CN110973553A - Propolis nutritional composition for enhancing immunity and application thereof - Google Patents
Propolis nutritional composition for enhancing immunity and application thereof Download PDFInfo
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- CN110973553A CN110973553A CN201911339925.8A CN201911339925A CN110973553A CN 110973553 A CN110973553 A CN 110973553A CN 201911339925 A CN201911339925 A CN 201911339925A CN 110973553 A CN110973553 A CN 110973553A
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- enhancing immunity
- nutritional composition
- shark cartilage
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Classifications
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- A—HUMAN NECESSITIES
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- 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
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a propolis nutritional composition for enhancing immunity and application thereof, and relates to the technical field of health-care products. The propolis nutritional composition for enhancing immunity comprises the following components in parts by weight: 10-20 parts of propolis, 60-70 parts of chitosan and 15-25 parts of shark cartilage powder. In the application, the propolis, the chitosan and the shark cartilage powder are compounded for use, so that the three components can be synergistic, and the aim of enhancing the immunity is fulfilled. The propolis, the chitosan and the shark cartilage powder are used as main components, and other conventional carriers or auxiliary materials can be added to prepare different formulations, so that the taking habits and requirements of different people are met; the process flow is simple, and the popularization and the achievement transformation are easy. The propolis nutritional composition for enhancing immunity can be widely applied to preparation of health-care food or functional food for enhancing immunity.
Description
Technical Field
The invention relates to the technical field of health care products, in particular to a propolis nutritional composition for enhancing immunity and application thereof.
Background
Immunity is the body's own defense mechanism, and is the body's ability to recognize and destroy any foreign body (virus, bacteria, etc.) that invades from the outside, to treat aged, damaged, dead, denatured self cells, and to recognize and treat mutant cells and virus-infected cells in the body.
Modern immunology considers that immunity is the physiological response of the human body to recognize and eliminate "isohexia". The immune system performs this function in the human body. For millions of years, human lives in an environment which is suitable for living and full of danger, so that the human can survive and also obtains extraordinary immunity. Immunity is therefore said to be a product of the biological evolution process.
The modern civilization brings material enjoyment to human beings, fundamentally changes the original life style and habits of the human beings, and simultaneously brings a great amount of negative effects to the human beings. Various metabolic disorders caused by overnutrition, obstructed toxin excretion in vivo and cardiovascular and cerebrovascular diseases induced; the indoor environment with temperature control in four seasons enables mites, aphids, spores, various bacteria and viruses to propagate in a large quantity, float in the air and enter human bodies through respiratory tracts. Modern furniture, indoor decoration and decorating materials carry harmful carcinogens such as formaldehyde and the like to erode the health of human beings.
The human body has the epidemic prevention capability for resisting various viruses and bacteria, creates a healthy body, and most importantly, enhances the natural immunity and the natural curative effect for various diseases, so that the physiological effect of hormones is vigorous.
At present, the medicines for clinically improving the immunity are divided into traditional Chinese medicines and western medicines, the traditional Chinese medicines comprise six-ingredient rehmannia pills, jade screen powder, middle-jiao-tonifying and qi-benefiting pills, bailing capsules, Jinshuibao and the like, and the immunity of a human body can be enhanced and the vital qi of the human body can be improved through the mode and theory of traditional Chinese medicines. The western medicine considers that the common medicines for improving the immunity are common albumen powder, thymopentin and medicines for improving the immunity by injection, such as human serum albumin, lentinan and the like. The existing medicine for improving the immunity has poor effect of enhancing the immunity.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a propolis nutritional composition for enhancing immunity, which has the effect of enhancing immunity and is suitable for people with low immunity.
The invention aims to provide application of the propolis nutrient composition for enhancing immunity in preparation of health-care food or functional food for enhancing immunity.
The invention is realized by the following steps:
in a first aspect, the embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components in parts by weight: 10-20 parts of propolis, 60-70 parts of chitosan and 15-25 parts of shark cartilage powder.
In an alternative embodiment, the components comprise, in parts by weight: 12-18 parts of propolis, 62-68 parts of chitosan and 17-23 parts of shark cartilage powder.
