphylactoid reaction by honeybee sting.
authors.affiliation: Health Service Center, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Nishikawa
authors.forename: Seiichi
authors.initials: S
authors.lastname: Kitani
mesh_terms: D000818:Animals; D001514:Bee Venoms / Q000037:antagonists & inhibitors* / Q000201:enzymology / Q000276:immunology / Q000633:toxicity; D002417:Cattle; D015550:Cell Degranulation / Q000187:drug effects / Q000276:immunology; D002460:Cell Line; D004285:Dogs; D005562:Formazans / Q000032:analysis; D005732:Gangliosides / Q000494:pharmacology*; D008407:Mast Cells / Q000187:drug effects* / Q000276:immunology; D008555:Melitten / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000633:toxicity; D021962:Membrane Microdomains / Q000378:metabolism; D064801:Phospholipase A2 Inhibitors; D054467:Phospholipases A2 / Q000276:immunology / Q000633:toxicity*; D013778:Tetrazolium Salts / Q000032:analysis; D001619:beta-N-Acetylhexosaminidases / Q000032:analysis
chemical_list: D001514:Bee Venoms; D005562:Formazans; D005732:Gangliosides; D064801:Phospholipase A2 Inhibitors; D013778:Tetrazolium Salts; D008555:Melitten; C079782:MTT formazan; D054467:Phospholipases A2; D001619:beta-N-Acetylhexosaminidases