Studies On The Anti-tumor Activity Of The Ganoderma Lucidum Polysaccharide Sulfate

In this paper, a more homogeneous composition of Ganoderma lucidum polysaccharides was extracted and purified from the fruit bodies of Ganoderma lucidum, using chlorosulfonic acid and pyridine as the agent for sulfation modification on the polysaccharides, and then the in vitro anti-tumor activity of the Ganoderma lucidum polysaccharide sulfate was detected by MTT method.Based on the previous studies, the effects of water consumption (A), extraction time (B) and extraction times (C) on the extraction yields of Ganoderma lucidum polysaccharides from Ganoderma lucidum fruit body were optimized with orthogonal experiment, and the extraction rate and yield of dry extract as the indexes were used to optimize the extraction process. It was indicated that the optimum extraction parameters were A3B3C3 that is 20 times of water was used to extract 3 times at 90℃with 2 hours with each time.In order to get more single components of Ganoderma lucidum polysaccharides, BS-Ⅱanion resin was used for decolorization of Ganoderma lucidum polysaccharides, and excepting protein with Sevag method. After that, different concentrations of ethanol and Sephadex G100 were used for further purification. Finally, the GLP-2 with average molecular weight is 6.5×104 was obtained. TLC assay showed that GLP-2 was consisting of the mannose, and infrared spectroscopy (IR) displayed GLP-2 was beta configuration pyranoid polysaccharide.To study the pharmacological activity of the Ganoderma lucidum polysaccharide sulfate, we decided to synthesize GLPS with chlorosulfonic acid and pyridine. With degree of substitution of sulfur (DS) as an index, the impact of the molar ratio of chlorosulfonic acid to sugar unit, reaction temperature, reaction time, and the volume ratio of chlorosulfonic acid and pyridine on the sulfation modification of the polysaccharide were studied by orthogonal design. The optimal sulfation modification condition was determined as below:1.0 g GLP - 2 and 20ml chlorosulfonic acid dissolved in 60ml pyridine, stirring for 4h at 90℃. Meanwhile, the application of infrared scanning technique for structural analysis of GLPS-2 confirmed that the sugar chain had been successfully introduced sulfate groups.In accordance with the optimum conditions, GLPS-2 was synthesized and its degree of substitution of sulfur was 1.36.And then the anti-tumor activity of GLPS-2 was tested in vitro by MTT method. The results showed that the inhibition of GLPS-2 to CaCo-2 cell was significantly higher than the GLP, when the drug concentration was 800μg/ml, the inhibition rate reached 66.87%. And this inhibitory effect showed a certain dose-effect relationship, that is, the higher the concentration, the stronger the inhibition. But the inhibitory effect of GLPS-2 to SMMC-7721 and MGC-803 cell was not obvious. This shows that there was certain selectivity when GLPS-2 effected on cancer cells.