In an alternative embodiment, the shark cartilage powder is prepared by the method comprising:
pulverizing shark cartilage, adding aqueous alkali, soaking and extracting for 13-15h, and separating extractive solution;
adding an acid solution into the extracting solution to adjust the pH value to 8-9, and then adding trypsin for enzymolysis;
decoloring and removing impurities from the enzymolysis liquid;
adding ethanol into the supernatant to perform primary crystallization, collecting precipitate, continuously adding hydrogen peroxide into the precipitate to perform oxidation reaction, adjusting the pH to 6-7 after the reaction is finished, then adding ethanol to perform secondary crystallization, and performing crystallization, filtration and drying;
preferably, the pulverizing of the shark cartilage comprises coarsely pulverizing the shark cartilage, followed by freeze-pulverization.
In an optional embodiment, the enzymolysis time is 1-2h, and the enzymolysis temperature is 50-60 ℃;
preferably, the time of the oxidation reaction is 5-6 h.
In an alternative embodiment, the propolis is propolis powder.
In an optional embodiment, the components further comprise 1-10 parts by weight of auxiliary materials;
preferably, the auxiliary materials comprise one or more of chitosan oligosaccharide, royal jelly, bee pollen, nucleotide, nacre powder, ganoderma lucidum extract and American ginseng extract.
In an optional embodiment, the propolis contains 8-18% of flavones by mass.
In an alternative embodiment, the propolis nutritional composition for enhancing immunity is in the form of any one of hard capsules, soft capsules and granules.
In an alternative embodiment, the composition further comprises a pharmaceutically acceptable carrier for preparing any of hard capsules, soft capsules, and granules.
In a second aspect, the embodiment provides a use of the propolis nutritional composition for enhancing immunity according to any one of the previous embodiments in preparing health food or functional food for enhancing immunity.
The invention has the following beneficial effects:
in the application, the propolis, the chitosan and the shark cartilage powder are compounded for use, so that the three components can be synergistic, and the aim of enhancing the immunity is fulfilled. The propolis, the chitosan and the shark cartilage powder are used as main components, and other conventional carriers or auxiliary materials can be added to prepare different formulations, so that the taking habits and requirements of different people are met; the process flow is simple, and the popularization and the achievement transformation are easy. The propolis nutritional composition for enhancing immunity can be widely applied to preparation of health-care food or functional food for enhancing immunity.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The application provides a propolis nutritional composition for enhancing immunity, which comprises the following components in parts by weight: 10-20 parts of propolis, 60-70 parts of chitosan and 15-25 parts of shark cartilage powder.
Wherein propolis contains flavonoids, terpenes, quinones, esters, alcohols, aldehydes, phenols, organic acids, amino acids, enzymes, vitamins, polysaccharides, trace elements, etc. Propolis has various biological characteristics, wherein bioactive substances playing an important role are flavonoid compounds including flavonoid, flavonol, dihydroflavonoid and the like, and account for about 4.13 percent of the propolis.
The propolis flavone can obviously increase the synthetic amount of total serum protein and antibody of animals to be tested, and obviously enhance the phagocytic capacity of macrophages, so that the propolis flavone can play a beneficial role in thymus, spleen and the whole immune system, and can improve the components of complement and the self-healing capacity of disease resistance of human bodies, so that the immune function of the organisms is in the optimal state of dynamic balance. Propolis has a wide regulating effect on the immune system of an organism, and a large number of reports are made at home and abroad, the propolis can enhance humoral immunity and promote cellular immunity, and can effectively improve the activity, phagocytic capacity and antioxidant capacity of macrophages, reduce the oxidative stress reaction of the macrophages and regulate the secretion of cell factors of the macrophages; increasing the activity of natural killer cells; regulating the activity of polymorphonuclear granulocytes, and controlling the proliferation thereof, thereby improving the immunity of the organism and keeping the immune function of the organism in an optimal state of dynamic balance. Propolis is said to be a natural and highly effective immunopotentiator. Can stimulate the immune function of the organism, enhance the activity of macrophage and improve the immunity of the organism. It has good effects in preventing common cold, cancer, aging, senile plaque and pigmentation.
Propolis can enhance nonspecific and specific immunity of organism, and has certain effect in regulating organism immunity. The propolis has the functions of enhancing the immune function, namely, the propolis has the physiological functions of three aspects of immune defense, immune self-stabilization and immune monitoring, namely, the propolis has the anti-infection capacity of organisms to pathogenic bacteria and viruses, the capacity of identifying and eliminating self-aged tissue cells, and the capacity of killing and eliminating abnormal mutant cells and inhibiting the growth of malignant tumors. Propolis is used as natural immunostimulant, and has effects of enhancing immune system and enhancing activity of immunocyte. For thymus gland, which is the central organ of immune system, propolis can promote the production of a large amount of T cells; propolis can promote spleen to produce a large amount of lymphocytes, has rich B cells, and can secrete special antibodies; has beneficial effects on bone marrow, lymph nodes, etc.
