CN115501294B - Preparation method and application of saccharomyces boulardii fermentation dioscorea opposita alcohol extract - Google Patents
Preparation method and application of saccharomyces boulardii fermentation dioscorea opposita alcohol extract Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/894—Dioscoreaceae (Yam family)
- A61K36/8945—Dioscorea, e.g. yam, Chinese yam or water yam
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
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- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/06—Antianaemics
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
A preparation method and application of a saccharomyces boulardii fermentation dioscorea opposita ethanol extract belong to the fields of food and medicine, and a specific scheme comprises the following steps: inoculating saccharomyces boulardii into a sterilized yam rhizome culture medium for fermentation to obtain a yam rhizome fermentation product; step two, collecting the supernatant of the Chinese yam fermentation product, concentrating and drying to obtain dry powder of the Chinese yam fermentation product, adding an alcohol solution into the dry powder of the Chinese yam fermentation product, leaching, carrying out solid-liquid separation to obtain an alcohol extract, concentrating the alcohol extract, and drying to obtain the alcohol extract of the Chinese yam fermented by the saccharomyces boulardii. The alcohol extract of the Chinese yam fermented by the saccharomyces boulardii has obvious protection and regulation effects on the hematopoietic system and the redox system of radiation experimental animals.
Description
Technical Field
The invention belongs to the field of foods and medicines, and particularly relates to a preparation method of a Chinese yam alcoholic extract fermented by saccharomyces boulardii and application of the Chinese yam alcoholic extract in radiation damage protection.
Background
Ionizing radiation is widely used in the fields of medical treatment and food industry, however, the ionizing radiation can cause certain damage to organisms and even induce canceration after long-term exposure. Traditional radioprotectors are mainly synthesized chemical substances, and are limited in application due to the factors of large side effects, high cost and the like. The natural radiation protective agent gradually becomes a research hot spot due to the advantages of safety, high efficiency, wide sources, low cost and the like. The Chinese yam is used as a medicinal and edible plant, contains rich nutritional ingredients and various functional factors, and the current comprehensive utilization degree of the Chinese yam and the related research on the fermented deep-processed product are still limited. The saccharomyces boulardii is a probiotic yeast, has good functions of antioxidation, immunoregulation and the like, and has gradually explored the application potential in the field of food fermentation in recent years. The development of radiation protection functional foods and medicines from natural products and microbial fermentation metabolites thereof to reduce the radiation oxidation injury degree and improve the body health is one of the new approaches of radiation protection at present.
Disclosure of Invention
The invention aims to provide a preparation method of a Chinese yam alcoholic extract fermented by saccharomyces boulardii and application of the Chinese yam alcoholic extract in the aspect of radiation damage protection.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
A preparation method of a Chinese yam alcohol extract by fermenting saccharomyces boulardii comprises the following steps:
inoculating saccharomyces boulardii into a sterilized yam rhizome culture medium for fermentation to obtain a yam rhizome fermentation product;
step two, collecting the supernatant of the Chinese yam fermentation product, concentrating and drying to obtain dry powder of the Chinese yam fermentation product, adding an alcohol solution into the dry powder of the Chinese yam fermentation product, leaching, carrying out solid-liquid separation to obtain an alcohol extract, concentrating the alcohol extract, and drying to obtain the alcohol extract of the Chinese yam fermented by the saccharomyces boulardii.
In the first step, the inoculation amount of the saccharomyces boulardii is 1-5% of the volume of the yam culture medium, the fermentation temperature is 30 ℃, and the fermentation time is 20-26 hours.
In the first step, the weight percentage of yam tuber powder in the formula of the yam culture medium is 2-6%, the weight percentage of peptone is 1%, and the balance is water, wherein the pH value is 5.5.
In the second step, the concentration temperature is 45 ℃, and the drying mode is vacuum freeze drying or spray drying.
In the second step, the mass volume ratio of the dry powder of the Chinese yam fermented product to the alcohol solution is 1 g:20-40 mL, wherein the alcohol in the alcohol solution is at least one of methanol, ethanol and isopropanol, and the volume ratio of the alcohol in the alcohol solution to the water is 5-7:3-5.
In the second step, the leaching temperature is 50-70 ℃ and the time is 40-100 min, and the leaching condition is water bath leaching.
The application of the ethanol extract of the dioscorea opposita in preparing foods or medicines for resisting radiation injury and preventing radiation injury.
