八种食用菌水溶性粗多糖的β-葡聚糖含量与体外免疫活性研究

β-Glucan Content and In Vitro Immunological Activity of Crude Water-soluble Polysaccharide Fractions from Eight Edible Fungi

采用水提醇沉法提取了8种不同食用菌子实体的水溶性粗多糖,测定了样品中多糖、葡聚糖和β-葡聚糖的含量,结果表明,刺芹侧耳(Pleurotus eryngii)、毛头鬼伞(Coprinus comatus)、香菇(Lentinula edodes)和桑黄(Phellinus baumii)粗多糖中的多糖含量较高,香菇和姬松茸(Agaricus blazei)粗多糖中葡聚糖含量较高;猴头菌(Hericium erinaceus)、刺芹侧耳、香菇和姬松茸粗多糖中β-葡聚糖含量较高。采用截留分子量8000～12000透析袋纯化8种食用菌粗多糖,将纯化产物用于体外刺激巨噬细胞RAW264.7释放NO实验和脾淋巴细胞增殖实验,8种样品均能刺激巨噬细胞释放NO,其中香菇、刺芹侧耳和毛头鬼伞粗多糖在低浓度(200μg/mL)时,刺激巨噬细胞释放NO量即可达到40μmol/L以上,高浓度(500μg/mL)时高于阳性对照;体外脾淋巴细胞增殖实验结果显示,8种供试样品均能刺激脾淋巴细胞增殖,以香菇、刺芹侧耳和猴头菌粗多糖效果较好。实验结果证明这8种食用菌粗多糖均具有体外免疫活性。

Crude polysaccharide fractions were prepared from the fruit bodies of eight different edible fungi（Hericium erinaceus,Pleurotus eryngii,Coprinus comatus,Lentinula edodes,Grifola frondosa,Phellinus baumii,Ganoderma lucidum,Agaricus blazei） by hot（100 ℃） water extraction followed by precipitation with ethanol,and total glucan and β-glucan levels were determined.Crude fractions from P.eryngii,C.comatus,L.edodes and P.baumii contained a higher percentage of polysaccharide,while L.edodes and A.blazei extracts contained a higher percentage of total glucan and extracts of H.erinaceus,P.eryngii,L.edodes and A.blazei contained a higher percentage of β-glucan.Solutions of the crude extracts（1 g in 50 mL ddH2O） were partially purified by exhaustive dialysis（72 h at 4 ℃ against 18 changes of ddH2O） using an 8000~12000 MW cut-off membrane.In vitro testing revealed that NO production by RAW264.7 macrophages was significantly enhanced by all the partially purified fractions（PPS） at 200 μg/mL and 500 μg/mL concentrations.NO production levels（55-65 μmol/L） generated by PPS（500 μg/mL） derived from L.edodes,P.eryngii and C.comatus were higher than that recorded with the lipopolysaccharide positive control.Similarly,all the PPS fractions stimulated splenocyte proliferation,with samples derived from L.edodes,P.eryngii,A.blazei and H.erinaceus the most effective.