The chitosan has the functions of regulating and improving the immunity of the organism, promoting the generation of antibodies and enhancing the disease resistance of the organism. The chitosan has resistance to pathogens, and can balance sympathetic and parasympathetic nerves. In addition, the chitosan can keep body fluid normal, thereby achieving the purpose of strengthening somatic cells and immunocytes and improving the immunity of human bodies.
The research of the inventor finds that the combination of the propolis and the chitosan has the function of enhancing the immunity, but the enhancing effect of the immunity of the propolis and the chitosan is still poor, so the shark cartilage powder is compounded on the basis of the propolis and the chitosan. Shark cartilage can inhibit chicken embryo vascularization and matrigel-induced angiogenesis, inhibit the growth of breast cancer and human malignant glioma, and prevent the metastasis of breast cancer and Lewis lung cancer. This effect of cartilage may be related to its inhibition of metalloproteases and serine proteases. The shark cartilage contains effective components with endothelial cell specificity, and can reduce the expression of vascular growth factor (VEGF) and its receptor on the surface of microvascular endothelial cell, and inhibit the action of VEGF.
Shark cartilage powder refers to powder made from shark cartilage. The shark cartilage powder has natural effects of promoting regeneration of human cartilage and inhibiting new blood vessel. Shark cartilage contains substances capable of promoting regeneration of human articular cartilage, such as glucosamine, chondroitin sulfate, collagen, etc. These substances are composed in a balanced ratio and can promote the regeneration of human cartilage. The shark cartilage component contains abundant mucopolysaccharide and high-efficiency antibacterial enzyme, so as to stimulate T, B cells and macrophages to attack cancer cells, and improve organism immunity.
In the application, the shark cartilage powder is prepared by the following preparation method:
s1, crushing shark cartilage, adding 1.5% aqueous alkali for soaking, extracting for 13-15h, and carrying out solid-liquid separation to obtain an extracting solution;
s2, adding an acid solution into the extracting solution to adjust the pH value to 8-9, and then adding trypsin for enzymolysis for 1-2 hours at 50-60 ℃;
s3, decoloring and removing impurities from the enzymatic hydrolysate;
s4, adding ethanol into the supernatant to crystallize, collecting the precipitate, continuously adding hydrogen peroxide into the precipitate to perform oxidation reaction for 5-6h, and then adjusting the pH value to 6-7. Then adding ethanol for secondary crystallization, and filtering and drying the crystals.
Wherein the alkaline solution includes but is not limited to NaOH, KOH, Na2CO3、K2CO3One or more of; acid solutions include, but are not limited to, HCl, H2SO4、HNO3One or more of (a).
Specifically, in the present application, the pulverization of shark cartilage comprises the steps of coarsely pulverizing shark cartilage and then freeze-pulverizing the pulverized shark cartilage. The freeze pulverization is carried out at a low temperature (for example, -20 ℃ to-45 ℃) and the pulverization is easier due to the reduced viscosity and elasticity of the shark cartilage under the freezing condition. In the application, the shark cartilage is crushed twice, and the obtained powdery material is easier to extract subsequently. Furthermore, in the application, the shark cartilage is soaked and extracted by the alkali solution, so that the shark cartilage can be fully dissolved for alkali extraction, and then the trypsin can be used for degrading proteins at the intercellular junctions of the shark cartilage, so that the cells are spherical under the tension of the cytoskeleton in the cells, and the cells are separated, thereby being more beneficial to the subsequent extraction process. In addition, the covalent bonding between the protein and the shark chondroitin sulfate can be destroyed by adding the hydrogen peroxide, so that the chondroitin sulfate in the protein is dissociated, impurities such as the protein can be effectively removed, and the purity and the content of the shark chondroitin sulfate sodium are improved. In the application, the purity of the shark cartilage powder can be obviously improved by adding ethanol twice for crystallization. The shark cartilage powder extracted by the specific extraction process has better purity, good stability among batches and better compounding effect with propolis and chitosan.
In addition, the preparation method of the shark cartilage powder is limited, the selection of the propolis is also limited, and the propolis with flavone accounting for 8-18% of the propolis by mass is preferably selected as a main component, so that the immunity is further enhanced.
