目的 優(yōu)化細柱五加莖多糖的閃式提取工藝，并研究其細胞毒性和免疫活性。方法 運用球面對稱設計試驗，綜合考慮料液比、提取溫度、提取時間3因素，優(yōu)化細柱五加莖多糖的閃式提取工藝；對閃式提取法與回流、超聲2種傳統(tǒng)提取方法進行比較。通過體外試驗對提取所得多糖的細胞毒性和免疫活性進行初步研究。結果 細柱五加莖多糖閃式提取最佳工藝為料液比1∶13.2，提取溫度80 ℃，提取時間420 s。在此條件下，細柱五加莖中多糖提取率為0.78%，且優(yōu)于回流和超聲法提取所得多糖提取率。體外活性實驗結果顯示，細柱五加莖多糖質量濃度在10～20 μg/mL對RAW 264.7細胞無明顯毒性，在10～40 μg/mL能夠促進RAW 264.7細胞NO的分泌。結論 建立了穩(wěn)定、簡便的細柱五加莖多糖的閃式提取工藝方法, 初步研究了其多糖的免疫活性，為合理的開發(fā)細柱五加資源提供理論依據(jù)。

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.

湖南省自然科學基金重點項目（11JJ2042）；湖南中醫(yī)藥大學藥物分析學“十二五”校級重點學科建設項目