Mechanism Of Hypoglycemic Effect By Rhizomorph Polysaccharide Of Armillaria Mellea

By different ways of abstracting,separating and purifying from the rhizomorph of Armillaria mellea, two homogeneous polysaccharides (AMP-1 and AMP-2) were obtained. In this paper, AMP-1 was studied furtherly. The main contents of the study were as follows:Abstraction,Separation and purification of the AMP:Armillaria mellea was cultured for some times till the excellent quality was reached,The collected rhizomoph of Armillaria mellea was extracted by aqueous extraction (70℃) depositing in alcohol (95%) and concentration,then purified by chromatography on DEAE-52 cellulose columns, and then recorded the optical density. From the absorption we could identify that AMP-1 was made of homogeneous and AMP-2 contained at least two components.Hypoglycemic and anti-apoptosis effects of AMP-1 in vivo:The hypoglycemic and anti-apoptosis effects of AMP-1 on INS-1 cells injured by alloxan showed that:the model of alloxan diabetic mice was establishmented in vivo. Then we studied the effects of AMP-1 on body weight, liver, spleen, pancreas and dried kidney weight of mouse; blood glucose values were detected by blood-glucose meter, insulin values were evaluated by biochemical analyser, and SOD activity, GSH-Px activity and MDA content in liver,spleen, pancreas and renal glands were measured by kits. The results show that:AMP-1 can antagonize weight reduction and atrophy of liver, pancreas, spleen and kidney gland caused by AXN;SOD, GSH-Px activity increased and MDA content decreased significantly,It was dose-dependable, AMP-1 can reduce blood glucose level and increase the number of islets.The effect of AMP-1 on INS-1 cells injured by alloxan:Different concentrations of AMP-1 (50,100,200,300,400,500mg-L-1) were added into the culture medium of INS-1 cells injured by alloxan. The survival rate was measured by MTT assay. The density of apoptotic cells was observed by PI fluorescence staining. The cell apoptosis rate was detected by flow cytometry. Results showed that:AMP-1 can protect INS-1 cells from AXN and increase the survival rat of INS-1 cells. AMP-1 between 50 mg-L-1 and 500 mg-L-1 could decrease the injury of INS-1 by the AXN, especially at 400mg-L-1. The density of the Apoptotic cells decreased with the increase of the concentration of AMP-1. The rate of the apoptosis of the AXN+AMP-1 groups was lower than the AXN group.Detection of the anti-apoptosis effect of AMP-1 and its mechanism:The cultured INS-1 cells were injured by AXN. We added different concentrations of AMP-1 (50,100,200,300,400,500 mg-L-1) into the culture fluid, then the cell apoptosis rate was detected by flow cytometry and the effect of AMP-1 on the apoptosis of INS-1 cells caused by AXN was studied by detecting the expression of bcl-2 and bax,both of them were measured by western blot. Finally we observed that: AMP-1 between 50 mg-L-1 and 500 mg-L-1 could inhibit the apoptosis of INS-1 cells caused by AXN obviously, this may be related to the raise of bcl-2 and decrease of bax.All the results have shown that:AMP-1 can decrease blood glucose and inhibit apoptosis obviously. The mechanism may be by regulating the levels of apoptosis protein, scavenging free radicals, inhibiting lipid peroxidation and enhancing immune function. Our study provided strong theoretical support to the development of food and medicine of AMP-1.