Preferably, the components comprise the following components in parts by weight: 12-18 parts of propolis, 62-68 parts of chitosan and 17-23 parts of shark cartilage powder. The inventor researches and discovers that when the proportion of the propolis, the chitosan and the shark cartilage powder is in the range, the compounding effect is better, and the synergistic effect is more obvious.
Further, the components in the application also comprise 1-10 parts by weight of auxiliary materials; preferably, the adjuvant comprises one or more of chitosan oligosaccharide, Lac Regis Apis, bee pollen, nucleotide, Concha Margaritifera powder, Ganoderma extract and radix Panacis Quinquefolii extract. When the auxiliary materials are a mixture, the proportion of the components is 1: 1.
The propolis nutritional composition for enhancing immunity provided by the application is in the form of any one of hard capsules, soft capsules and granules. It is to be understood that the above components also include a pharmaceutically acceptable carrier for preparing any of hard capsules, soft capsules, and granules. That is, when the above main ingredients (propolis, chitosan and shark cartilage powder) are prepared into different dosage forms, pharmaceutical excipients or carriers required in the preparation process of hard capsules, soft capsules or granules, and the like can be added according to the conventional process.
The preparation method of the propolis nutritional composition for enhancing immunity is simple, and the main components (propolis, chitosan and shark cartilage powder) are all powdery, so that the main components are directly mixed during preparation, and then the main components are used as main materials to be respectively prepared into any one of hard capsules, soft capsules and granules by a conventional process.
The propolis nutritional composition for enhancing immunity can be widely applied to preparation of health-care food or functional food for enhancing immunity.
The features and properties of the present invention are described in further detail below with reference to examples.
The shark cartilage powder in the following examples was prepared as follows:
firstly, roughly crushing shark cartilage to 60 meshes, then freezing and crushing the shark cartilage to 100 meshes, adding NaOH into the crushed shark cartilage for soaking and extracting for 14 hours, and separating an extracting solution; adding HCl into the extract to adjust pH to 8-9, and adding trypsin for enzymolysis at 55 deg.C for 1-2 hr; after enzymolysis is finished, decoloring and removing impurities from the enzymolysis liquid; and then adding 95% ethanol by mass into the supernatant to perform primary crystallization, collecting the precipitate, continuously adding hydrogen peroxide into the precipitate to perform oxidation reaction for 5-6h, adjusting the pH to 6-7 after the reaction is finished, then adding 95% ethanol by mass to perform secondary crystallization, and performing crystallization, filtration and drying.
Example 1
The embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components: 15 parts of propolis powder, 65 parts of chitin and 20 parts of shark cartilage powder.
Example 2
The embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components: 10 parts of propolis, 70 parts of chitosan and 20 parts of shark cartilage powder.
Example 3
The embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components: 20 parts of propolis powder, 65 parts of chitosan and 15 parts of shark cartilage powder.
Example 4
The embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components: 18 parts of propolis powder, 65 parts of chitosan and 17 parts of shark cartilage powder, wherein the mass percent of flavone in the propolis is 12%.
Example 5
The embodiment provides a propolis nutritional composition for enhancing immunity, which comprises the following components: 12 parts of propolis, 63 parts of chitosan, 17 parts of shark cartilage powder, 4 parts of royal jelly and 4 parts of American ginseng extract.
Experimental example 1, chitosan + propolis: test for enhancing immunity function (chitosan 82%, propolis 18%)
Experimental samples: the content of the hard capsule is earthy yellow powder, and the recommended amount for human body is 1600 mg/person/day of the net weight of the content. And (5) storing the mixture in a cool and dry place for experiments.
Experimental animals: a mouse antibody-producing cell test was conducted by selecting 60 healthy female mice (18.3-22.4 gF1, bred by Shanghai laboratory animals Co., Ltd.) and randomly dividing them into 4 groups (15 mice each) as group I. 60 healthy female mice, 18.0-22.6gF1, were randomly divided into 4 groups of 15 mice each, and used as group II for mouse clearance test. 18.8-22.1gF1 healthy female mice 40, randomly divided into 4 groups, each group of 10, as group III, mice ConA induced mice lymphocyte transformation, NK cell activity test.
