Ginseng polysaccharide as well as preparation method and application thereof

Abstract

The invention relates to ginseng polysaccharide as well as a preparation method and application thereof. The preparation method comprises the following steps: soaking ginseng slices in water, decocting, filtering, collecting filtrate, decocting filter residues in water, filtering, and collecting filtrate; combining the two filtrates, and concentrating to obtain a concentrated solution; adding 95% ethanol into the concentrated solution, standing and centrifuging, and collecting supernate; adding 95% ethanol into the supernate, standing and centrifuging, and collecting the precipitate; adding water into the precipitate for dissolving, then adding trichloromethane and n-butyl alcohol, shaking and standing, taking supernatant, adding trichloromethane and n-butyl alcohol into the supernatant again, shaking and standing, taking supernatant, repeating for many times, and taking final supernatant; adding 95% ethanol into the final supernatant, standing and centrifuging, and collecting the precipitate; and washing the precipitate with 95% ethanol, absolute ethyl alcohol, isopropanol and diethyl ether in sequence to obtain the ginseng polysaccharide. The ginseng polysaccharide prepared by the preparation method disclosed by the invention is brand new ginseng polysaccharide and has the effects of promoting cell proliferation and repairing cells damaged by ultraviolet rays.

Description

Technical Field

[0001] The invention relates to a preparation method of ginseng polysaccharide, ginseng polysaccharide prepared by the method, and application of the ginseng polysaccharide in promoting cell proliferation and repairing ultraviolet damaged cells, belonging to the technical field of medicine. Background Art

[0002] Ginseng polysaccharide (GP) is one of the important biologically active ingredients of ginseng, a plant of the Araliaceae family. It is a high-molecular-weight glucan with multiple pharmacological effects such as anti-tumor, immunomodulation, anti-radiation, and hypoglycemia. It also has anti-adhesion, anti-viral, anti-oxidation, anti-inflammatory response, and anti-septic effects. It is often used in the comprehensive treatment of various malignant tumors and to reduce the adverse reactions caused by chemotherapy and radiotherapy. At present, there are various preparation / extraction methods for ginseng polysaccharides reported, and the prepared / extracted ginseng polysaccharides and their applications are also different. For example, the invention patent with publication number CN117487035A discloses a ginseng polysaccharide and its preparation method and application. The preparation method of the ginseng polysaccharide includes: crushing ginseng, extracting it by hot water reflux, concentrating and centrifuging, precipitating the supernatant with an ethanol solution, dialyzing the obtained precipitate with distilled water, and freeze-drying the non-dialysate to obtain ginseng polysaccharide; the dialysis molecular weight cutoff is 3.5kDa. The ginseng polysaccharide prepared by this method has the effects of regulating intestinal flora, anti-depression, and anti-epileptic. The experimental results show that the ginseng polysaccharide prepared by the method can be used as a potential drug for treating depression and epilepsy. For another example, the patent with publication number CN113336866A discloses a composite extraction process of ginseng polysaccharide and its application, the composite extraction process comprises the following steps: (1) adding an ionic liquid aqueous solution to dry ginseng powder, the mass volume ratio of the ginseng powder to the ionic liquid aqueous solution being 1:20-40g / mL, and then transferring to a rapid solvent extractor for extraction to obtain an extract; (2) cooling the extract obtained in step (1) and centrifuging to obtain a supernatant, adding anhydrous ethanol 4 times the volume of the supernatant to the supernatant, precipitating for 24 hours, filtering to obtain a precipitate, washing and drying to obtain ginseng polysaccharide. The composite extraction process of ginseng polysaccharide uses a rapid solvent extractor, which has a high degree of automation and short time consumption, and has a higher extraction rate than using a single rapid solvent extractor or a single ionic liquid aqueous solution. The ginseng polysaccharide extracted by the extraction process has a moisturizing effect and can be used as an effective ingredient in cosmetics. However, there are no reports on the effects of ginseng polysaccharides on cell proliferation. Summary of the invention

[0003] The purpose of the present invention is to provide a novel ginseng polysaccharide and a preparation method and application of the ginseng polysaccharide.

