CN108567811B - Composition with anti-fatigue and immunoregulation functions and preparation method and application thereof - Google Patents
Composition with anti-fatigue and immunoregulation functions and preparation method and application thereof Download PDFInfo
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- CN108567811B CN108567811B CN201810725230.2A CN201810725230A CN108567811B CN 108567811 B CN108567811 B CN 108567811B CN 201810725230 A CN201810725230 A CN 201810725230A CN 108567811 B CN108567811 B CN 108567811B
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- ginseng
- water
- fatigue
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Abstract
The invention relates to a composition with anti-fatigue and immunoregulation functions and a preparation method thereof, wherein the composition comprises 30-80 parts of epimedium, 2-40 parts of ginseng, 5-40 parts of astragalus and 5-40 parts of dogwood. The composition provided by the invention improves the working capacity of the body, relieves fatigue, promotes appetite and promotes the health of the body through the synergistic effect of a plurality of active ingredients. The whole formula has the effects of tonifying the innate and acquired after the nourishing, the effect is concentrated, the combination of the tonics is converged, the medicines are compatible, the medicinal effect is synergistic, and the effect is doubled.
Description
Technical Field
The invention belongs to the technical field of medicines and health-care foods, and particularly relates to a composition with anti-fatigue and immunoregulation functions, and a preparation method and application thereof.
Background
Modern people have great living and working pressure, so that many people are in sub-health state and still feel tired frequently without strenuous physical activity or frequent mental activity. Fatigue is a state of non-organic change or undiagnosed disease, but the body shows functional changes, including physical and psychological discomforts, and shows vitality and decline of external adaptability. Long-term fatigue can not only cause substantial lesions in some organs and cause some related diseases, but also reduce immunity.
Immunity is the body's own defense mechanism, and is the body's ability to recognize, phagocytize and destroy foreign bacteria, viruses, senesced and dead cells, mutated cells and substances causing allergy, and is an important barrier to maintain the body's environment stable and to maintain the body's health. People with low immunity can have the conditions of frequent cold, difficult recovery, easy wound infection, multiple gastrointestinal diseases, easy infection or cancer, and the like, and the life quality and even the life span are obviously reduced. From about 30 years old, the immunity of the human body begins to decline slowly and continuously, the decline of the immunity is one of the most important reasons of aging, and fatigue accelerates the decline of the immunity, so that the human body enters a state of low immunity earlier.
Disclosure of Invention
Aiming at the technical problem that the human health is damaged due to fatigue and low immunity, the invention provides a composition with anti-fatigue and immunoregulation functions.
The invention also provides a preparation method of the composition with the functions of resisting fatigue and regulating immunity.
The invention also provides a preparation method of the second composition with the functions of resisting fatigue and regulating immunity.
The invention also provides a preparation method of the third composition with the functions of resisting fatigue and regulating immunity.
The invention also provides application of the composition with the functions of resisting fatigue and regulating immunity in preparation of medicines or health-care foods.
The invention also provides an oral preparation with the functions of resisting fatigue and regulating immunity.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a composition with anti-fatigue and immunoregulation functions comprises the following raw materials in parts by weight: 30-80 parts of epimedium, 2-40 parts of ginseng, 5-40 parts of astragalus and 5-40 parts of dogwood.
