[Effects of rHuEPO on cellular proliferation and endothelin-1 production in cultured endothelial cells].
To elucidate the effects of recombinant human erythropoietin (rHuEPO) on endothelial cell proliferation, protein synthesis and endothelin-1 (ET) production by endothelial cells were examined in a cultured endothelial cell (EC) system. EC, incubated with various concentrations of rHuEPO (0, 1000, 5000, 10,000mU/ml) for up to 3 days exhibited dose-dependent cellular proliferation that could be inhibited by anti-rHuEPO rabbit antiserum. DNA and protein synthesis, measured by the uptake of 3H-thymidine and 3H-leucine, respectively, showed dose-dependent increases after incubation of confluent EC in the resting phase of the cell cycle with various concentrations of rHuEPO (0, 1000, 2500, 5000, 10,000mU/ml) for 18 h. After an additional 6-h culture with rHuEPO at 5000mU/ml or more, the supernatant concentrations of immunoreactive ET showed significantly greater values than those without rHuEPO. Furthermore, rHuEPO increased the DNA synthesis by EC, which had been cultured in E-BM medium containing 0.5% or 2% FBS for 3 h, and which were recultured in E-BM medium containing 5% FBS or containing 2% FBS with 10 ng/ml EGF for 15 h. These results suggest that rHuEPO directly stimulates EC proliferation as a competence factor, and also accelerates endothelin-1 production in association with stimulation of DNA and protein synthesis by EC.
Nagai
T
T
Department of Internal Medicine, Showa University Fujigaoka Hospital, Yokohama, Japan.
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D016232:Endothelins / Q000096:biosynthesis*; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology*; D006801:Humans; D014176:Protein Biosynthesis; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology
D016232:Endothelins; D011994:Recombinant Proteins; D004921:Erythropoietin; D004247:DNA