[0004] The technical solution of the present invention to achieve the above object is: a method for preparing ginseng polysaccharide, comprising the following steps: (1) Soaking ginseng slices in water, boiling, filtering, collecting the filtrate, boiling the residue in water, filtering, and collecting the filtrate again; (2) combining the filtrates collected twice, concentrating and placing at room temperature to obtain a concentrated solution; (3) Add 95% ethanol to the concentrate, let stand, centrifuge, and collect the supernatant; (4) Add 95% ethanol to the supernatant, let stand, centrifuge, and collect the precipitate; (5) adding water to the precipitate to obtain a precipitate aqueous solution, adding chloroform and n-butanol to the precipitate aqueous solution, shaking (including shaking and other mixing methods commonly used), letting it stand, taking the upper layer liquid, adding chloroform and n-butanol to the upper layer liquid again, shaking, letting it stand, taking the upper layer liquid, repeating the process multiple times, and taking the final upper layer liquid; (6) Add 95% ethanol to the final supernatant, let stand, centrifuge, and collect the precipitate; (7) The precipitate is washed with 95% ethanol, anhydrous ethanol, isopropanol and ether in sequence, and the solvent is evaporated to obtain ginseng polysaccharide.

[0005] Preferably, the ginseng slices are ginseng decoction slices.

[0006] Preferably, in step (1), 1-3 parts by weight of ginseng slices are added with 8-15 parts by weight of water, the soaking time is 0.5-1.5 hours, the water is heated to boiling, the boiling time is 0.5-1.5 hours, the amount of water added to the filter residue is 8-15 parts by weight, and the boiling is carried out for 0.5-1 hour.

[0007] Preferably, in step (1), multiple layers of medical gauze stacked one above the other are used for filtration, and the number of stacked layers of the medical gauze may be three, four or five.

[0008] Preferably, in step (2), the amount of the concentrated solution obtained after the filtrate is concentrated is 2-4 parts by weight.

[0009] Preferably, the amount of 95% ethanol added in step (3) and step (4) is 0.23-0.29 times the volume of the concentrated solution in step (3).

[0010] Preferably, the standing time in step (3) is 10-12 hours.

[0011] Preferably, the centrifugal speed in step (3) is 3000 r / min and the centrifugal time is 10 min to remove the polysaccharide components with larger molecular weight in the concentrate.

[0012] Preferably, the standing time in step (4) is 12 hours.

[0013] Preferably, the centrifugal speed in step (4) is 3000 r / min and the centrifugal time is 10 min to remove the polysaccharide components with smaller molecular weight in the concentrate.

[0014] Preferably, in step (5), the precipitate is weighed, and the amount of water added to the precipitate is 10 times the weight of the precipitate.

[0015] Preferably, in step (5), the amount of chloroform added each time is 0.16-0.2 times the mass of the precipitate aqueous solution, and the amount of n-butanol added each time is 0.04-0.08 times the mass of the precipitate aqueous solution. For example, taking 5 parts by mass of the precipitate aqueous solution, the amount of chloroform added each time is 0.8-1 parts by mass, and the amount of n-butanol added each time is 0.2-0.4 parts by mass.

[0016] Preferably, in step (5), the standing time is 12 hours each time, the shaking speed is 140 r / min each time, and the shaking time is 10 minutes.

[0017] Preferably, in step (5), chloroform and n-butanol are added repeatedly, and the upper layer liquid is taken after shaking and standing for no less than 5 times.

[0018] Preferably, in step (6), the volume of the final upper layer liquid is measured, and the amount of 95% ethanol added to the final upper layer liquid is 0.7-0.76 times the volume of the final upper layer liquid.

[0019] Preferably, the standing time in step (6) is 12 h, the centrifugal speed is 3000 r / min, and the centrifugal time is 10 min.