Further, the radiation damage includes damage to the hematopoietic system and redox system of the body.
Further, the saccharomyces boulardii fermentation dioscorea opposita ethanol extract is used in foods or medicines for recovering the injury level of liver tissues and small intestine tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of liver tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of small intestine tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of serum after ionizing radiation and in foods or medicines for preparing the leucocyte level in peripheral blood after ionizing radiation.
Further, the antioxidant indicator comprises one or both of superoxide dismutase level and malondialdehyde level.
Compared with the prior art, the invention has the beneficial effects that:
The alcohol extract of the Chinese yam fermented by the saccharomyces boulardii has obvious protection and regulation effects on the hematopoietic system and the redox system of radiation experimental animals.
Drawings
FIG. 1 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on body weight of mice;
FIG. 2 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on liver tissue of mice after ionizing radiation;
FIG. 3 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on the small intestine tissue of mice after ionizing radiation;
FIG. 4 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on SOD in liver tissue of mice after ionizing radiation;
FIG. 5 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on MDA in liver tissue of mice after ionizing radiation;
FIG. 6 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on SOD in mouse intestinal tissue after ionizing radiation;
FIG. 7 is a graph showing the effect of fermented Dioscorea opposita ethanol extract on MDA in mouse intestinal tissue after ionizing radiation;
FIG. 8 is a graph showing the effect of fermented rhizoma Dioscoreae alcohol extract on SOD in mouse serum after ionizing radiation;
FIG. 9 is a graph showing the effect of fermented Dioscorea opposita ethanol extract on MDA in mouse serum after ionizing radiation.
Detailed Description
The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and examples, and it is apparent that the described examples are only some, but not all, of the examples of the invention, and all other examples obtained by those skilled in the art without making any inventive effort are within the scope of the present invention.
Detailed description of the preferred embodiments
A preparation method of a Chinese yam alcohol extract by fermenting saccharomyces boulardii comprises the following steps:
Inoculating activated saccharomyces boulardii into a sterilized yam culture medium for fermentation to obtain a yam fermentation product;
Step two, collecting the supernatant of the Chinese yam fermentation product, filtering, concentrating and drying to obtain dry powder of the Chinese yam fermentation product, adding an alcohol solution into the dry powder of the Chinese yam fermentation product, leaching, carrying out solid-liquid separation to obtain an alcohol extract, concentrating the alcohol extract, and drying to obtain the alcohol extract of the Chinese yam fermented by the saccharomyces boulardii.
Further, the mass volume ratio of the dry powder of the Chinese yam fermented product to the alcohol solution is 1 g:20-40 mL, preferably 1:30mL.
Further, the activating step of the saccharomyces boulardii comprises the following steps: inoculating the saccharomyces boulardii in a sand culture medium, and activating in a shaking table of 180 r/min; the activation temperature is 30 ℃, the time is 10-14 h, and the activation times are 1-3 times. Preferably, the activation time is 12 hours and the number of activations is 2.
In the first step, the inoculation amount of the saccharomyces boulardii is 1% -5% of the volume of the yam culture medium, preferably 2%; the fermentation temperature is 30 ℃, and the fermentation time is 20-26 hours, preferably 24 hours.
In the first step, the weight percentage of the yam tuber powder in the formulation of the yam culture medium is 2-6%, preferably 4%, the weight percentage of peptone is 1%, and the balance is water, with the pH value of 5.5.
Further, in the second step, the concentration temperature is 45 ℃, and the drying mode is freeze drying or spray drying, preferably vacuum freeze drying.
In the second step, the alcohol in the alcohol solution is at least one of methanol, ethanol and isopropanol, and the volume ratio of the alcohol to the water in the alcohol solution is 5-7:3-5, preferably 6:4.
Preferably, the alcohol solution is an ethanol solution.
Further, in the second step, the leaching is performed in a water bath kettle, and the leaching temperature is 50-70 ℃. Preferably 60℃for 40 to 100min, preferably 60min.
Further, in the second step, the leaching is performed while shaking or stirring, and the shaking or stirring rotation speed is 120-180 rpm.
In the second step, after solid-liquid separation, the precipitate is leached twice, and the solid-liquid separation is performed again, and the two ethanol extracts are combined.
In the second step, the solid-liquid separation mode is vacuum filtration.
