树莓多糖组分RPP-5的结构特征与免疫活性研究  

Structural characteristics and immunomodulatory activity of raspberry polysaccharide RPP-5

作　　者：尹星星 杨永晶 陆杰 王学红 晁沐 YIN Xingxing;YANG Yongjing;LU Jie;WANG Xuehong;CHAO Mu(School of Ecological and Environmental Engineering,Qinghai University,Xining 810016,China;Qinghai Raspberry Agricultural Industrialization Co.Ltd.,Xining 812100,China)

机构地区：[1]青海大学生态环境工程学院,青海西宁810016 [2]青海树莓农业产业化有限公司,青海西宁812100

出　　处：《食品与发酵工业》2024年第9期139-147,共9页Food and Fermentation Industries

基　　金：青海省科技厅基础研究项目(2019-ZJ-953Q);国家自然科学基金项目(81860619)。

摘　　要：利用DEAE-Sepharose Fast Flow和Sephadex G-200对树莓粗多糖进行分离纯化得到组分RPP-5,探究RPP-5的结构特征和免疫调节活性。采用高效凝胶渗透色谱、傅里叶红外光谱和气相色谱-质谱联用技术对RPP-5的结构进行表征。利用RAW264.7细胞研究RPP-5的免疫活性,通过CCK-8法、微板法、酶联免疫吸附测定和实时荧光定量PCR分别测定RPP-5对RAW264.7细胞的细胞活力、NO含量、炎症因子白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量以及炎症因子mRNA表达水平的影响。结果表明,RPP-5主要由物质的量比为44.5∶21.7∶5.9∶8.8∶5∶4.1的阿拉伯糖、葡萄糖、鼠李糖、半乳糖、甘露糖和木糖组成,其重均分子质量、数均分子质量和峰位分子质量分别为7598 Da、6183 Da和7492 Da,共存在12种单糖连接方式,糖链中→4)-Gal p-(1→、→4)-Xyl p-(1→、Ara f-(1→和→4,6)-Glc p-(1→的含量较高,分别为26.27%、16.50%、11.38%和10.41%。RPP-5可显著升高RAW264.7细胞的细胞活力,提高细胞内NO、IL-6、IL-1β和TNF-α的含量,促进IL-6、IL-1β和TNF-α的mRNA表达。综上,RPP-5是一种多支链的酸性杂多糖,具有较强的免疫增强活性,可作为功能性食品中的免疫调节剂,为树莓多糖的结构表征和免疫活性研究提供理论支持。In the present study,a raspberry polysaccharide named RPP-5 was isolated and purified using DEAE-Sepharose Fast Flow and Sephadex G-200.Its structural characteristics and immunomodulatory activity were also explored.The structural characteristics of RPP-5 were studied based on the results of high-performance gel permeation chromatography,Fourier transform infrared spectroscopy,and GC-MS.RAW264.7 cells were utilized to evaluate the immunomodulatory activity of RPP-5.The cell viability,contents of NO and inflammatory factors,and mRNA expression levels of inflammatory factors in RAW264.7 cells were measured by CCK-8 assay,microplate method,enzyme-linked immunosorbent assay assays,and real-time quantitative polymerase chain reaction,respectively.Results showed that RPP-5 was composed of arabinose,glucose,rhamnose,galactose,mannose,and xylose,with a molar ratio of 44.5∶21.7∶5.9∶8.8∶5∶4.1.The average molecular weight,peak molecular weight,and number average molecular weight were 7598 Da,7492 Da,and 6183 Da,respectively.RPP-5 contained 12 types of glycosyl linkages types,among which→4)-Gal p-(1→,→4)-Xyl p-(1→,Ara f-(1→,→4,6)-Glc p-(1→were the main connection types of sugar residues,accounted for 26.27%,16.50%,11.38%,and 10.41%,respectively.RPP-5 significantly increased the viability of RAW264.7 cells,increased the content of NO,IL-6,IL-1β,and TNF-α,and promoted the expression of IL-6,IL-1β,and TNF-αmRNA in RAW264.7 cells.In conclusion,RPP-5 was a homogeneous acidic heteropolysaccharide with a complex multi-branched structure and showed significant immune-enhancing activity.It could be used as an immunomodulator in functional foods,providing theoretical support for the structural characterization and immunological activity studies of raspberry polysaccharides.

关 键 词：树莓 多糖组分 结构特征 免疫活性 RAW264.7细胞 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]