CN1706280A - Health food capable of raising immunity and its production process - Google Patents

Abstract

The present invention discloses a health food capable of enhancing immunity and a manufacturing method thereof. The components of the health food capable of enhancing immunity of the present invention are prepared in parts by weight: Astragalus 600-900 parts, Rhodiola 400-600 parts 400-600 parts of matsutake, 100-200 parts of American ginseng, because the health food of the present invention is formulated scientifically by using the above-mentioned components, its formula is reasonable, and the joint use of each component can play an anti-hypoxia-resistant, Anti-cold and anti-tumor immune effects, and regulate the non-specific and specific immunity of the human body, so as to achieve a comprehensive and systematic enhancement of the body's immunity and enhance the immunity of people with low immune function. And they all adopt natural products or are prepared from natural products, so the health food is a kind of natural health food that is safe, completely without any toxic and side effects, and can be taken for a long time.

Description

A kind of health food capable of enhancing immunity and preparation method thereof

(1) Technical field:

The invention relates to a health food, especially a health food capable of enhancing immunity; the invention also relates to a preparation method of the health food.

(two) background technology:

The human body not only has a complete set of protective immune system, such as thymus, spleen, lymph nodes, intestinal respiratory submucosal lymphoid tissue, etc., but also has immune active cells that can perform immune functions, such as T cells, B lymphocytes, giant cells, etc. Phage cells, etc. These immune active cells, under the self-regulation of the nervous, humoral and immune systems, coordinate with each other to perform their own functions, so as to ensure that the human body is in a stable and normal state. Modern immunology believes that the immune function of the human body has three major functions: defense against infection, self-stabilization and immune surveillance. Recognition" and "removal of aliens" to achieve the physiological protection of self-stabilization. But when the immune function is abnormal, its own homeostasis will be broken, leading to a variety of autoimmune diseases, such as rheumatoid arthritis, lupus erythematosus, scleroderma, Behcet's disease, AIDS and other diseases , and is closely related to the aging of the human body and the occurrence of tumors, high blood pressure, and diabetes. The immunity referred to by modern medicine also has a similar meaning to the "righteousness" referred to in traditional Chinese medicine. The theory of traditional Chinese medicine believes that all pathogenic factors are evil qi, and all disease-resistant abilities are righteous qi. Righteous Qi can be further divided into Wei Qi, Yuan Qi, and Zang Fu Qi of major tissues and organs. The so-called Wei Qi is the healthy Qi that can defend the human body against external evils. Wei Qi travels outside the meridians and has the functions of moisturizing the skin, nourishing the skin, resisting external evils, and protecting the body. The reason why Wei Qi can play a role mainly depends on the regulation and distribution of lung Qi. Vitality is the driving force for maintaining life activities. Traditional Chinese medicine believes that the kidney is the innate foundation. Vitality is stored in the kidney. life. The generation of qi in the five internal organs and six internal organs also depends on the support of vitality. In addition, the spleen is the foundation of the acquired constitution, which has the function of transporting and transforming the essence of water and grain, and bears the important responsibilities of digesting food, transforming qi and blood, and then distributing them throughout the body to nourish other viscera. Therefore, the spleen is the source of qi and blood biochemistry, and it is also closely related to the immune function of the body. It can be seen that the strength of the body's righteousness is closely related to the lungs, spleen, and kidneys. The strength and decline of the human body's righteousness directly determines the occurrence, development and outcome of diseases. If the righteousness is strong, the body will be able to get rid of evils and go out to avoid illness. If righteous qi is deficient, evil qi can easily invade the body and cause disease. The so-called political disease is due to "when evil is beaten, its qi must be deficient." The development and outcome of the disease also depend on the ebb and flow of the evil and the positive. If the positive prevails, the evil will lose, and the disease will heal; Therefore, in the process of disease prevention and treatment, traditional Chinese medicine has put forward the basic principle of strengthening the body and eliminating pathogenic factors to consolidate the foundation, which is consistent with the health care method of enhancing the body's immunity. There are many types of immunity-enhancing drugs currently on the market. For example, the application number is: 03139711.5, and the name is "An Antiviral Drug for Improving Immunity and Preparation Method", which discloses a drug composed of inosine and p-acetamide The molecular ratio of dimethylamino isopropanol benzoate is 1:3. The medicine composed of starch-embedded microcapsules plays an antiviral effect by improving the immune function of the body. The product is stable and effective. The effect is remarkable, it not only has antiviral effect but also improves the immune function of the body. However, this type of immune-enhancing pharmaceutical product has relatively large side effects, and is not easy to take for a long time and in large doses, so it is impossible to achieve a sustained and systematic immune-enhancing effect.

(3) Contents of the invention:

The invention discloses a health food that is safe, has no side effects, and can comprehensively and systematically enhance human immunity, and also discloses a preparation method of the health food.

