Ganoderma lucidum fermentation product as well as preparation method and application thereof

Abstract

The invention relates to a ganoderma lucidum fermentation product and a preparation method and application thereof, and belongs to the field of medicine and health care products. The ganoderma lucidum fermentation product is prepared from total sugar, fat, total protein, polypeptide, vitamin A, vitamin E, vitamin C and total terpenoids according to the mass ratio of (2500 to 5000) to (100 to 500) to (1500 to 3000) to (100 to 500) to (1 to 50) to (10 to 50) to (20 to 100) to (200 to 500). The ganoderma lucidum fermentation product disclosed by the invention can improve lipid oxidation caused by radiation, so that the effects of maintaining body health and the like are achieved; the compound can be used for preparing medicines or health-care products for preventing and treating radiation.

Description

Technical Field

[0001] The present invention relates to the field of medicines and health care products, and in particular to a ganoderma lucidum fermentation product and a preparation method and application thereof. Background Art

[0002] As Ganoderma lucidum cultivation technology matures, a large amount of Ganoderma lucidum waste is discarded each year, polluting the surrounding environment and causing a significant waste of resources. Currently, Ganoderma lucidum waste is primarily used to replace part of the concentrate or coarse feed in various livestock and poultry feeds, as fertilizer, or to extract Ganoderma lucidum triterpenes and polysaccharides. It has no medical applications.

[0003] Chinese patent CN109096407A discloses a method for extracting total triterpenes and polysaccharides from Ganoderma lucidum fungus residue. The method comprises: crushing the Ganoderma lucidum fungus residue, adding ethanol, performing ultrasonic extraction using an ultrasonic cell disruptor, and centrifuging after ultrasonic extraction to obtain a filter residue and a clear liquid; concentrating the clear liquid, extracting it with chloroform, concentrating the chloroform layer by reduced pressure evaporation to a paste, and recovering the chloroform; washing the paste with ethanol, and then evaporating the ethanol to obtain the total triterpenes from the Ganoderma lucidum fungus residue; leaching the filter residue with water, filtering after the extraction to obtain a filtrate; concentrating the filtrate, adding ethanol to precipitate, desalting, drying, and crushing the precipitate to obtain Ganoderma lucidum fungus residue polysaccharides. This method requires complex processing of the Ganoderma lucidum fungus residue to obtain the total triterpenes and polysaccharides, and the remaining components are discarded, resulting in a waste of resources.

[0004] Jiang Hongmei et al. published in the Chinese Journal of Gerontology the protective effects of Ganoderma lucidum spore oil on radiation-damaged elderly mice. The oil improved the weight, peripheral blood leukocyte count, bone marrow DNA content, and micronucleus rate of the irradiated mice. As a protective agent, Ganoderma lucidum spore oil significantly increased peripheral blood leukocyte count, bone marrow DNA content, thymus / spleen coefficients, and blood / tissue SOD activity, while significantly reducing the bone marrow micronucleus rate. This suggests that an appropriate amount of Ganoderma lucidum spore oil can antagonize radiation-induced damage. Summary of the Invention

[0005] In response to the deficiencies in the prior art, the present invention provides a Ganoderma lucidum fermented product, a preparation method thereof, and an application thereof. The Ganoderma lucidum fermented product of the present invention has the function of preventing lipid oxidation and DNA damage caused by radiation, while increasing spleen weight and enhancing the body's immune function to alleviate the damage caused by radiation to the body. It can also effectively reduce resource waste and achieve high-value utilization of Ganoderma lucidum fungus bran.

[0006] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a Ganoderma lucidum fermentation product, comprising total sugar, fat, total protein, polypeptide, vitamin A, vitamin E, vitamin C and total terpenes in a mass ratio of 2500-5000: 100-500: 1500-3000: 100-500: 1-50: 10-50: 20-100: 200-500.

[0008] Preferably, the Ganoderma lucidum fermentation product comprises total sugar, fat, total protein, polypeptide, vitamin A, vitamin E, vitamin C and total terpenes in a mass ratio of 3080-3540:270-310:2010-2030:220-320:1-20:10-30:40-60:270-290.

