Armillaria mellea mycelium polysaccharide with hypoglycemic effect
A mycelium polysaccharide and blood sugar-lowering technology, which is applied to medical preparations containing active ingredients, polysaccharide/gum-containing food ingredients, organic active ingredients, etc., can solve problems such as lowering blood sugar
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Embodiment 1
[0019] Example 1: Preparation of Leucobacterium mycelium polysaccharide
[0020] 1. Inoculate 3-5g of Leucobacteria strains into 300-500mL glucose-potato liquid medium, and culture at 26°C for 20 days to obtain Leucobacteria mycelium.
[0021] 2. Add distilled water to the mycelia of Leucobacteria according to a liquid-to-solid ratio of 1:5, crush it with a flash extractor for 10 minutes, then extract it at 80°C for 3 hours, repeat the extraction three times, pass through a 60-mesh sieve, and combine the extracts;
[0022] 3. Concentrate the obtained extract to 1 / 250 of the original volume, add 4 times the volume of 95% ethanol to a final ethanol concentration of 75%, let stand at 4°C for 12 hours, centrifuge at 4500rpm / min, and collect the precipitate;
[0023] 4. Dissolve the precipitate obtained in step 3 with distilled water, then add an equal volume of Sevag reagent (n-butanol: chloroform = 1:4), stir magnetically for 2 hours, centrifuge to take the supernatant; repeat un...
Embodiment 2
[0024] Example 2: Determination of total sugar and monosaccharide composition of the polysaccharides of Leucobacterium mycelia
[0025] 1. The total sugar content of the polysaccharides in the mycelium of Leucobacterium was determined by the phenol-sulfuric acid method. Accurately draw 2mL of mycelia polysaccharide solution (50μg / ml) and glucose standard solution, add 1mL of 6% phenol and 5mL of concentrated sulfuric acid, set up three parallel groups, measure the absorbance at 490nm, and calculate according to the formula of the standard curve Total sugar content of polysaccharides in Leucobacterium mycelium.
[0026] 2. Analysis of the monosaccharide composition of polysaccharides in the mycelia of Leucobacterium spp. Acid hydrolysis-pre-column PMP derivatization was used to treat the samples, and the samples were analyzed and determined by high-performance liquid chromatography. Accurately weigh the mycelia polysaccharides (10 mg) of Leucobacteria, dissolve them in 5 mL of...
Embodiment 3
[0028] Example 3: Determination of hypoglycemic activity of polysaccharides from Leucobacterium mycelia
[0029] Using the prepared polysaccharides from the mycelium of Leucobacteria as raw materials, the hypoglycemic effect of the polysaccharides from Mycelia from Leucobacteria was evaluated experimentally by using a mouse model of type 2 diabetes.
[0030] 1. Preparation of diabetic mouse model
[0031] Select 32 SPF grade male C57BL / 6J mice, weighing 22±2g, and randomly divide them into 4 groups, 8 mice in each group. The blank group was fed with normal feed, and the other three groups were fed with high-sugar and high-fat diet (HFD). After feeding for 4 weeks, the HFD mice were fasted for 12 hours, and then injected intraperitoneally with 50 mg / kg of streptozotocin (STZ) in 4 small doses to establish a type 2 diabetes model; 72 hours after the model was established, blood was collected from the tail vein to measure the fasting blood sugar for 3 consecutive days Those wit...
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