【作者】 李瑞海；

【导师】 杨松松；

【作者基本信息】 辽宁中医药大学 ， 生药学， 2006， 博士

【摘要】 根据我们的立题目的，结合药效学的结果，首先确定活性部位，然后对蒺藜的具有活性的水溶性部位进行了系统的化学成分研究，以确定蒺藜的活性物质基础。同时对分离的皂苷和黄酮的单体化合物进性活性研究，确定其心血管活性的前提下，用以作为质量控制的指标成分，使得质量控制方法与活性有机结合，因此我们制定的质量控制方法对蒺藜的质量评价更具确切性和可靠性。具体研究内容如下： 首先对蒺藜水溶性主要成分进行初步研究。根据蒺藜药理方面的文献报道，蒺藜（全草）的水提取物是蒺藜的心脑血管系统主要活性部位（例如心脑舒通胶囊），因此本文对其具有活性的水溶性成分进行了初步研究。采用比色法和HPLC法分别对蒺藜的总皂苷类成分和总黄酮类成分分别进行含量测定，并对所制定的方法进行了方法学研究。之后，根据所确定的方法进行了含量测定，测定结果表明，蒺藜（全草）的水提取成分中，除了含有总皂苷类成分，还有含量较高的总黄酮类成分，提示蒺藜中总黄酮类成分也可能是蒺藜的活性部位。因此就蒺藜总黄酮在心脑血管系统方面的作用与蒺藜的总皂苷成分做了活性对比实验，药理实验研究结果提示，总黄酮与总皂苷均能显著提高小鼠耐缺氧能力，均能明显延长凝血时间，对离体家兔心脏的冠脉流量明显增加作用。实验结果说明，蒺藜中总黄酮类成分与蒺藜的总皂苷成分一样具有心脑血管系统方面的活性作用。本实验之前并无蒺藜中总黄酮类成分心脑血管系统方面的活性研究报道，蒺藜总黄酮类的活性研究，尚属首次。 由于大孔树脂对于水溶性成分具有良好的富集、纯化作用，因此，对蒺藜的水溶性成分进行有效成分大孔树脂的富集法研究。实验收集了9种常用大孔吸附树脂，对吸附容量进行了对比，确定采用树脂AB-8。又对AB-8树脂富集有效成分的工艺条件进行了考察，采用正交实验以总含量（总皂苷含量+总黄酮含量）为指标进行实验，结果确定富集蒺藜水溶性成分的最佳工艺条件（药液浓度0.5g／mL，流速8／1000BV／min；径高比1：40）。通过本实验得出的提取条件，为蒺藜的进一步研究提供了依据。 其次，化学分离实验主要对50Kg蒺藜，利用前述实验确定的条件，进行大孔树脂富集，对所得提取物（活性部位）进行化学分离。主要采用硅胶柱、葡聚糖凝胶色谱柱等分离手段进行了化学分离，共得到20个单体化合物，通过理化性质及波谱解析鉴定了其中的18个化合物的结构，分别是：terrestris A（TT1）；海柯皂苷元-3-O-β-D-木糖（1-2）-[β-D-葡萄糖-（1-3）]-β-D-葡萄糖（1-4）-β-D-甘露糖（TT2）；海柯皂苷元-3-O-β-D-木糖（1-2）-[β-D-木糖（1-3）]-β-D-葡萄糖（1-4）-[α-鼠李糖（1-2）]-β-D-甘露糖（TT3）；槲皮素-3-O-β-D-葡萄糖苷（TT4）；槲皮素-3-O-龙胆二糖苷（TT5）；山柰酚-3-O-龙胆二糖苷（TT6）；山柰酚-3-O-β-D-葡萄糖苷（TT7）；槲皮素（TT8）；海柯皂苷元（TT9）；25R-spirostan-4-ene-3，12-dione（TT10）；25R-spirostan-3，6，12-trione（TT11）；胡萝卜苷（TT12）；丁二酸（TT13）；β-sitosterol（TT14）；25R-spirostan-2,3-diol-4-ene-12-one（TT15）；7-羰基-△^5.23豆甾醇（TT16）；香兰素（TT17）；二十二烷酸（TT18）。其中terrestris A（TT1）是新化更多还原