[0009] In a second aspect, the present invention provides a method for preparing the above-mentioned Ganoderma lucidum fermented product, comprising the following steps:

[0010] (1) extracting and filtering Ganoderma lucidum fungus husk to obtain a filtrate;

[0011] (2) concentrating the filtrate obtained in step (1) to obtain a primary treated liquid;

[0012] (3) adding the primary treatment liquid obtained in step (2) to a buffer system for fermentation to obtain a fermentation liquid;

[0013] The fermentation liquid obtained in step (3) is homogenized and dried to obtain a Ganoderma lucidum fermentation product.

[0014] Specifically, the Ganoderma lucidum fungus residue in step (1) is the residue obtained after cultivating Ganoderma lucidum on the cultivation base material.

[0015] Specifically, the cultivation base material includes wood segments of broad-leaved trees.

[0016] Specifically, the extraction in step (1) is as follows: adding 10-20 times the weight of Ganoderma lucidum fungus dregs with water and / or ethanol and extracting for 1-3 times, each extraction taking 1-3 hours.

[0017] Specifically, the concentration in step (2) is: concentrating to a relative density of 1.01-1.25.

[0018] Specifically, the buffer system in step (3) contains 8-20 g / L of glucose, 2.5-8 g / L of peptone, 2-5 g / L of yeast extract, 1.5-4 g / L of phosphate and 10-45 mg / L of vitamin B1.

[0019] Specifically, the fermentation time in step (3) is 5-7 days, the temperature is 22-28° C., and the carbon dioxide concentration is 400 ppm-1800 ppm.

[0020] Specifically, the homogenization treatment in step (4) is: homogenizing and crushing the fermentation liquid at a pressure of 800 bar-1600 bar, and repeating 2-3 times.

[0021] In a third aspect, the present invention provides a use of the Ganoderma lucidum fermentation product or the Ganoderma lucidum fermentation product prepared by the above preparation method in the preparation of medicines or health products for alleviating or preventing radiation damage.

[0022] The radiation damage includes changes and damages to the structure and function of molecules, cells, tissues, and organs of a biological organism caused by radiation.

[0023] Preferably, the radiation damage includes lipid oxidation and DNA damage.

[0024] Specifically, the radiation is gamma ray radiation.

[0025] In a fourth aspect, the present invention provides a drug for alleviating and preventing radiation damage, comprising the above-mentioned Ganoderma lucidum fermentation product or the Ganoderma lucidum fermentation product prepared by the above-mentioned preparation method and pharmaceutically acceptable excipients.

[0026] In a fifth aspect, the present invention provides a health product for alleviating radiation damage, comprising the above-mentioned fermented Ganoderma lucidum product or the fermented Ganoderma lucidum product prepared by the above-mentioned preparation method and an auxiliary material acceptable for health products.

[0027] Definition and Description

[0028] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered ambiguous or unclear without a specific definition, but should be understood according to its ordinary meaning.

[0029] The definition of "total sugar" refers to monosaccharides such as mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose and fucose and the polysaccharides they constitute.

[0030] The definition of "total terpenes" refers to triterpenoid compounds such as ganoderlactone E, ganoderlactone D, ganoderic acid SZ, ganoderic acid DM, and ganoderic acid TR.

[0031] The technical effects achieved by the present invention are:

[0032] (1) The present invention adopts a preparation method of extracting and fermenting Ganoderma lucidum fungus husk to prepare a Ganoderma lucidum fermentation product with a specific composition and ratio, which has the effect of preventing lipid oxidation and DNA damage caused by radiation, and can also increase spleen weight and enhance the body's immune function to alleviate the damage caused by radiation to the body.

[0033] (2) The present invention uses Ganoderma lucidum fungus husk as raw material, which effectively reduces resource waste and realizes high-value utilization of resources.

[0034] (3) The preparation process of the present invention is simple, efficient and easy to promote. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is a chromatogram of the monosaccharide analysis of the Ganoderma lucidum fermentation product in Example 2, where the mixed standard is: Man (mannose), Rha (rhamnose), GlcA (glucuronic acid), GalA (galacturonic acid), Glc (glucose), Gal (galactose), Ara (arabinose), and Fuc (fucose).

[0036] Figure 2 H&E staining of mouse colon, scale bar 200 μm. DETAILED DESCRIPTION

[0037] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0038] Unless otherwise specified, the reagents used in the present invention were purchased from commercial channels. The following is an illustrative description.

