Extraction process of Grifola frondosa polysaccharide and its application in preparation of medicine for repairing gastric mucosal trauma
A technique of extracting polysaccharides from Grifola frondosa, which is applied in the direction of drug combinations, medical preparations containing active ingredients, and pharmaceutical formulas, to achieve the effects of promoting proliferation, promoting healing and repair, and avoiding hydrolysis loss
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Embodiment 1
[0031] Embodiment 1: Extraction of Grifola frondosa polysaccharide
[0032]Take the dry fruiting body of Grifola frondosa, pass through a 10-mesh sieve to obtain fruiting body particles of Grifola frondosa, add 10 times the amount (mass to volume ratio) of water and boil for 2 hours, filter, and concentrate the filtrate until Brix (soluble solids) ≥ 15 %, after centrifugation at 10000rpm for 10min, the supernatant is pumped successively into ultrafiltration separation modules with molecular weight cutoffs of 100kDa, 30kDa, and 5kDa, that is, the filtrate from the ultrafiltration separation module with a molecular weight cutoff of 100kDa continues to be pumped into an ultrafiltration separation module with a molecular weight cutoff of 30kDa , the filtrate of the ultrafiltration separation module with a molecular weight cut-off of 30kDa continues to be pumped into the ultrafiltration separation module with a molecular weight cut-off of 5kDa, and the retentate and filtrate of each...
Embodiment 2
[0035] Example 2: Effects of Grifola frondosa polysaccharides on the proliferation of human gastric mucosal epithelial cells
[0036] 1. Materials
[0037] Cell line: human normal gastric mucosal epithelial cells GES-1 were purchased from Beijing Institute of Cancer Prevention and Control.
[0038] Medium: The basal medium for GES-1 cell culture is DMEM medium (10% FBS, 100U penicillin, 100U streptomycin).
[0039] Grifola frondosa polysaccharide GFP (polysaccharide of Grifola frondosa): the Grifola frondosa polysaccharide prepared in Example 1 was used.
[0040] 2. Method
[0041] The effect of GFP on the proliferation activity of GES-1 was detected by MTS method. GES-1 in the logarithmic growth phase was digested with trypsin and its concentration was adjusted to 2.0×10 4 cells / mL, seeded in a 96-well plate, 100 μL per well, placed at 37°C, saturated humidity, 5% CO 2 After 24 hours of culture in a sterile incubator, the old culture medium was discarded. The experimenta...
Embodiment 3
[0047] Example 3: Effects of Grifola frondosa polysaccharides on human gastric mucosal wound healing and repair
[0048] 1. Material: Same as Example 2
[0049] 2. Method
[0050] Select GES-1 in the logarithmic growth phase, and use 6×10 per well 4 Cells were seeded in 24-well plates, 400 μL per well. Set at 37°C, 5% CO 2 Cultivate in the incubator for 24 hours. After the cells form a relatively tight connection to form a single layer, use a capillary tube with a diameter of 1.5 mm to draw a vertical trace in the center of each well of the 24-well plate, and the two sides of the wound can be seen under the microscope at the same time. side. Aspirate the original culture medium after scratching, wash each well twice with 1mL PBS to remove non-adherent cells, select the final concentration of GFP as 2, 0.4, 0.08mg / mL with reference to the results of the proliferation experiment, and culture medium 400μL, the control group only Add 400 μL of DMEM basal medium. For each con...
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