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  

Isolation, Structural Characterization, Immunoregulatory and AntioxidantActivity of Raspberry Polysaccharide RPP-6 in Vitro

作　　者：陆杰 杨永晶 尹星星 王学红 晁沐 LU Jie;YANG Yongjing;YIN Xingxing;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,西宁810016 [2]青海树莓农业产业化有限公司,西宁812100

出　　处：《西北农业学报》2024年第3期477-489,共13页Acta Agriculturae Boreali-occidentalis Sinica

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

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。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 Ara f-(1→,→4,6)-Glc p-(1→,→4)-Gal p-(1→and→4)-Xyl p-(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.

关 键 词：树莓 结构表征 免疫活性 抗氧化活性 

分 类 号：S663[农业科学—果树学] TS255.1[农业科学—园艺学]