Kaji T, Inada M, Yamamoto C, Fujiwara Y, Koizumi F
Department of Environmental Science, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Thromb Res. 1996 Jun 1;82(5):389-97. doi: 10.1016/0049-3848(96)00088-6.
We investigated the effect of agents which increase the intracellular level of cyclic AMP (cAMP) on the production of glycosaminoglycans (GAGs) by cultured bovine aortic endothelial cells. It was found that the incorporation of [3H] glucosamine into GAGs was significantly decreased by forskolin, 8-bromo cAMP, 3-isobutyl-1- methylxanthine and prostaglandin E1. The leakage of lactate dehydrogenase into the medium, a marker of nonspecific cell damage, were not changed by forskolin. Forskolin-induced decrease in the [3H]glucosamine incorporation occurred in both heparan sulfate and the other GAGs. The [3H]glucosamine incorporation into GAGs was suppressed by forskolin also in human aortic and human umbilical vein endothelial cells, bovine aortic smooth muscle cells, porcine kidney epithelial LLC-PK1 cells and human fetal lung fibroblastic IMR-90 cells. In conclusion, it was suggested that intracellular cAMP mediates the suppressive regulation of GAG production in vascular endothelial cells; as a result, the cell surface heparan sulfate was reduced. The cAMP-dependent pathway appears to be important as an intracellular mechanism by which endothelial production of anticoagulant heparan sulfate is regulated.
我们研究了提高细胞内环磷酸腺苷(cAMP)水平的试剂对培养的牛主动脉内皮细胞糖胺聚糖(GAGs)产生的影响。结果发现,毛喉素、8-溴化环磷酸腺苷、3-异丁基-1-甲基黄嘌呤和前列腺素E1可显著降低[3H]葡萄糖胺掺入GAGs的量。作为非特异性细胞损伤标志物的乳酸脱氢酶漏入培养基的情况,未因毛喉素而改变。毛喉素导致的[3H]葡萄糖胺掺入量减少在硫酸乙酰肝素和其他GAGs中均有发生。在人主动脉和人脐静脉内皮细胞、牛主动脉平滑肌细胞、猪肾上皮LLC-PK1细胞以及人胎儿肺成纤维IMR-90细胞中,毛喉素也抑制了[3H]葡萄糖胺掺入GAGs。总之,提示细胞内cAMP介导了血管内皮细胞中GAG产生的抑制性调节;结果,细胞表面硫酸乙酰肝素减少。cAMP依赖性途径似乎是调节内皮细胞产生抗凝硫酸乙酰肝素的重要细胞内机制。