Based on repeated tests and comprehensive Chinese and Western medicine theories, the inventor selected astragalus, rhodiola rosea, matsutake, and American ginseng that are safe, without any side effects, and can be taken directly to make the health food of the present invention that can enhance human immunity. When selecting the various raw materials of the health food of the present invention for combination and matching, it is necessary to consider their respective functions and the coordinated effect that they can produce. First of all, astragalus is selected because it has the function of regulating the body's immunity, and the total flavonoids of astragalus contained in it have antioxidant properties and free radical scavenging effects, which can prevent immune cells from lipid peroxidation damage and change the energy of immune cells. Metabolism and promotion of lymphocyte division, so it has a certain protective effect on immune system damage during radiotherapy, such as decreased lymphocyte transformation rate, weakened neutrophil bactericidal activity, etc., and can effectively prevent tissue damage caused by radiotherapy in lung cancer surgery. Cell damage reduces the MDA content of the tissue after irradiation, protects the SOD activity, relieves lung tissue congestion, consolidation, pleural adhesions, and esophageal mucosal erosion, and enhances the immune function of the human body. It can also improve the anti-tumor immune effect. Rhodiola rosea has anti-hypoxia resistance and protects hypoxic myocardium. It can increase the level of hemoglobin and red blood cells in the blood, increase arterial blood oxygen partial pressure and blood oxygen saturation, and improve the oxygen supply of tissues and organs in the body. Based on this, Rhodiola rosea It can achieve its antioxidant effect by reducing oxygen consumption, scavenging free radicals, increasing the activity of antioxidant enzymes, inhibiting or reducing lipid peroxidation, and inhibiting hemorheological changes caused by hypoxia. In addition, Rhodiola can also regulate the non-specific and specific immune function of normal human body under low temperature conditions, and enhance the ability to resist cold and fatigue. The matsutake contained in the formula can improve the human body's ability to resist hypoxia, and can improve the low immune function of the human body to keep it normal, and the polysaccharide components of the matsutake extract show enhanced effects on non-specific immunity and specific immunity. Has anti-stress effect. The American ginseng root crude polysaccharide contained in the American ginseng in the formula has the effect of significantly antagonizing the reduction in the weight of the body's white blood cells and immune organs caused by cyclophosphamide, can promote the transformation of the body's lymphocytes with low immune function, and can protect the body's non-specific immunity. and cellular immune function play a role in enhancing. In the formula of the health food of the present invention, the selected astragalus, rhodiola, matsutake, and American ginseng all have a good effect of enhancing immunity, and rhodiola and matsutake not only have anti-hypoxia resistance, but also have It can enhance the function of non-specific and specific immunity of the human body, plus the regulation of American ginseng on the body's non-specific immunity and cellular immunity, as well as the effect of astragalus on the anti-tumor immune effect of the human body and Rhodiola scavenging free radicals in the body When used in combination, they can promote and influence each other, play a coordinated role, and achieve a comprehensive and systematic effect of enhancing immunity. In addition, the theory of traditional Chinese medicine believes that Huang's nature is sweet and slightly warm, which has the functions of invigorating the temper, benefiting lung qi, and strengthening the body; while American ginseng is bitter, cold, and slightly sweet, which has the effect of nourishing qi and nourishing yin and invigorating vitality; Rhodiola It has the effects of invigorating the spleen and replenishing qi, promoting blood circulation and removing blood stasis, and matsutake, as an edible fungus, has the functions of strengthening the body, benefiting the stomach, nourishing deficiency and strengthening the body. Combination of all the products plays the functions of invigorating the spleen and benefiting the lungs, invigorating vitality, and regulating qi and blood, so as to achieve the purpose of invigorating deficiency, strengthening vital energy and eliminating pathogenic factors. Therefore, no matter from the theory of traditional Chinese medicine or modern pharmacology, it can be demonstrated that the comprehensive use of each component in this formula can achieve a better effect of enhancing immunity.

The ratio of each component of the health food that can enhance immunity in the present invention has been tested by the applicant many times, and various indicators such as the intake of the four components of astragalus, rhodiola rosea, matsutake, and American ginseng. According to the formulation, the parts by weight of each component are: 600-900 parts of astragalus, 400-600 parts of rhodiola, 400-600 parts of matsutake, 100-200 parts of American ginseng. The ratio in parts by weight can be: 700-800 parts of astragalus, 450-550 parts of rhodiola rosea, 450-550 parts of matsutake, and 130-180 parts of American ginseng. The optimum ratio of each component in the health food of the present invention is in parts by weight: 750 parts of astragalus, 500 parts of rhodiola, 500 parts of matsutake, and 150 parts of American ginseng.

The preparation method of the health food capable of enhancing immunity of the present invention can be as follows:

1) Take astragalus, rhodiola rosea, matsutake and American ginseng for later use;

2) Rhodiola rosea is soaked in ethanol that is 4 to 5 times its weight and has a weight concentration of 45 to 95%, and then extracted and filtered to obtain an alcohol extract; then the ethanol in the alcohol extract is recovered, and then concentrated , to obtain Rhodiola thick paste; wherein the Rhodiola soaking time is generally 3 to 5 hours, the extraction time is generally 1 to 2 hours, and the extraction times are generally 1 to 3 times;

3) Soak matsutake in water 6 to 8 times its weight, decoct to obtain a water extract; then filter the water extract, and then concentrate the filtrate to obtain matsutake thick paste; the soaking time of matsutake is generally 3 to 5 hours, the extraction time is generally 1 to 2 hours, and the extraction frequency is generally 1 to 3 times;

4) Soak Astragalus membranaceus in water 6 to 8 times its weight, decoct to obtain a water extract; then filter the water extract, and concentrate the filtrate to obtain a thick paste of Astragalus membranaceus; the soaking time of Astragalus membranaceus is generally 3 to 5 hours, the extraction time is generally 1 to 2 hours, and the extraction frequency is generally 1 to 3 times;

5) Wash the American ginseng, dry it at 50-70°C, and crush it into fine powder of American ginseng;

6) Mix the obtained rhodiola thick paste, matsutake thick paste, astragalus thick paste and American ginseng fine powder evenly, dry and pack to obtain a finished product.