[0020] Preferably, in step (7), the mass of the precipitate is weighed, and the amount of 95% ethanol, anhydrous ethanol, isopropanol and ether used to wash the precipitate is the same as the mass of the precipitate.

[0021] The centrifugation in step (3), step (4) and step (6) can be performed using any suitable centrifugation device (e.g., a centrifuge) in the prior art, and the shaking in step (5) can be performed using any suitable shaking device (e.g., a shaker) in the prior art.

[0022] The ginseng polysaccharide is prepared by any of the ginseng polysaccharide preparation methods disclosed in the invention.

[0023] The peak molecular weight, number average molecular weight and weight average molecular weight of the ginseng polysaccharide are 4646 g / mol, 19895 g / mol and 246356 g / mol, respectively.

[0024] The invention discloses an application of any ginseng polysaccharide in cell proliferation.

[0025] The invention discloses an application of any ginseng polysaccharide in repairing cells damaged by ultraviolet rays.

[0026] The beneficial effects of the present invention are as follows: experiments and analysis show that the ginseng polysaccharide prepared by the ginseng polysaccharide preparation method of the present invention is a brand-new ginseng polysaccharide different from the existing ginseng polysaccharide, which is composed of glucose, and has a peak molecular weight, a number average molecular weight and a weight average molecular weight of 4646 g / mol, 19895 g / mol and 246356 g / mol, respectively, and has the effects of promoting cell proliferation and repair of ultraviolet-damaged cells. The experimental results show that it can be used as a potential drug for clinical applications related to cell proliferation and repair of ultraviolet-damaged cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is an overlay graph (mixed spectrum) of the HPLC analysis results of the monosaccharide reference substance PMP derivatization product in the monosaccharide composition analysis test of the present invention; Figure 2 This is a diagram showing the results of monosaccharide composition analysis of ginseng polysaccharides of the present invention; Figure 3 It is a comparison chart of the effects of ginseng polysaccharide solutions of different concentrations on fibroblast proliferation; Figure 4 It is a comparison chart of the effects of ginseng polysaccharide solutions of different concentrations on the proliferation of ultraviolet-damaged fibroblasts. DETAILED DESCRIPTION

[0028] The invention discloses a method for preparing ginseng polysaccharide. The ginseng polysaccharide prepared by the method is a brand-new ginseng polysaccharide.

[0029] Preparation of Ginseng Polysaccharide Embodiment 1: (1) Take 1 kg of ginseng slices, add 10 kg of water, soak for 1.5 h, heat to boiling, boil for 0.5 h, filter through four layers of medical gauze stacked up and down, collect the filtrate, add 15 kg of water to the residue, boil for 0.8 h, filter through four layers of medical gauze stacked up and down, and collect the filtrate again; (2) Combine the filtrates collected twice, concentrate to 2 kg, and place at room temperature to obtain a concentrated solution; (3) Measure the volume of the concentrate, add 0.25 times the volume of 95% ethanol to the concentrate, let it stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the supernatant; (4) Add 0.25 times the volume of the concentrate to the supernatant, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (5) Weigh the precipitate, add 10 times the mass of water to obtain an aqueous solution of the precipitate, take 5 kg of the aqueous solution of the precipitate, add 0.8 kg of chloroform and 0.3 kg of n-butanol, shake at a shaking speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, add 0.8 kg of chloroform and 0.3 kg of n-butanol to the absorbed upper layer liquid again, shake at a shaking speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, repeat this operation 5 times, and finally absorb the upper layer liquid to obtain the final upper layer liquid; (6) Measure the volume of the final supernatant, add 0.7 times the volume of 95% ethanol to the final supernatant, let it stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (7) Weigh the weight of the precipitate, and wash the precipitate with 95% ethanol, anhydrous ethanol, isopropanol and ether in the same amount as the precipitate. After washing, evaporate the solvent to obtain ginseng polysaccharide.

