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
汤敏 1陆姗姗 2谷巍 1胡守芳3
作者信息
- 1. 南京中医药大学 药学院,江苏 南京 210023
- 2. 南京中医药大学 药学院,江苏 南京 210023;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
- 折叠
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1 452 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25～400 μg/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.
Abstract
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 compo-nent D1 with a relative molecular weight of 1 452 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 mono-saccharide 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 25～400 μg/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 iN-OS 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Key words
Lu Dangshen/Oligosaccharide/Separation and purification/Physicochemical property/Anti-inflammatory activity引用本文复制引用
基金项目
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)
泰州市社会发展项目(SSF20230025)
大学生创新创业训练项目(202313981013Y)
大学生创新创业训练项目(202313981008Y)
江苏省高等学校自然科学面上项目(23KJD360003)
出版年
2024
NETL
NSTL
万方数据