Certainly also can directly adopt the extract of astragalus root, rhodiola rosea, matsutake and American ginseng together to prepare the health food that can enhance immunity of the present invention, and its each component can be by weight: Astragalus extract 300～400 parts, 300-400 parts of rhodiola extract, 300-400 parts of matsutake extract, 100-200 parts of American ginseng; and our preferred ratio of each component is: 320-380 parts of astragalus extract, 320-380 parts of rhododendron 320 to 380 parts of fenugreek extract, 320 to 380 parts of matsutake extract, and 120 to 180 parts of American ginseng; 350 parts of Tianyan extract, 350 parts of matsutake extract, and 150 parts of American ginseng.

Wherein, the extracts of Astragalus, Rhodiola, and matsutake can be commercially available extracts of Astragalus, Rhodiola, and matsutake, such as the extract of Astragalus produced by Zhejiang Kangenbei Pharmaceutical Co., Ltd., or extracted with water or other The astragalus, rhodiola and matsutake extracts extracted by the method can also be prepared by the above-mentioned method.

The health food capable of enhancing immunity of the present invention is scientifically prepared from astragalus, rhodiola rosea, matsutake, and American ginseng. Tumor immune effect, as well as regulating the non-specific and specific immunity of the human body, so as to achieve a comprehensive and systematic enhancement of the body's immunity and enhance the immunity of the immunocompromised population. And the raw material of the health food of the present invention adopts natural products or is prepared from natural products, so the health food of the present invention is a kind of natural health food that is safe, completely free from any toxic and side effects, and can be taken for a long time.

(4) Specific implementation methods:

Example 1:

1) Prepare astragalus extract:

Soak astragalus in water 8 times its weight for 4 hours, decoct for 1.5 hours, filter to obtain water extract A, then soak it in water 6 times its weight for 4 hours, decoct for 1.5 hours, Filtrate to obtain water extract B; combine A and B, perform microfiltration; and then concentrate under reduced pressure to obtain thick paste of Astragalus membranaceus for later use;

2) Prepare Rhodiola rosea extract:

Rhodiola rosea is placed in 5 times its weight and soaked in ethanol with a weight concentration of 70% for 4 hours, and extracted for 1.5 hours, filtered, and the filtrates are combined twice; then the ethanol is recovered under reduced pressure, and then Concentrate under reduced pressure to obtain a rhodiola thick ointment for subsequent use;

3) Prepare matsutake extract:

Soak matsutake in water six times its weight for 3 hours, decoct for 2 hours, filter to obtain a water extract; then microfilter the water extract, and then concentrate under reduced pressure to obtain matsutake thick paste for use;

4) Take by weighing 300 grams of Astragalus thick paste, 300 grams of Rhodiola thick paste, 300 grams of matsutake thick paste prepared by the above method, and 100 grams of dried and pulverized American ginseng, mix well, then concentrate under reduced pressure, Promptly get the health food ointment of the present invention that can enhance immunity.

Example 2:

1) Prepare astragalus extract:

Soak Astragalus membranaceus in water 7 times its weight for 5 hours, decoct 2 times for 2 hours each time, filter, combine the two water extracts, then microfilter the water extracts, and then carry out Concentrate under reduced pressure to obtain thick paste of Astragalus membranaceus for subsequent use;

2) Take by weighing 350 grams of Astragalus thick paste prepared by the above method, 350 grams of Rhodiola rosea extract prepared by water extraction method, and 350 grams of matsutake extract produced by Guilin Rhine Biological Products Co., Ltd., dried, 150 grams of pulverized American ginseng are uniformly mixed together, then dried under reduced pressure, pulverized, and packaged to obtain the health food capsule capable of enhancing immunity of the present invention.

Example 3:

1) Weigh 600 grams of astragalus, 400 grams of rhodiola, 500 grams of matsutake, and 150 grams of American ginseng for later use;

2) Mix the above-mentioned components uniformly, then dry under reduced pressure, pulverize, and package to obtain the health food granule capable of enhancing immunity of the present invention.

Example 4:

1) Weigh 600 grams of astragalus, 400 grams of rhodiola rosea, 400 grams of matsutake, and 100 grams of American ginseng.

