Ganoderma lucidum polysaccharide with light color and high molecular weight, and preparation method thereof
A technology of high molecular weight and Ganoderma lucidum polysaccharide, which is applied in the direction of antineoplastic drugs, drug combinations, allergic diseases, etc., can solve the problems of unfavorable large-scale production of product quality, difficult to control product quality, and harmful organic solvent residues, etc., to achieve enhanced anti-tumor The effect of high capacity, uniform distribution and high content of active ingredients
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Embodiment 1
[0021] 1. Extraction of Ganoderma lucidum crude polysaccharides: Ganoderma lucidum fruiting body (Ganoderma lucidum strain with the strain number 2442 preserved by the Culture Collection Center of Edible Fungi Research Institute, Shanghai Academy of Agricultural Sciences, obtained through artificial cultivation) as raw material, crushed into coarse particles , Weigh 2kg, add 20L of water, soak at room temperature for 30min, heat to boiling, keep slightly boiling for 90min, and filter. Repeat the above steps for the filter residue, extract twice more, and combine the filtrate.
[0022] 2. Concentration of the crude extract of Ganoderma lucidum: the filtrate is coarsely filtered with a 120-mesh filter (purchased from Shanghai Xinhua Wire Mesh Store), and then filtered with a 100-micron membrane (purchased from Shanghai Freet Company), and the filtrate is concentrated under reduced pressure to 3L.
[0023] 3. Ethanol precipitation and washing: slowly add absolute ethanol to the conce...
Embodiment 2
[0028] 1. Extraction of Ganoderma lucidum crude polysaccharides: Ganoderma lucidum fruiting body (strain number of Ganoderma 2442 preserved by the Culture Collection Center of Edible Fungi Research Institute, Shanghai Academy of Agricultural Sciences, obtained by artificial cultivation) as raw material, crushed into coarse particles , Weigh 1.5kg, add 20L of water, soak for 30min at room temperature, heat to boiling, keep slightly boiling for 60min, and filter. Repeat the above steps for the filter residue, extract once more, and combine the filtrate.
[0029] 2. Concentration of the crude extract of Ganoderma lucidum: the filtrate is coarsely filtered with a 120-mesh filter (purchased from Shanghai Xinhua Wire Mesh Store), and then filtered with a 100-micron membrane (purchased from Shanghai Freet Company), and the filtrate is concentrated under reduced pressure to 2L.
[0030] 3. Ethanol precipitation and washing: slowly add absolute ethanol to the concentrated solution, stirrin...
Embodiment 3
[0035] Determination of the activity of Ganoderma lucidum polysaccharide to stimulate macrophages to release NO in vitro:
[0036] 1. Sample preparation: accurately weigh the Ganoderma lucidum polysaccharide samples prepared in Example 1 and Example 2 into a sterilized eppendorf tube, and prepare a sample solution with a concentration of 5 mg / mL with sterile PBS. After fully dissolved, centrifuge at 15000r / min for 30min, transfer the supernatant to a new sterile eppendorf tube under aseptic conditions, and dilute the sample to 2mg / ml, 0.5mg / ml for later use.
[0037] 2. Cell culture: Take the logarithmic growth phase RAW264.7 macrophage cell line (purchased from the Institute of Cells, Chinese Academy of Sciences), and use DMEM complete medium (purchased from Gibco) at 37°C with 5% CO 2 Subculture under conditions, digest with 0.05% trypsin or 5% EDTA solution, and collect the cells after centrifugation at 1000 rpm / min for 3 minutes, and count for use.
[0038] 3. Reagent preparation...
PUM
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