Ku Sae-Kwang, Bae Jong-Sup
Department of Anatomy and Histology, College of Korean Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
College of Pharmacy, CMRI, Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
Vascul Pharmacol. 2014 Mar;60(3):120-6. doi: 10.1016/j.vph.2014.01.009. Epub 2014 Feb 15.
Withaferin A (WFA), an active compound from Withania somnifera, is widely researched for its anti-inflammatory, cardioactive and central nervous system effects. However, antiplatelet, anticoagulant, and profibrinolytic properties of WFA have not been studied. In this study, the anticoagulant activities of WFA were measured by monitoring activated partial thromboplastin-time (aPTT), prothrombin time (PT), fibrin polymerization, platelet aggregation, thrombus formation, and the activities of cell-based thrombin and activated factor X (FXa). The effects of WFA on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-α (TNF-α) activated human umbilical vein endothelial cells (HUVECs). Our data showed that WFA inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation, FeCl3-induced thrombus formation, prolonged aPTT and PT significantly and inhibited the activities and production of thrombin and FXa. WFA prolonged in vivo and ex vivo bleeding time and inhibited TNF-α induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by WFA. Collectively, these results indicate that WFA possesses antithrombotic activities and suggest that the current study could provide bases for the development of new anticoagulant agents.
印度人参中的活性化合物睡茄内酯A（WFA）因其抗炎、心脏活性和对中枢神经系统的作用而受到广泛研究。然而，WFA的抗血小板、抗凝和纤维蛋白溶解特性尚未得到研究。在本研究中，通过监测活化部分凝血活酶时间（aPTT）、凝血酶原时间（PT）、纤维蛋白聚合、血小板聚集、血栓形成以及基于细胞的凝血酶和活化因子X（FXa）的活性来测定WFA的抗凝活性。还在肿瘤坏死因子-α（TNF-α）激活的人脐静脉内皮细胞（HUVECs）中测试了WFA对1型纤溶酶原激活物抑制剂（PAI-1）和组织型纤溶酶原激活物（t-PA）表达的影响。我们的数据表明，WFA抑制凝血酶催化的纤维蛋白聚合和血小板聚集、FeCl3诱导的血栓形成，显著延长aPTT和PT，并抑制凝血酶和FXa的活性及产生。WFA延长体内和体外出血时间，并抑制TNF-α诱导的PAI-1产生。此外，WFA使PAI-1/t-PA比值显著降低。总体而言，这些结果表明WFA具有抗血栓形成活性，并表明当前研究可为新型抗凝剂的开发提供依据。