Compared with the prior art, the composition with the functions of resisting fatigue and regulating immunity provided by the invention improves the working capacity of the body, relieves fatigue and promotes appetite through the synergistic effect of a plurality of active ingredients, thereby being capable of promoting the health of the body, particularly the old and the weak and promoting recovery after illness. In the formula, the ginseng and the astragalus root are good products for traditional invigoration, and the ginseng and the astragalus root contain various saponins and flavonoid active ingredients, so that the endurance of the organism can be improved, the fatigue can be reduced, and the defense capability and the adaptability of the organism can be enhanced. Ginseng is sweet and warm in nature and tonifying, has powerful and powerful effect, can strengthen the primordial qi of the kidney, strengthen the qi of the spleen and lung to nourish blood and promote the production of body fluid, and also enters the heart meridian to help essence and spirit, so it has the efficacy of invigorating primordial qi, tonifying spleen and kidney. Epimedium has the functions of nourishing liver and kidney, strengthening muscles and bones, dispelling wind and eliminating dampness and strengthening heart, and icariin is used as a main effective component contained in epimedium, can obviously enhance T cell function, generates immune activation effect, promotes the increase of spleen cell number, and has wide physiological effect in various aspects such as anti-stress, immune regulation system, cardiovascular system and the like. Cornus officinalis sour is warm and moist in nature, warm but not dry in nature, and tonic but not greasy, and has the effects of strengthening primordial qi, replenishing essence and tonifying. In the formula, the ginseng and the astragalus root have the effects of tonifying qi and spleen to strengthen the spleen and stomach, promote the transportation and transformation of the spleen and stomach, and generate qi and blood, and have the effect of naturally generating qi after the cultivation; herba Epimedii is pungent, sweet and warm, and has effects of nourishing liver and kidney, and strengthening tendons and bones; cornus officinalis is sour and tonifying but not greasy, astringes and astringes, has the tonifying efficacies of strengthening primordial qi and replenishing essence, and can make the yang qi of the other three tonifying flavors not disperse and converge so as to exert the maximum efficacy. The whole formula has the effects of tonifying the innate and acquired after the nourishing, the effect is concentrated, the combination of the tonics is converged, the medicines are compatible, the medicinal effect is synergistic, and the effect is doubled.
Preferably, the composition comprises the following raw materials in parts by weight: 35-75 parts of epimedium, 2-30 parts of ginseng, 5-30 parts of astragalus and 5-30 parts of dogwood.
Preferably, the composition comprises the following raw materials in parts by weight: 40-70 parts of epimedium, 2-25 parts of ginseng, 10-30 parts of astragalus and 10-30 parts of dogwood.
Preferably, the composition comprises the following raw materials in parts by weight: 52 parts of epimedium, 5 parts of ginseng, 15 parts of astragalus and 14 parts of dogwood.
Preferably, the composition comprises the following raw materials in parts by weight: 45 parts of epimedium, 23 parts of ginseng, 23 parts of astragalus and 22 parts of dogwood.
The embodiment of the invention also provides a preparation method of the composition with the functions of resisting fatigue and regulating immunity, which comprises the following steps:
weighing the raw materials according to the raw material ratio of the composition, respectively extracting the astragalus, the dogwood, the epimedium and the ginseng with water, or extracting any two of the raw materials together with water, separately extracting the rest components with water, and mixing the extracts to obtain the composition.
Preferably, the preparation method comprises the following specific implementation modes: extracting herba Epimedii, radix astragali, Corni fructus and Ginseng radix with water respectively to obtain herba Epimedii extract, radix astragali extract, Corni fructus extract and Ginseng radix extract, and mixing.
The embodiment of the invention also provides a preparation method of a second composition with the functions of resisting fatigue and regulating immunity, which comprises the following steps:
weighing the raw materials according to the raw material proportion of the composition, and respectively extracting the astragalus, the dogwood and the epimedium with water, or extracting any two or more of the three components with water together, and independently extracting the rest components with water to prepare a mixed extract; pulverizing Ginseng radix, and mixing with the mixed extract.
The method extracts radix astragali, Corni fructus and herba Epimedii with water, concentrates and aggregates effective components to obtain product containing effective components with higher concentration per unit weight, and directly adds pulverized Ginseng radix to make the effective components in Ginseng radix fully utilized and fully exert its effects of invigorating qi and tranquilizing mind.