Detailed description of the preferred embodiments
The application of the ethanol extract of the dioscorea opposita in preparing foods or medicines for resisting radiation injury and preventing radiation injury. The radiation damage includes radiation damage to cancer patients and radiation damage to normal persons; radiation damage to a cancer patient is radiation damage to normal tissue organs in radiation therapy of the cancer patient; radiation damage to normal persons is radiation damage to the staff engaged in the radiation.
Further, the radiation damage includes damage to the hematopoietic system and redox system of the body.
Further, the saccharomyces boulardii fermentation dioscorea opposita ethanol extract is used in foods or medicines for recovering the injury level of liver tissues and small intestine tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of liver tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of small intestine tissues after ionizing radiation, in foods or medicines for preparing the oxidation resistance index level of serum after ionizing radiation and in foods or medicines for preparing the leucocyte level in peripheral blood after ionizing radiation.
Further, the antioxidant indicator includes one or both of a superoxide dismutase (SOD) level and a Malondialdehyde (MDA) level.
Preferably, the saccharomyces boulardii fermented dioscorea opposita ethanol extract is prepared according to the method in the first specific embodiment.
The saccharomyces boulardii is a probiotic yeast and has good functions of antioxidation, immunoregulation and the like. The Chinese yam is used as a medicinal and edible plant and contains rich nutritional ingredients and various functional factors, however, the comprehensive utilization degree and the related research of the fermented deep-processed product are still limited at present. The embodiment of the invention is found through intensive researches, and provides the preparation of the ethanol extract of the Chinese yam fermented by the saccharomyces boulardii and the application of the ethanol extract in foods or medicines for resisting radiation injury and preventing radiation injury. Experiments of the embodiment of the invention show that the saccharomyces boulardii fermented dioscorea opposita ethanol extract has obvious protection effect on the hematopoietic system and the redox system of radiation experimental animals after 60 Co-gamma rays (dose rate of 2 Gy/min) with 6Gy dose are irradiated on a Kunming mouse.
Example 1
A preparation method of a Chinese yam alcohol extract by fermenting saccharomyces boulardii comprises the following steps:
1. Preparing raw materials: cleaning fresh rhizoma Dioscoreae, peeling, slicing, sun drying, pulverizing with 40 mesh sieve to obtain sample powder, packaging, and sealing.
2. Preparing a saccharomyces boulardii seed solution: inoculating Saccharomyces boulardii to sterilized Saccharomycetes culture medium (121deg.C, 20 min), and culturing at 30deg.C in 180r/min shaking table for 12 hr. And then performing secondary activation according to the same method to obtain the saccharomyces boulardii seed liquid.
3. Inoculating: inoculating activated Brevibacterium strain seed solution into sterilized rhizoma Dioscoreae culture medium (culture medium-liquid ratio is 1:25, sterilization temperature is 121deg.C, sterilization time is 20 min), fermenting at 30deg.C for 24 hr at 180r/min, and inoculating 2% of the volume of rhizoma Dioscoreae culture medium as fermentation group. The unfermented control group was inoculated at 0% inoculum size, and the other steps remained consistent with the fermented group. The formula of the Chinese yam culture medium comprises the following components: (in terms of mass fraction) yam tuber powder 4%, peptone 1%, balance water, pH 5.5.
4. Vacuum freeze drying: collecting supernatant of the fermented and unfermented products, concentrating, and drying to obtain dry powder of rhizoma Dioscoreae ferment and dry powder of unfermented rhizoma Dioscoreae.
5. Leaching: taking fermented Chinese yam and unfermented Chinese yam powder according to the following ratio of 1:30 The (g/mL) feed liquid ratio is added into ethanol solution, and the leaching conditions are that the ethanol extract of the fermented Chinese yam and the ethanol extract of the unfermented Chinese yam are obtained by water bath oscillation at 60 ℃ for 60min (150 rpm). The volume ratio of ethanol to water in the ethanol solution was 6:4.
6. And (3) filtering: and (3) carrying out rough filtration on the alcohol extract by using gauze, and carrying out suction filtration to obtain fermented Chinese yam alcohol extract and unfermented Chinese yam alcohol extract respectively.
7. Concentrating: and (3) carrying out rotary evaporation concentration on the two groups of alcohol extract solutions at 45 ℃ to obtain fermented Chinese yam concentrated solution and unfermented Chinese yam concentrated solution.
8. Vacuum freeze drying: drying the concentrated solutions of the two groups by a vacuum freeze dryer to obtain fermented rhizoma Dioscoreae ethanol extract and unfermented rhizoma Dioscoreae ethanol extract.
