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党参多糖不同组分的抗炎活性及机制研究
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| 篇名： | 党参多糖不同组分的抗炎活性及机制研究 |
| TITLE： | Study on the Anti-inflammatory Activity and Mechanism of Different Components from Codonopsis Radix Polysaccharides |
| 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DEAESepharoseFastFlow、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、100μg/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、48h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[1μg/mL脂多糖（LPS）]和3种多糖的低、中、高质量浓度组（1μg/mLLPS+25、50、100μg/mLCP、CP1-2-1、CP3-1-1），加药培养24h。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25.8kDa；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49.5kDa。3种多糖分别作用24、48h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-αmRNA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、100µg/mLCP组细胞中TLR4、NF-κB、TNF-α、IL-6mRNA表达水平均显著降低（P＜0.05或P＜0.01），25µg/mLCP1-2-1组细胞中TLR4、NF-κBmRNA表达水平以及50、100µg/mLCP1-2-1组细胞中TLR4、NF-κB、TNF-α、IL-6mRNA表达水平均显著降低（P＜0.05或P＜0.01），25µg/mLCP3-1-1组细胞中IL-6mRNA表达水平和50µg/mLCP3-1-1组细胞中NF-κB、TNF-α、IL-6mRNA表达水平以及100µg/mLCP3-1-1组细胞中TLR4、NF-κB、TNF-α、IL-6mRNA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。 |
| 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 CP 1-2-1 and CP 3-1-1 were obtained and then characterized. The survival rate of RAW 264.7 cell was determined by MTT method after cultured with CP ，CP1-2-1 and CP 3-1-1（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）％. CP 1-2-1 was a single neutral polysaccharide composed of fructose with a relative molecular weight of 25.8 kDa. CP 3-1-1 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 TLR 4，IL-6，NF-κB and TNF-α in cells were increased significantly in model group （P＜0.05 or P＜ . Compared with model group ， NO content in cell 2016CFB412） culture solution of each administration group was significantly reduced，mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 50，100 μg/mL CP group were significantly decreased ong1991@163.com （P＜0.05 or P＜0.01）；mRNA expressi ons of TLR 4，NF-κB in cells in 25 µg/mL CP 1-2-1 group and mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 50，100 µg/mL CP 1-2-1 group were significantly decreased （P＜0.05 or P＜0.01）；mRNA expressionsof IL- 6 in cells in 25 µg/mL CP 3-1-1 group，mRNA expressions of NF-κB，TNF-α，IL-6 in cells in 50 µg/mL CP 3-1-1 group and mRNA expressions of TLR 4，NF-κB，TNF-α，IL-6 in cells in 100 µg/mL CP 3-1-1 group were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：CP，CP 1-2-1 and CP 3-1-1 all have certain anti-inflammatory activities. The mechanism may be related to inhibiting the generation and releasing of inflammatory cytokines such as TNF-α，IL-6 by inhibiting the activation of TLR 4/NF-κB pathway. |
| 期刊： | 2020年第31卷第11期 |
| 作者： | 孟燕，徐玉洁，张宝徽，常聪，郑国华，吴勇 |
| AUTHORS： | MENG Yan，XU Yujie，ZHANG Baohui ，CHANG Cong，ZHENG Guohua ，WU Yong |
| 关键字： | 党参多糖；粗多糖；多糖组分；抗炎活性；机制 |
| KEYWORDS： | Codonopsis Radix polysaccharides；Crude polysaccharide ；Polysaccharide component ；Anti-inflammatory activity ； |
| 阅读数： | 700 次 |
| 本月下载数： | 15 次 |
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