| The Chinese herbal medicine is the most bright pearl in medicine treasure-house in the world, is the most precious cultural inheritance of Chinese nation. People have discovered a group of compound which has a living creature activity from the traditional Chinese herbal medicine in recent years, successfully develop a several ten kinds of new drugs. The heart- cerebral disease has already line up in the diseases of harming the mankind health of first, and present to the trend of increasing year by year .The formation of Thrombosis is the main pathology foundation of resulting in the heart- cerebral paroxysm with high deformity rate and death rate like CHD, atherosclerosis, cerebral ischemic stroke etc. The development of the new drugs of repressing the formation of thrombosis has important meaning to mankind health business .This research starts from the purpose of investigating the mechanism of Flemingia Flavonoid repressing the formation of thrombosis , firstly to research the extracting condition selecting of Flemingia extract By separation and identification of Flemingia extract compounds with water extraction and alcohol precipition method, to define that the compounds wereβ-sitosterol and genistein flavonoid compound, build up animal thrombosis model, establish new manufacturing animal thrombosis model method, contribute to the tested thrombosis model realm. Also to investigate the function and mechanism of Flemingia Flavonoid repressing the formation of thrombosis. It proved that Flemingia Flavonoid had the function of repressing the formation of thrombosis, bringing into play the function by repressing blood platelets to activate and promoting the celulose to dissolve, to lay theories foundation and to provide science basis for the treatment of the heart- cerebral disease by Flemingia Flavonoid, to develop Flemingia of new realms which pull out a medicine to use, contributed to the development of the precious motherland ancient Chinese herbal medicine and new drugs.This text took the root of Flemingia as raw material, through the water extraction and alcohol precipition method with material liquefied, water extraction time, water extraction temperature and water extraction times, has conducted the three elements- four levels- orthogonal experiment. By analyzing the extracting rate directly, from the result of analysis of variance, finally defined the optimum extraction of Flemingia with water extraction and alcohol precipition method was A1B2C3D3 :the rate of solid and water in 1:20 , leaching temperature is 90℃for 2 h, and extracting times are 4; through the alcohol extraction method with different alcohol density, different alcohol dosage, different extraction time and different extraction times, has conducted the three elements- four levels- orthogonal experiment, By analyzed the extracting rate directly, then from the result of analysis of variance, finally defined the optimum extraction of Flemingia with the alcohol extraction method was A1B2C3D3 : the rate of 55% alcohol in 15 , extracting for 3h, and extracting times are 2. Through the excellent project to verify that, the extraction condition which had the optimum selected rate of extract was the water extraction and alcohol precipition method. This extracting method is simple and has low cost, high extracting rate , has a certain reference value to the industrialization productions of Flemingia. To do a TLC examination to the extract by the water extraction and alcohol precipition method. Degreased extracting the extract through the water extraction and alcohol precipition method by n-hexane, then used diethyl ether again to extract further to the remnant, continue to extract with the chloroform, decompressed and concentrated by water layer as for back up.This experiment chose a wet method to pack pillar and hung silicon gum in chloroform.Placed it in ultrasonic wave instrument to remove some air bubble. Used the silicon gum pillar tochromatography, to go on the elution with several steps degree by different developing chemical. Separated to 7 kinds of mixture. Took a little amount sample which is dissolved in chloroform, joined with dense sulphuric acid, then the sulphuric acid layer and the chloroform layer present a different color, the chloroform layer present a blood-red color, the reaction between the sample and the alcohol solution with 25% three chlorine- acetic acid shew the positive, the sample was dissolved in the ice acetic acid, joined with sulphuric acid-acetic anhydride(1:20) then produced green-deeply green etc. color, that color reaction was proved initially that the compound I was sitosterol material. Hydrochloric acid zinc powder respond, it was thus clear that the color of the solution changed from yellow to tiny red, the AlCl3 respond was thus clear that yellow fluorescence, it was proved initially that the compoundâ… ,â…£was flavonoid material; To carry on a TLC comparison with developing system between the gained single compound I,â…£andβ-sitosterol,genistein standard article separately, the shape, size, color of the spot,the RF value of the compound I and which ofβ-sitosterol were all consistent , So it was proved initially that the compound I wasβ-sitosterol . Samely the compoundâ…£was defined initially as genistein, through the character and melting point measurement, we were able to make sure that the compound I wasβ-sitosterol, the compoundâ…£was genistein. It was confirmed that the valid components of the Flemingia extract was flavonoid compounds, the result has provided the science bases for the developments and application of the Chinese herbal medicine—Flemingia at molecular level, have important meaning at the new medicine developing realm. To investigate the mechanism of Flemingia Flavonoid repressing the formation of thrombosis, firstly have to build up animal thrombosis model. This research established a new method which was silver needle puncturing—pliers clipping method to construct an experimental thrombosis model, compared with traditional methods just like injuring blood vessel tunica intima method ,the thrombosis success rate has already shown significant difference(P<0.01), and gained pathoanatomy support, it is able to illustrated that this method of thrombosis model is dependable, scientific and believable, it has innovated the experimential thrombosis model, has contribute to the experimental thrombosis model realm. Through examining various blood rheology indicators about Flemingia flavonoid, the ADP inducement of the blood platelets coming together of the medicine group was obviously lower than the control group; The blood platelets glueing attached a rate to obviously descending; Flemingia flavonoid has the function of enhancing the red cell's transforming ability and repressing red cells to coming together; The length, wet heavy, dry heavy of the thrombosis in the medicine group were all lower than the control group, it was confirmed that Flemingia flavonoid had a function of repressing the formation of thrombosis; and trough the measurement of the MGP-140, tPA, PAI-1 content of big rat blood plasma in front and back the experiment , tPA of medicine group and matched control both increase in different degree, but the increase of the medicine set was obvious(P<0.05);The GMP-140 group keeps foundation level and the matched control increases ,and it has already shown significant difference;(P<0.05) PAI-1 of medicine set is lower, while PAI-1 of the matched control blood plasma increases, two sets relatively have significant difference(P<0.05) and prove the mechanism of Flavonoid Flemingia repressing the formation of thrombosis is to repress blood platelets to activate and promote the celulose to dissolve. That experiment result lays theories foundation and provides science basis for the treatment of the heart- cerebral disease by Flemingia Flavonoid, and inaugurate new field for Flavonoid .Developing a new medicine by withdrawing valid single body compound from the Chinese herbal medicine and other ways have vast development space,huge social value and latent and economic performance. Using the valid composition of natural herbal grants to treat blood bolt sex disease has become a research hotspot in both domestic and international academic circles and large medicine group .This research is on the neck of time and contributes to the research and development of new medicine which can repress the formation of thrombosis ,and to the development of Flavonoid with abundant natural medicine and make contributions for our country to become a strong medicine country from a big one.
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