Changes of main functional components of rhizoma Dioscoreae probiotic Saccharomyces boulardii before and after fermentation
The fermented dioscorea opposita alcohol extract of example 1 was used to compare the total sugar, protein, total phenol, total flavone content in the unfermented dioscorea opposita alcohol extract and the fermented dioscorea opposita alcohol extract.
The method for detecting the total sugar content comprises the following steps: determining the total sugar content in the sample by adopting a phenol-sulfuric acid method;
the protein detection method comprises the following steps: measuring the protein content in the sample by adopting a Coomassie brilliant blue method;
the method for detecting the total phenols comprises the following steps: measuring the total phenol content in the sample by adopting a Fu Lin Fen method;
The detection method of total flavonoids comprises the following steps: determining the content of total flavonoids in the sample by adopting a sodium nitrite-aluminum chloride method;
the experimental results are shown in table 1:
TABLE 1
Note that: the data are expressed as mean ± standard deviation (n=3), wherein the statistical difference from the unfermented dioscorea opposita ethanol extract is expressed as P <0.05, P <0.01, P <0.001.
Table 1 shows that the fermented rhizoma Dioscoreae ethanol extract contains four common functional active components including polysaccharide, protein, polyphenol and flavone, wherein the main composition changes in saccharide, phenols and flavone substances.
Experimental example 2 Effect of fermented Dioscorea opposita alcohol extract on mouse body weight
1. Medicaments and agents
Test drug: the fermented dioscorea opposita alcohol extract of example 1;
positive drug: likejun tablet.
2. Experimental animals: male Kunming mice raised in animal grade SPF barrier system, 6-8 weeks old, and body weight 18-20 g. Placing in a dry and clean plastic cage, and feeding standard experimental animal feed with free water.
3. The experimental method comprises the following steps: animals were randomly grouped, 12 per group. The experiment is carried out by setting a blank control group, a radiation control group, a positive control group and a high dose group, a medium dose group and a low dose group of the fermented Chinese yam ethanol extract and a high dose group of the unfermented Chinese yam ethanol extract.
Blank control (NC): 10mL/kgbw physiological saline;
radiation control group (MC): 10mL/kgbw physiological saline;
positive control group (PC): 12mg/kgbw Likejun tablet;
low dose group of fermented dioscorea opposita ethanol extract (FCYE-L): 50mg/kgbw;
dose group (FCYE-M) of fermented rhizoma Dioscoreae alcohol extract: 100mg/kgbw;
high dose group of fermented dioscorea opposita ethanol extract (FCYE-H): 200mg/kgbw;
High dose group of unfermented dioscorea opposita ethanol extract (UCYE-H): 200mg/kgbw.
After continuous gavage for 30 days in the above groups, the change in body weight of the mice was recorded. The detection results are shown in FIG. 1.
As can be seen from FIG. 1, the mice taking the fermented Dioscorea opposita ethanol extract had a normal tendency to gain body weight. I.e. without causing obesity and without toxic side effects.
Experimental example 3 Effect of fermented Dioscorea opposita alcohol extract on liver tissue of mice after ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after the stomach was irrigated for 30 days, radiation treatment was performed under the condition of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min, and the total radiation dose of mice was 6Gy. After the irradiation is finished for 24 hours, the mice are dissected, the livers are washed clean by pre-cooled normal saline, and after the filter paper is sucked dry, the livers are placed into 4% paraformaldehyde tissue fixing solution. After 24H fixation, the tissue wax blocks were cut into slices and H & E stained, and finally observed under an optical microscope and images were acquired.
As shown in figure 2, the liver of the mice is damaged after ionizing radiation, and the fermented dioscorea opposita ethanol extract can recover the damage caused by the liver of ionizing radiation.
Experimental example 4 Effect of fermented Dioscorea opposita alcohol extract on mouse intestinal tissue after mouse ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after the stomach was irrigated for 30 days, radiation treatment was performed under the condition of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min, and the total radiation dose of mice was 6Gy. After the irradiation is finished for 24 hours, the mice are dissected, the small intestine is washed clean by pre-cooled normal saline, and after the filter paper is sucked dry, the small intestine is placed into 4% paraformaldehyde tissue fixing solution. After 24H fixation, the tissue wax blocks were cut into slices and H & E stained, and finally observed under an optical microscope and images were acquired.
As shown in figure 3, the small intestine of the mouse is damaged after the ionizing radiation, and the fermented dioscorea opposita ethanol extract can recover the damage caused by the small intestine of the ionizing radiation.