[0039] Ganoderma lucidum fungus bran comes from the Ganoderma lucidum planting base of Zhejiang Shouxiangu Pharmaceutical Co., Ltd.

[0040] Peptone was purchased from Jinhua Southeast Chemical Instrument Co., Ltd.

[0041] 1. Composition Analysis of Examples and Comparative Examples

[0042] Example 1

[0043] Using Ganoderma lucidum chaff as raw material, add 20 times the amount of 50% ethanol to extract for 2 hours, filter, take the filter residue and continue to add 10 times the amount of 50% ethanol to extract for 1.5 hours, filter, combine the two filtrates, and concentrate the filtrate under reduced pressure to a relative density of 1.01 to obtain the primary treated liquid.

[0044] The primary treatment liquid was added into a buffer system, which was a mixed solution of 10 g / L glucose, 2.5 g / L peptone, 2.5 g / L yeast extract, 2 g / L phosphate and 10 mg / L vitamin B, for secondary fermentation. The secondary fermentation liquid was homogenized and dried to prepare a Ganoderma lucidum fermentation product.

[0045] Example 2

[0046] Using Ganoderma lucidum chaff as raw material, add 20 times the amount of 50% ethanol to extract for 2 hours, filter, take the filter residue and continue to add 15 times the amount of 50% ethanol to extract for 1.5 hours, filter, combine the two filtrates, and concentrate the filtrate under reduced pressure to a relative density of 1.15 to obtain the primary treated liquid.

[0047] The primary treatment liquid was added into a buffer system, which was a mixed solution of 20 mg / L glucose, 8 mg / L peptone, 5 g / L yeast extract, 4 g / L phosphate and 45 mg / L vitamin B, for secondary fermentation. The secondary fermentation liquid was homogenized and dried to prepare a Ganoderma lucidum fermentation product.

[0048] Example 3

[0049] Using Ganoderma lucidum chaff as raw material, add 20 times the amount of 50% ethanol to extract for 2 hours, filter, take the filter residue and continue to add 20 times the amount of 50% ethanol to extract for 1.5 hours, filter, combine the two filtrates, and concentrate the filtrate under reduced pressure to a relative density of 1.25 to obtain the primary treated liquid.

[0050] The primary treatment liquid was added into a buffer system, which was a mixed solution of 8 mg / L glucose, 2.5 mg / L peptone, 2 g / L yeast extract, 1.5 g / L phosphate and 10 mg / L vitamin B, for secondary fermentation. The secondary fermentation liquid was homogenized and dried to prepare a Ganoderma lucidum fermentation product.

[0051] Comparative Example 1

[0052] Ganoderma lucidum fruiting body was used as raw material, 20 times the amount of 50% ethanol was added and extracted for 2 hours, filtered, and the residue was taken and further 16 times the amount of 50% ethanol was added and extracted for 1.5 hours, filtered, and the two filtrates were combined and concentrated under reduced pressure to a relative density of 1.01-1.25 to obtain Ganoderma lucidum fruiting body extract.

[0053] Comparative Example 2

[0054] Using Ganoderma lucidum chaff as raw material, add 20 times the amount of 50% ethanol to extract for 2 hours, filter, take the filter residue and continue to add 15 times the amount of 50% ethanol to extract for 1.5 hours, filter, combine the two filtrates, and concentrate the filtrate under reduced pressure to a relative density of 1.15 to obtain the primary treated liquid, which is concentrated and dried.

[0055] Comparative Example 3

[0056] Using Ganoderma lucidum chaff as raw material, add 20 times the amount of 50% ethanol to extract for 2 hours, filter, take the filter residue and continue to add 15 times the amount of 50% ethanol to extract for 1.5 hours, filter, combine the two filtrates, and concentrate the filtrate under reduced pressure to a relative density of 1.15 to obtain the primary treated liquid.

[0057] The primary treatment liquid is added to a buffer system comprising a mixed solution of 25 mg / L glucose, 10 mg / L peptone, 6 g / L yeast extract, 4.5 g / L phosphate and 50 mg / L vitamin B, and a secondary fermentation is performed. The secondary fermentation liquid is homogenized and dried to obtain a Ganoderma lucidum fermentation product, and a secondary fermentation is performed. The secondary fermentation liquid is homogenized and dried to obtain a Ganoderma lucidum fermentation product.

