潞党参中低聚糖的分离纯化及其抗炎活性研究  

Study on Separation,Purification and Anti-inflammatory Activity of Oligosaccharides from Lu Dangshen

作　　者：汤敏 陆姗姗[1,2] 谷巍 胡守芳[3] Tang Min;Lu Shanshan;Gu Wei;Hu Shoufang(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China;Nanjing University of Chinese Medicine Hanlin College,Taizhou 225300,China;Nanjing Pukou District Hospital of Traditional Chinese Medicine,Nanjing 211800,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京中医药大学翰林学院,江苏泰州225300 [3]南京市浦口区中医院,江苏南京211800

出　　处：《中国野生植物资源》2024年第7期9-16,共8页Chinese Wild Plant Resources

基　　金：国家中医药管理局道地药材生态种植及质量保障项目(012072003039-1);江苏省林业科技创新与推广项目(LYKJ-常州[2020]04);泰州市社会发展项目(SSF20220031,SSF20230025);大学生创新创业训练项目(202313981013Y,202313981008Y);江苏省高等学校自然科学面上项目(23KJD360003)。

摘　　要：目的:分析低聚糖的成分构成及其在潞党参中的抗炎作用。方法:采用DEAE-52阴离子纤维素和Sepha‐dex G-15葡聚糖凝胶色谱柱分离纯化低聚糖组分。利用体外细胞实验、Griess法、Western blot检测法检测其抗炎活性。结果:得到了相对分子量为1452 Da的低聚糖组分D1,得率为0.13%±0.05%,蛋白杂质含量为(31.08±1.25)μg/mg,纯度为96.89%。单糖组成研究表明低聚糖D1主要由葡萄糖(63.40%)、阿拉伯糖(30.06%)和甘露糖(6.54%)组成。体外细胞实验表明,低聚糖D1样品在25~400μg/mL浓度范围对RAW 264.7巨噬细胞无细胞毒作用。在LPS诱导的RAW 264.7巨噬细胞炎症模型中,低聚糖D1能有效地抑制炎症因子NO的释放。Western blot检测发现,低聚糖D1样品的共孵育处理能剂量依赖性抑制iNOS蛋白的表达。结论:潞党参中低聚糖组分D1能通过抑制炎症介质NO的释放,起到良好的抗炎作用。Objective:Exploring the composition and anti-inflammatory activity of oligosaccharides in Lu Dangshen.Methods:The oligosaccharide components were separated and purified using DEAE-52 anionic cellulose and Sephadex G-15 dextran gel column.The anti-inflammatory activity was detected by in vitro cellular assay,Griess method,and Western blot assay.Results:The oligosaccharide component D1 with a relative molecular weight of 1452 Da was isolated and purified.The yield was 0.13%±0.05%,the protein impurity content was(31.08±1.25)μg/mg,and the purity was 96.89%.The monosaccharide composition study showed that D1 was mainly composed of glucose(63.40%),arabinose(30.06%)and mannose(6.54%).The anti-inflammatory activity of D1 was explored,and it was found that D1 sample had no cytotoxic effect on RAW 264.7 cells in the concentration range of 25~400μg/mL.In LPS-induced RAW 264.7 cell inflammation model,D1 could inhibit the release of NO.Western blot assay revealed that co-incubation treatment of D1 samples dose-dependently inhibited iNOS protein expression.Conclusion:Oligosaccharide component D1 of Lu Dangshen could inhibit the release of inflammatory factor NO and played a good anti-inflammatory role.

关 键 词：潞党参 低聚糖 分离纯化 理化性质 抗炎活性 

分 类 号：R284.1[医药卫生—中药学]