灵芝β-葡寡糖的分离纯化和体外抗炎活性

β-glucooligosaccharide separated and purified from Ganoderma lingzhi and its in vitro anti-inflammation activities

为阐释抗炎活性与灵芝葡寡糖聚合度之间的关系,将灵芝β-葡寡糖组分GLPW-A(DP2–14)通过Bio-Gel P-2凝胶色谱柱分离,分别从洗脱液、流速以及每管接样量3个方面优化分离条件。通过阴离子色谱和基质辅助激光解析串联飞行时间质谱(MALDI-TOF-MS)分析测定其聚合度,并进行体外抗炎活性评价。当洗脱液为0.1 mol/L的NH_(4)HCO_(3)溶液,流速为0.1 mL/min,接样量为2 mL时分离效果最佳,根据出峰时间先后收集得到了8个组分(F1–F8)。聚合度分析表明,F8和F7分别为DP2和DP3的葡寡糖组分,其余6个组分F6–F1均为主要含有2–3种聚合度的葡寡糖。体外抗炎活性表明,主要含有DP4–DP6的F5和F6组分在浓度为0.5–10μg/mL的范围内无明显抗炎活性,而其余各组分均在一定浓度下表现出不同程度的抗炎作用,且活性优于GLPW-A组分,说明灵芝β-葡寡糖的抗炎活性与寡糖片段的聚合度有关。

Theβ-glucooligosaccharide fraction GLPW-A(DP2–14)from Ganoderma lingzhi was separated by Bio-Gel P-2 chromatography column.The eluting conditions including eluent,flow rate and collection volume per tube were optimized.The polymerization degree of isolated fractions were analyzed by matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS),and the anti-inflammatory activity of these fractions were evaluated in vitro.Under the optimal separation conditions of 0.1 mol/L NH_(4)HCO_(3)solution eluted at a flow rate of 0.1 mL/min and 2 mL collection volume per tube,eight fractions(F1–F8)were successively obtained.The polymerization degree analysis showed that F8 and F7 mainly comprised oligosaccharides with DP2 and DP3,respectively,and other isolated fractions(F6–F1)mainly contained components with 2–3 kinds of different polymerization degrees.In vitro anti-inflammatory activity showed that F5 and F6 mainly contained DP4–DP6 had no obvious anti-inflammatory activity in the concentration of 0.5–10μg/mL,while the other fractions showed different level of anti-inflammatory activity at specific concentrations.The anti-inflammatory activity ofβ-glucooligosaccharide was related to the degree of polymerization of oligosaccharide fragments.