桃红四物汤各分离部位对体外ADP诱导血小板聚集、凝血酶活性的影响及效应物质基础研究(1)
[摘要]评价桃红四物汤各分离部位对体外二磷酸腺苷(ADP)诱导血小板聚集及凝血酶活性的影响，探讨其效应物质基础。采用比浊法测定体外ADP诱导家兔血小板聚集率；采用改进凝血酶时间法测定抑制凝血酶活性；采用UPLC-Q-TOF-MS对各分离部位进行定性分析鉴定。桃红四物汤醇沉沉淀部位、醇沉上清液部位及20%～30%醇洗脱部位对ADP诱导的血小板聚集具有显著的抑制作用；水不溶部位、醇沉上清液部位及40%～70%醇洗脱部位具有显著的抑制凝血酶活性的作用；芳香酸类成分、苷类成分和苯酞类成分是醇沉沉淀部位、醇沉上清液部位及20%～40%醇洗脱部位的主要成分。桃红四物汤抑制ADP诱导家兔血小板聚集及抑制凝血酶活性的主要效应物质基础是芳香酸类成分、苷类成分和苯酞类成分。该实验为进一步探讨活血化瘀方桃红四物汤治疗血瘀证的物质基础及作用机制提供了科学依据。
[关键词]桃红四物汤；血小板聚集；凝血酶活性；物质基础
[Abstract]To evaluate the effect of different fractions of Taohong Siwu decoction on ADP-induced platelet aggregation and thrombin activity， and to exploit the bioactive constituents， ADP-induced platelet aggregation rate in rabbits was determined by using the method of turbidity method. A bioassay called thrombin time was developed for determining anti-thrombin activities. UHPLC-Q-TOF-MS method was used to qualitatively analyze the chemical constituents of different parts. Alcohol precipitation deposition fraction， alcohol precipitation supernatant fraction and 20% to 30% alcohol elution fraction could significantly inhibit ADP-induced platelet aggregation. Alcohol precipitation supernatant fraction， water insoluble fraction and 40% to 70% alcohol elution fraction could significantly inhibit thrombin activity. The main components of alcohol precipitation deposition fraction， alcohol precipitation supernatant fraction and 20% to 40% alcohol elution fraction were analyzed and identified as aromatic acids， glycosides and phthalides. The bioactive constituents of Taohong Siwu decoction for inhibiting ADP-induced platelet aggregation and thrombin activity include aromatic acids， glycosides and phthalides. This experiment provides scientific basis to further explore the bioactive constituents and mechanism of Taohong Siwu decoction in treating blood stasis syndrome.
[Key words]Taohong Siwu decoction； platelet aggregation； thrombin activity； bioactive constituents
桃红四物汤出自清·吴谦的《医宗金鉴》，由四物汤配伍活血祛瘀药对桃仁-红花衍化而来，由桃仁、红花、当归、白芍、熟地黄、川芎6味中药组成。方中桃仁活血祛瘀、润肠通便，红花活血通经、祛瘀止痛，川芎活血祛瘀、行气开郁、祛风止痛，当归补血、活血、调经止痛、润燥滑肠，熟地黄补血滋阴、益精填髓，白芍养血和营、缓急止痛、敛肝平肝，具有活血化瘀、养血补血的双重功效，在现代中医临床被广泛应用于妇科血瘀证的治疗，且有较好疗效[1]。
中医学对血瘀证的阐述很多，如“血凝而不行”，“血泣则不通”，“凝血蕴里而不散”等，都说明血在脉中不能“如水之流”而发生瘀滞的状态。中医认为血瘀证的形成与“血行”、“血脉”皆有关，“血行失度”或“血脉不通”是产生血瘀证的根本原因。而现代医学认为，血瘀证的本质是血液循环和微循环障碍、血栓形成、血液流变性异常等。其中血小板活化在各个环节中起重要作用[1]。活化的血小板参与血栓形成，而血小板聚集性增强是血栓形成的重要因素。血小板活化后有自身放大作用，可释放二磷酸腺苷(ADP)。ADP是血小板聚积和释放的主要介质，能促进血小板聚集和血栓形成。因此，血瘀证中存在着血小板活化现象，血小板活化各项指标增高与血瘀证密切相关。血小板活化参与了血瘀证的发生与发展，血小板活化是血瘀证产生的重要病理生理基础[2-3]。 (刘立 段金廒 宿树兰 刘培 唐于平 钱大玮)
[关键词]桃红四物汤；血小板聚集；凝血酶活性；物质基础
[Abstract]To evaluate the effect of different fractions of Taohong Siwu decoction on ADP-induced platelet aggregation and thrombin activity， and to exploit the bioactive constituents， ADP-induced platelet aggregation rate in rabbits was determined by using the method of turbidity method. A bioassay called thrombin time was developed for determining anti-thrombin activities. UHPLC-Q-TOF-MS method was used to qualitatively analyze the chemical constituents of different parts. Alcohol precipitation deposition fraction， alcohol precipitation supernatant fraction and 20% to 30% alcohol elution fraction could significantly inhibit ADP-induced platelet aggregation. Alcohol precipitation supernatant fraction， water insoluble fraction and 40% to 70% alcohol elution fraction could significantly inhibit thrombin activity. The main components of alcohol precipitation deposition fraction， alcohol precipitation supernatant fraction and 20% to 40% alcohol elution fraction were analyzed and identified as aromatic acids， glycosides and phthalides. The bioactive constituents of Taohong Siwu decoction for inhibiting ADP-induced platelet aggregation and thrombin activity include aromatic acids， glycosides and phthalides. This experiment provides scientific basis to further explore the bioactive constituents and mechanism of Taohong Siwu decoction in treating blood stasis syndrome.
[Key words]Taohong Siwu decoction； platelet aggregation； thrombin activity； bioactive constituents
桃红四物汤出自清·吴谦的《医宗金鉴》，由四物汤配伍活血祛瘀药对桃仁-红花衍化而来，由桃仁、红花、当归、白芍、熟地黄、川芎6味中药组成。方中桃仁活血祛瘀、润肠通便，红花活血通经、祛瘀止痛，川芎活血祛瘀、行气开郁、祛风止痛，当归补血、活血、调经止痛、润燥滑肠，熟地黄补血滋阴、益精填髓，白芍养血和营、缓急止痛、敛肝平肝，具有活血化瘀、养血补血的双重功效，在现代中医临床被广泛应用于妇科血瘀证的治疗，且有较好疗效[1]。
中医学对血瘀证的阐述很多，如“血凝而不行”，“血泣则不通”，“凝血蕴里而不散”等，都说明血在脉中不能“如水之流”而发生瘀滞的状态。中医认为血瘀证的形成与“血行”、“血脉”皆有关，“血行失度”或“血脉不通”是产生血瘀证的根本原因。而现代医学认为，血瘀证的本质是血液循环和微循环障碍、血栓形成、血液流变性异常等。其中血小板活化在各个环节中起重要作用[1]。活化的血小板参与血栓形成，而血小板聚集性增强是血栓形成的重要因素。血小板活化后有自身放大作用，可释放二磷酸腺苷(ADP)。ADP是血小板聚积和释放的主要介质，能促进血小板聚集和血栓形成。因此，血瘀证中存在着血小板活化现象，血小板活化各项指标增高与血瘀证密切相关。血小板活化参与了血瘀证的发生与发展，血小板活化是血瘀证产生的重要病理生理基础[2-3]。 (刘立 段金廒 宿树兰 刘培 唐于平 钱大玮)