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党参多糖不同组分的抗炎活性及机制研究 Δ
孟 燕 ,徐玉洁 ,张宝徽 ,常 聪 ,郑国华 ,吴 勇 (1.湖北中医药大学药学院,武汉 430065;2.湖北中医
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药大学中药资源与中药复方教育部重点实验室,武汉 430065)
中图分类号 R285 文献标志码 A 文章编号 1001-0408(2020)11-1348-05
DOI 10.6039/j.issn.1001-0408.2020.11.11
摘 要 目的:筛选出党参多糖中具有较好抗炎效果的组分,并探索其抗炎机制。方法:以党参药材为样品,采用水提醇沉法得党
参粗多糖(CP)。将CP通过DEAE Sepharose Fast Flow、SephadexG-150等凝胶柱分离纯化后得到组分CP1-2-1、CP3-1-1,并对其进
行表征。采用MTT法测定0.01、0.1、1、10、100 μg/mL的CP、CP1-2-1、CP3-1-1分别对小鼠腹腔巨噬细胞RAW264.7作用24、48 h后
的细胞存活率。然后将细胞分为空白组(空白培养基)、模型组[1 μg/mL脂多糖(LPS)]和3种多糖的低、中、高质量浓度组(1 μg/mL
LPS+25、50、100 μg/mL CP、CP1-2-1、CP3-1-1),加药培养24 h。分别测定细胞培养液中一氧化氮(NO)含量以及细胞中Toll样受
体4(TLR4)、白细胞介素6(IL-6)、核因子-κB(NF-κB)、肿瘤坏死因子α(TNF-α)mRNA表达水平。结果:CP的糖含量高达(92.20±
0.73)%;CP1-2-1是由果糖组成的单一中性多糖,相对分子量为25.8 kDa;CP3-1-1是由阿拉伯糖、鼠李糖、半乳糖以及半乳糖醛酸
等组成的酸性杂多糖,相对分子量为49.5 kDa。3种多糖分别作用24、48 h后,细胞的存活率均大于99%。与空白组比较,模型组
细胞培养液中NO含量以及细胞中TLR4、IL-6、NF-κB、TNF-α mRNA表达水平均显著升高(P<0.05或P<0.01)。与模型组比较,
各给药组细胞培养液中NO含量均显著降低;50、100 µg/mL CP组细胞中TLR4、NF-κB、TNF-α、IL-6 mRNA表达水平均显著降低
(P<0.05 或 P<0.01),25 µg/mL CP1-2-1 组细胞中 TLR4、NF-κB mRNA 表达水平以及 50、100 µg/mL CP1-2-1 组细胞中 TLR4、
NF-κB、TNF-α、IL-6 mRNA 表达水平均显著降低(P<0.05 或 P<0.01),25 µg/mL CP3-1-1 组细胞中 IL-6 mRNA 表达水平和 50
µg/mL CP3-1-1组细胞中NF-κB、TNF-α、IL-6 mRNA表达水平以及100 µg/mL CP3-1-1组细胞中TLR4、NF-κB、TNF-α、IL-6 mRNA
表达水平均显著降低(P<0.05或P<0.01)。结论:党参CP与均一多糖组分CP1-2-1、CP3-1-1均具有一定的抗炎活性;其机制可能
与抑制TLR4/NF-κB途径的活化来抑制TNF-α、IL-6等炎症因子的生成与释放有关。
关键词 党参多糖;粗多糖;多糖组分;抗炎活性;机制
Study on the Anti-inflammatory Activity and Mechanism of Different Components from Codonopsis Radix
Polysaccharides
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MENG Yan ,XU Yujie ,ZHANG Baohui ,CHANG Cong ,ZHENG Guohua ,WU Yong(1.College of Pharmacy,
Hubei University of Chinese Medicine,Wuhan 430065,China;2.Key Laboratory of Chinese Medicine Resource
and Compound Prescription,Ministry of Education,Hubei University of Chinese Medicine,Wuhan 430065,China)
ABSTRACT OBJECTIVE:To screen the components with better anti-inflammatory effect from Codonopsis Radix polysaccharide
and explore the anti-inflammatory mechanism. METHODS:Using Codonopsis Radix as sample,the crude polysaccharide(CP)
was obtained by water extraction and alcohol precipitation. After CP was separated and purified by Fast Flow DEAE Sepharose,
SephadexG-150 gel column and so on,the components CP1-2-1 and CP3-1-1 were obtained and then characterized. The survival
rate of RAW264.7 cell was determined by MTT method after cultured with CP,CP1-2-1 and CP3-1-1(0.01,0.1,1,10,100
μg/mL)for 24 and 48 h respectively. The cells were divided into blank group(blank culture medium),model group(1 μg/mL
LPS)and low-,medium- and high-concentration groups of 3 kinds of Codonopsis Radix polysaccharide(1 μg/mL LPS+25,50,
100 μg/mL CP,CP1-2-1,CP3-1-1 solution). After cultured for 24 h,NO content in cell culture solution,mRNA expressions of
TLR4,IL-6,NF-κB and TNF-α in cells were determined. RESULTS:The sugar content of CP reached(92.20±0.73)%. CP1-2-1
was a single neutral polysaccharide composed of fructose with a relative molecular weight of 25.8 kDa. CP3-1-1 was an acidic
heteropolysaccharide composed of arabinose,rhamnose,galactose and galacturonic acid and so on,with a relative molecular
weight of 49.5 kDa. Cell survival rates were higher than 99%,after cultured with 3 kinds of polysaccharide for 24 and 48 h.
Compared with blank group,NO content in cell culture solution and mRNA expressions of TLR4,IL-6,NF-κB and TNF-α in cells
were increased significantly in model group(P<0.05 or P<
Δ 基金项目:湖北省知识创新专项(自然科学基金)项目(No.
0.01). Compared with model group, NO content in cell
2016CFB412)
culture solution of each administration group was significantly
*讲师,博士。研究方向:中药新制剂与新剂型。E-mail:yan-
reduced,mRNA expressions of TLR4,NF-κB,TNF-α,IL-6
meng2016@126.com
# 通信作者:副教授,博士。研究方向:中药药理学。E-mail:wuy- in cells in 50,100 μg/mL CP group were significantly decreased
ong1991@163.com (P<0.05 or P<0.01);mRNA expressions of TLR4,NF-κB
·1348 · China Pharmacy 2020 Vol. 31 No. 11 中国药房 2020年第31卷第11期