[0058] The components of Examples 1-3 and Comparative Examples 1-3 were analyzed as follows:

[0059] Total sugar content was determined using the phenol-sulfuric acid method. First, accurately weigh 20 mg of glucose into a 500 ml volumetric flask and add water to the mark. Pipet 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6, and 1.8 ml of glucose, respectively. Each flask was made up to 2 ml with distilled water, with a 2 ml blank. Then, add 1.0 ml of 6% phenol and 5.0 ml of concentrated sulfuric acid, shake well, and let stand for 30 minutes. Measure absorbance at 490 nm. A standard curve was constructed with glucose concentration as the horizontal axis and absorbance as the vertical axis. Next, take 1 ml of the Ganoderma lucidum fermentation sample, add 1 ml of distilled water, 1 ml of 6% phenol, and 5.0 ml of concentrated sulfuric acid, vortex, and let stand for 30 minutes. Measure absorbance at 490 nm and substitute the value into the standard curve to calculate total sugar concentration.

[0060] Fat content was determined using Soxhlet extraction. A 2g sample of Ganoderma lucidum fermentation product was weighed, wrapped in filter paper, and placed in an extraction cylinder connected to a receiving flask. Anhydrous ether or petroleum ether was added through the upper end of the extractor's condenser tube to a volume two-thirds of the flask's internal volume. The sample was heated in a water bath, with the anhydrous ether or petroleum ether continuously refluxed (6-8 times / hour). The extraction period was typically 6-10 hours. At the end of the extraction, one drop of the extract was collected with a frosted glass rod. The absence of oil stains on the rod indicated completion.

[0061] Remove the receiving flask and recover the anhydrous ether or petroleum ether. When 1 mL to 2 mL of solvent remains in the receiving flask, evaporate it to dryness on a water bath. Dry it at 100°C ± 5°C for 1 hour. Cool it in a desiccator for 0.5 hour, then weigh it. Repeat the above steps until a constant weight is achieved.

[0062] The fat content in the sample is calculated according to the following formula:

[0063] X=(m1-m0) / m2*100.

[0064] Where X is the fat content in the sample, m0 is the weight of the extractor, m1 is the weight after constant weight, and m2 is the weight of the Ganoderma lucidum fermentation product.

[0065] Total protein was determined using the Kjeldahl method. A 2g sample of Ganoderma lucidum fermentation product was weighed and placed in a digestion tube. 0.4g of copper sulfate, 6g of potassium sulfate, and 20mL of sulfuric acid were then added to the digestion incubator. After the incubator temperature reached 420°C, digestion was continued for 1 hour. The liquid in the digestion tube became green and transparent. After cooling, 50mL of water was added. The sample was then automatically added, distilled, titrated, and the titration data recorded using an automated Kjeldahl nitrogen analyzer (pre-use). Sodium hydroxide solution, hydrochloric acid or sulfuric acid standard solution, and boric acid solution containing mixed indicator A or B were added.

[0066] The protein content in the sample is calculated according to the following formula:

[0067] X=[(V1-V2)*c*0.0140 / m*(V3 / 100)]*F*100.

[0068] Where X is the protein content in the sample, V1 is the volume of sulfuric acid or hydrochloric acid standard titrant consumed by the test solution, V2 is the volume of sulfuric acid or hydrochloric acid standard titrant consumed by the reagent blank; C is the concentration of sulfuric acid or hydrochloric acid standard titrant, 0.0140 is the mass of nitrogen equivalent to 1 ml of hydrochloric acid [c(HCl) = 1.000 mol / L] standard titrant, m is the mass of the sample, V3 is the volume of the digestive fluid absorbed, F is the coefficient for converting nitrogen to protein, and 100 is the conversion factor.

[0069] To determine peptides using the biuret method, take 0, 0.2, 0.4, 0.6, 0.8, and 1.0 ml of the standard protein solution, make up to 1 ml with water, and then add 4 ml of biuret reagent. After thorough shaking, incubate at room temperature (20-25°C) for 30 minutes. Perform colorimetric analysis at 540 nm. Use the first test tube without protein solution as a blank control. Plot a standard curve using protein content as the horizontal axis and absorbance as the vertical axis.

