Hurley M M, Gronowicz G, Kream B E, Raisz L G
Department of Medicine, University of Connecticut Health Center, Farmington.
Calcif Tissue Int. 1990 Mar;46(3):183-8. doi: 10.1007/BF02555042.
To assess the effects of heparin on bone formation we measured [3H]proline incorporation into collagenase-digestible (CDP) and noncollagen protein (NCP), [3H]thymidine (TdR) incorporation into DNA, and DNA content in 21-day-old fetal rat calvaria cultured in BGJ medium with bovine serum albumin for 24-96 hours. Heparin at 5-125 micrograms/ml decreased TdR incorporation by 26-51% at 24 and 96 hours. At 96 hours, heparin 5, 25, and 125 micrograms/ml decreased [3H]proline incorporation into CDP by 41, 48, and 32%, respectively, with no significant change in NCP. To evaluate the possible role of PGE2 in these inhibitory responses, media PGE2 concentration was measured and the effects of heparin on CDP labeling and DNA synthesis were tested in the presence of indomethacin, piroxicam, and flurbiprofen to inhibit endogenous prostaglandin E2 (PGE2) production and in the presence of a high concentration (10(-7) M) of exogenous PGE2. Heparin did not alter PGE2 production at 24 hours but at 48 hours there was a significant reduction. At 96 hours, indomethacin (10(-6) M) inhibited [3H]-proline incorporation into CDP by 38% but had no effect on the labeling of NCP. Heparin had no further significant inhibitory effect in the presence of indomethacin. Piroxicam and flurbiprofen did not alter DNA content and had a smaller inhibitory effect than indomethacin on the labeling of CDP. Moreover, addition of heparin produced a further inhibition of CDP and DNA content and finally, heparin decreased CDP labeling by 71% in the presence of PGE2.(ABSTRACT TRUNCATED AT 250 WORDS)
为评估肝素对骨形成的影响，我们测定了在添加牛血清白蛋白的BGJ培养基中培养24 - 96小时的21日龄胎鼠颅骨中，[3H]脯氨酸掺入胶原酶可消化蛋白（CDP）和非胶原蛋白（NCP）的情况、[3H]胸腺嘧啶核苷（TdR）掺入DNA的情况以及DNA含量。5 - 125微克/毫升的肝素在24小时和96小时时使TdR掺入量降低了26 - 51%。在96小时时，5、25和125微克/毫升的肝素分别使[3H]脯氨酸掺入CDP的量降低了41%、48%和32%，而NCP无显著变化。为评估前列腺素E2（PGE2）在这些抑制反应中的可能作用，我们测定了培养基中PGE2的浓度，并在吲哚美辛、吡罗昔康和氟比洛芬存在的情况下测试了肝素对CDP标记和DNA合成的影响，以抑制内源性前列腺素E2（PGE2）的产生，同时也测试了在高浓度（10(-7) M）外源性PGE2存在的情况下肝素的作用。肝素在24小时时未改变PGE2的产生，但在48小时时有显著降低。在96小时时，吲哚美辛（10(-6) M）使[3H]脯氨酸掺入CDP的量降低了38%，但对NCP的标记无影响。在吲哚美辛存在的情况下，肝素没有进一步的显著抑制作用。吡罗昔康和氟比洛芬未改变DNA含量，且对CDP标记的抑制作用比吲哚美辛小。此外，添加肝素进一步抑制了CDP和DNA含量，最后，在PGE2存在的情况下，肝素使CDP标记降低了71%。（摘要截断于250字）