[0030] Embodiment 2: (1) Take 2 kg of ginseng slices, add 15 kg of water, soak for 0.5 h, heat to boiling, boil for 1 h, filter through three layers of medical gauze stacked up and down, collect the filtrate, add 8 kg of water to the residue, boil for 1 h, filter through three layers of medical gauze stacked up and down, and collect the filtrate again; (2) Combine the filtrates collected twice, concentrate to 3 kg, and place at room temperature to obtain a concentrated solution; (3) Measure the volume of the concentrate, add 0.29 times the volume of 95% ethanol to the concentrate, let it stand for 10 h, centrifuge at 3000 r / min for 10 min, and collect the supernatant; (4) Add 0.29 times the volume of the concentrate to the supernatant, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (5) Weigh the precipitate, add 10 times the mass of water to obtain a precipitate aqueous solution, take 5 kg of the precipitate aqueous solution, add 0.9 kg of chloroform and 0.4 kg of n-butanol, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, add 0.9 kg of chloroform and 0.4 kg of n-butanol to the absorbed upper layer liquid again, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, repeat this operation 6 times, and finally absorb the upper layer liquid to obtain the final upper layer liquid; (6) Measure the volume of the final supernatant, add 0.72 times the volume of the final supernatant to the final supernatant, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (7) Weigh the weight of the precipitate, and wash the precipitate with 95% ethanol, anhydrous ethanol, isopropanol and ether in the same amount as the precipitate. After washing, evaporate the solvent to obtain ginseng polysaccharide.

[0031] Embodiment three: (1) Take 3 kg of ginseng slices, add 8 kg of water, soak for 1 hour, heat to boiling, boil for 1.5 hours, filter through five layers of medical gauze stacked up and down, collect the filtrate, add 10 kg of water to the residue, boil for 0.5 hour, filter through five layers of medical gauze stacked up and down, and collect the filtrate again; (2) Combine the filtrates collected twice, concentrate to 4 kg, and place at room temperature to obtain a concentrated solution; (3) Measure the volume of the concentrate, add 0.23 times the volume of 95% ethanol to the concentrate, let it stand for 11 h, centrifuge at 3000 r / min for 10 min, and collect the supernatant; (4) Add 0.23 times the volume of the concentrate to the supernatant, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (5) Weigh the precipitate, add 10 times the mass of water to obtain a precipitate aqueous solution, take 5 kg of the precipitate aqueous solution, add 1 kg of chloroform and 0.2 kg of n-butanol, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, add 1 kg of chloroform and 0.2 kg of n-butanol to the absorbed upper layer liquid again, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, repeat this operation 7 times, and finally absorb the upper layer liquid to obtain the final upper layer liquid; (6) Measure the volume of the final supernatant, add 0.76 times the volume of the final supernatant to the final supernatant, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (7) Weigh the weight of the precipitate, and wash the precipitate with 95% ethanol, anhydrous ethanol, isopropanol and ether in the same amount as the precipitate. After washing, evaporate the solvent to obtain ginseng polysaccharide.

[0032] Embodiment 4: (1) Take 2 kg of ginseng slices, add 30 L of water, soak for 1 hour, heat to boiling, boil for 1.5 hours, filter through four layers of medical gauze stacked up and down, collect the filtrate, add 20 L of water to the residue, boil for 1 hour, filter through four layers of medical gauze stacked up and down, and collect the filtrate again; (2) Combine the filtrates collected twice, concentrate to 4 kg, and place at room temperature to obtain a concentrated solution; (3) Add water to the concentrate to a total volume of 2.6 L, add 0.676 L of 95% ethanol, let stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the supernatant; (4) Add 0.676 L of 95% ethanol to the supernatant, let it stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (5) Weigh the precipitate, add 10 times the mass of water to obtain a precipitate aqueous solution, take 2 L of the precipitate aqueous solution, add 0.32 L of chloroform and 0.8 L of n-butanol, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, add 0.32 L of chloroform and 0.8 L of n-butanol to the absorbed upper layer liquid again, shake at a speed of 140 r / min for 10 min, let it stand for 12 h, absorb the upper layer liquid, repeat this operation 5 times, and finally absorb the upper layer liquid to obtain the final upper layer liquid; (6) Take 1.9 L of the final upper layer liquid, add 95% ethanol until the alcohol content reaches 40%, let it stand for 12 h, centrifuge at 3000 r / min for 10 min, and collect the precipitate; (7) Weigh the weight of the precipitate, and wash the precipitate with 95% ethanol, anhydrous ethanol, isopropanol and ether in the same amount as the precipitate. After washing, evaporate the solvent to obtain ginseng polysaccharide.

