| As people working pressure increased, there are more and more patients withcancer, which present a younger trend. Now cancer has become the second killer thathazard to human health after cardiovascular disease.Current treatment of malignanttumors is still given priority to with chemotherapy,radiotherapy,surgical. Anticancerdrugs in clinical all have different degrees of side effects,such as chemotherapydrugs can kill tumor cells,and destroy the normal tissue cells, undermine the body’simmune system at the same time.It has become a popular development ofanti–cancer research at home and abroad that to seek antitumor active ingredient ofefficacy and less side effects from the traditional Chinese medicine,especially plantresources.Long–term animal and clinical experiments showed that Inonotus obliquuspolysaccharide has no obvious side effects and has been shown to have anti–tumoreffect.Based on the extraction, deproteinization and decolorization methods ofInonotus obliguus polysaccharide, monosaccharide composition and anti-tumoractivity were studied.Firstly,the experiment extracted Inonotus obliquus polysaccharide,and theoptimal conditions for extraction by orthogonal array were as follows:solid/waterratio was1:40(W/V),the extracting time was2.5h,the extracting temperature was90℃.Under this condition, the repeated experiment derived polysaccharide averageextraction rate of1.82%,good reproducibility,the purity of polysaccharide reached21.1%.Secondly,the deproteinization and decolorization methods of Inonotus obliguuspolysaccharide were studied.To investigate the depmteinization methods fromInonotus obliguus polysaccharide,the experiment compared the deproteinization of Sevage, trichloroacetic acid (TCA), trichloroacetic acid–n–Butanol(TCA–NBA),hydrochloric acid and enzymatic auxiliary method according topolysaccharide retention rate and deproteinization rate.The results showed that theeffectiveness of enzymatic auxiliary TCA–NBA method was the best for Inonotusobliguus polysaccharide with the deproteinization rate of82.3%and a higherpolysaccharide retention rate of76.1%,the purity of polysaccharide reached81.1%.Decolorization effect of active carbon powder,H2O2,chitosan,and polyamidechromatography was researched according to decolorization rate and polysaccharideretention rate. The results showed that the effectiveness of polyamidechromatography was the best for Inonotus obliguus polysaccharide. Thedecolorization rate was89.3%, and the polysaccharide retention rate was91.7%, thepurity of polysaccharide reached85.8%.Thirdly,the monosaccharide composition of Inonotus obliguus polysaccharidewas studied.High performance liquid chromatographic (HPLC) method ofprecolumn–derivatization with1–phenyl–3–methyl–5–pyrazolone (PMP) were usedfor detection of the monosaccharide composition. The result showed thatInonotus obliguus polysaccharide was composed of mannose,rhamnose,glucose,galactose,xylose,arabinose,whose mole ratio was2.13：1.36：7.01：2.98：1：1.78.An unknown peak that m/z was164by HPLC–MS analysis was found in thechromatogram. With a certain amount of fucose,the peak area of polysaccharidehydrolyzed sample has increased dramatically in the corresponding time of unknowncomposition by HPLC method,the unknown composition was proved to be fucose.Finally,the inhibition to the proliferation of human cervical cancercell(Hela),mouse myeloma cells(SP2) and human Chang liver cells(Chang) wasstudied by MTT method.Inonotus obliguus polysaccharide can inhibit theproliferation of Hela,SP2and Chang cells with growth inhibition rate from4.78%to4.78%,from2.89%to44.90%,from3.56%to22.56%with aconcentration–dependent relation,when its concentration was in the range of10～1000μg/mL,and there were significant differences between each concentration(p<0.01).The anti–tumor activity of Inonotus obliguus polysaccharide was significantcompared with5–fluorouracil,and was anti–tumor substances with a developmentprospect. |
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