桦褐孔菌纯化多糖体外降血糖活性研究

Hypoglycemic Activity of Purified Polysaccharides from Inonotus obliquus in Vitro

采用水提醇沉法得到桦褐孔菌发酵粗多糖,将粗多糖过DEAE-52纤维素柱分离纯化得到两种多糖(EIOP1、EIOP2),本文以α-葡萄糖苷酶抑制活性、正常HepG2细胞及胰岛素抵抗HepG2细胞的单位细胞葡萄糖消耗量为主要指标,探究桦褐孔菌两种纯化多糖不同浓度(10、20、40、80、160和320μg/mL)的降血糖活性,其中α-葡萄糖苷酶抑制活性以阿卡波糖作为阳性对照,葡萄糖消耗实验以二甲双胍作为阳性对照。结果表明,EIOP1、EIOP2的α-葡萄糖苷酶抑制活性均高于阳性对照组阿卡波糖与粗多糖,IC50值分别为39.18、29.87μg/mL。EIOP1在浓度为80μg/mL时,对HepG2细胞的葡萄糖消耗量极显著高于对照组,促进效果最好,比对照组提高了30.62%(P<0.01);EIOP2在浓度在40μg/mL时,葡萄糖消耗量极显著高于对照组,比对照提高了75.99%(P<0.01);对胰岛素抵抗HepG2细胞的葡萄糖消耗实验发现EIOP1在浓度为40μg/mL时,促进效果最好,比胰岛素抵抗组提高了30.00%,EIOP2在浓度为80μg/mL时,促进效果最好,比胰岛素抵抗组提高了90.49%,高于Met组(P<0.01)。因此,桦褐孔菌纯化多糖对正常HepG2细胞和胰岛素抵抗HepG2细胞的葡萄糖消耗均具有促进作用,对α-葡萄糖苷酶的抑制活性显著高于粗多糖。

The crude polysaccharides from Inonotus obliquus were obtained by water extraction and alcohol precipitation.And the crude polysaccharides were separated and purified from DEAE-52 cellulose column to obtain two kinds of polysaccharides(EIOP1 and EIOP2).A-Glucosidase inhibitory activity,glucose consumption of normal HepG2 cells,and insulin-resistant HepG2 cells were used as the main indicators to investigate the hypoglycemic activity of different concentrations of two purified polysaccharides from Inonotus obliquus(10,20,40,80,160,and 320μg/mL).α-glucosidase inhibitory activity used acarbose as a positive control,and glucose consumption experiment was used with metformin as a positive control.The results showed that theα-glucosidase inhibitory activity of EIOP1 and EIOP2 were higher than that of the positive control group acarbose and crude polysaccharide,and the IC50 values were 39.18 and 29.87μg/mL,respectively.The concentration of EIOP1 on HepG2 cells was extramly significant higher than that of the control group at a concentration of 80μg/mL(P<0.01).The promotion effect was the best,which was 30.62%higher than that of the control group.When the concentration of EIOP2 was 40μg/mL,the glucose consumption was extramly significant higher than that of the control group,which was 75.99%higher than the control(P<0.01).The glucose consumption experiment of IR-HepG2 cells showed that EIOP1 had the best promotion effect at the concentration of 40μg/mL,which was 30.00%higher than that of the IR group.The concentration of EIOP2 was the best at the concentration of 80μg/mL.The IR group increased by 90.49%,which was higher than that of the Met group(P<0.01).Therefore,the purified polysaccharide of Inonotus obliquus would had a promoting effect on glucose consumption in normal HepG2 cells and insulin-resistant HepG2 cells,and better inhibitory activity againstα-glucosidase than the crude polysaccharides.