澳门大学硕士学位论文 Abstract The root of Panax notoginseng (Burk.) F. H. Chen (Sanqi in Chinese) is a valuable and commonly used Chinese medicine. It is known for its efficacy in promoting blood circulation, removing blood stasis, inducing blood clotting, relieving swelling, and alleviating pain. It has been widely used for treatment of cardio- and cerebro-vascular diseases. But the compounds which are mainly contribute to the beneficial effects such as antiplatelet aggregation on cardiovascular and cerebrovascular systems is still unknown. A conventional procedure for finding bioactive components is extraction of traditional Chinese medicines (TCMs) followed by pharmacological assay of the purified compounds. In addition, screen of bioactive compounds carried out on animal models are time-consuming, labor-intensive, expensive and low efficient for directly screening bioactive compounds from TCMs. Modern pharmacological studies have shown that combining with some receptors or channels on cell membrane is the first step of drug action. In present study, a method, human platelet biospecific extraction and HPLCDAD-MS/MS, was developed for screening anti-platelet agents from Paпаx notoginseng. The thesis includes: 1. Human platelet biospecific extraction and HPLC/ESI-MS/MS was developed and applied for screening potential antiplatelet agents in Sanqi. Two nucleosides, guanosine and adenosine, and 3 saponins were found in P. notoginseng. 2. The anti-platelet aggregation of adenosine, guanosine and three extracts from P. notoginseng were also investigated and compared in vitro. Guanosine and adenosine had strong inhibition of platelet aggregation induced by ADР, AA and THR. 澳门大学硕士学位论文 3. Simultaneous quantification of guanosine and adenosine in different parts of P. notoginseng from different locations was performed by using HPLC-DAD. The contents of the two nucleosides from the same parts were similar, but varied in the samples from the different locations. The data suggest that human platelet biospecific extraction and HPLCDAD-MS/MS is a useful method for screening antiplatelet agents from Chinese medicines. Keywords: Human platelet biospecific extraction; Panax notoginseng; HPLCMS; Antiplatelet aggregation; Nucleosides; Saponins

三七为五加科植物 Panax notoginseng (Burk.) F.H. Chen 的干燥根及根茎, 为我国名贵中药,具有散瘀止血、消肿定痛之功效,近年来多用于治疗冠心 病、心绞痛等心脑血管系统疾病。血小板聚集在心血管疾病发生发展中具有重 要作用,但三七对其的影响及活性成分还不尽清楚。 传统从中药中筛选活性成分的方法是先采用植物化学的方法分离得到纯的 化合物,再进行药理实验,但是这样的方法费时费力并且成功率低。现代药理 研究表明,药物要产生效应一般先要与细胞膜上的受体或通道结合。本研究采 用血小板生物特异性萃取结合 HPLC-MS 筛选三七抗血小板活性成分。主要内 容包括: 1. 采用人血小板生物特异性萃取结合高效液相色谱-电喷雾质谱法筛选三七水 提醇沉上清和三七总皂苷中能与血小板特异性结合的成分,从水提醇沉上清中 发现两个活性成分,分别为鸟苷和腺苷;从总皂苷中发现三个活性成分,其中 之一为人参皂苷 Rh1. 2. 对筛选出来的活性成分鸟 、腺苷及三种三七提取物进行了体外抗兔血小板 聚集试验。利用光电比浊法测定了五个样品体外抑制血小板聚集率并进行比 较。结果显示鸟 和腺苷具有很强的抗血小板聚集活性,三种提取物具有一定 的抗血小板聚集作用。 3. 利用 HPLC-DAD 方法,对不同产地和部位的三七药材中5种核苷类成分, 即尿 啶、尿苷、胞 、鸟苷和腺苷进行了含量测定。结果显示三七同一部位 的核苷含量变化不大,但是不同部位中核苷含量差异显著。 关键词:人血小板生物特异性萃取;三七;HPLC-MS;抗血小板聚集;核苷; 皂苷