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树莓多糖组分RPP-6的分离、结构表征及其体外免疫和抗氧化活性研究
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引用本文:陆 杰,杨永晶,尹星星,王学红,晁 沐.树莓多糖组分RPP-6的分离、结构表征及其体外免疫和抗氧化活性研究[J].西北农业学报,2024,(3):477~489
DOI:10.7606/j.issn.1004-1389.2024.03.010
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作者单位
陆 杰,杨永晶,尹星星,王学红,晁 沐 (1.青海大学 生态环境工程学院西宁 8100162.青海树莓农业产业化有限公司西宁 812100) 
基金项目:国家自然科学基金(81860619);青海省科技厅基础研究项目(2019-ZJ-953Q)。
中文摘要:旨在对从树莓粗多糖中分离纯化出的树莓多糖组分RPP-6进行结构表征及体外免疫活性和抗氧化活性研究。采用DEAE-Sepharose Fast Flow和Sephadex G-200对树莓粗多糖进行分离纯化。结合高效凝胶渗透色谱法(High-Performance Gel Permeation Chromatography,HPGPC)、傅里叶红外光谱法(Fourier Transform Infrared Spectroscopy,FT-IR)、气相色谱-质谱联用(Gas Chromatography and Mass Spectrometry,GC-MS)和甲基化等方法对RPP-6的结构进行表征。利用小鼠巨噬细胞RAW264.7和自由基清除试验研究RPP-6的体外免疫与抗氧化活性。结果表明,RPP-6是一种酸性杂多糖,由阿拉伯糖、葡萄糖、半乳糖、鼠李糖和甘露糖组成,摩尔比为37.8∶30.8∶21.4∶6.0∶3.9 。RPP-6的峰位分子质量为7.645 ku、重均分子质量为7.769 ku、数均分子质量为6.310 ku。RPP-6中存在15种单糖连接方式,其中Araf-(1→、→4,6)-Glcp- (1→、→4)-Galp-(1→和→4)-Xylp-(1→为主要连接方式,含量分别为20.64%、8.59%、24.04%和 16.92%。RPP-6能明显提高小鼠巨噬细胞RAW264.7的活力和吞噬能力,并促进细胞中TNF-α(Tumor Necrosis Factor,TNF)、IL-6(Interleukin-6,IL-6)、IL-1β(Interleukin-1β,IL-1β)和NO(Nitric Oxide,NO)释放。RPP-6对ABTS·+具有较高的清除作用,对DPPH·的清除作用不明显。表明RPP-6是一种多支链的均一酸性杂多糖,具有显著的免疫增强活性和一定的抗氧化活性。
中文关键词:树莓  结构表征  免疫活性  抗氧化活性
 
Isolation, Structural Characterization, Immunoregulatory and Antioxidant Activity of Raspberry Polysaccharide RPP-6 in Vitro
Abstract:In the present study, the raspberry polysaccharide RPP-6 was isolated from raspberry pulp,and its structural characteristics,immunomodulatory and antioxidant activities were studied in vitro. RPP-6 was isolated and purified by DEAE-Sepharose Fast Flow and Sephadex G-200 chromatography and thestructural characteristics of RPP-6 were studied by high-performance gel permeation chromatography (HPGPC), Fourier-transform infrared spectroscopy (FT-IR), gas chromatography-mass spectroscopy (GC-MS) and methylation. RAW264.7 macrophages and free radical scavenging tests were conducted to evaluate the immunomodulatory and antioxidant activities of RPP-6.The results showed that RPP-6 was composed of arabinose, glucose, galactose, rhamnose and mannose at a molar ratios of 37.8∶30.8∶21.4∶6.0∶3.9,respectively. The peak molecular mass, average molecular mass and number average molecular mass were 7.645 ku,7.769 ku, and 6.310 ku, respectively. RPP-6 was composed of 15 glycosyl linkages patterns, among which Araf-(1→, →4,6)-Glcp-(1→, →4)-Galp-(1→ and →4)-Xylp-(1→ were the main connection modes of sugar residues, accounting for about 20.64%, 8.59%, 24.04%, and 16.92%, respectively. RPP-6 significantly promoted the proliferation and phagocytosis of RAW264.7 cells and increased the release of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and nitric oxide (NO) in RAW264.7 cells. RPP-6 exhibited strong ABTS·+ radical scavenging ability,but exhibited no DPPH· free radical scavenging ability. In conclusion, RPP-6 is a homogeneous acidic heteropolysaccharide with a complex multi-branched structure, and has significant immune-enhancing activity and certain antioxidant activity.
keywords:Raspberry polysaccharide  Structural characterization  Immune activity  Antioxidant activity
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