【作者】 刘旭；

【导师】 徐江平；

【作者基本信息】 第一军医大学 ， 药理学， 2004， 硕士

【摘要】 目的：为寻求更好的治疗心绞痛的中药活性成分有效部位群，制备符合现代标准的中成药，我们设计了该实验——分离提纯护心灵(HXL)四味药的有效活性部位，使之达到疗效确切、质量可控、工艺先进的目的，为心肌缺血性疾病提供新的治疗药物。 方法：HXL的含量测定采用了高效液相和气相色谱，紫外分光光度法等研究方法，其仪器化程度高、灵敏度高、重现性好，适合中药复杂成分的分离分析。对HXL的制备工艺采用正交设计的方法优化四个工艺条件的三个水平，筛选出最佳提取工艺条件，并利用大孔吸附树脂对提取物进行分离纯化，使有效活性成分达到了较好的分离。采用结扎冠状动脉左前降支的方法制备犬和大鼠心肌缺血模型，在成功制备该模型后，通过检测心电图以及血清中CK、LDH等的含量改变对HXL的药效进行了研究。 结果：分别建立了三七提取物中三七总皂苷和川芎提取物中阿魏酸的含量测定方法，建立了HXL中有效成分的含量检测方法。本研究建立的方法可对该产品的质量进行控制。用该方法对按优化工艺条件制备的三批成品进行了含量测定，其有效成分含量在50％左右。以三七总皂苷和阿魏酸的含量为指标，分别筛选出了HXL的最佳提取工艺条件：选用热浸法对三七、川芎进行提取，以13倍80％的乙醇在温度65℃时提取9h的方法提取三七总皂苷，用10倍量75％的乙醇在60℃时热浸提取9h的方法提取川芎中的阿魏酸，二者均可获得较高的提取效率。以三七总皂苷和阿魏酸含量为指标，分别考察了大孔树脂的吸附容量，流速、样液浓度等对大孔树脂吸附的影响，优化出了最佳纯化工艺条件。即按三七提取液:制流速为树脂体积(BV)=l:1.5上样装入大孔树脂柱中，控IBV瓜，洗脱后75%乙醇洗脱部分减压浓缩;取液:树脂体积(BV)=2:1上样装入大孔树脂柱中，按川芍提控制流速为1.SBV爪，洗脱后60%乙醇洗脱部分减压浓缩，按比例合并即得。采用结扎冠状动脉左前降支的方法制备大鼠心肌缺血模型，与假手术组相比，心电图s一T段发生变化，LDH及CK活力均明显增加，MDA含量明显增加，SOD的活力显著降低，以上指标与假手术组相比有非常显著性差异(P<0.01)，说明造模成功。实验结果表明，HXL可以改变缺血再灌注损伤大鼠心电图S一T段的变化幅度，降低大鼠急性心肌梗死面积，抑制血清中cK、LDH的活性，提高SOD的活力，同时降低MDA的含量。采用麻醉犬冠状动脉左前降支结扎造成心肌梗塞模型，观察HXL的治疗作用。实验结果显示，HXL可显著降低犬冠脉结扎后心肌缺血的程度，显著减小梗塞范围，作用强度与剂量有一定的关系。HXL还能显著降低犬冠脉结扎后血清乳酸、肌酸激酶及游离脂肪酸升高的程度，显著减缓全血ATP降低的程度。结论:优化工艺后的HXL是一种质量可控、疗效确切、使用安全的新型中成药复方制剂，对心肌缺血性疾病的治疗有着广阔的应用前景。更多还原

【Abstract】 Objective:The aim of this study was to scan for a better Chinese traditional medicine for the treatment of angina pectoris. In addition, an effective Chinese traditional medicine according with modern standards was designed to provide a new drug for the treatment of angina pectoris. Methods:The contents of HXL were determined by many methods such as HPLC, GC, UV, et al. It was proved that these methods were accurate, sensitive, reliable, specific and suitable for the determination of herbal drugs. The optimum extract conditions were selected by using L9 (34) orthogonal experimental design, and the chief available composition was refined with macroporous resin.Acute ischemia/reperfusion model was performed by ligating of the left coronary artery in rats and dogs. The electrocardiogram (ECG), myocardial infarct size (MIS), change of enzymes in serum were simultaneously monitered. After preparing the ischemia/reperfusion model successful, the Pharmacodynamics of HXL was studied. Results:The methods for determination of total notoginseng saponin (PNS) in Panax notoginseng (Burk.) F.H.Chen. (PN), ferulic acid (FA) in Ligusticum chuanxiong Hort. (LCH), and for quality control of HXL were established respectively.These methods can be used for routine quality control of HXL and three batches of the product weredetected.The contents of the active components were about 50%. The optimum conditions for the extraction were investigated according to the contents of PNS and FA. PN and LCH were extracted by decoction method. The optimum conditions for the extraction of PNS were to soak the drug in 13 times amount of 80% alcohol for nine hours at 65 . The more effective way to extract FA from LCH was extracting with 10 times amount of 75% alcohol under reflux for nine hours at 60 .Considered the contents of PNS and FA as criterions of evaluation, the effects of adsorptive capacities of macropore resin, velocity of flow, content on adsorption of macropore resin column were investigated. The optimum conditions for the purification were investigated. The extract from PN was loaded in macropore resin column in which the volume of PN’s extract is 1.5 times to that of the resin. The flow rate was controlled to IBV/h, and the eluting parts of 75% alcohol were concentrated decompressly. In the same way, the extract from LCH was loaded in macropore resin column and the volume of the extract is 2 times to that of the resin. The flow rate was controlled to 1.5BV/h, and at last the eluting parts of 60% alcohol were concentrated decompressly. Acute ischemia/reperfusion model was performed by ligating of the left coronary artery in rats. We found that the S-T of ECG was elevated, the content of malondialdehyde (MDA), lactic dehydrogenase (LDH), and creatine phosphokinase (CK) were increased significantly, and the activity of superoxide dismutase (SOD) was decreased in the model group, which means this model is successful. The results showed that HXL could significantly alleviate S-T segment elevation, reduce myocardial infarct size, both the LDHand CK activity in serum were decreased, and HXL improved the activity of SOD, decreased the MDA contents, respectively.The parameters of coronary circulation, the degree and range of myocardial ischemia, myocardial infarct size, the activity of serum CK, the content of FFA were determined by using the AMI model of ligating the left coronary artery in the anaesthetized openchest dogs. In dogs treated with HXL, degree and range of myocardial ischemia, myocardisl infarct size, the activity of serum CK and the content of FFA were decreased. At the same time, the content of ATP was improved significantly. Conclusion:The HXL produced with optimized technic is a safe effective Chinese patent medicine with a good quality standard; its application perspective is wide for the treatment of myocardial ischemia reperfusion injury disease.更多还原