The Extraction And Separation Of The Silkworm Chrysalis Polysaccharide And Research On The Effects Of Proliferation Of HeLa

ObjectiveCervical cancer is one of the most serious diseases threatening the healthof women, in the worldr, The disease to increase at the rate of500000about ayear.The study found that silkworm chrysalis polysaccharide can obviouslyenhance the immunity of the mice, the aim of this experimental study is theoptimization of silkworm chrysalis polysaccharide extraction process and thestudy of silkworm chrysalis polysaccharide decoloring conditions, purification ofsilkworm chrysalis polysaccharide and research influence on HeLa proliferation,In order to prevent and treat of cervical cancer to provide an experimental basisfor new drug research.MethodsThis study with the traditional process method and microwave extractionmethod to extract silkworm chrysalis polysaccharide, the traditional processmethod to extract silkworm chrysalis polysaccharide relate to the extractiontemperature, extraction time, ratio of feed liquid three factors; Microwaveextraction to extract silkworm chrysalis polysaccharide relate to the microwavepower, microwave time, and solid-liquid ratio three factors, Using orthogonalexperiment in two ways respectively, calculate the polysaccharide yield, Theyield indicators to silkworm chrysalis polysaccharide, Optimize the extractionprocess of silkworm chrysalis polysaccharide.On the basis of the comparison,this research through the effect of differentresin decolorization of silkworm chrysalis polysaccharide, The static adsorptionand dynamic adsorption experiments were used to decolorizing of polysaccharide respectively. Static adsorption experiments used orthogonal,Study the best conditions of static adsorption method. Then on the basis ofstatic adsorption experiment carried out dynamic experiment, Respectively toinvestigate on the column of fluid velocity, pH and temperature on the elutioneffect,Research silkworm chrysalis polysaccharide suitable bleachingconditions.This study do the dialysis treatment to silkworm chrysalis polysaccharide ofdecoloration and deproteinization, intercept silkworm chrysalis polysaccharidemolecular weight greater than5000, let after dialysis treatment of silkwormchrysalis polysaccharide made DEAE-52and SephadexG-150columnchromatography experiment, further on separation and purification of silkwormchrysalis polysaccharide.This study will separation and purification of silkworm chrysalisconcentration of polysaccharide PSP1is divided into four groups, The fourconcentration groups on HeLa respectively,determined by MTT test experimentwas done on HeLa,And under the fluorescence microscope observe themorphology of HeLa.ResultsRespectively with two kinds of extraction method optimum processconditions of extracting silkworm chrysalis polysaccharide, through comparingresearch, the traditional method of polysaccharide extraction of silkwormchrysalis average yield was4.05%, the average yield of microwave method toextract the silkworm chrysalis polysaccharide is4.49%, found that themicrowave method to extract yield of silkworm chrysalis polysaccharide wasobviously higher than that of traditional method.Through static adsorption orthogonal test results of research, selected thebest process conditions of HZ-803resin, pH4.0, temperature45℃. On the basisof the static adsorption experiments do the dynamic experiment, obtains theoptimum process conditions for the velocity1BV/h, pH4, temperature of35℃.Through to the dialysis treatment of silkworm chrysalispolysaccharide,further purified using DEAE-52, anion exchange columnchromatography respectively with distilled water and different concentrations of NaCl on phase gradient elution, isolated PSP1, PSP2, PSP3threecomponents.PSP1performed on SephadexG-150column chromatographyexperiments, It was found PSP1elution peak shape the basic symmetry, shouldbelong to the homogeneity of the components.Experiments found that silkworm chrysalis polysaccharide on theproliferation of HeLa has certain inhibition, and In a certain range in atime-dependent doses.ConclusionsThrough the optimum process conditions of two extraction methodspolysaccharide extraction of silkworm chrysalis, found that the microwavemethod to extract yield of silkworm chrysalis polysaccharide was obviouslyhigher than that of traditional method. Respectively by using the static methodand dynamic method decolorization rate of silkworm chrysalis polysaccharide,polysaccharide retention rate, protein removal rate are measured, found that thedynamic method for determining the decolorization rate of silkworm chrysalis ofpolysaccharide, polysaccharide retention a static method is higher than staticmethod, protein removal rate lower than static method,so the later of silkwormchrysalis polysaccharide can consider the two methods used in experimentalstudy. Through to the silkworm chrysalis polysaccharide further purification, thestudy found that the isolated PSP1belongs to homogenous composition, can beused in the next cell experiment. Through the cell experiment found thatsilkworm chrysalis polysaccharide on the proliferation of Hela has certaininhibition, and In a certain range in a time-dependent doses.