2) Rhodiola rosea is placed in 5 times its weight and soaked in 95% ethanol for 5 hours, extracted 2 times, each extraction time is 1.5 hours, filtered, and combined with twice alcohol extracts; then recovered Concentrate the ethanol in the ethanol extract to obtain the rhodiola thick paste;

3) Soak matsutake in water 8 times its weight for 4 hours, decoct for 1.5 hours, filter to obtain water extract A, then soak it in water 6 times its weight for 5 hours, decoct for 1 hour, filtered to obtain water extract B; combined two water extracts; then filtered the water extract, and concentrated the filtrate to obtain matsutake thick paste;

4) Soak Astragalus membranaceus in water 8 times its weight for 5 hours, decoct for 2 hours, filter to obtain a water extract; then fine filter the water extract, and concentrate the filtrate to obtain a thick paste of Astragalus membranaceus;

5) Wash the American ginseng, dry it at 50°C, and crush it into fine powder of American ginseng;

6) Mix the obtained rhodiola thick paste, matsutake thick paste, astragalus thick paste, and American ginseng fine powder evenly, dry, and pack to obtain the health food powder capable of enhancing immunity of the present invention.

Example 5:

1) Weigh 350 grams of Astragalus extract, 380 grams of Rhodiola rosea extract, and 400 grams of matsutake extract produced by Guilin Laiyin Biological Products Co., Ltd. for subsequent use;

2) Wash 150 grams of American ginseng, dry it at 70°C, and crush it into fine powder of American ginseng;

3) Mix the above-mentioned astragalus extract, rhodiola rosea extract, matsutake extract and American ginseng fine powder evenly, perform tableting treatment, and pack to obtain the health food tablet capable of enhancing immunity of the present invention.

Experimental example:

1, the consumption of health food: the health food capsule of the present invention described in embodiment 2 that can enhance immunity, 0.5g/grain, content is gray powder, seals, puts shady and cool, dry place to preserve. The recommended dosage for human oral administration is 2 times a day, 2 capsules each time, and the adult body weight is calculated as 60kg, the equivalent dose is 0.0333g/kg·bw. Take the capsule contents for testing.

2. Experimental animals and grouping: 120 clean-grade Kunming female mice, weighing 18g to 22g, provided by the experimental animal farm of the College of Animal Science and Technology, Hunan Agricultural University, and the experimental animal production license number is SCXK (Xiang) 2003-0003 . Every 40 were divided into 1 group, a total of three groups. In the immune group I, the carbon clearance test was carried out; in the immune group II, the viscera ratio determination, the mouse peritoneal macrophage phagocytosis experiment of chicken red blood cells, the delayed type allergic reaction test, the determination of the half hemolysis value (HC ₅₀ ) and the number of antibody-producing cells were carried out Determination of immunization group III, ConA-induced mouse lymphocyte transformation experiment, NK cell activity determination.

3. Health food dose selection and treatment during the experiment: According to the recommended oral dose of the human body, the low, medium and high doses of the health food capsules of the present invention that can enhance immunity are respectively 0.167g/kg bw and 0.333g/kg bw , 0.999g/kg·bw (respectively equivalent to 5, 10, 30 times the recommended human dose). Get respectively the health food capsule content 1.67g, 3.33g, 9.99g of the present invention described in embodiment 2 and add distilled water to mix to 200ml, the animal that does not take medicine gives the distilled water of equal volume, give test animal respectively The stomach was administered once a day, with a volume of 0.2ml/10g·bw, for 30 consecutive days.

4. Experimental method:

(1) Determination of organ/body weight ratio: the mice were sacrificed after weighing, the spleen and thymus were taken out, weighed on an electronic analytical balance, and the organ/body ratio was calculated.

(2) Delayed type hypersensitivity (DTH) (foot plantar thickening method): 4 days after mice were sensitized by intraperitoneal injection of 2% (v/v) SRBC (0.2ml/mouse), measure the thickness of the left hind foot plantar , and then subcutaneously inject 20% (v/v) SRBC (20ul/mouse) at the measurement site, measure the thickness of the left hind foot sole 24 hours after the injection, measure the same site three times, and get the average value. The degree of DTH was expressed by the difference in the thickness of the paw (paw swelling) before and after the attack.

(3) ConA-induced mouse lymphocyte transformation experiment (MTT method): the spleen was aseptically removed, placed in a small plate filled with an appropriate amount of sterile Hank's solution to make a cell suspension, and filtered through a 200-mesh sieve. Wash twice with Hank's solution and centrifuge for 10 minutes each time (1000r/min). Then the cells were suspended in 1 mL of complete culture medium, the number of viable cells was counted, and the cell concentration was adjusted to 3×10 ⁶ cells/mL with RPMI1640 culture medium. Add the cell suspension into two wells of a 24-well culture plate, 1 mL per well, add 75 μL of ConA solution (equivalent to 7.5 μg/mL) to one well, and the other well as a control, place 5% carbon dioxide, and incubate at 37°C for 72 hours . 4 hours before the end of the culture, gently aspirate 0.7 mL of the supernatant from each well, add RPMI1640 culture solution without calf serum, and add 50 μL/well of MTT (5 mg/mL) at the same time, and continue to culture for 4 hours. After culturing, add 1mL of acidic isopropanol to each well, and mix well by pipetting to completely dissolve the purple crystals. Then it was divided into 96-well culture plates, and each well was made into three parallel wells, and the optical density value was measured at a wavelength of 570 nm with a microplate reader. The proliferative ability of lymphocytes was expressed by subtracting the optical density value of the wells without ConA from the optical density value of the wells with ConA added.

