Studies On Sulfation Of Polysaccharide From Arillus Longan And Antitumor Activities Of The Polysaccharide And Its Sulfated Derivative In Vitro

Objective: To sulfat the purified polysaccharide from Arillus Longan and investigate the antitumor activities of the primitive polysaccharide and its sulfated derivative in vitro.Methods:?The polysaccharide was extracted with hot water from Arillus longan. The total sugar content of crud polysaccharide was determined by using the phenol-sulfuric acid method. The polysaccharide was purified by the deproteinizing of enzyme-sevag method and the decolouring by H2O2.?The polysaccharide was further purified by DEAE-cellulose laminar analysis. The purity of polysaccharide-A was detected by testing with cellulose acetate membrance electrophoresis and polarimetry. The polysaccharide-A was sulfated with concentrated sulfuric acid. The content of LYRP4 was investigated by opacity analysis of permanent white sulfate precipitation. Reagent proportion, reaction temperature and reaction time were selected with substitute degree and yield as index by L9(34) orthogonal test. The LYRP4 was further purified by DEAE-cellulose so as to gain LYRP4^-1. Both the molecuar weights of polysaccharide-A and LYRP4-1 were respectively determined through GPC. The antitumor activities of polysaccharide-A and LYRP4-1 were assayed by MTT method in vitro. The apoptotic cells were observed by using the Hematoxylin-eosin staining and FCM.Results: The yield of hot-water extraction method was 6.30% and the total sugar content of Arillus Longan was 58.72%. The deproteinate rate of polysaccharide from Arillus Longan was 52.30%. The yield of polysaccharide-A by DEAE-cellulose laminar analysis was 64.13%, the yield of polysaccharide-B was 4.85% and polysaccharide-C was 15.43%. The polysaccharide-A was a homogeneous polysaccharide. When the reaction time was 3h, the reaction temperature was 10℃, and the reagent ratio between butyl alcolho and concentrated sulfuric acid was 3:1, the concentrated sulfuric acid esterification could get a high degree from substitution of sulfation which was 2.03. The pH of LYRP4 was 4.63. The LYRP4-1's growth inhibition ratio of nasopharyngeal carcinoma HONE1 cells was gradually increasing in the concentration range of 5-80μg·ml^-1, the inhibition ratio could reach to 31.07% when its concentration was 80μg·ml^-1, while the polysaccharide-A's growth inhibition ratio of nasopharyngeal carcinoma HONE1 cells at the same concentration was 16.32%. Hematoxylin-eosin staining showed that nasopharyngeal carcinoma HONE1 cells appeared apoptosis phenomenon with the LYRP4-1 concentration elevating. The apoptosis rate in high dose group of LYRP4-1 was 31.98% and the polysaccharide-A's was 11.63%, detected by FCM.Conclusion: The polysaccharide-A can get the polysaccharide-A's sulfated derivative by using the concentrated sulfuric acid esterification. The LYRP4-1's growth inhibition ratio of nasopharyngeal carcinoma HONE1cells is higher than the polysaccharide-A's. The LYRP4-1 has apoptotic effect on the nasopharyngeal carcinoma HONE1 cells.