【Abstract】 Tribulus terrestris L. are a famous traditional Chinese medicine and has been used for over hundreds years. It can restore the depressed liver for the treatment of fullness in the chest and mastits and also used to dispel the wind and clear the eyes for the treatment of an acute conjunctivitis, headache and vertigo. It has been used for a variety of liver, kidney, dizziness, psoriasis, eczema, premature ejaculation and cardiovascular diseases. The Greeks used the medicinal herb Tribulus terrestris L.as a diuretic and a mood-enhancer. Eastern Indians used it as a diuretic, antiseptic, and anti-inflammatory. The people of Bulgaria used Tribulus terrestris L.as a sex enhancer and to treat infertility.Tribulus terrestris L.is used in various herbal formulas to treat headaches eye problems such as itching, conjunctivitis and weak vision, pain relief, enhancement of the immune system, improved mood and sense of well-being, cholesterol reduction, and relief of premenstrual （PMS）, menopausal symptoms in women, and nervousness, making it a great menopause herb. In women, hormonal imbalances can lead to the typical unpleasant symptoms associated with PMS and menopause. It is also used to treat high blood pressure and rib pain. Recently, eastern European athletes and strength champions have used it as well. Many athletes and bodybuilders have found Tribulus terrestris L. a safe and natural alternative to anabolic steroids.Tribulus terrestris L.contains three groups of active physiochemicals:Dioscin, protodioscin, diosgenin and similar, these substances have effect on sexual performance and may treat various sexual disorders, they regulate sexual energy level and strength by increasing the percentage of free testosterone level for men and they effect pregnenolone, progesterone and estrogen. The hormone balancing effects of Bulgarian Tribulus terrestris L.for women makes this herb suitable for premenstrual syndrome and menopausal syndrome. Sterols like betasitosterols or stigma. These substances protect the prostate from swelling and in combination with the steroidal saponins, protect the prostate from cancer. Steroidal saponins currently referred to as steroidal saponins. These steroidal saponins effect the complete immune system. They have been demonstrated to possess anti-bacterial and anti-viral effects. Bulgarian Tribulus terrestris L.may be used internally and externally to treat herpes, and virus infections such as influenza and the common cold.In this studies, firstly, in order to do more studying on Tribulus terrestris L., we investigated its water-solubility constituents by using UV-spectral method to determine the content of the total saponins and HPLC method to to determine the total flavonoid glycosides. Methods studying results shows that the two methods were convenient, rapid and accurate, and could be used for quality control of quality of Tribulus terrestris L. According these two methods, we took study on Tribulus terrestris L. The 3 samples results show that the total Saponins in Tribulus terrestris L. are 1.55、1.59、1.67, average is 1.60%, and the total flavonoid glycosides are0.97、 1.12、1.23, average 1.11%. The resins are more proper used to exacting water-solubility constituents in herbs these days. We compared nine kinds of different resin. AB-8, performed a good reputation, was being adapted. The optimum condition of using AB-8 were concentration: 0.5g/mL , velocity of flow:8/1000BV/min, ratio of diameter and high: 1: 40, and were development by using orthogonal tests.Secondly, We investigated Tribulus terrestris L.chemical constituents of the fruits of Tribulus terrestris L by using SiO₂, polyamide, gel, LH-20. Chemical constituents from Tribulus terrestris L. 20compounds were isolated from it and 17 of them were structurally elucidated. With the compounds name: terrestrisA（TTl）; hecogenin-3-O-.(3-D-xylopyransyl（1-2）-[β-D-glucopyranosyl更多还原