(4) Detection of antibody-producing cells (Jerne's modified glass slide method): take sheep blood, wash it with normal saline for 3 times, centrifuge (2000r/min) for 10min each time, and mix the packed SRBC with normal saline to make 2% (v/ For the cell suspension in v), 0.2 ml was injected intraperitoneally into each mouse. The mice 4 days after immunization were sacrificed, the spleen was taken out, shredded gently, and the cell suspension was prepared with Hanks solution, filtered through a 200-mesh screen, washed and centrifuged twice, and finally the cells were suspended in 5ml Hanks solution. Heat and dissolve the surface medium, mix it with the same amount of Hanks solution with pH 7.4 and 2 times the concentration, divide into small test tubes, 0.5ml per tube, and then add 50ul (v/v) of 10% SRBC prepared with SA solution into the tube , 20ul splenocyte suspension, mix quickly and pour it on the glass slide that has been brushed with thin layer of agarose. Complement (1:10) diluted with SA solution was added to the groove of the glass slide and incubated for 1.5 h, and then the number of hemolytic plaques was counted.

(5) Determination of half hemolysis value (HC ₅₀ ): sheep blood was collected, washed 3 times with normal saline, and each mouse was injected intraperitoneally with 2% (v/v, prepared with normal saline) RBC 0.2mL for immunization. After 5 days, the eyeballs were removed and the blood was collected in a centrifuge tube for about 1 hour. The coagulated blood was separated from the tube wall, and the serum was fully separated and centrifuged at 2000rpm for 10 minutes to collect the serum. Dilute the serum to 200 times with SA buffer, take 1 mL and put it in a test tube, add 10% (v/v, prepared with SA buffer) successively to pack 0.5Ml of SRBC, and complement 1 mL (dilute 1:10 with SA buffer) ). Set up a control tube without serum (replaced with SA buffer). Place it in a constant temperature water bath at 37°C for 30 minutes, and then stop the reaction in an ice bath. Centrifuge at 2000rpm for 10min, take 1mL of supernatant, add 3mL of Douglas reagent. At the same time, take 10% (v/v, prepared with SA buffer solution) pressurized SRBC 0.25mL, add Douglas reagent to 4mL in another test tube, mix well, leave it for 10min, and use the control tube as a blank at 540nm, respectively Measure the optical density of each tube. The amount of hemolysin is expressed by the half hemolysis value (HC ₅₀ ), calculated according to the following formula:

Half hemolysis value (HC ₅₀ ) = sample optical density value/optical density value of SRBC half hemolysis × dilution factor

(6) Carbon clearance experiment in mice: Indian ink diluted 4 times with normal saline was injected into the tail vein of mice, 0.1 mL per 10 g of body weight, timed immediately after the injection of the ink, and 2 and 10 minutes after the injection of the ink, respectively, from the inner canthus 20ul of blood was collected from the venous plexus, added to 2ml of Na ₂ CO ₃ solution as a blank control, and the optical density (OD) was measured colorimetrically at a wavelength of 600nm with a 722-type spectrophotometer. The mice were killed, and the liver and spleen were taken out, weighed, and the phagocytosis index a was calculated.

(7) Mouse peritoneal macrophage phagocytosis experiment of chicken red blood cells (semi-in vivo method): intraperitoneal injection of 20% (v/v, prepared with physiological saline) packed chicken red blood cell (2000rpm, 10min) suspension 1mL, interval After 30 minutes, they were sacrificed by cervical dislocation, and 1 mL of peritoneal macrophage washing solution was taken, dropped on a glass slide, placed in an enamel box lined with wet gauze, and incubated in a 37°C incubator for 30 minutes. After incubation, rinse in saline to remove unattached cells. After drying, fix with methanol: acetone (1:1), stain with 4% (v/v) Giemsa-phosphate buffer solution, rinse with distilled water and dry. Count 100 macrophages per slice under the oil immersion microscope, and calculate the phagocytosis rate and phagocytosis index according to the following formula:

Phagocytosis rate% = the number of macrophages that phagocytized chicken red blood cells/the number of counted macrophages × 100

Phagocytosis index = the total number of phagocytized chicken red blood cells/the number of counted macrophages

(8) Determination of the prevalence of NK cells (isotope ³ H-TdR assay): Take the well-growing YAC-1 cells (survival rate > 95%) 24 hours after passage, and use 1×10 ⁶ /mL YAC-1 cell suspension Add 10 μCi of ³ H-TdR for labeling, culture in a 5% carbon dioxide incubator at 37°C for 2 hours, shake once every 30 minutes, wash the labeled cells with culture medium for 3 times, resuspend in culture medium, and make the cell concentration 1 ×10 ⁵ cells/mL. The tested mice were killed by cervical dislocation, and the spleen was aseptically removed to make a spleen cell suspension, washed 3 times with Hank's solution, centrifuged at 1000rpm for 10min each time, suspended in 2mL of RPMI1640 complete culture medium containing 10% calf serum, and used Trypan blue staining and counting (the number of viable cells should be above 95%), adjusted the cell concentration to 1×10 ⁷ cells/mL. Add 100 μL triton X-100 to each well of a 96-well plate. Set up 3 duplicate wells for each palm, set 5% carbon dioxide, and culture at 37°C for 4h. The cells were collected on glass fiber filter paper with a multi-head cell collector, and the pulses per minute (cpm) were measured with a liquid scintillation meter.

