牛蒡低聚果糖的羧甲基化修饰及对RAW264.7细胞免疫活性提升  

Carboxymethylation modification of fructooligosaccharide from burdock and its enhancement on immune activity of RAW264.7 cells

作　　者：赵志刚 金丹 黄慧丹 李贵文 单振伟 ZHAO Zhigang;JIN Dan;HUANG Huidan;LI Guiwen;SHAN Zhenwei(School of Pharmacy,Wannan Medical College,Wuhu 241002,Anhui,China;Anhui Provincial Engineering Research Center for Polysaccharide Drugs,Wuhu 241002,Anhui,China)

机构地区：[1]皖南医学院药学院,安徽芜湖241002 [2]安徽省多糖药物工程技术研究中心,安徽芜湖241002

出　　处：《精细化工》2024年第7期1542-1549,1624,共9页Fine Chemicals

基　　金：国家自然科学青年基金资助项目(82204238);安徽省大学生创新训练项目(S202210368067、S202210368073);皖南医学院重点项目科研基金(WK2021Z03)。

摘　　要：采用水媒法对牛蒡低聚果糖(BFO)进行了羧甲基化修饰,制备了不同羧甲基化取代度(DS)的修饰产物(CM-BFO),通过单因素和响应面实验考察了反应条件对DS的影响,以及各因素的交互作用。采用GC、FTIR、^(1)HNMR、^(13)CNMR对BFO及CM-BFO的结构进行了表征,以RAW264.7细胞为模型测试了不同DS的CM-BFO体外细胞实验的免疫活性。结果表明,0.5 g BFO用30 mL质量分数为20%的Na OH水溶液碱化处理1h,以30 mL质量分数为2.5%的氯乙酸水溶液为羧甲基化试剂,在反应温度65℃、反应时间3.3 h的最佳条件下,BFO的DS最高,为1.06。BFO和CM-BFO均能提升RAW264.7细胞的免疫活性,3种DS为0.82、0.93和0.96的CM-BFO免疫活性较BFO明显提升,而低DS(0.73)和高DS(1.06)的CM-BFO则影响较小。CM-BFO的免疫活性在一定范围内随其DS的升高呈先增强后减弱的趋势。Carboxymethylated burdock fructooligosaccharide(CM-BFO) with different degrees of carboxymethyl substitution(DS) was prepared by aqueous method.BFO and CM-BFO characterized by GC,FTIR,^(1)HNMR,and ^(13)CNMR.The effect of reaction conditions on DS and the interaction between each factor were analyzed by single factor and response surface experiments,while the influence of CM-BFO with different DS on cell immunity were further evaluated using RAW264.7 cells as model in vitro.The results showed that,under the optimal conditions of 0.5 g BFO alkalized in 30 m L mass fraction of 20%NaOH aqueous solution for 1 h,using 30 mL mass fraction of 2.5% chloroacetic acid aqueous solution as carboxymethylation reagent,reaction temperature 65 ℃,and reaction time 3.3 h,BFO obtained the highest DS degree of 1.06.Moreover,both BFO and CM-BFO exhibited enhancement on the immune activity of RAW264.7 cells in vitro.CM-BFO with DS of 0.82,0.93 and 0.96 displayed significant improvement on the cell immune activity compared with BFO,while the ones with low DS(0.73) and high DS(1.06) only showed slight enhancement.The immune activity increased first and then decreased with the increase of DS in a certain range.

关 键 词：牛蒡低聚果糖 羧甲基化修饰 取代度 免疫活性 中药现代化技术 

分 类 号：R285[医药卫生—中药学]