[0070] Then take an appropriate amount of Ganoderma lucidum fermentation sample and use the same method as above to substitute into the standard curve to calculate the protein concentration;

[0071] Reverse phase high performance liquid chromatography was used to determine vitamins A and E. The samples were saponified, extracted, washed, concentrated, and filtered through a 0.22 μm organic filter membrane before testing. The chromatographic reference conditions are listed below: Column: C 30 Column (column length 250 mm, inner diameter 4.6 mm, particle size 3 μm); column temperature: 20°C; mobile phase: A: water; B: methanol, elution gradient: 0-12 min, A: 4%, B: 96%; 13-24 min, B: 100%; 24-30 min, A: 4%, B: 96%; flow rate: 0.8 ml / min; UV detection wavelength: 325 nm for vitamin A, 294 nm for vitamin E; injection volume: 10 μL;

[0072] Vitamin C was determined by high performance liquid chromatography. The sample was dissolved and reduced, and then passed through a 0.45 μm water filter membrane for testing. The chromatographic reference conditions are listed as follows: Chromatographic column: C 18 Column (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); column temperature: 25°C; mobile phase: A: 6.8 g potassium dihydrogen phosphate and 0.91 g hexadecyltrimethylammonium bromide, dissolved in water and diluted to 1 L (pH adjusted to 2.5-2.8 with phosphoric acid); B: methanol, mixed in an A:B ratio of 98:2, filtered through a 0.45 μm water filter, and ultrasonically degassed; flow rate: 0.7 ml / min; UV detection wavelength: 245 nm; injection volume: 20 μL.

[0073] Total terpenes were determined spectrophotometrically. Oleanolic acid was used as the standard, and standard solutions of varying concentrations (8, 16, 24, 32, and 40 μg / mL) were prepared in anhydrous ethanol. Color development was then performed with a 5% vanillin-glacial acetic acid solution and perchloric acid. The absorbance of the standard solutions was measured at 540 nm, and a standard curve was plotted with oleanolic acid concentration as the abscissa and absorbance as the ordinate. An appropriate amount of Ganoderma lucidum fermentation product was then taken, subjected to a color development reaction, and its absorbance was measured. The absorbance was then substituted into the standard curve to calculate the total triterpenoid content (as oleanolic acid).

[0074] The assay results of the Ganoderma lucidum fermented products prepared in Example 1 and Comparative Example 1 are shown in Table 1.

[0075] Table 1

[0076]

[0077] It can be seen that the ingredients of Examples 1-3 of the present invention are significantly different from those of Comparative Examples 1-3.

[0078] The triterpenoid components of Example 1 were analyzed using a quadrupole time-of-flight mass spectrometer (Q-TOF MS). Chromatographic conditions were: mobile phase A: 0.1% formic acid in water; mobile phase B: acetonitrile. Gradient elution conditions were: 0 min, 20% B; 0-10 min, 20% → 30% B; 10-20 min, 30% → 35% B; 20-30 min, 35% → 60% B; 30-40 min, 60% → 90% B; 40-60 min, 100% B. Flow rate: 0.45 mL / min, column temperature: 25°C, injection volume: 1 μL. Results are shown in Table 2.

[0079] Table 2 Identification results of triterpenoids in Ganoderma lucidum fermentation products

[0080]

[0081]

[0082] The monosaccharide composition of Example 2 was determined using a pre-column derivatization method. 30 mg of Example 2 was added to 3 ml of water and thoroughly shaken, sonicated, and heated to dissolve. The mixture was allowed to stand, centrifuged, and the supernatant collected. 12 ml of 75% ethanol was added to the supernatant, shaken, and allowed to stand at 4°C for 12 h. Centrifuged, the precipitate was collected and used for monosaccharide composition analysis. 4 mg of the sample was weighed into a hydrolysis tube, 1 ml of 2 M trifluoroacetic acid (TFA) solution was added, and hydrolysis was performed at 121°C for 3 h. After drying with nitrogen purging, 1 ml of methanol was added to mix thoroughly. The nitrogen purging was continued, and this was repeated three times. To each of the dried samples and mixed standards (mannose (Man), glucose (Glc), galactose (Gal), rhamnose (Rha), arabinose (Ara), fucose (Fuc), glucuronic acid (GlcA), and galacturonic acid (GalA)), 400 μL of aqueous ammonia solution was added to dissolve the samples. After dissolution, 400 μL of a 0.3 M methanolic solution of PMP was added. The samples were incubated in a 70°C water bath for 30 minutes and dried under nitrogen. Subsequently, 1 mL of 1% glacial acetic acid was added to each sample tube to dissolve the samples. 1 mL of chloroform was then added, mixed, allowed to stand, and the chloroform was removed. The samples were then extracted with chloroform three times to remove excess PMP. The samples were filtered through a 0.22 μm microporous membrane and analyzed using a liquid chromatography-mass spectrometer equipped with a PDA detector and an LXQ mass spectrometer. The chromatographic column was a Silgreen ODS C18 column (250×4.6 mm, 5 μm); the column temperature was 30°C; the injection volume was 10 μL; the mobile phase was 20 mM ammonium acetate-acetonitrile (78:22, v / v); and the flow rate was 1 mL / min.

