Liu Li, Duan Jin-Ao, Su Shu-Lan, Liu Pei, Tang Yu-Ping, Qian Da-Wei
School of Pharmacy, Guangdong Medical College, Dongguan 523808, China.
Jiangsu Key Laboratory for High Technology Research of Traditional Chinese Medicine Formulae Research, Nanjing University of Chinese Medicine, Nanjing 210046, China.
Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):716-721. doi: 10.4268/cjcmm20160429.
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.
为评价桃红四物汤不同部位对二磷酸腺苷(ADP)诱导的血小板聚集及凝血酶活性的影响,并探寻其生物活性成分,采用比浊法测定家兔ADP诱导的血小板聚集率,建立一种名为凝血酶时间的生物测定法来测定抗凝血酶活性,运用超高效液相色谱-四极杆-飞行时间质谱(UHPLC-Q-TOF-MS)法对不同部位的化学成分进行定性分析。醇沉沉淀部位、醇沉上清部位及20%~30%乙醇洗脱部位可显著抑制ADP诱导的血小板聚集;醇沉上清部位、水不溶部位及40%~70%乙醇洗脱部位可显著抑制凝血酶活性。对醇沉沉淀部位、醇沉上清部位及20%~40%乙醇洗脱部位的主要成分进行分析鉴定,为芳香酸、糖苷及苯酞类。桃红四物汤抑制ADP诱导的血小板聚集及凝血酶活性的生物活性成分包括芳香酸、糖苷及苯酞类。本实验为进一步探究桃红四物汤治疗血瘀证的生物活性成分及作用机制提供了科学依据。