Preferably, the preparation method comprises the following specific embodiments:
extracting herba Epimedii, radix astragali and Corni fructus with water, drying, respectively making into herba Epimedii extract, radix astragali extract and Corni fructus extract, mixing, adding Ginseng radix powder, and mixing;
or extracting herba Epimedii, radix astragali and Corni fructus with water respectively to obtain herba Epimedii extractive solution, radix astragali extractive solution and Corni fructus extractive solution, mixing, adding Ginseng radix powder, and mixing;
or extracting herba Epimedii, radix astragali and Corni fructus with water respectively, concentrating into fluid extract, making into herba Epimedii fluid extract, radix astragali fluid extract and Corni fructus fluid extract, adding Ginseng radix powder, mixing, and drying;
or extracting herba Epimedii, radix astragali and Corni fructus with water, drying, making into mixed extract, adding Ginseng radix powder, and mixing.
Or extracting radix astragali and Corni fructus with water, drying, extracting herba Epimedii with water, drying, making into mixed extract, adding Ginseng radix powder, and mixing.
The embodiment of the invention also provides a preparation method of a third composition with the functions of resisting fatigue and regulating immunity, which specifically comprises the following steps:
weighing the raw materials according to the raw material ratio of the composition, and respectively extracting the astragalus, the dogwood, the epimedium and part of the ginseng with water, or extracting any two or more of the components with water together, and independently extracting the rest components with water to prepare a mixed extract; pulverizing the rest part of Ginseng radix, and mixing with the mixed extract.
Preferably, the preparation method comprises the following specific embodiments:
extracting herba Epimedii, radix astragali, Corni fructus and part of Ginseng radix with water, drying, making into herba Epimedii extract, radix astragali extract, Corni fructus extract and Ginseng radix extract, respectively, adding the rest Ginseng radix powder, and mixing;
or mixing radix astragali, Corni fructus, herba Epimedii and part of Ginseng radix with water, extracting, drying, making into mixed extract, adding the rest Ginseng radix powder, and mixing;
the three preparation methods and the optimal selection method are simple in production flow and easy for mass production.
The embodiment of the invention also provides application of the composition with the functions of resisting fatigue and regulating immunity in preparation of medicines or health-care foods.
After the composition is prepared into a medicine or a health-care food, the composition is more suitable for people with fatigue and low immunity to improve sub-health state, resist fatigue and improve immunity.
The embodiment of the invention also provides an oral preparation with the functions of resisting fatigue and regulating immunity, which comprises the composition prepared by the first preparation method and an excipient; the oral preparation can be in the form of tablet, oral liquid, hard capsule, soft capsule, granule, powder, pill or teabag.
After the composition prepared by the preparation method is prepared into an oral preparation, the use is more convenient and the compliance is better.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further illustrated with reference to the following specific embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 52g of epimedium herb, 5g of ginseng, 15g of astragalus root and 14g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, cutting the astragalus and the epimedium into segments, and crushing the dogwood; placing radix astragali and Corni fructus in extraction tank, decocting in water for 3 times (290 g water for each time), extracting for the first time for 2 hr, extracting for the second time for 2 hr, extracting for the third time for 1 hr, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 3 times (water amount is 520g each time), extracting for the first time for 2 hr, extracting for the second time for 2 hr, extracting for the third time for 1 hr, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, vacuum drying, and making into extract powder b; mixing the extract powder a and the extract powder b to prepare a mixed extract;
and step b, grinding the ginseng, sieving the ground ginseng with a 300-mesh sieve, and uniformly mixing the ground ginseng and the mixed extract.
Example 2
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 45g of epimedium, 23g of ginseng, 23g of astragalus root and 22g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, and cutting the astragalus and the epimedium into segments; placing radix astragali and Corni fructus in extraction tank, decocting in water for 2 times (water amount is 360g each time), extracting for 3 hr for the first time and 2 hr for the second time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and spray drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 2 times (water amount is 360g each time), extracting for 3 hr for the first time and 2 hr for the second time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and spray drying to obtain extract powder b; mixing the extract powder a and the extract powder b to prepare a mixed extract;
and step b, grinding the ginseng, sieving the ground ginseng with a 100-mesh sieve, and uniformly mixing the ground ginseng and the mixed extract.
