Extraction Polysaccharides Of Longan Shell And Preliminary Activity Research

Objective: To extract and purify polysaccharides from longan shell for chemical analysis and preliminary immune activity and anti-tumor activity.Method: Orthogonal experiment to determine the optimal extraction conditions of longan shell and extract polysaccharides from longan shell. The polysaccharides content was determined by phenol-sulfuric acid method. And polysaccharides of longan shell was purified by deproteinizing,decolouring,DEAE-cellulose column and Sephacryl S-300 HR chromatographies. The purity of LP1-2A were detected by column chromatography , cellulose acetate membrane electrophoresis and specific rotation. Monosaccharide component in LP1-2A was determined by complete acid hydrolysis. The molecular weight of LP1-2A was determined by GPC.Lymphocyte proliferation and ConA-induced lymphocyte proliferation were detected by MTT,secretion of IL-2 and ConA-induced secretion of IL-2 were detected by ELISA and NK cell activity were detected by LDH release in vitro immune activity experiments.The LP1-2A or LP1-2B was given by gavage. Lymphocyte proliferation and ConA-induced lymphocyte proliferation were detected by MTT,secretion of IL-2 and ConA-induced secretion of IL-2 were detected by ELISA and NK cell activity were detected by LDH release in vivo immune activity experiments.The inhibitory rate of tumor cells Bel-7404,SKVO3,MCF-7 with LP1-2A and LP1-2B were detected by MTT in the vitro anti-tumor experiments.The mice inoculated in abdominal cavity with S180 cell were treated with LP1-2A or LP1-2B respectively by gavage and calculate the life-prolongation rate in the vitro anti-tumor experiments.Results: The optimal condition of longyan shell polysacchrides of hot-water extraction method were respectively temperature of extracttion was 90℃,extracttion ratio between waster and material was 1:30,extraction frequency was 3 times and extraction time taken 3.5h. The yield of crude polysaccharides was 1.31% by above conditions of extraction method and polysaccharides content of the crude polysaccharides was 27.33%. IR spectrum of LP1-2A indicated that the LP1-2A was pyranose with the glycoside conFigureuration ofα-D-hemiacetal hydroxyl. The molecular weight of LP1-2A was 1.14×104u. The monosaccharide component of LP1-2A was glucose only.The vitro immune activity results of the LP1-2A and the LP1-2B showed that the stimulate index and the 300～4.7mg/L concentration range was dose-dependence in lymphocyte proliferation and ConA-induced lymphocyte proliferation experiments. The IL-2 levels and the 300～4.7mg/L concentration range was dose-dependence in secretion of IL-2 and ConA-induced secretion of IL-2 experiments. NK cells acivity and the 300～4.7mg/L concentration range was dose-dependence.The vivo immune activity results of the LP1-2A and the LP1-2B showed that the stimulate index and the 250～62.5mg/L concentration range was dose-dependence in lymphocyte proliferation and ConA-induced lymphocyte proliferation experiments. The IL-2 levels and the 250～62.5mg/L concentration range was dose-dependence in secretion of IL-2 and ConA-induced secretion of IL-2 experiments. NK cells acivity and the 250～62.5mg/L concentration range was dose-dependence.The vitro anti-tumor experiments results of LP1-2A and LP1-2B showed that the inhibitory rate on liver cancer cell Bel-7404,ovarian cancer cell SKVO3,breast cancer cell MCF-7 and the 400～50mg/L concentration range was dose-dependence.The vivo anti-tumor experiments results of LP1-2A and LP1-2B showed that the life-prolongation rate of S180 mice and the 320～80mg/L concentration range was dose-dependence.Conclusion: LP1-2A is pyranose with the glycoside configuration ofα-D-hemiacetal hydroxyl and its molecular weight is 1.14×104u. The monosaccharide component of LP1-2A is glucose only. LP1-2A and LP1-2B can enhance the immune system functions. LP1-2A and LP1-2B can inhibit the growth of tumor cell of Bel-7404,SKVO3 and MCF-7. LP1-2A and LP1-2B can prolong the survival time of S180 mice.