Experimental example 5 Effect of fermented Dioscorea opposita alcohol extract on antioxidant index in liver after mouse ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after the stomach was irrigated for 30 days, radiation treatment was performed under the condition of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min, and the total radiation dose of mice was 6Gy. After 24 hours from the end of irradiation, the mice were dislocation killed, the livers were immediately removed, fat was removed, blood was washed off with sterile ice-cold saline, and the surface water was blotted dry by filter paper to homogenize tissues. The results of experiments obtained by measuring the activity of SOD and the MDA content in the liver of mice are shown in FIG. 4 and FIG. 5. Fig. 4 shows the effect of fermented dioscorea opposita alcohol extract on the level of SOD enzyme activity in the liver of mice after ionizing radiation, and fig. 5 shows the effect of fermented dioscorea opposita alcohol extract on the MDA content in the liver of mice after ionizing radiation, wherein ### P <0.001 compared to NC group, P <0.05 compared to MC group, P <0.01 compared to MC group, P <0.001 compared to MC group (n=8).
As shown in fig. 4, the SOD enzyme activity level in the liver of the mice after the ionizing radiation is obviously reduced, and the fermented dioscorea opposita ethanol extract can obviously improve the SOD enzyme activity level in the liver of the mice after the ionizing radiation, and is better than the action effect of the unfermented dioscorea opposita ethanol extract. As shown in fig. 5, the MDA content in the liver of the mice after the ionizing radiation is obviously increased, and the fermented dioscorea opposita ethanol extract can obviously reduce the MDA content in the liver of the mice after the ionizing radiation, and is better than the action effect of the unfermented dioscorea opposita ethanol extract.
Experimental example 6 Effect of fermented Dioscorea opposita alcohol extract on antioxidant index in small intestine of mice after ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after the stomach was irrigated for 30 days, radiation treatment was performed under the condition of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min, and the total radiation dose of mice was 6Gy. After 24 hours from the end of irradiation, the mice were dislocation killed, the small intestine was immediately removed, fat was removed, blood was washed off with sterile ice-cold saline, and the surface water was blotted dry with filter paper to perform tissue homogenization. The results of the experiments obtained by measuring the activity level of SOD and MDA content in the small intestine tissue of the mice are shown in FIG. 6 and FIG. 7. Fig. 6 is the effect of fermented dioscorea opposita alcohol extract on the level of SOD enzyme activity in the small intestine of mice after ionizing radiation, fig. 7 is the effect of fermented dioscorea opposita alcohol extract on the MDA content in the small intestine of mice after ionizing radiation, wherein ## P <0.01 is compared with NC group, ### P <0.001 is compared with NC group, P <0.05 is compared with MC group, P <0.001 is compared with MC group (n=8).
As shown in fig. 6, the SOD enzyme activity level in the small intestine of the mice after the ionizing radiation is obviously reduced, and the fermented dioscorea opposita ethanol extract can obviously improve the SOD enzyme activity level in the small intestine of the mice after the ionizing radiation, and is better than the action effect of the unfermented dioscorea opposita ethanol extract. As shown in fig. 7, the MDA content in the small intestine of the mice after the ionizing radiation is obviously increased, and the fermented dioscorea opposita ethanol extract can obviously reduce the MDA content in the small intestine of the mice after the ionizing radiation, and is better than the action effect of the unfermented dioscorea opposita ethanol extract.
Experiment 7 Effect of fermented Dioscorea opposita alcohol extract on antioxidant index in serum after mouse ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after continuous gastric lavage for 30 days, radiation treatment was performed under the conditions of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min and total radiation dose of 6Gy in mice. After 24 hours of irradiation, blood is taken from eyeballs of the mice, the blood is placed in a refrigerator at 4 ℃, after 4 hours, the blood is coagulated, supernatant is taken and preserved at-80 ℃ for standby after centrifugation at low temperature (4000 rpm) for 10 minutes, SOD enzyme activity and MDA content in serum of the mice are detected, and the obtained experimental results are shown in figures 8 and 9. Fig. 8 is the effect of yam fermented alcohol extract on SOD enzyme activity level in mouse serum after ionizing radiation, fig. 9 is the effect of yam fermented alcohol extract on MDA content in mouse serum after ionizing radiation, wherein # P <0.05 is compared with NC group, ### P <0.001 is compared with NC group, P <0.05 is compared with MC group, P <0.01 is compared with MC group (n=8).
