Kaji T, Akai T, Hayashi T, Sakuragawa N
Department of Environmental Science, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Thromb Res. 1991 Feb 15;61(4):375-84. doi: 10.1016/0049-3848(91)90651-c.
The effect of thrombin on the metabolism of glycosaminoglycans (GAG) in cultured bovine aortic endothelial cells was investigated. Incubation of confluent endothelial cell cultures with thrombin at 1.0 NIH U/ml for 8 h and above caused a significant decrease in the accumulation of [35S]sulfate-labeled GAG (35S-GAG) in the cell layer. After a 24 h incubation, thrombin at 0.5 U/ml and above decreased the accumulation of 35S-GAG in both cell layer and medium. The percentage of 35S-GAG released into the medium during the last 3 h of 24 h incubation was significantly increased by thrombin. In thrombin-treated cell layer, both heparan sulfate and the other GAG was significantly decreased at the same degree. The incorporation of both [3H]thymidine and [14C]leucine was significantly increased by thrombin. Gabexate mesilate, a serine protease inhibitor, suppressed the thrombin inhibition. From these results, it was suggested that thrombin decreased 35S-GAG in the endothelial cell layer without a nonspecific damage of the cells through mainly an inhibition of GAG production and additionally a stimulation of GAG release.
研究了凝血酶对培养的牛主动脉内皮细胞中糖胺聚糖（GAG）代谢的影响。将汇合的内皮细胞培养物与1.0 NIH U/ml的凝血酶孵育8小时及以上，导致细胞层中[35S]硫酸盐标记的GAG（35S-GAG）积累显著减少。孵育24小时后，0.5 U/ml及以上的凝血酶降低了细胞层和培养基中35S-GAG的积累。在24小时孵育的最后3小时内，凝血酶显著增加了释放到培养基中的35S-GAG的百分比。在凝血酶处理的细胞层中，硫酸乙酰肝素和其他GAG均以相同程度显著减少。凝血酶显著增加了[3H]胸腺嘧啶核苷和[14C]亮氨酸的掺入。丝氨酸蛋白酶抑制剂甲磺酸加贝酯抑制了凝血酶的抑制作用。从这些结果表明，凝血酶通过主要抑制GAG产生以及额外刺激GAG释放，在不造成细胞非特异性损伤的情况下减少了内皮细胞层中的35S-GAG。
