Effect of YM598, a selective endothelin ETA receptor antagonist, on endothelin-1-induced bone formation.
We investigated the effect of endothelin-1 on bone formation in vitro and in vivo, and the effect of YM598, a novel selective endothelin ET(A) receptor antagonist, on endothelin-1-induced responses. In in vitro studies, the effect of endothelin-1 on cellular responses was investigated by measuring intracellular Ca(2+) levels, cell growth and alkaline phosphatase activity in the mouse osteoblast-like cell line MC3T3-E1. In in vivo studies, the effect of endothelin-1 on bone morphogenetic protein-2-induced ectopic bone formation in rats was investigated. A carrier containing bone morphogenetic protein-2 with or without endothelin-1 was subcutaneously implanted over the thorax, and the tissue (carrier) calcium content 3 weeks after implantation was evaluated. The inhibitory effect of YM598 on these responses was also investigated. In the in vitro studies, endothelin-1 (10(-13) to 10(-6) M) significantly increased intracellular Ca(2+) concentration, DNA synthesis and cell number in a concentration-dependent manner, while significantly decreasing alkaline phosphatase activity. YM598 (10(-12) to 10(-4) M) significantly inhibited these increases, as well as the decrease in alkaline phosphatase activity, in a concentration-dependent manner. In the in vivo studies, the tissue calcium content 3 weeks after carrier implantation was significantly higher in the group that received both bone morphogenetic protein-2 and endothelin-1 than in the group receiving bone morphogenetic protein-2 alone. Chronically administered YM598 (1 mg/kg/day) marginally inhibited this endothelin-1-potentiated ectopic bone formation. These results suggest that endothelin-1 may induce bone formation via endothelin ET(A) receptors in vitro and in vivo.
Someya
Akiyoshi
A
Pharmacology Laboratories, Institute for Drug Discovery Research, Astellas Pharma, Inc. 21, Miyukigaoka, Tsukuba-shi, Ibaraki 305-8585, Japan. akiyoshi.someya@jp.astellas.com
Yuyama
Hironori
H
Fujimori
Akira
A
Ukai
Masashi
M
Fukushima
Shinji
S
Sasamata
Masao
M
D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000008:administration & dosage; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004343:Drug Implants; D065130:Endothelin A Receptor Antagonists; D019332:Endothelin-1 / Q000008:administration & dosage / Q000378:metabolism* / Q000494:pharmacology; D008297:Male; D051379:Mice; D009999:Ossification, Heterotopic / Q000378:metabolism / Q000517:prevention & control*; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D051381:Rats; D020318:Rats, Long-Evans; D044022:Receptor, Endothelin A / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology* / Q000627:therapeutic use; D016212:Transforming Growth Factor beta / Q000008:administration & dosage
C525719:Bmp2 protein, mouse; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D004343:Drug Implants; D065130:Endothelin A Receptor Antagonists; D019332:Endothelin-1; C449179:N-(6-methoxy-5-(2-methoxyphenoxy)-2-(pyrimidin-2-yl)pyrimidin-4-yl)-2-phenylethenesulfonamide; D011743:Pyrimidines; D044022:Receptor, Endothelin A; D011994:Recombinant Proteins; D013449:Sulfonamides; D016212:Transforming Growth Factor beta; D000469:Alkaline Phosphatase; D002118:Calcium