Experimental dose: the recommended daily amount of each person (measured by a weight of 60 kg) is 1600mg, which is equivalent to 26.7mg/d/kg · bw. Three dose groups, namely 133mg/kg · bw, 267mg/kg · bw and 800mg/kg · bw, are designed according to 5 times, 10 times and 30 times of the recommended intake of a human body respectively, a 0mg/kg · bw group is additionally arranged, water (disinfected) is used for replacing a test object, the test object with the corresponding dose is orally given to the mouse once a day, and various immunity enhancing function indexes are tested after continuous intragastric administration for 30 days. The gavage amount of the mice is 10 mL/kg-bw.
The test method comprises the following steps: antibody-producing cell detection (Jerne modified slide method), mouse clearance assay, NK cell activity assay (lactate dehydrogenase assay), ConA-induced mouse spleen lymphocyte transformation assay (MTT method).
Feeding conditions are as follows: mice are raised in a barrier system at a temperature of 18-22 ℃ and a relative humidity of 40-70%.
And (3) test results:
1. effect of samples on mouse body weight
Mice were given different doses of sample 30d orally and the weight gain of the mice was not affected.
2. Effect of samples on thymus and spleen organs
Mice were given different doses of sample 30d orally, and the three dose groups were compared, with no significant difference.
3. Effect of the sample on antibody-producing cells (number of hemolytic plaques)
As can be seen from the results in Table 1, the number of hemolytic plaques in mice in the 267 mg/kg-bw and 800 mg/kg-bw groups was significantly higher than that in the 0 mg/kg-bw group.
TABLE 1 Effect of samples on hemolytic plaques
4. Effect of samples on carbon purge Effect
As can be seen from the results in Table 2, the phagocytic index of mice in the groups of 133 mg/kg-bw and 800 mg/kg-bw was significantly higher than that in the group of 0 mg/kg-bw.
TABLE 2 Effect of samples on mouse carbon clearance
5. Effect of samples on NK cell Activity
As can be seen from the results in Table 3, the NK cell activity of the mice in the 800 mg/kg-bw group was significantly higher than that in the 0 mg/kg-bw group.
TABLE 3 Effect of samples on NK cell Activity
6. Effect of samples on ConA-induced splenic lymphocyte transformation in mice
As can be seen from the results in Table 4, the difference in absorbance between the mice in the 800 mg/kg-bw group with ConA wells and those without ConA wells was significantly higher than that in the 0 mg/kg-bw group.
TABLE 4 Effect of samples on ConA-induced splenic lymphocyte transformation in mice
And (4) conclusion: the samples are continuously fed for 30 days at 133 mg/kg-bw, 267 mg/kg-bw and 800 mg/kg-bw, and the results show that the samples can promote the formation of hemolytic plaques of mouse splenocytes; enhancing the carbon clearance capacity of the mice; increasing NK cell activity; can enhance the proliferation ability of spleen lymphocyte. Therefore, the sample is considered to have the function of enhancing immunity.
Experimental example 2: chitosan + propolis + shark cartilage powder: test for enhancing immunity function (chitosan 65.6%, propolis 14.4%, shark cartilage powder 20%)
Experimental samples: the recommended amount for human bodies in the hard capsules is 2.0 g/person/day of the net weight of the contents, which corresponds to 0.033g/kg · bw/d (the weight of human bodies is 60 kg). The samples were formulated with normal edible salad oil to the concentrations required for the test.
Experimental animals and groups: male Kunming mouse, clean grade, weight 18-22 g; female BALB/c mice, clean grade, weight 18-22 g. Mice were randomly divided into a negative control group and three low, medium and high dose groups, and were administered with common edible salad oil and samples of 165 mg/kg-bw/d, 330 mg/kg-bw/d, 660 mg/kg-bw/d, respectively (three dose groups were equivalent to 5 times, 10 times and 20 times of the recommended intake of human body, respectively). All experimental animals eat complete nutrition compound feed and freely eat drinking water. In the ConA-induced splenic lymphocyte transformation test and NK cell activity test of mice, BALB/c mice are used as experimental animals, and Kunming mice are used as experimental animals in other tests.
The experimental method comprises the following steps:
1. ConA-induced splenic lymphocyte transformation assay (MTT method) in mice
2. Delayed allergy test induced by dinitrofluorobenzene (ear swelling method)
3. Antibody-producing cell detection (Jerne modified slide method)
4. Serum hemolysin assay (hemagglutination)
5. Mouse carbon clearance test
6. Mouse abdominal macrophage phagocytic chicken erythrocyte test (half internal method)
7. NK cell Activity assay (lactate dehydrogenase assay)
The experimental results are as follows:
1. body weight, thymus/body weight ratio and spleen/body weight ratio
The weight, thymus ratio and spleen/body ratio of the sample to mice of each dose group are not significant compared with those of a negative control group (p is more than 0.05).