NK cell activity=(1-experimental well cpm/(blank control well cpm-maximum release well cpm))×100%

5. Experimental results:

(1) The impact of the health food capsules of the present invention that can enhance immunity on the body weight of mice is shown in Tables 1, 2, and 3:

Table 1 The health food capsule of the present invention that can enhance immunity enhances immunity function experiment (I) group mouse (female) body weight

Dose group Number of animals Initial body weight Mid-term body weight Final body weight Weight gain

(g/kg BW) (only) x±s(g) x±s(g) x±s(g) x±s(g)

0.000 10 19.28±1.15 28.07±1.68 35.76±2.19 16.48±1.74

0.167 10 19.27±1.18 28.12±1.76 35.50±2.10 16.23±1.76

0.333 10 19.31±1.08 28.27±1.77 35.82±2.17 16.51±1.85

0.999 10 19.87±1.45 28.42±1.88 36.37±1.55 16.50±1.69

Table 2 The health food capsules of the present invention that can enhance immunity can enhance immunity function experiment (II) group mice (female) body weight

Dose group Number of animals Initial body weight Mid-term body weight Final body weight Weight gain

(g/kg BW) (only) x±s(g) x±s(g) x±s(g) x±s(g)

0.000 10 20.26±1.47 28.86±1.99 36.56±2.23 16.30±1.42

0.167 10 20.25±1.40 28.66±1.59 35.43±1.85 15.18±1.38

0.333 10 20.33±1.18 28.17±1.37 35.81±1.64 15.48±1.72

0.999 10 20.38±1.35 29.07±1.87 36.09±1.69 15.71±1.70

Table 3 The health food capsules of the present invention that can enhance immunity can enhance immunity function experiment (III) group mice (female) body weight

Dose group Number of animals Initial body weight Mid-term body weight Final body weight Weight gain

(g/kg BW) (only) x±s(g) x±s(g) x±s(g) x±s(g)

0.000 10 19.95±1.27 28.77±1.66 36.19±1.67 16.24±1.42

0.167 10 19.88±1.21 28.49±1.85 35.95±2.32 16.07±1.46

0.333 10 19.90±1.09 28.60±1.84 35.89±1.55 15.99±1.02

0.999 10 19.99±1.24 28.04±1.86 35.79±1.92 15.80±1.22

As can be seen from Table 1-3, at the beginning of the experiment, in the middle of the experiment, and at the end of the experiment, the body weight of the mice in each dose group and the weight increase of the mice during the experiment compared with the control group, and the difference was not significant (P>0.05)

(2) The impact of the health food capsules of the present invention that can enhance immunity on the mouse immune organ viscera/body weight ratio is shown in Table 4:

Table 4 The impact of the health food capsules of the present invention that can enhance immunity on the mouse immune organ viscera/body weight ratio

Dose group Number of animals Spleen/body weight Thymus/body weight

(g/kg BW) (only) x±s(%) P-value x±s(%) P value

0.000 10 0.479±0.028 --- 0.284±0.043 ---

0.167 10 0.492±0.032 0.752 0.302±0.052 0.806

0.333 10 0.507±0.036 0.199 0.315±0.075 0.447

0.999 10 0.516±0.043 0.063 0.311±0.039 0.542

As can be seen from Table 4, oral administration of different doses of the health food capsule content of the present invention that can enhance immunity to mice has no significant impact on mouse spleen/body weight ratio and thymus/body weight ratio (P>0.05)

(3) The influence of the health food capsule capsule of the present invention that can enhance immunity on mouse cellular immune function, see table 5,6:

A: The health food capsule of the present invention can enhance immunity to the impact of mouse delayed type hypersensitivity (DTH), see Table 5:

Table 5 The present invention can strengthen the influence of the health food capsule of immunity on mouse delayed type hypersensitivity (DTH)

Dosage group Number of animals Paw swelling 24 hours after injection

(g/kg BW) (only) x±s(mm) P value

0.000 10 0.275±0.066 ---

0.167 10 0.313±0.064 0.315

0.333 10 0.340±0.054 0.036

0.999 10 0.351±0.036 0.012

As can be seen from Table 5, different doses of the health food capsule content of the present invention that can enhance immunity were given orally to mice for 30 days, and the paw swelling degree of the middle and high dose groups was significantly higher than that of the control group, and the difference was significant (P> 0.05).

B: The impact of the health food capsules of the present invention that can enhance immunity on the mouse lymphocyte transformation ability experiment induced by mouse ConA, see Table 6:

Table 6 The impact of the health food capsule that can enhance immunity of the present invention on mouse lymphocyte transformation ability experiment

Dose group Number of animals Lymphocyte proliferation ability (OD difference)

(g/kg BW) (only) x±s P-value

0.000 10 0.053±0.017 ---

0.167 10 0.080±0.054 0.271

0.333 10 0.074±0.029 0.461

0.999 10 0.070±0.036 0.620

As can be seen from Table 6, different dosages of the health food capsule contents of the present invention that can enhance immunity were given to mice orally for 30 days, and the lymphocyte transformation ability of mice in each dosage group was compared with the control group, and the difference was not significant (P>0.05 ).

