The Preparation And Antitumor Activity Of Polysaccharide From Pholiota Adiposa

Polysaccharide is the main active ingredient of Pholiota Adiposa,which has many biological activitiessuch as anti-oxidation,anti-tumor,lowering blood lipids,antibacterial,immune regulation etc.Pholiota Adiposa was chosen as the experimental raw materials,because of the few research on the anti-tumor in vitro of polysaccharide from it.This paper studied the effects of different solvent pH and protein removal of the anti-tumor activity of polysaccharide from Pholiota adiposa in vitro and the apoptosis of tumor cells.The main conclusions are as follows:This paper studied the effects of different solvent pH and protein removal on the yield of polysaccharides from Pholiota Adiposa,the normal somatic cells and the tumor cells.The results showed that the yield of polysaccharides from Pholiota Adiposa prepared by 0.lmol/L sodium hydroxide solution(11.02%)was significantly higher than that prepared by 0.1mol/L hydrochloric acid solution(9.26%)and water(9.08%).In addition,the deproteinization process can significantly improve the purity of polysaccharide from Pholiota Adiposa,the purity of the deproteinized acid-soluble polysaccharide from Pholiota Adiposa was increased by 73.91%,followed by deproteinized alkali-soluble polysaccharide from Pholiota Adiposa(66.23%),and the lowest was deproteinized water-soluble polysaccharide from Pholiota Adiposa(45.11%).The results showed that the toxicity of acid-soluble polysaccharide from Pholiota Adiposa was the weakest on the cervical cancer cells,colon cancer cells,and hepatocellular carcinoma cells,followed by water-soluble polysaccharide from Pholiota Adiposa,the toxicity of polysaccharide from Pholiota Adiposa were significantly reduced after deproteinization.The results showed that the inhibitory effect of the deproteinized acid-soluble polysaccharide from Pholiota Adiposa on the proliferation of the cervical cancer cells,colon cancer cells,and hepatocellular carcinoma cells was significantly higher than that of the non-deproteinized acid-soluble polysaccharide from Pholiota Adiposa,non-deproteinized and deproteinized alkali-soluble and water-soluble polysaccharide from Pholiota Adiposa,which inhibitory effect on the proliferation of cervical cancer cells was 38.1±1.75%when the concentration was 1mg/mL,considering the safe concentration of normal somatic cells.This paper infrared spectroscopy analysis the six kinds of polysaccharide from Pholiota Adiposa.The results showed that deproteinized acid-soluble polysaccharide from Pholiota Adiposa has the strongest absorption peak in 1078±4cm-1 and 759.83cm-1.The results showed that the content of C-O-H or C-O-C and epoxy group was higher than other five kinds of polysaccharide from Pholiota Adiposa.It could speculated that the inhibitory effect of the polysaccharide from Pholiota Adiposa on the proliferation of human tumor cells may berelated to the functional group.Selected the deproteinized polysaccharide extracted by acid-soluble and cervical cancer cellsfor subsequent research.This paper purified two fractions named SPAP2-1 and SPAP2-2 by applying DEAE-cellulose column chromatography on deproteinized polysaccharides extracted by acid soluble from Pholiota adiposa.The yield of two polysaccharides were 62.41%and 3.82%.This paper investigated the toxicity of SPAP2-2 and SPAP2-1 on three kinds of normal somatic cells to hepatocytes,myoblasts and fibroblasts.It showedthatthe toxicity of SPAP2-1 on three normal somatic cells was weaker than SPAP2-2;This paper investigatedthe inhibitory effect of SPAP2-1 and SPAP2-2 on the proliferation of cervical cancer cells.On the premise of taking into account the safety concentration of normal somatic cells,it showed that the inhibitory effect of SPAP2-1 on the proliferation of cervical cancer cells was significantly higher than SPAP2-2 and it achieved best for 73.52± 1.36%in 4mg/mL concentration and 48h actuation duration.Selected SPAP2-1 for subsequent research.By investigating the effect of SPAP2-1 on the growth cycle of cervical cancer cell using PI single staining method founded that SPAP2-1 could effectively block the cervical cancer cells in G1 phase.Through investigating the inhibitory effect of SPAP2-1 on cervical cancer cells using Annexin V-FITC and PI double staining found that SPAP2-1 could effectively induce apoptosis of cervical cancer cells.Pholiota Adiposa is widely distributed and rich in resources.Its main active substance polysaccharide has good antitumor activity,which provides a theoretical basis for the further development and utilization of Pholiota Adiposa resources.