The Preparation Of Oral Persimmion Leaf Flavonoids And The Research Of Pharmacodynamics In Vivo

[Object]At present,the persimmon leaf flavonoid as the active ingredient in the treatment of atherosclerosis has been widely used in the clinic.But because of its poor liposoluble,the treatment effect of oral persimmon leaf flavonoid is not ideal and the treatment cycle is too long.With the continuous improvement of the pharmaceutics basic theory and the continuous emergence of new preparation technologies,oral liposome as the carrier of small molecule drugs have been attracting more and more attention.Therefore,this paper mainly studies the preparation of oral persimmon leaf flavonoid liposome and investigates the appearance,particle size distribution,zeta potential,encapsulation efficiency and drug loading of the oral liposome.By constructing the rat model of atherosclerosis,we investigate the treatment effect of the oral persimmon leaf flavonoid liposome on the atherosclerosis and compare with the common clinical medicines.[Method]The oral persimmon leaf flavonoid liposome is prepared by thin film dispersion-incubation method,and get the optimal process of preparation which is based on the results of single factor experiment and the principle of orthogonal test.The results are validate by entrapment efficiency and drug loading.The liposome and free drug are separated by low temperature overspeed centrifugation,and the content of drug is determined by High Performance Liquid Chromatography and Ultraviolet Spectrophotometry.The particle size distribution,and zeta potential of oral liposome are tested by Laser Particle Size Analyzer,and we use Transmission Electron Microscope to observe the appearance of oral liposome.Construct the rat model of atherosclerosis by high fat feeding method,and investigate the treatment effect of the Oral Persimmon Leaf Flavonoid Liposome,NAOXINQING Tablet,Persimmon Leaf Flavonoid Phospholipid Complex and Simvastatin on the atherosclerosis.Meanwhile,analyse the advantages and disadvantages of this several drugs.[Result]In this study,we made flavonoid from persimmon leaves with strong pharmacological activity as index components and established a test method for the identification and quantitative analysis of the flavonoid from the leaves of persimmon leaves.The results of the method study show that the analytical method has high sensitivity,good reproducibility and high recovery rate,which meets the requirements of the experimental analysis.The Preparation of diospyrin leaf flavonoid was studied by membrane dispersion method,the encapsulation efficiency and drug loading were used as the inspection indexes.The optimized formulation was obtained through single factor and orthogonal experiment design in order to get the best preparation process:Persimmon leaf flavonoid:10 mg,ratio of drug to fat:1:20,phospholipids:cholesterol:10:1,hydration agent:deionized water(pH=7).The prepared persimmon leaf flavonoid liposome suspension showed light yellow with light blue opalescence and good stability.Transmission Electron Microscopy was used to observe the homogenous spherical or spheroidal vesicle structure of the leaves of Persimmon leaf flavonoid,the appearance of roundness.The average particle size and potential were measured by laser Malvin persimmon.The average particle size of the flavonoid liposome of Persimmon leaf was 163.8nm,and the distribution was uniform with a potential of-46.4mV.Basing on Preparation of Oral Persimmon Leaf Flavonoid Liposome by Incubation,the index of encapsulation rate,change of drug loading,physical stability and relative coating rate are used as the inspection indexes,we determined to use 5g/L carbomer solution as coating material,suspension of carbopol coated persimmon flavone liposome was pale yellow,the color of the liposome is slightly lighter than that of the uncoated liposome,the physical stability is good,and relatively high rate of coating.There was no significant difference in the entrapment efficiency and drug loading of liposome before and after coating.Transmission Electron Microscopy was used to observe the homogenous spherical or spheroidal vesicle structure of coated persimmon leaf flavonoid liposome the,the appearance of roundness.The average particle size and potential were measured by laser Malvin.The average particle size of the flavonoid liposome of Persimmon leaf was 257.2nm,and the distribution was uniform with a potential of-47.1 mV.This study successfully established a rat model of atherosclerosis by high-fat feeding.The weight of rats,changes in blood lipids,and pathological changes in the aorta were used as indicators,and taking simvastatin,NAOXINQING tablets and Persimmon leaf flavonoid phospholipid complexes as positive control drugs,in order to compare the therapeutic effect of oral lipase with diospyrin leaf flavonoid and three kinds of positive drugs on atherosclerosis in rats.After the seventh week,the body weight of the blank group increased steadily,the blood lipid test results were normal,and the histological examination of the aorta showed no abnormality.The weight of the model group rats was 637.9±6.63g,which was significantly higher than that of the blank group,and Hyperlipidemia was found in the model group.A large number of foam cells and atherosclerotic plaques were observed in the aorta.There was no significant difference in the body weight and blood lipid test results between the medium-dose group and the blank group,and there was no atherosclerotic plaque formation in the aorta.Although the weight and blood lipid test results of Persimmon leaf flavonoid phospholipid complex,the simvastatin group,and the NAOXINQING group were also similar to those of the blank group,the histological examination of the three groups of blood vessels showed different degrees of changes smooth muscle cell structure.This indicates that the medium dose of persimmon leaf flavonoid liposome has a better therapeutic effect on atherosclerosis.[Conclusion]In this experiment,we used the film dispersion method to successfully prepare a suspension of flavonoid from persimmon leaves.The method has the advantages of simple operation,high stability,good reproducibility,small average particle size,uniform particle distribution,stable potential,and no burst release phenomenon.The encapsulation efficiency and drug loading amount of the flavonoid of persimmon leaves are high.Using the incubation method and carbomer to modify the persimmon leaf flavonoid liposome successfully to obtain the oral suspension of flavonoid liposome of persimmon leaf,which effectively increased the stability of the flavonoid liposome of persimmon leaves and significantly improved therapeutic effects of Persimmon leaf flavonoid on atherosclerosis.