2. ConA-induced splenic lymphocyte transformation assay (MTT method) in mice
The mean of the absorbance differences between each group of ConA plus and no ConA plus wells was analyzed for one-way anova, with significant differences between groups (F ═ 5.59, P < 0.01). The Dunnett's t test showed that the difference between the high dose group and the negative control group in the sample was significant (P < 0.05). The results show that the sample has the function of enhancing the cellular immune function at the dosages of 330 mg/kg-bw/d and 660 mg/kg-bw/d (which are equivalent to 10 times and 20 times of the recommended intake of a human body).
TABLE 5 Effect of samples on ConA-induced splenic lymphocyte transformation assays in mice
| Group (mg/kg) | Animal number (only) | Difference between absorbance of wells with and without ConA |
| 0 | 10 | 0.049±0.015 |
| 165 | 10 | 0.056±0.010 |
| 330 | 10 | 0.067±0.013* |
| 660 | 10 | 0.071±0.015* |
Dunnett's t test showed significant differences compared to the negative control group (P < 0.05)
3. Antibody-producing cell detection (Jerne modified slide method)
The mean value of the number of the hemolytic plaques of each group is subjected to one-way anova, and the difference between the groups has significance (F is 3.54, and P is less than 0.05). The Dunnett's t test showed significant differences between the sample high dose group and the negative control group (P < 0.05). The results show that the sample has the function of enhancing the humoral immune function at the dosage of 660 mg/kg-bw/d (which is equivalent to 20 times of the recommended intake of human bodies).
TABLE 6 influence of samples on the number of hemolytic plaques
| Group (mg/kg) | Animal number (only) | Number of hemolytic plaques (number/10)6Spleen cell) |
| 0 | 10 | 234±32.7 |
| 165 | 10 | 261±39.2 |
| 330 | 10 | 269±42.0 |
| 660 | 10 | 289±38.7* |
Dunnett's t test showed significant differences compared to the negative control group (P < 0.05)
4. Serum hemolysin assay (hemagglutination)
The mean value of the antibody product number of each group is subjected to one-way anova, and the difference between groups has significance (F is 4.38, and P is less than 0.01). The Dunnett's t test showed significant differences between the sample high dose group and the negative control group (P < 0.05). The results show that the sample has the function of enhancing the humoral immune function at the dosage of 660 mg/kg-bw/d (which is equivalent to 20 times of the recommended intake of human bodies).
TABLE 7 sample anti-mouse serum hemolysinInfluence of volume number
| Group (mg/kg) | Animal number (only) | Number of antibody products |
| 0 | 10 | 32.0±8.92 |
| 165 | 10 | 39.2±10.15 |
| 330 | 10 | 42.8±12.38 |
| 660 | 10 | 49.3±11.80* |
Dunnett's t test showed significant differences compared to the negative control group (P < 0.01)
5. Mouse carbon clearance test
The mean value of the phagocytosis indexes of each group is subjected to one-way anova, and the difference between groups has significance (F is 4.64, and P is less than 0.01). The Dunnett's t test showed significant differences between the sample high dose group and the negative control group (P < 0.05). The results show that the sample has the effect of enhancing the phagocytic function of mouse macrophages at the dosage of 660 mg/kg-bw/d (which is equivalent to 20 times of the recommended human intake).
TABLE 8 Effect of samples on the phagocytic index of mice
| Group (mg/kg) | Animal number (only) | Ear weight difference (mg) |
| 0 | 10 | 4.79±0.62 |
| 165 | 10 | 5.11±0.83 |
| 330 | 10 | 5.35±0.64 |
| 660 | 10 | 5.92±0.71* |
Dunnett's t test showed significant differences compared to the negative control group (P < 0.01)
6. Mouse abdominal macrophage phagocytic chicken erythrocyte test (half internal method)
The mean values of the phagocytosis rate and the phagocytosis index of each group are subjected to one-factor anova, and the difference between groups has significant significance (F is 4.05, P is less than 0.05, F is 4.20, and P is less than 0.05). The Dunnett's t test showed significant differences between the sample high dose group and the negative control group (P < 0.05). The results show that the sample has the effect of enhancing the phagocytic function of mouse macrophages at the dosage of 660 mg/kg-bw/d (which is equivalent to 20 times of the recommended human intake).
