李溢真¹, 于志洋¹, 田欣², 宋琳^2,3
1.青岛农业大学生命科学学院, 山东 青岛 266109;2.青岛科技大学海洋科学与生物工程学院, 山东 青岛 266071;3.无穷食品有限公司, 广东 饶平 515726

摘要:
采用水提醇沉法制备海蒿子多糖，利用红外光谱法（FT-IR）和PMP柱前衍生化法研究海蒿子多糖的理化特性及糖元组成，并采用实时荧光定量PCR法和ELISA法，研究免疫相关因子和细胞凋亡相关因子的mRNA表达。结果表明，海蒿子多糖总糖含量为24.32%，硫酸根含量为5.39%，具有α糖苷键。单糖分析表明，海蒿子多糖主要由半乳糖、岩藻糖、甘露糖组成。进一步体外免疫实验结果表明，海蒿子多糖具有促进小鼠巨噬细胞RAW264.7增殖的作用，能够提高巨噬细胞系免疫因子IL-1β、IL-6、iNOS和TNF-α的mRNA表达量。此外，海蒿子多糖还能抑制肺癌细胞A549的增殖，提高Bax/Bcl-2的比例，说明其具有抑制肿瘤的效果。ELISA分析表明，海蒿子粗多糖能显著提高RAW264.7分泌NO、IL-1β、TNF-α和IL-6细胞因子。因此，海蒿子多糖不仅具有免疫增强活性，还具有一定的抗肿瘤活性。
关键词:海蒿子多糖理化性质免疫活性
DOI：10.11759/hykx20200418002
分类号:R932
基金项目:山东省自然基金（ZR201702170401）；青岛科技大学2019年创新训练计划项目（201910426018）

Physicochemical properties and immunomodulatory activities of crude polysaccharides isolated from Sargassum pallidum
LI Yi-zhen¹, YU Zhi-yang¹, TIAN Xin², SONG Lin^2,3
1.College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China;2.College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao 266071, China;3.Wuqiong food Co., Ltd, Raoping 515726, China
Abstract:
Sargassum pallidum polysaccharides (SPPS), a group of natural polysaccharides, were isolated from Sargassum pallidum through water extraction and alcohol precipitation. Infrared spectroscopy (FT-IR) and 1-phenyl-3-methyl-5-pyrazolone (PMP) pre-column derivatization were employed to study the physical and chemical properties, as well as the monosaccharide composition of SPPS. Quantitative real-time Polymerase Chain Reaction (qRT-PCR) and enzyme linked immunosorbent assay (ELISA) techniques were employed to study the mRNA expression of immune-related and apoptosis-related factors. The total sugar content of SPPS was 24.32%, while the sulfate content was 5.39%. The FT-IR spectra analysis indicated that SPPS contain sulfuric acid groups and has α-glycosidic bonds. In regard to the monosaccharide composition, the most abundant components in SPPS were galactose, fucose, and mannose. Bioassays indicated that SPPS significantly enhance the proliferation of RAW264.7 macrophage cells and also stimulate the corresponding mRNA expression of iNOS and interleukins (IL-6, IL-1β, and TNF-α) in a dose-dependent manner. In addition, SPPS effectively suppressed the proliferation of A549 cells and increase the ratio of Bax/Bcl-2, indicating its anti-proliferative activity against A549 cells. The ELISA analysis revealed that the crude SPPS significantly increased the production of NO, IL-1β, TNF-α, and IL-6 cytokines in RAW264.7 cells. These results indicate that SPPS demonstrated both immunomodulatory and anti-tumor activities.
Key words:Sargassumpallidumpolysaccharidesphysicochemical propertiesimmunomodulatory activity





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