黑木耳多糖结构分析及其保护肝细胞氧化损伤活性

Structural Analysis of Auricularia heimuer Polysaccharide and Its Activity in Protecting Liver Cells against Oxidative Damage

为确定黑木耳多糖的结构特征及其体外保护肝细胞氧化损伤活性,通过破壁提取法、Sevag法、透析法对黑木耳子实体多糖(AHP)进行了分离纯化,采用高效凝胶渗透色谱(HPGPC)、离子色谱(IC)和红外光谱(IR)法对黑木耳多糖进行了分子质量、单糖组成及特征官能团分析;建立肝细胞氧化损伤模型,对AHP的肝保护活性进行了研究。结果表明:AHP糖含量为83.879%,分子质量410.3 kD,主要由岩藻糖(Fuc)、阿拉伯糖(Ara)、半乳糖(Gal)、葡萄糖(Glc)、甘露糖(Man)按2.240∶0.692∶3.994∶22.754∶65.696摩尔比组成的β型吡喃糖。H2O2损伤人肝癌细胞HepG2细胞中加入AHP后发现,AHP可抑制受损细胞发生的细胞核皱缩和边缘羽化现象;提高损伤细胞的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)等抗氧化酶活性;减少一氧化氮(NO)等氧化产物生成量。AHP具有保护肝细胞氧化损伤的活性,具有将其进一步开发为保肝健康食品的潜力。

The structural characteristics of Auricularia heimuer polysaccharides(AHP)and their activity to protect liver cells from oxidative damage in vitro were studied.The total polysaccharide of A.heimuer were separated and purified by breaking cell wall,Sevag technique,and dialysis method.Highperformance gel permeation chromatography(HPGPC),ion chromatography(IC),and infrared spectroscopy(IR)methods were used.The molecular weight,monosaccharide composition and characteristic functional groups were analyzed.The model of oxidative damage to hepatocytes was established to investigate the hepatoprotective activity of AHP.The results show that the AHP sugar content is 83.879%,and the relative molecular weight is 410.3 kD.It is composed of fucose(Fuc),arabinose(Ara),galactose(Gal),glucose(Glc),xylose(Xyl),and mannose(Man),galacturonic acid(GalAc),glucuronic acid(GlcAc)according to the ratio of 2.240∶0.692∶3.994∶22.754∶65.696βtype pyranose composition.After AHP was added into H2 O2 damaged HepG2 cells,it was found that AHP can inhibit nuclear shrinkage and edge feathering of damaged cells,increase the antioxidant capacity of damaged cells such as superoxide dismutase(SOD)and catalase(CAT)Enzyme activity and reduce the production of oxidation products such as nitric oxide(NO).AHP has the activity of protecting liver cells from oxidative damage and has the potential use as liverhealthy food.