As shown in fig. 8, the SOD enzyme activity level in the serum of the mice after the ionizing radiation is obviously reduced, and the fermented alcohol extract of the yam can obviously improve the SOD enzyme activity level in the serum of the mice after the ionizing radiation and is better than the action effect of the unfermented alcohol extract of the yam. As shown in fig. 9, the MDA content in the serum of the mice after the ionizing radiation is obviously increased, and the yam fermentation alcohol extract can obviously reduce the MDA content in the serum of the mice after the ionizing radiation, and is better than the action effect of the unfermented yam alcohol extract.
Experimental example 8 Effect of fermented Dioscorea opposita alcohol extract on peripheral blood of mice after ionizing radiation
Animal experiments were performed by using the fermented dioscorea opposita ethanol extract of example 1 in the manner of experimental example 2, and after the stomach was irrigated for 30 days, radiation treatment was performed under the condition of 60 Co-gamma-ray disposable whole body irradiation at a dose rate of 2Gy/min, and the total radiation dose of mice was 6Gy. After 24 hours from the end of irradiation, the mice were subjected to eyeball blood collection, and the numbers of white blood cells, red blood cells, platelets and the content of hemoglobin in the peripheral blood were detected.
The experimental results are shown in table 2:
TABLE 2
Note that: the data are expressed as mean ± standard deviation (n=8), wherein there is a statistical difference compared to NC group expressed as ### P <0.001; statistical differences compared to the MC group are expressed as P <0.05.
As shown in Table 2, the white blood cells in the peripheral blood of the mice after the ionizing radiation are obviously reduced, and the fermented dioscorea opposita alcohol extract can obviously improve the content of the white blood cells in the peripheral blood of the mice after the ionizing radiation.
Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.
Claims (9)
1. An application of a saccharomyces boulardii fermentation dioscorea opposita ethanol extract in preparing foods or medicines for resisting radiation injury and preventing radiation injury, wherein the preparation method of the dioscorea opposita ethanol extract comprises the following steps:
inoculating saccharomyces boulardii into a sterilized yam rhizome culture medium for fermentation to obtain a yam rhizome fermentation product;
And step two, collecting supernatant of the Chinese yam fermentation product, concentrating and drying to obtain dry powder of the Chinese yam fermentation product, adding an ethanol solution into the dry powder of the Chinese yam fermentation product, leaching, wherein the volume ratio of alcohol to water in the ethanol solution is 5-7:3-5, carrying out solid-liquid separation to obtain an alcohol extract, concentrating the alcohol extract, and drying to obtain the ethanol extract of the Chinese yam fermented by the saccharomyces boulardii.
2. The use according to claim 1, characterized in that: in the first step, the inoculation amount of the saccharomyces boulardii is 1% -5% of the volume of the Chinese yam culture medium, the fermentation temperature is 30 ℃, and the fermentation time is 20 h% -26 h.
3. The use according to claim 1, characterized in that: in the first step, the weight percentage of yam tuber powder in the formula of the yam culture medium is 2% -6%, the weight percentage of peptone is 1%, the balance is water, and the pH is 5.5.
4. The use according to claim 1, characterized in that: in the second step, the concentration temperature is 45 ℃, and the drying mode is vacuum freeze drying or spray drying.
5. The use according to claim 1, characterized in that: in the second step, the mass volume ratio of the dry powder of the Chinese yam fermented product to the alcohol solution is 1 g:20-40 mL.
6. The use according to claim 1, characterized in that: in the second step, the leaching temperature is 50-70 ℃ and the time is 40-100 min, and the leaching condition is water bath leaching.
7. The use according to claim 1, characterized in that: the radiation damage includes damage to the hematopoietic system and redox system of the body.
8. The use according to claim 1, wherein the saccharomyces boulardii fermented dioscorea opposita alcohol extract is used for at least one of the following applications: the liver tissue and small intestine tissue injury level after the ionizing radiation is recovered, the oxidation resistance index level in the liver tissue after the ionizing radiation is recovered, the oxidation resistance index level in the small intestine tissue after the ionizing radiation is recovered, the oxidation resistance index level in serum after the ionizing radiation is recovered, and the leucocyte level in peripheral blood after the ionizing radiation is recovered.
9. The use according to claim 8, characterized in that: the antioxidant index comprises one or two of superoxide dismutase level and malondialdehyde level.
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