TABLE 9 Effect of samples on phagocytosis rate and phagocytosis index of chicken erythrocytes by macrophages in mouse peritoneal cavity
| Group (mg/kg) | Animal number (only) | Phagocytosis ratio (%) | Phagocytic index |
| 0 | 10 | 23.8±5.06 | 0.271±0.041 |
| 165 | 10 | 25.7±3.80 | 0.293±0.050 |
| 330 | 10 | 28.2±4.38 | 0.313±0.043 |
| 660 | 10 | 30.1±4.01* | 0.337±0.039* |
Dunnett's t test showed significant differences compared to the negative control group (P < 0.05)
Experimental example 3:
experimental animals and groups: male Kunming mouse, clean grade, weight 18-22 g; female BALB/c mice, clean grade, weight 18-22 g. Mice were randomly divided into negative control groups and experimental groups 1-4, 10 mice per group. The recommended dose for humans is 2 g/person/day.
Control group (0mg/kg bw group with water instead of test substance)
Experimental group 1 (chitosan 65% + propolis 15% + pregelatinized starch 20%) 333.3mg/kg bw
Experimental group 2 (shark cartilage powder 20% + pregelatinized starch 80%) 333.3mg/kg bw
Experimental group 3 (Chitosan 65% + propolis 15% + shark cartilage powder 20%, i.e. example 1)333.g/kg bw
Experimental group 4 (Chitosan 65% + propolis 15% + shark cartilage powder 20%; wherein shark cartilage powder is commercially available) 333.3mg/kg bw
(experimental groups 1, 2, 3 and 4 correspond to 10 times the recommended intake for humans). All experimental animals eat complete nutrition compound feed and freely eat drinking water.
The experimental method comprises the following steps:
1. serum hemolysin assay
2. Mouse carbon clearance test
3. Antibody-producing cell assay
The experimental results are as follows:
1. body weight, thymus/body weight ratio and spleen/body weight ratio
The weight, thymus ratio and spleen/body ratio of the sample to mice of each dose group are not significant compared with those of a negative control group (p is more than 0.05).
2. Effect of samples on mouse serum hemolysin formation
TABLE 10 Effect of samples on mouse serum hemolysin formation
| Group of | Animal number (only) | Half maximal hemolysis value |
| Control group | 10 | 29.23±4.66 |
| Experimental group 1 | 10 | 36.06±6.16* |
| Experimental group 2 | 10 | 35.17±6.95* |
| Experimental group 3 | 10 | 118.0±15.5** |
| Experimental group 4 | 10 | 75.78±9.78** |
Significant difference compared with the control group (P < 0.05)
The difference between the two groups has very significant meaning (P < 0.01)
The results show that the experimental groups all have the effect of promoting the formation of the mouse serum hemolysin, and the difference of the experimental group 3 and the control group is very obvious.
3. Effect of samples on mouse carbon clearance
TABLE 11 Effect of samples on mouse carbon clearance
| Group of | Animal number (only) | Phagocytic index |
| Control group | 10 | 6.60±1.00 |
| Experimental group 1 | 10 | 7.69±0.76* |
| Experimental group 2 | 10 | 7.58±0.68* |
| Experimental group 3 | 10 | 7.99±0.87** |
| Experiment ofGroup 4 | 10 | 7.70±0.75* |
Significant difference compared with the control group (P < 0.05)
The difference between the two groups has very significant meaning (P < 0.01)
The results show that the experimental groups have the capability of enhancing the carbon clearance of the mice, wherein the difference of the experimental group 3 is very obvious compared with the control group.
4. Effect of samples on mouse antibody-producing cells
TABLE 12 Effect of samples on mouse antibody-producing cells
| Group of | Animal number (only) | Number of hemolytic plaques (number/10)6Spleen cell) |
| Control group | 10 | 49±20.3 |
| Experimental group 1 | 10 | 68±22.6* |
| Experimental group 2 | 10 | 69±21.4* |
| Experimental group 3 | 10 | 99±33.2* |
| Experimental group 4 | 10 | 71±19.2* |
Significant difference compared with the control group (P < 0.05)
The results show that the experimental groups all have the effect of promoting the formation of hemolytic plaques of mouse splenocytes, wherein the effect of the experimental group 3 is the most obvious.
And (4) conclusion: experimental example 3 shows that experimental groups 1-4 can promote the immunization of mice. The experimental group 3 is stronger than the experimental groups 1, 2 and 4, and stronger than the control group. The propolis, the chitosan and the shark cartilage powder provided by the application are proved to have a synergistic effect.
