凝血酶及凝血因子Xa诱导血管平滑肌细胞中植物凝集素样氧化型低密度脂蛋白受体-1表达的实验研究  

Tyrosine kinase dependent lectin-like oxidized LDL receptor-1 upregulation by thrombin and factor Xa in vascular smooth muscle cells

作　　者：苏琳[1] 苗懿德[1] 孙立新[1] 孙宁玲[2] 

机构地区：[1]北京大学人民医院老年科,100044 [2]北京大学人民医院心内科,100044

出　　处：《中华心血管病杂志》2006年第3期262-266,共5页Chinese Journal of Cardiology

摘　　要：目的研究血管平滑肌细胞中凝血酶及凝血因子Xa对新型的氧化型低密度脂蛋白的清道夫受体,即植物凝集素样氧化型低密度脂蛋白受体-1(LOX-1)表达的影响。方法培养的牛主动脉平滑肌细胞,予凝血酶及凝血因子Xa刺激后,用鼠抗LOX-1单克隆抗体,对细胞裂解液和浓缩的培养基进行Western blot分析,观察LOX-1表达的变化。结果在凝血酶2·0U/ml及凝血因子Xa50nmol/L时,可观察到细胞膜结合型LOX-1表达明显增加。凝血酶3·0U/ml和凝血因子Xa100nmol/L刺激14h后,细胞培养基中可溶型LOX-1表达明显升高。对平滑肌细胞给予1·0U/ml凝血酶及100nmol/L凝血因子Xa刺激,4h后LOX-1表达开始增加,12h后达高峰。AG1478是表皮生长因子受体相关酪氨酸激酶抑制剂。用指定浓度AG1478预刺激后,再予凝血酶和凝血因子Xa,然后对细胞裂解液进行Western blot分析。AG1478可显著抑制凝血酶及凝血因子Xa导致的LOX-1表达增加。结论凝血酶及凝血因子Xa可诱导LOX-1表达增加,此作用由表皮生长因子受体介导。Objective Thrombin and factor Xa are key players in the process of arterial thrombi formation and lectin-like oxidized low density lipoprotein receptor-1 （ LOX-1 ） is a cell surface endocytosis receptor for atherogenic oxidized LDL （ox-LDL）. Here we investigate whether thrombin and factor Xa can induce LOX-1 protein expressions in cell-associated forms and soluble forms in cultured bovine aortic smooth muscle cells （ BSMCs ）. Methods BSMCs were treated with thrombin or factor Xa in the presence or absence of AG1478, an epidermal growth factor （EGF） receptor-associated tyrosine kinase inhibitor. Total cell lysates and concentrated culture medium were then analyzed by Western blot using a mouse anti-LOX-1 monoclonal antibody. Results LOX-1 protein levels in cell lysates and culture medium were significantly increased by thrombin and factor Xa in a concentration- and time-dependent manner. Upregulatlon of LOX-1 protein expressions in cell ]ysates and concentrated culture medium was observed at concentrations above 2.0 and 3.0 U/m[ of thrombin and 50 and 100 nmol/L of factor Xa, respectively. Increased LOX-1 protein expressions in cell lysates and cell culture medium were detectable as early as 4 h and peaked at 12 h after treatment with thrombin or factor Xa. LOX-1 expression induced by thrombin and factor Xa could be blocked by pretreatment with AG1478. Conclusions Thrombin and factor Xa can act as LOX-1 inducers via tyrosine kinase activation.

关 键 词：动脉硬化 肌 平滑 血管 受体 LDL 凝血酶 因子xa 

分 类 号：R543[医药卫生—心血管疾病]