论文部分内容阅读
目的:制备分枝杆菌多糖并考察其对小鼠淋巴细胞增殖的影响。方法:用苏通培养基培养分枝杆菌,菌体脱脂后,用5种方法提取,比较多糖得率,并用正交试验优化超声提取工艺,再用稀碱提取残渣,粗多糖经Sevag法除蛋白、DEAE-Sepharose Fast Flow层析柱纯化后得到分枝杆菌多糖;用MTT法检测分枝杆菌多糖对小鼠脾淋巴细胞增殖的影响。结果:超声后热水提取多糖得率最高,超声提取的优化工艺是提取时间40 min,固液比为1∶150,超声功率600 W;单因素试验优化的Sevag法除蛋白条件为体积比1∶5,萃取时间为20 min,萃取6次;提取得到的4种多糖组分在6.25~50 mg.L-1范围内均能显著促进脾淋巴细胞的增殖。结论:用本实验工艺成功制备了分枝杆菌多糖,制备物具有刺激小鼠脾淋巴细胞增殖的作用。
Objective: To prepare Mycobacterium polysaccharides and investigate its effect on lymphocyte proliferation in mice. Methods: Mycelia were cultured with Sutong medium. The mycelia were degreased and extracted by five methods. The yield of polysaccharides was compared. The optimum extraction conditions were determined by orthogonal test. The residue was extracted with dilute alkali. The crude polysaccharides were separated by Sevag method Mycobacterium polysaccharide was purified by DEAE-Sepharose Fast Flow chromatography. The effect of mycobacterium polysaccharide on the proliferation of mouse splenic lymphocytes was detected by MTT assay. Results: The yield of polysaccharides extracted by hot water was the highest after ultrasound extraction. The optimum extraction conditions were extraction time 40 min, solid-liquid ratio 1: 150 and ultrasonic power 600 W. The single-factor experiments optimized Sevag method for protein removal : 5, the extraction time was 20 min and the extraction was performed 6 times. The four polysaccharides extracted could significantly promote the proliferation of splenic lymphocytes in the range of 6.25-50 mg.L-1. Conclusion: Mycobacterium polysaccharide was successfully prepared by this experiment. The preparation has the effect of stimulating the proliferation of mouse splenic lymphocytes.