[0083] Example 2 Monosaccharide composition analysis results are shown in Figure 1 , it was found that the monosaccharide with the highest proportion was glucose, 85%, mannose 7%, and also contained small amounts of glucuronic acid, rhamnose, arabinose and xylose.

[0084] 2. Animal Experiments

[0085] (1) Examples 1-3 were used in an experiment to intervene in radiation damage in mice. The specific operation was as follows:

[0086] Fifty SPF male KM mice, 3 weeks old, weighing 18-22 g, were housed 10 per cage. The mice were maintained at a temperature of 20-25°C and a humidity of 40-60%. Lights were kept on a 12h:12h day / night cycle, and food and water were freely available. Laboratory-prepared ultrapure water was used. The mice were randomly divided into five groups: 10 control mice; 10 radiation model mice; and 30 experimental mice: 10 mice receiving the treatment in Example 1; 10 mice receiving the treatment in Example 2; and 10 mice receiving the treatment in Example 3. The experiments were conducted after 7 days of acclimatization. The gavage doses were shown in Table 3, and gavage samples were administered for 30 days. On day 31, the irradiation and experimental groups were irradiated with 60Co radiation to a total dose of 5 Gy. Three days after irradiation, the mice were weighed and sacrificed by cervical dislocation. Blood was collected from the eyeballs, and organs such as the spleen, liver, thymus, colon, and femur were removed. Blood was aspirated, and organ weights were measured. The mice were stored at -80°C for subsequent analysis. The spleen index was calculated as: spleen weight / (mouse weight×10).

[0087] Table 3 Oral dose for mice

[0088] Group Number of mice feed Test samples and dosage Blank group 10 Maintenance feed Normal saline Radiation Group 10 Maintenance feed Normal saline Experimental group 30 Maintenance feed Ganoderma lucidum fermentation product (800mg / kg·BW)

[0089] (2) H&E staining

[0090] Fresh colon tissues of mice in the blank group, radiation group and Example 2 were taken and cut into tissue blocks of appropriate size. The blocks were immersed in 4% paraformaldehyde fixative and fixed for more than 24 hours. After washing the samples to remove the fixative, they were dehydrated with an ethanol solution and transparentized with an equal volume mixture of anhydrous ethanol and xylene. Afterwards, the tissues were embedded and sectioned with a thickness of 6 μm. After the sections were stained with hematoxylin and eosin (H&E), the tissue structure was observed with an optical microscope and photographed, as shown in Figure 2. Figure 2 .

[0091] (3) Data processing

[0092] SPSS 24.0 statistical software was used for data statistics. The data results were expressed as mean ± standard deviation. The significant differences between the groups were compared using variance analysis, and the significance level was set at p < 0.05.

[0093] (4) Experimental results

[0094] ① Effects of Ganoderma lucidum fermentation on the physical signs of mice after irradiation

[0095] through 60 After Co ray irradiation, mice in the radiation group died and moved slightly slower than those in the blank group. Their fur became dull and light yellow, accompanied by severe hair loss. They also had difficulty defecating and had small amounts of stool. During autopsy, their internal organs were dark red.

[0096] The behavior of the mice in each example was not significantly different from that of the blank group. They were more energetic, had a slightly yellowish coat with some hair loss, had normal bowel movements, and no mortality occurred. This indicates that radiation damages the body functions of the mice and that the fermented Ganoderma lucidum produced by the present invention has a certain repair effect on radiation damage.