Example 3
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 52g of epimedium herb, 15g of ginseng, 15g of astragalus root and 14g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, and crushing the astragalus, the dogwood and the epimedium; placing radix astragali and Corni fructus in extraction tank, decocting in water for 1 time (water amount is 174g), extracting for 3 hr, filtering, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 1 time (water amount is 312g), extracting for 3 hr, filtering, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder b;
and step b, grinding the ginseng, sieving the ground ginseng by a 80-mesh sieve, and uniformly mixing the ground ginseng with the extract powder a and the extract powder b to obtain the ginseng extract.
Example 4
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 60g of epimedium, 10g of ginseng, 20g of astragalus root and 25g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, and crushing the astragalus, the dogwood and the epimedium; placing radix astragali and Corni fructus in an extraction tank, decocting in water for 2 times (water amount is 225g each time), extracting for 2 hr for the first time and 1 hr for the second time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 2 times (water amount is 600g each time), extracting for 2 hr for the first time and 1 hr for the second time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder b; mixing the extract powder a and the extract powder b to prepare a mixed extract;
and step b, grinding the ginseng, sieving the ground ginseng with a 80-mesh sieve, and uniformly mixing the ground ginseng and the mixed extract.
Example 5
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 65g of epimedium, 20g of ginseng, 30g of astragalus root and 15g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, and crushing the astragalus, the dogwood and the epimedium; placing radix astragali and Corni fructus in an extraction tank, decocting in water for 3 times (water consumption is 180g each time), extracting for 3 hr for the first time, 2 hr for the second time, and 1 hr for the third time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 3 times (water amount is 520g each time), extracting for 3 hr for the first time, 2 hr for the second time, and 1 hr for the third time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder b; mixing the extract powder a and the extract powder b to prepare a mixed extract;
and step b, grinding the ginseng, sieving the ground ginseng with a 100-mesh sieve, and uniformly mixing the ground ginseng and the mixed extract.
Example 6
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 50g of epimedium, 12g of ginseng, 25g of astragalus and 20g of dogwood.
The preparation method comprises the following steps:
step a, weighing the raw materials according to the raw material proportion of the composition, and crushing the astragalus, the dogwood and the epimedium; placing radix astragali and Corni fructus in extraction tank, decocting in water for 1 time (water amount is 180g), extracting for 2 hr, filtering, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder a; placing herba Epimedii in extraction tank, decocting in water for 1 time (water amount is 300g), extracting for 2 hr, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and vacuum drying to obtain extract powder b; mixing the extract powder a and the extract powder b to prepare a mixed extract;
and step b, grinding the ginseng, sieving the ground ginseng by a 120-mesh sieve, and uniformly mixing the ground ginseng and the mixed extract.
Example 7
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 30g of epimedium, 2g of ginseng, 5g of astragalus root and 5g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material proportion of the composition, crushing the astragalus, the dogwood, the epimedium and the ginseng, performing ultrasonic extraction on the astragalus, the dogwood and the epimedium for 1 time and 2 hours respectively by using water with the weight being 10 times of that of the raw materials, filtering, performing reduced pressure concentration to obtain extract, mixing the extract, the astragalus extract and the dogwood extract, adding ginseng powder, uniformly mixing, and performing reduced pressure drying to obtain the composition.
Example 8
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 35g of epimedium, 2g of ginseng, 5g of astragalus root and 5g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material ratio of the composition, respectively cutting the astragalus, the epimedium and the ginseng into sections, crushing the dogwood, respectively adding water which is 10 times of the weight of each raw material, performing ultrasonic extraction for 1 time, extracting for 1 hour, filtering, concentrating under reduced pressure to obtain clear paste, and performing boiling drying to obtain an epimedium extract, an astragalus extract, a dogwood extract and a ginseng extract; mixing the above mixtures.