In conclusion, the propolis, the chitosan and the shark cartilage powder are compounded for use, so that the three components can be synergistic, and the aim of enhancing the immunity is fulfilled. The propolis, the chitosan and the shark cartilage powder are used as main components, and other conventional carriers or auxiliary materials can be added to prepare different formulations, so that the taking habits and requirements of different people are met; the process flow is simple, and the popularization and the achievement transformation are easy. The propolis nutritional composition for enhancing immunity can be widely applied to preparation of health-care food or functional food for enhancing immunity.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The propolis nutritional composition for enhancing immunity is characterized by comprising the following components in parts by weight: 10-20 parts of propolis, 60-70 parts of chitosan and 15-25 parts of shark cartilage powder.
2. The propolis nutritional composition for enhancing immunity according to claim 1, wherein the components comprise, in parts by weight: 12-18 parts of propolis, 62-68 parts of chitosan and 17-23 parts of shark cartilage powder.
3. The propolis nutritional composition for enhancing immunity according to claim 1, wherein the shark cartilage powder is prepared by the following steps:
pulverizing shark cartilage, adding aqueous alkali, soaking and extracting for 13-15h, and separating extractive solution;
adding an acid solution into the extracting solution to adjust the pH value to 8-9, and then adding trypsin for enzymolysis;
decoloring and removing impurities from the enzymolysis liquid;
adding ethanol into the supernatant to perform primary crystallization, collecting precipitate, continuously adding hydrogen peroxide into the precipitate to perform oxidation reaction, adjusting the pH to 6-7 after the reaction is finished, then adding ethanol to perform secondary crystallization, and performing crystallization, filtration and drying;
preferably, the pulverizing of the shark cartilage comprises coarsely pulverizing the shark cartilage, followed by freeze-pulverization.
4. The propolis nutrient composition for enhancing immunity according to claim 3, wherein the enzymolysis time is 1-2h, and the enzymolysis temperature is 50-60 ℃;
preferably, the time of the oxidation reaction is 5-6 h.
5. The propolis nutrient composition for enhancing immunity according to claim 1, wherein the propolis is propolis powder.
6. The propolis nutritional composition for enhancing immunity according to any one of claims 1 to 5, which is characterized by further comprising 1 to 10 parts by weight of auxiliary materials;
preferably, the auxiliary materials comprise one or more of chitosan oligosaccharide, royal jelly, bee pollen, nucleotide, nacre powder, ganoderma lucidum extract and American ginseng extract.
7. The propolis nutritional composition for enhancing immunity according to any one of claims 1 to 5, wherein the propolis flavone accounts for 8 to 18 mass percent of the propolis.
8. The immunity-enhancing propolis nutritional composition according to any one of claims 1 to 5, wherein the immunity-enhancing propolis nutritional composition is in the form of any one of a hard capsule, a soft capsule and a granule.
9. The propolis nutrient composition for enhancing immunity according to claim 8, wherein the components further include a pharmaceutically acceptable carrier for preparing any one of hard capsules, soft capsules and granules.
10. Use of the propolis nutritional composition for enhancing immunity according to any one of claims 1 to 9 in the preparation of health food or functional food for enhancing immunity.
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| KR20000034717A (en) * | 1998-11-30 | 2000-06-26 | 김해용 | Process for preparing health food by mixing propolis extract with chitin, chitosan, or shark cartilage |
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| CN101358220A (en) * | 2008-09-25 | 2009-02-04 | 王清荣 | Method for extracting calcium chondroitin sulfate in shark cartilage |
| CN102283865A (en) * | 2010-06-17 | 2011-12-21 | 陆兴艳 | Compound chitosan preparation |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20000034717A (en) * | 1998-11-30 | 2000-06-26 | 김해용 | Process for preparing health food by mixing propolis extract with chitin, chitosan, or shark cartilage |
| CN1981783A (en) * | 2005-12-16 | 2007-06-20 | 武兴战 | Nano-bee productf for adjusting immune function and its production |
| CN101358220A (en) * | 2008-09-25 | 2009-02-04 | 王清荣 | Method for extracting calcium chondroitin sulfate in shark cartilage |
| CN102283865A (en) * | 2010-06-17 | 2011-12-21 | 陆兴艳 | Compound chitosan preparation |
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Application publication date: 20200410 |