[0097] ②Effects of Ganoderma lucidum fermentation on the spleen of irradiated mice

[0098] The spleen index of each mouse group is shown in Table 4. Spleens were removed by dissection, and after blotting with filter paper to remove any residual blood, they were accurately weighed and recorded, and the organ index (spleen) was calculated. The organ index was calculated as follows: Organ index (%) = Organ mass (g) / Animal mass (g) × 100%. Compared to the blank control group, the spleen index of all irradiated mice groups was significantly lower than that of the blank control group. The spleen index of mice in the Example group was significantly higher than that of the irradiated group, also showing a significant difference. This suggests that Ganoderma lucidum fermentation products promote spleen growth in mice after irradiation.

[0099] Table 4 Spleen index of each group of mice

[0100]

[0101] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0102] ③ Effects of Ganoderma lucidum fermentation products on DNA content in bone marrow of irradiated mice

[0103] The femur was peeled off, and a certain volume of buffer solution was drawn into the syringe to flush out all the bone marrow cells in the femur. The cell suspension was fully dispersed and the absorbance (OD) value was measured at a wavelength of 260 nm. The OD260 value is equivalent to 50 μg / mL DNA. The DNA content of bone marrow cells (μg / mL) = dilution factor × 50 (μg / mL) × OD260. The DNA content of the bone marrow of each group of mice is shown in Table 5. After irradiation with 60Co rays, the DNA content of the bone marrow of mice decreased; compared with the irradiation group, the DNA content of the bone marrow of mice in the experimental group increased significantly, which was significantly different from the irradiation group, and Example 2 was even higher than the blank group. This shows that Ganoderma lucidum fermentation has a certain protective effect on the body's DNA.

[0104] Table 5 Mouse bone marrow DNA content

[0105]

[0106] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0107] ④ Effect of Ganoderma lucidum fermentation on SOD activity in the blood of irradiated mice

[0108] The serum was separated and assayed using a SOD kit. The results are shown in Table 6. 60 After Co irradiation, SOD activity in the irradiated group was significantly reduced. However, serum SOD activity in the experimental group was significantly higher than in the irradiated group. This suggests that Ganoderma lucidum fermentation enhances SOD activity in the body after irradiation.

[0109] Table 6 SOD activity in mouse blood

[0110]

[0111]

[0112] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0113] ⑤ Effect of Ganoderma lucidum fermentation on MDA content in the blood of irradiated mice

[0114] The serum was separated and the MDA content in the blood of mice was determined using an MDA kit, as shown in Table 7. 60 After Co irradiation, the MDA levels in the irradiated group increased significantly. Compared with the irradiated group, the serum MDA levels in the experimental group decreased, showing a significant difference. This suggests that Ganoderma lucidum fermentation products have an inhibitory effect on MDA levels in the body after irradiation.

[0115] Table 7 MDA content in mouse blood

[0116]

[0117] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0118] ⑥ Effect of Ganoderma lucidum fermentation on blood GSH content in mice after irradiation

[0119] The serum was separated and the GSH content in the mouse blood was determined using a GSH kit, as shown in Table 8. 60 After Co irradiation, GSH levels in the irradiated group decreased significantly. However, serum GSH levels in the experimental group increased significantly compared to the irradiated group. This suggests that Ganoderma lucidum fermentation enhances GSH levels in the body after irradiation.

[0120] Table 8 GSH content in mouse blood

[0121]

[0122] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0123] ⑦Effects of Ganoderma lucidum fermentation products on T-AOC in the blood of irradiated mice

[0124] The serum was separated and T-AOC kit was used to determine the T-AOC of mouse blood, see Table 9. 60 After Co irradiation, T-AOC in the irradiated group was significantly reduced. Compared with the irradiated group, serum T-AOC in the treated group was significantly increased, showing a significant difference from the irradiated group. This suggests that Ganoderma lucidum fermented products enhance T-AOC in the body after irradiation.

[0125] Table 9 T-AOC in mouse blood

[0126]

[0127] Wherein, # represents P < 0.05 compared with the blank group, and * represents P < 0.05 compared with the radiation group.