[0033] Analysis of Chemical Characteristics of Ginseng Polysaccharides 1. Molecular weight distribution analysis test: Chromatographic conditions Column: TSK ® Gel chromatography column guard column PW XL (40mm × 6.0mm) Series TSK ® G5000PW XL Gel chromatography column (300mm × 7.8mm) connected in series with TSK ® G3000PW XL Gel chromatography column (300mm × 7.8mm); mobile phase: 0.01% NaCl solution; column temperature: 30°C; detector: differential refractometer; flow rate: 0.6mL / min; injection volume: 50μL.

[0034] Preparation of reference solution Weigh 2 mg of Dextran reference substances (1000, 5000, 80000, 410000, 1100000 Da) respectively, add 1 mL of 0.01% NaCl solution to make a solution, filter with a 0.45 μm microporous filter membrane, and take the filtrate to obtain the solution.

[0035] Preparation of test solution Weigh 2 mg of the ginseng polysaccharide prepared in Example 4, add 1 mL of 0.01% NaCl solution to prepare a solution, filter with a 0.45 μm microporous filter membrane, and take the filtrate to obtain the solution.

[0036] Sample measurement and results 50 μL of the reference solution and the test solution were accurately drawn, respectively, and injected into the liquid chromatograph for determination. The GPC software was calibrated using the weight average molecular weight (y) and retention time (x) of the series of reference substances. The GPC software was used to statistically analyze the chromatogram of the test sample and fit the molecular weight distribution of the polysaccharide. The peak molecular weight, number average molecular weight and weight average molecular weight of the ginseng polysaccharide prepared by the preparation method of the ginseng polysaccharide of the present invention were 4646 g / mol, 19895 g / mol and 246356 g / mol, respectively.

[0037] 2. Monosaccharide composition analysis test: Chromatographic conditions Chromatographic column: C18 column (250 mm × 4.6 mm, 5 μm); column temperature: 30 ℃; mobile phase: acetonitrile-4.5% NaAc solution (18:82); flow rate: 1 mL / min; detector: UV detector (detection wavelength is 245 nm); injection volume: 10 μL.

[0038] Preparation of reference solution Weigh 1 mg of arabinose and place it in a stoppered test tube. Add 0.3 mL of water to dissolve it, add 0.4 mL of 0.3 mol / L NaOH solution and 0.3 mol / L PMP methanol solution, mix well, stopper tightly, place in an oven, and keep at 70°C for 1 hour; take out, cool, add 0.4 mL of 0.3 mol / L HCl solution, and mix well. Wash with chloroform 3 times, 0.6 mL each time, and discard the chloroform solution; filter the water layer with a 0.45 μm microporous filter membrane, and take the filtrate to obtain. Prepare galactose, glucose, rhamnose, mannose, glucuronic acid, and galacturonic acid reference solutions in the same way.

[0039] Preparation of test solution Weigh 2 mg of ginseng polysaccharide prepared in Example 4, accurately weigh it, and place it in a stoppered test tube. Add 2 mL of 2 mol / L TFA solution, stopper it, and place it in an oven at 115°C for 6 hours. Reduce the pressure to recover the solvent from the hydrolyzate until it is dry, add 1 mL of water to the residue to dissolve it, and reduce the pressure to recover the solvent again until it is dry. Repeat the above operation until the residue has no obvious sour taste. Add 0.3 mL of water to the residue to dissolve it, and prepare the test solution according to the method for preparing the reference solution, starting from the step of "adding 0.4 mL of 0.3 mol / L NaOH solution and 0.3 mol / L PMP methanol solution".

