| Objective: In this paper,the extract of Trichosanthus rosthornii Harms was prepared on the basis of screening the best extraction solvent,making clear the reasonable ratio of the peel to seed of Trichosanthus and optimizing the process of alcohol sink.Prepare Trichosanthin extract,research its character,determination of substance content and effects on myocardial ischemia reperfusion injury rats.Methods: 1)Establish a method for quantitative analysis of total effective components in Tricholoma.According to the preparation methods of the Tricholoma of the Pharmacopoeia(35% peel and 65% seed),the extraction of water,60% ethanol and dichloromethane were used to study the content of effective components in different solvents and the ability to confrontation myocardial ischemia in rats.2)The content of effective components in different ratio extracts and the ability to confrontation myocardial ischemia in rats were investigated.The optimum proportion of peel-seed was selected by comparing the similarity between 6 different extracts and the common pattern of chemical fingerprint.3)According to the principle of the response surface method,selecting the optimum alcohol-sink process.4)Preparation of Trichosanthin extract.The characters,examination,fingerprint and content determination of Trichosanthin extract were studied by high performance liquid chromatography(HPLC-PDAD).5)Danshen pills were used as control medicine,saline as blank control,and Trichosanthin extract as experimental group.A model of myocardial ischemia was made by ligation of coronary artery anterior descending left branch.Determination of rat plasma myocardial myoglobin(Myoglobin,MYO-MB),myocardial troponin(Troponin-T,c Tn T)and creatine kinase isoenzyme mass(Cretine Kinase-MB,CK-MB);The myocardial changes were examined by light microscope and the area of myocardial infarction was determined.Results: 1)The total amino acid content was high in water and 60% alcohol extract(P<0.01)and the total flavone content was high(P<0.01)in dichloromethane extract.Compared with the model group,water extract significantly decreased the st segment of electrocardiogram(P<0.01 or P<0.05).The levels of CK-MB,c Tn T and MYO in rat plasma were reduced(P<0.05).2)Compared with normal control group,the effects of six ratio trichosanthin extracts on plasma levels of MYO,CK-MB and c Tn T in myocardial ischemia rats,P<0.01.The similarity of peel-seed ratio of 50:50 was 0.9363±0.0082,higher than that of other groups(P<0.05 or 0.01).3)The relative density of drug liquid had a great influence on the comprehensive score(P<0.01).With the increase of alcohol sink time,the comprehensive score increased,but the effect was smaller.The concentration and the time of alcohol sink increased,the comprehensive score increased.When the relative density was 1.20,the ethanol concentration was 80%,and the alcohol sink time was 36 hour.4)Trichosanthin extract is a brownish-yellow powder substance.It is sweet,bitter and soluble in water.The extract of Trichosanthin contains many nucleoside components,of which adenosine content is the highest,0.82‰.5)Compared with the model group,the high dose of Trichosanthin extract could obviously decreased the st segment of electrocardiogram(P<0.01 or P<0.05).And can significantly reduce the levels of MYO and CK-MB in rat plasma(P<0.05).The high dose group of Trichosanthin extract could protect rat myocardial tissue from damage and reduce the area of infarct and danger area,P<0.05,the difference is statistically significant.Conclusion: The solvent was water,the ratio of peel to seed was 50:50,and alcohol precipitation method could be used for the refining of this extract.The extract of Trichosanthin has a certain effect on myocardial ischemia reperfusion injury. |
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