| Codonopsis pillosula is a joint name of the roots of perennial herbs in Campanulaceae. Codonopsis pillosula is a traditional chinese rein-forcing medicine which benefits qi, strengthens spleen and tonifies lung. Codonopsis pillosula has many effective constituents such as polysaccharides, phenols, sterols, saponins, etc. It has been reported that polysaccharides which is one of major component of Codonopsis pillosula have many bioactivities including immuno-enhancement, anti-tumor, antioxidant, hypolipidemic, etc. However, most of the samples applied to the study of pharmacological activity are crude polysaccharides. There is still a lack of understanding of the components and structures of all the Codonopsis pillosula polysaccharides. To solve this problem, we carried out the research on the totally systemic fraction of Codonopsis pillosula polysaccharides in this paper. Fistly, the water-soluble polysaccharides were extracted by hot water and precipitated by ethanol. Then, the polysaccharides were gradually fractionated and purified by ion-exchange and gel permeation chromatography to obtain four fractions. Further, the homogeneity, molecular weight and sugar composition were determinated. The research results are as follows:The water-soluble Codonopsis pillosula polysaccharides, refered to as CPP, were extracted by hot water, precipitated by80%ethanol and deproteinated by the Sevage method. The yield of CPP is7.84%. CPP contains89.1%total carbohydrates,16.2%uronic acid and0.6%protein. Sugar composition by HPLC showed that CPP consisted of rhamnose (3.1%), glacturonicacid (38.8%), arabinose (16.6%), galactose (25.0%), glucose (13.1%) and mannose (2.9%).CPP was fractionated on DEAE-Cellulose ion-exchange column into three fractions by eluted with water,0.2M NaCl and0.3M NaCl respectively. CPP-1is a neutral fraction. CPP-2, CPP-3are acidic fractions. According to the molecular weight distribution, the acidic fractions CPP-2and CPP-3were further separated on Sepharose CL-6B into four fractions CPP-2-1, CPP-2-2, CPP-3-1, CPP-3-2. The molecular weight distribution of the four fractions on Sepharose CL-6B showed that CPP-2-1, CPP-2-2and CPP-3-2are homogenous with molecular weight of1.7×105Da,6.0×105Da and1.4×105Da respectively. Based on the results that all these four fractions are mainly composed of rhamnose, galacturonic acid, arabinose, galactose., We speculated that they all contain rhamnogalacturonan (RG) domins and homogalacturoman (HG) domains. CPP-3-1contains67.2%AG domins while CPP-2-2is homogenous homogalacturomans with high amount of galacturonic acid (81.3%). Summarizing the results from our experiments, we preliminary conclude that the fraction CPP-2-2is HG pectin and the fraction CPP-3-1is AG pectin while both CPP-2-1and CPP-3-2are RG-I pectin.Tumor necrocis factor (TNF-)-related apoptosis-inducing ligand (TRAIL), is a potent cancer cell-specific apoptosis-inducing agent with little to no effect on normal tissues. However, the potential application of TRAIL in cancer therapy is limited, as many human tumors, are partially or completely resistant to apoptotic effects induced by TRAIL. Therefore, combination TRAIL with other agents to overcome the low sensitivity and resistance of cancer cells to TRAIL has been a promising strategy to potentiate the therapeutic applications of TRAIL. In this research, the synergistic effect of CPP, CPP-2, CPP-2-1, CPP-2-2, CPP-3, CPP-3-1and CPP-3-2with TRAIL were investigated. The results showed that the crude polysaccharide CPP and TRAIL had significant synergistic effect in suppressing the proliferation of human colon cancer HCT116cells. The synergistic of TRAIL with CPP-2, CPP-2-1, CPP-2-2, CPP-3, CPP-3-1and CPP-3-2were moderate. All these results suggest that the polysaccharides might have synergistic effect themselves in synergizing with TRAIL... |
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