Melittin cardiotoxicity in cultured mouse cardiac myocytes and its correlation with calcium overload.
Venom from the honey bee Apis mellifera induces cardiovascular dysfunction. We studied which constituent(s) of the venom induces cardiotoxicity and how, using cultured cardiac myocytes from mouse fetuses. Among the venom constituents, only melittin caused contractile and morphological effects; other peptides, such as apamin and mastparan; enzymes, such as phospholipase A2; and low-molecular-weight compounds, such as histamine and dopamine, did not. Treatment with 4.5 micrograms/ml melittin, which accounts for about half the dry weight of the venom, induced the same cardiotoxic effects as treatment with 9.0 micrograms/ml whole venom; these effects were a transient increase in the spontaneous beating rate, then a decrease, then cessation of beating, and finally, morphological degeneration. The cardiotoxicity of whole bee venom was completely destroyed by pretreatment of the venom with antimelittin antibody. These results suggest that bee venom cardiotoxicity is attributable to melittin. When spontaneous beating ceased following the addition of melittin or whole venom, an increase in systolic [Ca2+]i, was observed. On further incubation with melittin or bee venom, morphological injury, such as balloon degeneration, occurred concomitant with a further increase in the [Ca2+]i. An extracellular Ca2+ concentration of more than 10(-6) M was necessary for morphological injury. Melittin depolarized the maximum diastolic potentials, inhibited the generation of action potentials, and induced an increase in [Na+]i. Cells were protected against the melittin-induced increase in [Ca2+]i by pretreatment with bepridil, an inhibitor of Na(+)-Ca2+ exchange, but not by Ca2+ channel blockers such as verapamil. These observations suggest that the melittin-induced increase in [Ca2+]i was due to entry of extracellular Ca2+ via the sarcolemmal Na(+)-Ca+ exchange system.
Okamoto
T
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Japan.
Isoda
H
H
Kubota
N
N
Takahata
K
K
Takahashi
T
T
Kishi
T
T
Nakamura
T Y
TY
Muromachi
Y
Y
Matsui
Y
Y
Goshima
K
K
D000818:Animals; D001514:Bee Venoms / Q000633:toxicity; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D005318:Fetal Heart / Q000166:cytology / Q000187:drug effects; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D008555:Melitten / Q000633:toxicity*; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008813:Mice, Inbred ICR; D009200:Myocardial Contraction / Q000187:drug effects; D011247:Pregnancy; D012964:Sodium / Q000378:metabolism
D001514:Bee Venoms; D008555:Melitten; D012964:Sodium; D002118:Calcium