| Objective Areca is the seed of areca,a plant of the genus areca,in the palm family,which are mainly planted in Hainan,Fujian,Yunnan and other regions.areca nut contains phenolic substances about 30%,with various compositions,mainly flavonoids.As the main active substance of betel nut,polyphenols have bacteriostatic and anti-aging effects.Among them,the strong antioxidant effect can effectively prevent a variety of chronic diseases.Hypoxia at high altitude will cause hypoxic damage in tissue and cells,among which oxidative stress damage to tissues and organs is the pathophysiological basis of most diseases.According to the preliminary experiments of the research group,the strong antioxidant properties of areca nut can effectively alleviate the oxidative damage induced by hypoxia in plateau.Therefore,in order to clarify the role of betel nut polyphenols in anti-plateau hypoxia,areca nut was extracted and purified to increase the content of polyphenols.Wistar rats are used as research objects to conduct pharmacodynamic experiments.Into a suitable dosage form,it is developed into a new anti-hypoxia drug,which plays a very important role in combating the development of plateau hypoxia resources.Methods The extraction of polyphenols from areca nut and the optimized of the conditions:Solvent extraction method was used to extract betel nut polyphenols.The content of polyphenols was measured using ferrous tartrate method.The effect of ethanol concentration,ratio of solid and liquid,extraction temperature and extraction time on the extraction rate was studied through the Single Factor Experiment.The conditions of extraction process were optimized by the orthogonal experiment on the basis of above results.The purification of crude polyphenols of areca nut:Macroporous Resin Adsorption was adopted to purify the polyphenols of areca nut.The types of macroporous resins suitable for purification of areca nut polyphenols were determined through static adsorption and analytical experiments.The dynamic adsorption experiments were used to examine the loading concentration of macroporous resins to areca nut polyphenols.At the same time,the effect of the concentration of the analytical solution on the resolution of the macroporous resin was studied through dynamic analytical experiments to determine the optimal process conditions for dynamic adsorption and analysis.Anti-hypoxic effect of areca nut polyphenols:A hypoxic model of Wistar rats was established on the rapid advance plateau with positive control group and the areca nut polyphenol dose group（400mg/kg,800mg/kg,1600mg/kg）.The experimental content includes the measurement of blood gas indicators of arterial blood in rats,the observation of pathological changes of liver tissue,lung tissue,brain tissue and myocardial tissue,and determination of oxidative stress indicators in various tissues in order to study the protective effect of different polyphenol doses on hypoxia in rats.Preparation technology of areca nut polyphenol capsules:In the preparation process,the angle of repose and moisture absorption are used as indicators for selection,and the amount of filler and auxiliary materials is selected.Select the appropriate wetting agent according to the granulation situation,particle yield and molding rate to determine the capsule volume.The relevant tests of the content,disintegration time and moisture of betel nut polyphenol capsules were made to determine the best preparation process.Results The extraction of polyphenols from areca nut and the optimized of the conditions:The ferrous tartrate method has good accuracy and precision for the determination of areca nut polyphenols.It has the advantages of good repeatability,convenient operation and high sensitivity,which is suitable for the determination of polyphenols content.The result showed that the optimum conditions of extraction were temperature 65℃,ratio of solid and liquid1:30（g/mL）and concentration of ethanol 80%for 2h with the extraction rate of 33.25%.The purification of crude polyphenols of areca nut:The experimental results show that AB-8 macroporous resin has good adsorption and desorption properties for phenol compounds,and is suitable for purification of areca polyphenols.The suitable conditions for the purification of phenols from AB-8 macroporous resin are as follows:the loading concentration is about 40mg/mL,and after the resin adsorption reaches saturation,using concentration of ethanol 95%as the eluent,the extraction rate of Areca polyphenols is increased to 45.03%.Anti-hypoxic effect of areca nut polyphenols:The results of the hypoxia experiment on the rapid plateau showed that the blood oxygen saturation SatO2 of the areca nut low,medium and high dose groups increased by 3.14%,4.51%,and 5.69%（P<0.05）.The content of SOD and GSH increased significantly（P<0.05）,and the content of MDA decreased significantly（P<0.05）.Preparation technology of areca nut polyphenol capsules:The best molding process is:the main medicine and microcrystalline cellulose are mixed at a ratio of 1:1,70%ethanol is used to make a soft material,granulated through a 24 mesh sieve,dried for 4 hours,and filled into No.0 capsule after granulation.The angle of repose of the particles is 29.05°,the moisture absorption is 11.38%,and the bulk density is 0.5717g/mL.The obtained molding process is reasonable and feasible.Conclusion Through optimization and purification of areca nut polyphenol extraction process,the extraction rate of polyphenol increased from 33.25% to 45.03%.The preferred process is stable and reliable,has good reproducibility,and has practicality and operability.In pharmacodynamic experiments,the polyphenols of areca nut（400mg/kg,800mg/kg）improved hypoxemia in hypoxic rats,increased tissue antioxidant enzyme activity,relieved lipid peroxidation damage,and had significant resistance to plateau Hypoxia.Areca nut polyphenol preparation was prepared to developed as a new anti-hypoxia drug on the basis of purification,which has very important significance in combating the development of plateau hypoxia resources. |
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