Active Ingredient Of Black Bone Vine Polysaccharide

Marsdenia tenacissima (Roxb.)Wight et Arn.,a member of Asclepiadaceae family,is a kind of herb.Which is mainly produced in Yunnan, Guizhou, etc.It contained in the"South Yunnan Materia Medica"firstly. In this paper, we separated and purfied the raw Marsdenia tenacissima(Roxb.)Wight et Arn polysaccharide and obtained three homogeneous polysaccharide. we studied the structure and pharmacological action preliminarily. The main content of the papers are as follows:1. Use the method of single factors to determine the extraction of water after ertracted by alcohol between the four methods of flooding by water,extracting by water,flooding by water after alcohol extracted,extracting by water after alcohol extracted as an indicator of total polysaccharides.Orthogonal design method is used to optimize the extracting conditions.Considering the extraction efficiency and quality of polysaccharides, the final optimization extracting method is 8 times the amount of water, 100℃extracted 3 times,each time 2 hours.Orthogonal design method was used to optimize the alcohol precipitation conditions,the final optimization extracting method is added 6 times the volume of 95% ethanol ,then precipitated for 12 hours.2. Use deae-cellulose column to separated and decolor the purified polysaccharides, water and sodium chloride solution eluting,UV under 490 nm and phenol-sulfuric acid method detect component.Then three polysaccharides fractions were got:CMP-1,CMP-2,CMP-3.CMP-1,CMP-2,CMP-3 separated by sephadex G-100 column,collected every fraction; dialysis; then freeze drying again. We got the fine purification Marsdenia tenacissima Polysaccharides: CMP-â… ,CMP-â…¡,CMP-â…¢. UV spectrum and Superdex G-100 column chromatography showed that the fine purification Marsdenia tenacissima Polysaccharides(CMP-â… ,CMP-â…¡,CMP-â…¢) were homogeneous polysaccharides.IR showed that the composition of the three preliminary .3. Use colorimetry to determine the total polysaccharide content of Marsdenia tenacissima. The linear relationship was good, precision, stability repeatability and recovery were up to standard.This method is stable and feasible.Be scientific and objectibe to evaluate the quality of medicine. 4. Antitumor effect in vivo test results that the total polysaccharide content in different doses had inhibitory effort on S180 in mice.While the total polysaccharide in 200mg/kg had remarkable inhibitory effort on S180 in mice with the inhibitory ratio of 37.5% respectively.The experimental results show that the total polysaccharide content in high,medium and low dose all can regulate the immune function.High dose of the total polysaccharide content has the best effect.The thymus index was 2.79,and the spleen index was 5.16.5. Make a research of the total polysaccharide content and CMP-â… ,CMP-â…¡,CMP-â…¢on proliferation effect of HepG2 in vitro with MTT method.The results are as follows:the total polysaccharide content and CMP-â… ,CMP-â…¡,CMP-â…¢all inhibite the growth of hepatoma cells. The total polysaccharide content in the range of 0.25~0.45mg/ml,CMP-â… in the range of 0.175~0.225mg/ml and CMP-â…¡å'ŒCMP-â…¢in the range of 0.125~0.225mg/ml,showed good dose-effect relationship.Summing up the above,a method of determined the total polysaccharide content of Marsdenia tenacissima which established in this paper can control the quality of medicines.The total polysaccharide content of Marsdenia tenacissima and CMP-â… ,CMP-â…¡,CMP-â…¢inhibit the tumer growth.And the total polysaccharide content can regulate the immune function.