四川小金县野生羊肚菌多糖的结构解析及体外抗肿瘤活性初探

Structural identificatin and in vitro antitumor activity of wild Morchella esculenta polysaccharide from Xiaojin County of Sichuan Province

本研究通过热水浸提法从四川小金县的野生羊肚菌中提取多糖(Morchella esculenta polysaccharide,ME-P),利用傅里叶红外光谱(FT-IR)、高效液相色谱(HPLC)、气相质谱连用(GC-MS)和核磁共振(NMR)波谱等技术鉴定ME-P的结构,采用CCK-8法检验ME-P的药物毒性并探寻ME-P体外抗肿瘤活性。结果表明,ME-P中含有吡喃糖环,单糖组成为甘露糖、葡萄糖、半乳糖,比例为4∶4∶1;具有1,2,3,4,6-D-甘露糖、2,3,4,6-D-甘露糖交替连接为主链,1,6-D-葡萄糖和1,2,6-D-半乳糖作为支链,4-D-葡萄糖与1,2,6-D-半乳糖连接作为末端糖的重复单位的结构新颖的多糖。ME-P对巨噬细胞(RAW 264.7)无毒害,且有极显著(P<0.01)增殖作用,在质量浓度为1.25μg/mL时对RAW 264.7的增殖率可高达79.5%。同时,对小鼠结肠癌细胞(CT26.WT)和胃癌细胞(MFC)均有抑制作用,且均为低浓度时抑制效果较好,在ME-P为1.25μg/mL时,ME-P对CT26.WT和MFC的抑制率分别为28.20%和34.23%。本研究解析出四川小金县羊肚菌多糖(ME-P)为甘露糖、葡萄糖、半乳糖组成的结构新颖的多糖,无毒且对肿瘤细胞显示出显著的抑制作用,该结果为羊肚菌多糖的开发与应用提供了一定的科学依据。

In this study,Morchella esculenta polysaccharide(ME-P)were extracted by hot water extraction from wild M.esculenta in Xiaojin County,Sichuan Province.Fourier transform infrared spectroscopy(FT-IR),high performance liquid chromatography(HPLC),gas chromatography mass spectrometry(GC-MS)and nuclear magnetic resonance(NMR)were used to identify the structure of ME-P.CCK-8 method was used to detect the drug toxicity of ME-P and to explore the antitumor activity of ME-P in vitro.The results showed that ME-P contained pyranose ring,and the monosaccharides were composed of mannose,glucose and galactose with a ratio of 4∶4∶1.The polysaccharides are characterized by a novel structure,which is characterized by the alternating linkage of 1,2,3,4,6-D-mannose and 2,3,4,6-D-mannose as the main chain,1,6-D-glucose and 1,2,6-D-galactose as the branch chain,and the connection of 4-D-glucose and 1,2,6-D-galactose as a repeating unit of terminal monosaccharide.ME-P is non-toxic to macrophages(RAW 264.7)and has a very significant effect(P<0.01)on proliferation.The proliferation rate of RAW 264.7 can be as high as 79.5%at a mass concentration of 1.25μg/mL.At the same time,it had inhibitory effect on mouse colon cancer cells(CT26.WT)and gastric cancer cells(MFC),and the inhibition effect was better at low concentration.When ME-P was 1.25μg/mL,the inhibition rates of ME-P on CT26.WT and MFC were 28.20%and 34.23%,respectively.This study shows that M.esculenta polysaccharides(ME-P)in Xiaojin County,Sichuan Province is a novel polysaccharide composed of mannose,glucose and galactose,which is non-toxic and has significant inhibitory effect on tumor cells.The results provide a scientific basis for the development and application of Morchella polysaccharides.