[0040] Sample measurement and results Accurately pipette 10 μL of the reference solution and the test solution respectively, inject into the liquid chromatograph, and measure. Superimpose the chromatograms of each monosaccharide reference substance to obtain the reference substance mixed spectrum, such as Figure 1 As shown in the figure, 1 is D-mannose, 2 is L-rhamnose, 3 is D-glucuronic acid, 4 is D-galacturonic acid, 5 is D-glucose, 6 is D-galactose, and 7 is L-arabinose. The chromatogram of the test sample is compared with the mixed spectrum of the monosaccharide reference substance, and the monosaccharide composition of the ginseng polysaccharide prepared by the preparation method of the ginseng polysaccharide of the present invention is determined by the retention time of each chromatographic peak. The results show that the ginseng polysaccharide prepared by the preparation method of the ginseng polysaccharide of the present invention is composed of glucose, such as Figure 2 As shown in the figure, 5 is D-glucose.

[0041] 3. Specific rotation test Weigh 30 mg of ginseng polysaccharide prepared in Example 4, add 30 ml of water, and prepare a polysaccharide solution. Use SGW®-1 automatic polarimeter to detect the specific rotation of ginseng polysaccharide sample. Use D sodium light for measurement, the detection wavelength is 589.4 nm, the test tube length is 20 dm, and the sample temperature is 25.7°C. First use purified water to zero the automatic polarimeter, and then measure the specific rotation of the sample. Specific rotation [α] = +158.3.

[0042] Analysis of Pharmacological Activity of Ginseng Polysaccharide 1. Preparation of ginseng polysaccharide solution Weigh 1 mg of the ginseng polysaccharide prepared in Example 4, add 1 mL of water to prepare a mother solution with a concentration of 1 mg / mL, and store it at -20°C for later use.

[0043] 2. Experiment on the effect of ginseng polysaccharides on cell proliferation (taking fibroblast NIH3T3 as an example) Culture cells routinely. When the cells grow to 80% confluence under microscope, digest and resuspend the cells and add 5× 10 4 The cells were inoculated in 96-well plates at a density of 100 μL / mL. The cells were starved for 24 h and randomly divided into a (blank) control group and a drug-treated group (low, medium, and high concentration groups), with 6 wells in each group.

[0044] When the cells in the 96-well plate grew to 40% confluence under the microscope, the culture medium was aspirated and discarded. The (blank) control group was only added with culture medium, and the drug groups were given with concentrations of 6.4×10 -4 g / L、1.6×10 -2 g / L, 4.0×10 -1g / L ginseng polysaccharide solution, 100μL per well, culture for 24h. Add 10μL CCK-8 solution to each well, incubate in the cell culture incubator for 2h, and measure the absorbance at 450nm with an enzyme marker. Cell proliferation rate = (OD value of the drug group - OD value of the blank control group) / OD value of the blank control group × 100%. Comparison of cell survival rates in each group Figure 3 shown.

[0045] 3. Experiment on the effect of ginseng polysaccharide on the repair of UV-damaged cells (taking fibroblast NIH3T3 as an example) When the cells grew to 80% confluence under microscope, they were digested and resuspended, and 5 × 10 4 The cells were seeded in 96-well plates at a density of 100 μL / mL. The cells were starved for 24 h and randomly divided into a (blank) control group, a damaged control group (UVA group), and a drug-treated group (low, medium, and high concentration groups), with 6 wells in each group.

