Heparin specifically inhibits the inositol 1,4,5-trisphosphate-induced Ca2+ release from skinned rat aortic smooth muscle cells in primary culture.
By measuring the 45Ca2+ movement in saponin-skinned primary cultured rat aortic smooth muscle cells, we examined the specificity of the inhibitory effect of heparin on the IP3-induced Ca2+ release. IP3 (100 mumol/l) markedly (98%) decreased the MgATP-dependent 45Ca2+ content in the non-mitochondrial Ca2+ stores in the presence of 1 mumol/l free Ca2+. Heparin (1-100 micrograms/ml) dose-dependently inhibited this Ca2+ release by IP3. In Ca2(+)-free solution, heparin (100 micrograms/ml) inhibited the increases in 45Ca2+ efflux rate evoked by 10 mumol/l IP3. De-N-sulfated heparin did not inhibit the IP3-induced Ca2+ release. Hyaluronic acid, heparan sulfate, chondroitin sulfate A, chondroitin sulfate B, chondroitin sulfate C and 2,6-disulfated D-glucosamine had no inhibitory effects on the IP3-induced Ca2+ release. High concentrations (over 1 mg/ml) of heparin inhibited the 45Ca2+ influx and decreased the 45Ca2+ content in skinned cells. These results suggest that heparin (1-100 micrograms/ml) specifically inhibits the IP3-induced increase in Ca2+ permeability of Ca2+ stores and that three sulfate groups at different locations on the molecule of heparin, two at the D-glucosamine and one at the iduronic acid, may be important for this action, in skinned vascular smooth muscle cells, in culture.
Yamamoto
H
H
Medical Education Center, Kyushu University, Fukuoka, Japan.
Kanaide
H
H
Nakamura
M
M
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002132:Calcium Radioisotopes; D002478:Cells, Cultured; D006025:Glycosaminoglycans / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology*; D007292:Inosine Nucleotides / Q000037:antagonists & inhibitors*; D007293:Inosine Triphosphate / Q000037:antagonists & inhibitors*; D008931:Mitochondria, Muscle / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D051381:Rats
D002132:Calcium Radioisotopes; D006025:Glycosaminoglycans; D007292:Inosine Nucleotides; D007293:Inosine Triphosphate; D002110:Caffeine; D006493:Heparin; D002118:Calcium