(4) The health food capsule of the present invention that can strengthen immunity is to the influence of humoral immunity, see table 7,8:

A: The impact of the health food capsules of the present invention that can enhance immunity on the number of antibody-producing cells in mice is shown in Table 7:

Table 7 The influence of the health food capsules capable of enhancing immunity of the present invention on the number of antibody-producing cells in mice

Dose group Number of animals Number of hemolytic plaques

(g/kg BW) (only) x±s(×10 ³ /whole spleen) P value

0.000 10 22.6±8.5  …

0.167 10 30.4±11.8 0.404

0.333 10 38.9±10.6 0.022

0.999 10 46.6±18.8 0.001

As can be seen from Table 7, different doses of the health food capsule contents of the present invention capable of enhancing immunity were orally administered to mice for 30 days, and the number of antibody-producing cells in mice in the middle and high dose groups was significantly higher than that in the control group, and the difference was significant ( P<0.05 or P<0.01).

B: the impact of the health food capsules of the present invention that can enhance immunity on the half hemolysis value (HC ₅₀ ) of mice, see Table 8:

Table 8 The impact of the health food capsules of the present invention that can enhance immunity on the half hemolysis value (HC50) of mice

Dose group Number of animals Half hemolysis value

(g/kg BW) (only) x±s P-value

0.000 10 181.05±28.73  …

0.167 10 197.28±18.63 0.334

0.333 10 201.99±18.23 0.159

0.999 10 220.11±30.46 0.003

As can be seen from Table 8, oral administration of different doses of the health food capsule content of the present invention that can enhance immunity to mice for 30 days, the half hemolysis value of mice in the high dose group compared with the control group, the difference is significant (P＜0.01)

(5) The influence of the health food capsule of the present invention that can enhance immunity on the phagocytic function of mouse monocyte-macrophage, see Table 9, 10, 11:

A: The effect of the health food capsules of the present invention that can enhance immunity on mouse monocyte-macrophage carbon clearance is shown in Table 9:

Table 9 The influence of the health food capsule of the present invention that can enhance immunity on mouse monocyte-macrophage carbon clearance

Dose group Number of animals Phagocytosis index (a)

(g/kg BW) (only) x±s P-value

0.000 10 5.538±0.851 ---

0.167 10 6.055±0.550 0.304

0.333 10 6.411±0.769 0.037

0.999 10 6.534±0.818 0.015

As can be seen from Table 9, the different doses of the health food capsule content of the present invention that can enhance immunity were given to mice orally for 30 days, and the phagocytosis index of mice in the middle and high dose groups was significantly higher than that of the matched group, and the difference was significant (P< 0.05).

B: The influence of the health food capsules of the present invention that can enhance immunity on the ability of mouse macrophages to phagocytize chicken red blood cells, see Tables 10 and 11:

Table 10 The health food capsule that can enhance immunity of the present invention is to the impact of mouse macrophage phagocytosis chicken erythrocyte phagocytosis rate

Dose group Number of animals Phagocytosis rate Square root of phagocytosis rate

(g/kg BW) (only) x±s(%) Inverse sine conversion value x±s P value

0.000 10 28.2±7.6 31.9±4.8 ---

0.167 10 29.6±7.5 32.8±4.7 0.950

0.333 10 34.0±8.5 35.5±5.2 0.227

0.999 10 34.9±7.0 36.1±4.2 0.133

Table 11 The health food capsule that can enhance immunity of the present invention is on the impact of mouse macrophage phagocytosis chicken erythrocyte phagocytosis index

Dose group Number of animals Phagocytic index

(g/kg BW) (only) x±s P-value

0.000 10 0.929±0.184 ---

0.167 10 0.988±0.212 0.868

0.333 10 1.075±0.257 0.300

0.999 10 1.086±0.186 0.247

It can be seen from Table 10-11 that when mice were orally given different doses of the contents of the health food capsules of the present invention capable of enhancing immunity for 30 days, each dose group had no significant effect on the ability of mouse macrophages to phagocytize chicken erythrocytes.

(6) The influence of the health food capsule of the present invention that can enhance immunity on mouse NK cell activity is shown in Table 12:

Table 12 The influence of the health food capsule that can enhance immunity of the present invention on mouse NK cell activity

Dose group Number of animals NK cell activity NK cell activity squared

(g/kg BW) (only) x±s(%) root arcsine conversion value x±s P value

0.000 10 34.0±11.3 35.4±7.1 ---

0.167 10 38.0±7.9 37.9±5.0 0.792

0.333 10 38.2±12.6 38.0±7.5 0.771

0.999 10 46.4±16.3 42.9±9.7 0.079

As can be seen from Table 12, the different doses of the health food capsule contents of the present invention that can enhance immunity were orally administered to mice for 30 days, and each dose group had no significant difference in activity of NK cells in mice compared with the control group (P>0.05 ).

