通关藤多糖的提取工艺及免疫调节作用研究

目的:优化通关藤多糖的提取工艺,测定通关藤多糖的免疫活性。方法:采用正交试验,以粗多糖得率和总含糖量为指标考察通关藤多糖的最佳提取工艺;采用环磷酰胺腹腔注射复制免疫低下小鼠模型,通过ELISA法测定血清中IL-2,IL-6的含量和流式细胞术检测小鼠外周血中CD4+,CD8+表达水平等,考察最佳提取工艺得到的多糖对机体免疫功能的影响。结果:以粗多糖得率为指标时各因素影响主次顺序为煎煮次数>煎煮温度>煎煮时间>加水量;以总含糖量为指标时各因素影响主次顺序为煎煮温度>煎煮次数>加水量>煎煮时间。与模型组比较,通关藤多糖高、中剂量组脾脏指数均有显著升高,血清中IL-2,IL-6含量显著升高,CD4+/CD8+比值显著升高(P<0.05),结论:最终确定最佳提取条件为加8倍量水,煎煮3次,煎煮温度100℃,煎煮时间为1.5 h;通关藤多糖具有增强细胞免疫和体液免疫的作用。

Study on extraction and immune regulation of polysaccharides from Marsdenia tenacissima

Objective:To optimize the extraction technology and determine the immune activity of polysaccharides from Marsdenia tenacissima.Method:The optimum extraction technology of polysaccharide from M.tenacissima was detected by orthogonal experiments with the extraction rate of polysacchride and the total sugar content as indicators.The immunocompromised mice model was established by intraperitoneal injection cyclophosphamide to detected the content of IL-2,IL-6 in serum,CD4+,CD8+ in the peripheral blood by ELISA and flow cytometry,respectively.Result:By the extraction rate of polysaccharide,the sequence of seriousness of all affecting factors from high to low was extracting times,temperature,heating time and water ratio.By the total sugar content,the sequence was temperature,extracting times,water ratio and heating time.Compared with the model group,the pleen index,IL-2,IL-6,CD4+/CD8+ were increased significantly(P<0.05) in the 0.14 g·kg-1 group and 0.28 g·kg-1 group.Conclusion:The optimum extraction condition was as follows:extraction three times/1.5 hours at 100 ℃ with 1∶8 ratio of M.tenacissima to water.The polysaccharide of M.tenacissima can enhance the cellular immune and humoral immune.