天府花生叶多糖AHL-P结构解析及体外抗肿瘤和免疫活性

Structural Identification,in Vitro Immunomodulatory and Antitumor Activities of Polysaccharides AHL-P from Arachis hypogaea Leaves

本研究利用热水浸提法制备天府花生叶片多糖(Arachis hypogaea leaves polysaccharide,AHL-P),分别利用高效凝胶渗透色谱(high performance gel permeation chromatography,HPGPC)、傅里叶变换红外光谱(Fourier transform infrared spectroscopy,FTIR)、核磁共振波谱(nuclear magnetic resonance spectroscopy,NMR)、气相色谱-质谱联用(gas chromatography-mass spectrometry,GC-MS)等技术对AHL-P进行结构鉴定,并采用CCK-8法探索AHL-P对腹水癌细胞(S180)、小鼠胃癌细胞(MFC)、免疫B细胞(Raji)、T细胞(Jurkat)和巨噬细胞(RAW 264.7)体外增殖的影响。结果表明,AHL-P分子质量为12.84 kDa,其单糖组成为葡萄糖、木糖、半乳糖和阿拉伯糖,物质的量比为7∶2∶1∶1,是一种主链由(1→4,6)-葡萄糖和(1→4)-葡萄糖连接,(1→4)-木糖、(1→4)-阿拉伯糖和(1→4)-半乳糖作为支链,→1)-葡萄糖作为末端糖的重复结构单位的新颖多糖。体外抗肿瘤活性研究显示,AHL-P在质量浓度为5μg/mL时对S180的抑制率最高,为39.07%,2.5μg/mL时对MFC的抑制率最高,为52.22%。体外免疫活性研究显示,当AHL-P的质量浓度为10μg/mL时,对Raji和RAW 264.7增殖率分别为104.39%和32.25%,但对Jurkat没有显著增殖效果。本研究解析出了AHL-P的结构并探寻了其体外抗肿瘤及免疫活性,可为天府花生的开发利用提供一定的科学依据。

In this study,a polysaccharide(AHL-P)was obtained from Arachis hypogaea leaves by hot water extraction.High performance gel permeation chromatography(HPGPC),Fourier transform infrared spectroscopy(FTIR),nuclear magnetic resonance spectroscopy(NMR),and gas chromatography mass spectrometry(GC-MS)were used to identify the structure of AHL-P.The effects of AHL-P on the proliferation of MFC cells,S180 cells,B cells(Raji),T cells(Jurkat)and RAW 264.7 cells in vitro were also tested by cell counting kit-8(CCK-8).The results indicated that the molecular mass of AHL-P was 12.84 kDa and its monosaccharide composition consisted of glucose,xylose,galactose and arabinose at a ratio of 7:2:1:1.AHL-P was a novel polysaccharide having a backbone chain composed of(1→4,6)-Glc,(1→4)-Xyl and(1→4)-Glc,with(1→4)-Ara and(1→4)-Gal as branches and→1)-Glc as a terminal monosaccharide of the repeating unit.The antitumor activity assay in vitro indicated that the highest inhibitory rate of AHL-P on S180 and MFC cells of 39.07%and 52.22%was observed at a concentration of 5 and 2.5μg/mL,respectively.The immunomodulatory activity assay in vitro indicated that at 10μg/mL AHL-P,the proliferation rate of B cells(Raji)and RAW 264.7 cells was 104.39%and 32.25%,respectively.However,AHL-P had no significantly proliferative effect on T cells(Jurkat).This study could provide a scientific basis for the development and utilization of Arachis hypogaea leaves.