Wu Junwen, Zhou Xuefei, Zhang Min, Yao Yun, Han Juanjuan, Liu Kehai
Department of Biopharmaceutics, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Mar Drugs. 2017 May 18;15(5):140. doi: 10.3390/md15050140.
In this study, the extraction conditions of the crude polysaccharide from were optimized by response surface methodology. The optimum extraction conditions were: a ratio of raw material to water volume of 1:80 (g/mL); an extraction temperature of 72 °C; and an extraction time of 3 h. Then, a purified polysaccharide named polysaccharide-1 (CSP-1) was obtained from the crude polysaccharide by the Diethylaminoethyl cellulose-52 (DEAE-52) cellulose chromatography column and Sephadex G-100 column. The molecular weight and monosaccharide composition of CSP-1 was determined through Gel Permeation Chromatography (GPC) and Gas Chromatography-Mass Spectrometer (GS-MS), respectively. The results showed that CSP-1 with an average molecular weight of 56,335 Da was composed of l-(-)-Fucose, d-(+)-Mannose, d-Glucose and mainly possessed 1→2, 1→2, 6, 1→4, and 1→4, 6 of glycosyl linkages. The immunomodulatory activities of CSP-1 were also evaluated using lipopolysaccharide (LPS)-induced human monocytic (THP-1) cells. The results demonstrated that CSP-1 dose-dependently protected against LPS-induced toxicity, and CSP-1 significantly inhibited the Toll-like receptor 4 (TLR-4) mRNA, myeloid differentiation factor 88 (MyD88) mRNA and tumour necrosis factor receptor-associated factor-6 (TRAF-6) mRNA expression of the LPS-induced THP-1 cells, as well as suppressing reactive oxygen species (ROS) generation.
在本研究中，采用响应面法对从[具体来源未给出]中提取粗多糖的条件进行了优化。最佳提取条件为：原料与水体积比为1:80（g/mL）；提取温度为72℃；提取时间为3小时。然后，通过二乙氨基乙基纤维素-52（DEAE-52）纤维素色谱柱和葡聚糖凝胶G-100柱从粗多糖中获得了一种纯化多糖，命名为[具体名称未给出]多糖-1（CSP-1）。分别通过凝胶渗透色谱法（GPC）和气相色谱-质谱联用仪（GS-MS）测定了CSP-1的分子量和单糖组成。结果表明，平均分子量为56335 Da的CSP-1由L-（-）-岩藻糖、D-（+）-甘露糖、D-葡萄糖组成，主要具有1→2、1→2,6、1→4以及1→4,6的糖苷键连接。还使用脂多糖（LPS）诱导的人单核细胞（THP-1）细胞评估了CSP-1的免疫调节活性。结果表明，CSP-1对LPS诱导的毒性具有剂量依赖性保护作用，并且CSP-1显著抑制LPS诱导的THP-1细胞中Toll样受体4（TLR-4）mRNA、髓样分化因子88（MyD88）mRNA和肿瘤坏死因子受体相关因子-6（TRAF-6）mRNA的表达，同时抑制活性氧（ROS）的产生。
