Inhibition of histamine release from RBL-2H3 cells by protein synthesis inhibitors.
Effects of cycloheximide, an inhibitor of protein synthesis, on histamine release from RBL-2H3 cells were examined. RBL-2H3 cells sensitized by rat antiserum to ascaris extract were challenged by the antigen, and histamine release during a period of 30 min was measured. Pretreatment with cycloheximide (1 microgram/ml) for 1 h significantly inhibited the antigen-induced histamine release (36% inhibition). The cycloheximide-induced inhibition of histamine release was abolished when the cells were further incubated in the absence of cycloheximide for 2 h. Pretreatment with puromycin (3 and 10 micrograms/ml), an inhibitor of protein synthesis, or actinomycin D (0.1-1 microgram/ml), an inhibitor of DNA-dependent RNA synthesis, also inhibited the antigen-induced histamine release in a concentration-dependent manner. Both ionomycin- and thapsigargin-induced histamine release were also inhibited by pretreatment with cycloheximide. Measurement of intracellular Ca2+ levels using quin 2 revealed that cycloheximide inhibits the increase in Ca2+ levels induced by the antigen, ionomycin or thapsigargin. These results suggest that histamine release induced by the antigen, ionomycin and thapsigargin in RBL-2H3 cells is mediated by protein(s) which is newly synthesized and inactivated rapidly, and the newly synthesized protein(s) is involved in the increase of intracellular Ca2+ levels induced by these stimulants.
Hirasawa
N
N
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Miyagi, Japan.
Kawamura
R
R
Matsui
N
N
Katoh
M
M
Yomogida
S
S
Mue
S
S
Watanabe
M
M
Ohuchi
K
K
D000818:Animals; D000941:Antigens / Q000494:pharmacology; D002118:Calcium / Q000032:analysis; D002273:Carcinogens / Q000494:pharmacology; D017077:Culture Media, Conditioned / Q000737:chemistry; D003513:Cycloheximide / Q000494:pharmacology*; D003609:Dactinomycin / Q000494:pharmacology; D006636:Histamine Release / Q000187:drug effects* / Q000276:immunology; D015759:Ionomycin / Q000494:pharmacology; D015471:Leukemia, Basophilic, Acute; D014176:Protein Biosynthesis; D011506:Proteins / Q000037:antagonists & inhibitors; D011691:Puromycin / Q000494:pharmacology; D051381:Rats; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin; D014407:Tumor Cells, Cultured / Q000378:metabolism
D000941:Antigens; D002273:Carcinogens; D017077:Culture Media, Conditioned; D011506:Proteins; D013729:Terpenes; D003609:Dactinomycin; D011691:Puromycin; D015759:Ionomycin; D019284:Thapsigargin; D003513:Cycloheximide; D002118:Calcium