北苍术多糖的酶法辅助超声提取工艺的优化及其体外抗肿瘤活性研究

目的通过生物酶解技术优化北苍术多糖的酶辅助超声提取方法,并考察其体外抗肿瘤活性。方法选用纤维素酶和果胶酶水解北苍术药材,酶解后采用超声法提取北苍术药材中多糖。以多糖得率为指标,在单因素试验的基础上采用4个因素（酶用量、酶解时间、酶解pH值、酶解温度）3个水平L9（34）正交设计法对酶解条件进行优选,并选用苯酚–浓硫酸法测定多糖,采用MTS、台盼蓝法对北苍术粗多糖体外抗肿瘤活性进行初探。结果最佳酶解条件为酶用量为药材1.2%、在60℃、pH 5条件下酶解50 min,再以超声提取40 min,北苍术多糖的得率为32.29%,所提粗多糖对HeLa细胞增殖的抑制率为64.42%。结论酶解辅助超声提取北苍术中多糖简便易行,可以提高多糖得率,多糖活性较好。

Optimization of enzymatic auxiliary ultrasonic extraction of polysaccharide from Atractylodes chinensis and its antitumor activity in vitro

Objective To optimize the enzymatic auxiliary ultrasonic extraction of polysaccharide from Atractylodes chinensis by biological enzyme technology and to study its antitumor activity in vitro. Methods Cellulase and pectinase were used for hydrolysis and then ultrasonic extracting method was chosen to extract the polysaccharide from roots of A. chinensis. The yield of polysaccharide purity from crude polysaccharide was taken as indicators. Based on single-factor test, the ultrasonic-assisted extracting technology of A. chinensis polysaccharide was optimized by L9（34） orthogonal test, and four factors included enzymolysis temperature, enzyme content, enzymolysis pH value, and enzymolysis time with three levels. The phenol-sulphuric acid method was applied for the determination of polysaccharide, and then using MTS and trypan blue staining to study antitumor activity on the polysaccharides. Results The optimal enzymolysis extracting conditions of A. chinensis polysaccharide were as follows： temperature was 50 ℃, content was 1.2%, pH value was 5, and time was 50 min. Under the conditions, the purity of polysaccharide was 32.29%, and inhibitory rate of polysaccharide on the proliferation of HeLa cells was 64.42%. Conclusion Enzymatic auxiliary for ultrasonic extraction process has the advantages of short time, simple operation, and higher extract rate, which could provide the reference for development and utilization of A. chinensis.