[0128] ⑧Effects of Ganoderma lucidum fermentation products on colon tissue of irradiated mice

[0129] like Figure 2 Shown are histological sections of the colon tissue of each group of mice after hematoxylin-eosin (H&E) staining. The results of tissue section analysis revealed that the experimental group had reduced inflammatory cell infiltration and a decreased number of goblet cells.

[0130] The fermented ganoderma lucidum prepared by the present invention can improve the antioxidant capacity of mice by gavage, and has a certain therapeutic effect on the oxidative damage caused by radiation, thereby reducing the oxidative damage and protecting the body.

[0131] In summary, the fermented Ganoderma lucidum prepared by the present invention can achieve the purpose of resisting radiation damage by promoting spleen growth, protecting the body's DNA, and improving the body's antioxidant capacity.

[0132] The present invention has been described in detail above. Specific examples have been used in the present invention to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only intended to help understand the present invention and its core concepts. It should be noted that those skilled in the art may make various improvements and modifications to the present invention without departing from the principles of the present invention, and such improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A fermented product of Ganoderma lucidum, characterized in that: It includes total sugar, fat, total protein, polypeptide, vitamin A, vitamin E, vitamin C and total terpenes in a mass ratio of 2500-5000: 100-500: 1500-3000: 100-500: 1-50: 10-50: 20-100: 200-500.

2. The fermented Ganoderma lucidum product according to claim 1, characterized in that: It includes total sugar, fat, total protein, polypeptide, vitamin A, vitamin E, vitamin C and total terpenes with a mass ratio of 3080-3540: 270-310: 2010-2030: 220-320: 1-20: 10-30: 40-60: 270-290.

3. A method for preparing the fermented Ganoderma lucidum product according to any one of claims 1 to 2, characterized in that: The following steps are involved: (1) extracting and filtering the bran of Ganoderma lucidum to obtain a filtrate; (2) concentrating the filtrate obtained in step (1) to obtain a primary treatment solution; (3) adding the primary treatment liquid obtained in step (2) into a buffer system for fermentation to obtain a fermentation liquid; (4) homogenizing and drying the fermentation liquid obtained in step (3) to obtain a Ganoderma lucidum fermentation product.

4. The preparation method according to claim 3, characterized in that: The Ganoderma lucidum husk in step (1) is the residue obtained after cultivating Ganoderma lucidum on a cultivation base material, wherein the cultivation base material comprises wood segments of a broad-leaved tree; The extraction in step (1) is as follows: adding 10-20 times the weight of Ganoderma lucidum bran with water and / or ethanol and extracting for 1-3 times, each extraction taking 1-3 hours.

5. The preparation method according to claim 3, characterized in that: The concentration in step (2) is: concentrating to a relative density of 1.01-1.

25.

6. The preparation method according to claim 3, characterized in that: The buffer system in step (3) contains 8-20 g / L glucose, 2.5-8 g / L peptone, 2-5 g / L yeast extract, 1.5-4 g / L phosphate and 10-45 mg / L vitamin B1; The fermentation time of step (3) is 5-7 days, the temperature is 22-28° C., and the carbon dioxide concentration is 400 ppm-1800 ppm; The homogenization treatment in step (4) is as follows: the fermentation liquid is homogenized and crushed at a pressure of 800 bar-1600 bar, and the process is repeated 2-3 times.

7. Use of the fermented Ganoderma lucidum product according to any one of claims 1 to 2 or the fermented Ganoderma lucidum product prepared by the preparation method according to any one of claims 3 to 6 in preparing medicines or health products for alleviating or preventing radiation damage.

8. The use according to claim 7, characterized in that: The radiation damage includes changes and damages to the structure and function of molecules, cells, tissues, and organs of biological organisms caused by radiation; The radiation is gamma ray radiation.

9. A drug for alleviating and preventing radiation damage, characterized in that: The invention comprises the fermented ganoderma lucidum according to any one of claims 1 to 2 or the fermented ganoderma lucidum prepared by the preparation method according to any one of claims 3 to 6 and pharmaceutically acceptable excipients.

10. A health product for alleviating radiation damage, characterized in that: The invention comprises the fermented ganoderma lucidum according to any one of claims 1 to 2 or the fermented ganoderma lucidum prepared by the preparation method according to any one of claims 3 to 6 and auxiliary materials acceptable to health care products.

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