6. Experimental conclusion:

Oral administration of 0.167g/kg bw, 0.333g/kg bw, 0.999g/kg bw doses of the health food capsule content of the present invention that can enhance immunity to mice for 30 days, 0.333g/kg bw, 0.999 The dose of g/kg bw can enhance the delayed allergic reaction of mice, increase the number of antibody-producing cells in mice, and improve the ability of mouse monocyte-macrophage carbon clearance, and the dose of 0.999g/kg bw can increase the serum concentration of mice half hemolysis value. Compared with the control group, P<0.05 or P<0.01. It has no significant effect on mouse weight gain, spleen/body weight ratio, thymus/body weight ratio, ConA-induced mouse lymphocyte transformation ability, mouse NK cell activity, and mouse macrophage chicken red blood cell ability. It is suggested that the health food capable of enhancing immunity of the present invention has the function of enhancing immunity.

Claims (10)

1. A health food capable of enhancing immunity, characterized in that each component is calculated in parts by weight: 600-900 parts of astragalus, 400-600 parts of rhodiola, 400-600 parts of matsutake, and 100-200 parts of American ginseng. 2. The health food capable of enhancing immunity according to claim 1, characterized in that each component is calculated in parts by weight: Astragalus 700-800 parts, Rhodiola rosea 450-550 parts, matsutake 450-550 parts, American ginseng 130-180 copies. 3. The health food capable of enhancing immunity according to claim 2, characterized in that each component is calculated in parts by weight: 750 parts of astragalus, 500 parts of rhodiola rosea, 500 parts of matsutake, and 150 parts of American ginseng. 4. The preparation method of the health food capable of enhancing immunity according to any one of claims 1-3, the steps are as follows: 1) Take astragalus, rhodiola rosea, matsutake and American ginseng for later use; 2) Rhodiola rosea is soaked in ethanol that is 4 to 5 times its weight and has a weight concentration of 45 to 95%, and then extracted and filtered to obtain an alcohol extract; then the ethanol in the alcohol extract is recovered, and then concentrated , get rhodiola thick paste; 3) Soak matsutake in water 6 to 8 times its weight, decoct to obtain a water extract; then filter the water extract, and concentrate the filtrate to obtain matsutake thick paste; 4) Soak Astragalus membranaceus in water 6 to 8 times its weight, and decoct to obtain a water extract; then filter the water extract, and concentrate the filtrate to obtain a thick paste of Astragalus membranaceus; 5) Wash the American ginseng, dry it at 50-70°C, and crush it into fine powder of American ginseng; 6) Mix the obtained rhodiola thick paste, matsutake thick paste, astragalus thick paste and American ginseng fine powder evenly, dry and pack to obtain a finished product. 5. The preparation method of health food capable of enhancing immunity according to claim 4, characterized in that: In step 2), the soaking time of Rhodiola rosea is 3 to 5 hours, the extraction time is 1 to 2 hours, and the extraction times are 1 to 3 times; In step 3), the matsutake soaking time is 3 to 5 hours, the extraction time is 1 to 2 hours, and the extraction times are 1 to 3 times; In step 4), the soaking time of Radix Astragali is 3-5 hours, the extraction time is 1-2 hours, and the extraction frequency is 1-3 times. 6. A health food capable of enhancing immunity, characterized in that each component is calculated in parts by weight: 300-400 parts of Astragalus extract, 300-400 parts of Rhodiola rosea extract, 300-400 parts of matsutake extract, 100-200 parts of American ginseng. 7. The health food capable of enhancing immunity according to claim 7, characterized in that each component is calculated in parts by weight: 320-380 parts of astragalus extract, 320-380 parts of rhodiola extract, matsutake extract 320-380 parts, 120-180 parts of American ginseng. 8. The health food capable of enhancing immunity according to claim 8, wherein the components are in parts by weight: 350 parts of Astragalus extract, 350 parts of Rhodiola rosea extract, 350 parts of matsutake extract, American ginseng 150 servings. 9. The health food capable of enhancing immunity according to any one of claims 6-8, characterized in that: Wherein the preparation method of Rhodiola rosea extract, such as the following steps: 1) Rhodiola rosea is soaked in ethanol that is 4 to 5 times its weight and has a weight concentration of 45 to 95%, extracted, filtered, and an alcohol extract is obtained; 2) recovering the ethanol in the ethanol extract and concentrating it to obtain the rhodiola rosea extract. Wherein the preparation method of matsutake extract, such as the following steps: 1) Soak matsutake in water 6 to 8 times its weight, decoct to obtain water extract; 2) filtering the water extract; 3) Concentrate the filtrate to obtain matsutake extract. Wherein the preparation method of Radix Astragali extract, as following steps: 1) soak astragalus in water 6 to 8 times its weight, and decoct to obtain a water extract; 2) filtering the water extract; 3) Concentrate the filtrate to obtain the Radix Astragali extract. 10. The health food capable of enhancing immunity according to claim 9, characterized in that: In step 1) of the preparation method of Rhodiola rosea extract, the soaking time of Rhodiola rosea is 3 to 5 hours, the extraction time is 1 to 2 hours, and the extraction times are 1 to 3 times; In step 1) of the preparation method of the matsutake extract, the soaking time of the matsutake is 3 to 5 hours, the extraction time is 1 to 2 hours, and the extraction times are 1 to 3 times; In the step 1) of the preparation method of the astragalus extract, the soaking time of the astragalus is 3-5 hours, the extraction time is 1-2 hours, and the extraction times are 1-3 times.