[0046] When the cells in the 96-well plate grew to 40% confluence under the microscope, UVA was used to establish the model in the injury control group and the drug-treated group. The culture medium was aspirated and discarded, and only the culture medium was added to the blank control group and the injury control group. The drug-treated group was given a concentration of 2.0×10 -4 g / L, 1.0×10 -4 g / L, 5.0×10 -5 g / L ginseng polysaccharide solution, 100μL per well, culture for 24h. Add 10μL CCK-8 solution to each well, incubate in a cell culture incubator for 2h, and measure the absorbance at 450nm using an enzyme marker. Cell proliferation rate = (OD value of the injury control group or drug administration group - OD value of the blank control group) / OD value of the blank control group × 100%. Comparison of cell survival rates in each group Figure 4 shown.

[0047] Conclusion The pharmacological activity of ginseng polysaccharide prepared by the preparation method of ginseng polysaccharide of the present invention was analyzed, and it was found that ginseng polysaccharide solution had a promoting effect on cell proliferation, and when the concentration of ginseng polysaccharide solution was 6.4×10 -4 g / L, the cell proliferation promoting effect was the strongest. It was found that ginseng polysaccharide solution had a promoting effect on fibroblasts after UV killing, and when the concentration of ginseng polysaccharide solution was 1.0×10 -4 g / L, it has the strongest effect on promoting the proliferation of fibroblasts killed by ultraviolet rays.

Claims

1. A method for preparing ginseng polysaccharide, characterized in that The following steps are involved: (1) Soaking ginseng slices in water, boiling, filtering, collecting the filtrate, boiling the residue in water, filtering, and collecting the filtrate again; (2) combining the filtrates collected twice, concentrating and placing at room temperature to obtain a concentrated solution; (3) Add 95% ethanol to the concentrate, let stand, centrifuge, and collect the supernatant; (4) Add 95% ethanol to the supernatant, let stand, centrifuge, and collect the precipitate; (5) Add water to the precipitate to obtain a precipitate aqueous solution, add chloroform and n-butanol to the precipitate aqueous solution, shake, let stand, take the upper layer, add chloroform and n-butanol to the upper layer again, shake, let stand, take the upper layer, repeat several times, and take the final upper layer; (6) Add 95% ethanol to the final supernatant, let stand, centrifuge, and collect the precipitate; (7) The precipitate is washed with 95% ethanol, anhydrous ethanol, isopropanol and ether in sequence, and the solvent is evaporated to obtain ginseng polysaccharide.

2. The method for preparing ginseng polysaccharide according to claim 1, characterized in that The amount of 95% ethanol added in step (3) and step (4) is 0.23-0.29 times the volume of the concentrated solution in step (3).

3. The method for preparing ginseng polysaccharide according to claim 1, characterized in that The amount of water added to the precipitate in step (5) is 10 times the amount of the precipitate.

4. The method for preparing ginseng polysaccharide according to claim 1, characterized in that In the step (5), the amount of chloroform added each time is 0.16-0.2 times the mass of the precipitate aqueous solution, and the amount of n-butanol added each time is 0.04-0.08 times the mass of the precipitate aqueous solution.

5. The method for preparing ginseng polysaccharide according to claim 1, characterized in that The amount of 95% ethanol added in step (6) is 0.7-0.76 times the volume of the final upper layer liquid.

6. The method for preparing ginseng polysaccharide according to claim 1, characterized in that The amounts of 95% ethanol, anhydrous ethanol, isopropanol and ether used to wash the precipitate in step (7) are the same as the mass of the precipitate.

7. The method for preparing ginseng polysaccharide according to claim 1, characterized in that The centrifugal speed in step (3), step (4) and step (6) is 3000 r / min, and the centrifugal time is 10 min. The shaking speed in step (5) is 140 r / min, and the shaking time is 10 min.

8. Ginseng polysaccharide, characterized in that The ginseng polysaccharide is prepared by the preparation method of any one of claims 1 to 7.

9. Use of the ginseng polysaccharide according to claim 8 in cell proliferation.

10. Use of the ginseng polysaccharide according to claim 8 in repairing cells damaged by ultraviolet rays.

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