[关键词]
[摘要]
目的 优化细柱五加茎多糖的闪式提取工艺,并研究其细胞毒性和免疫活性。方法 运用球面对称设计试验,综合考虑料液比、提取温度、提取时间3因素,优化细柱五加茎多糖的闪式提取工艺;对闪式提取法与回流、超声2种传统提取方法进行比较。通过体外试验对提取所得多糖的细胞毒性和免疫活性进行初步研究。结果 细柱五加茎多糖闪式提取最佳工艺为料液比1∶13.2,提取温度80 ℃,提取时间420 s。在此条件下,细柱五加茎中多糖提取率为0.78%,且优于回流和超声法提取所得多糖提取率。体外活性实验结果显示,细柱五加茎多糖质量浓度在10~20 μg/mL对RAW 264.7细胞无明显毒性,在10~40 μg/mL能够促进RAW 264.7细胞NO的分泌。结论 建立了稳定、简便的细柱五加茎多糖的闪式提取工艺方法, 初步研究了其多糖的免疫活性,为合理的开发细柱五加资源提供理论依据。
[Key word]
[Abstract]
Objective To optimize the extraction process for polysaccharides from the stems of Acanthopanax gracilistylus by smashing tissue extraction (STE) and to further investigate their cytotoxicity and immunological activities. Methods Extraction process for the polysaccharides from the stems of A. gracilistylus was optimized by using the spherical symmetrical design test, and three factors were considered (material-liquid ratio, extraction temperature, and extraction time). The test compared STE with two traditional extraction methods, reflux extraction and ultrasonic extraction. The bioassay tests of cytotoxicity and immunological activities on polysaccharides extracted were studied in vitro. Results The optimal extraction conditions for polysaccharides from the stems of A. gracilistylus were as follows: material-liquid ratio was 1∶13.2, the extraction temperature was 80 ℃, and the extraction time was 420 s. With the best extraction conditions, the yield of polysaccharides from the stems of A. gracilistylus was 0.78%. The yield by STE was higher than those by reflux extraction and ultrasonic extraction in this research. In addition, the bioassay results showed the polysaccharides had no significant toxicity on RAW 264.7 cells at the dose of 10-20 μg/mL and facilitated the secretion of nitric oxide (NO) in the cells supernatant at the dose of 10-40 μg/mL in vitro. Conclusion These results establish a steady and convenient extraction technology for polysaccharides from the stems of A. gracilistylus using STE method, indicating the polysaccharides have immunological activity to some extent, which provides a theoretical basis for the reasonable development of A. gracilistylus resources.
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[基金项目]
湖南省自然科学基金重点项目(11JJ2042);湖南中医药大学药物分析学“十二五”校级重点学科建设项目