Example 9
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 75g of epimedium, 30g of ginseng, 30g of astragalus root and 30g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material proportion of the composition, respectively cutting the astragalus, the epimedium and half of the ginseng into segments, crushing the dogwood, respectively adding water with the weight 8 times of that of each raw material, decocting and extracting for 1 time and 3 hours, filtering, concentrating under reduced pressure to obtain clear paste, and boiling and drying to obtain an epimedium extract, an astragalus extract, a dogwood extract and a ginseng extract; pulverizing the rest Ginseng radix, sieving with 100 mesh sieve, and mixing with the above mixture.
Example 10
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 40g of epimedium, 2g of ginseng, 10g of astragalus root and 10g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material proportion of the composition, respectively crushing the astragalus, the dogwood and the epimedium, respectively adding water which is 7 times of the weight of each raw material, decocting and extracting for 2 times, extracting for 2 hours each time, filtering, combining extracting solutions, concentrating under reduced pressure to obtain clear paste, and boiling and drying to obtain an epimedium extract, an astragalus extract and a dogwood extract; pulverizing Ginseng radix, sieving with 100 mesh sieve, and mixing with the above mixture.
Example 11
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 70g of epimedium, 25g of ginseng, 30g of astragalus root and 30g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material ratio of the composition, respectively pulverizing radix astragali, Corni fructus, herba Epimedii and Ginseng radix, mixing, decocting in water for 2 times (water amount is 1200g each time), extracting for 3 hr each time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and boiling and drying.
Example 12
The embodiment of the invention provides a composition with anti-fatigue and immunoregulation functions, which comprises the following raw materials in parts by weight: 80g of epimedium, 40g of ginseng, 40g of astragalus root and 40g of dogwood.
The preparation method comprises the following steps:
weighing the raw materials according to the raw material ratio of the composition, respectively pulverizing radix astragali, Corni fructus, herba Epimedii and half of Ginseng radix, mixing, decocting in water for 3 times (water amount is 1200g each time), extracting for 2 hr each time, filtering, mixing extractive solutions, concentrating under reduced pressure to obtain fluid extract, and boiling and drying to obtain extract powder; pulverizing the rest Ginseng radix, sieving with 100 mesh sieve, and mixing with the extract powder.
Example 13
The embodiment of the invention provides a capsule with the functions of fatigue resistance and immunoregulation: and adding a proper amount of dextrin and magnesium stearate into the composition obtained in the example 1, mixing uniformly, and filling into capsules to obtain the capsule.
Example 14
The embodiment of the invention provides a soft capsule with the functions of fatigue resistance and immunoregulation: adding a proper amount of vegetable oil and beeswax into the composition obtained in the embodiment 1, mixing uniformly, and pressing into soft capsules to obtain the soft capsules.
Example 15
The embodiment of the invention provides an oral liquid with the functions of fatigue resistance and immunoregulation: and (3) adding a proper amount of water, a flavoring agent and a preservative into the composition obtained in the example 2, and mixing to obtain the composition.
Example 16
The embodiment of the invention provides granules with the functions of resisting fatigue and regulating immunity: and (3) adding a proper amount of dextrin into the composition obtained in the embodiment 2, mixing uniformly, and granulating to obtain the composition.
Example 17
The embodiment of the invention provides a pill with the functions of resisting fatigue and regulating immunity: adding a proper amount of starch and honey into the composition obtained in the example 1, mixing uniformly, and making into pills.
Example 18
The embodiment of the invention provides powder with the functions of resisting fatigue and regulating immunity: and (3) mixing the composition obtained in the example 5 uniformly, and packaging in small bags to obtain the composition.
Example 19
The embodiment of the invention provides a tablet with anti-fatigue and immunoregulation functions, which comprises the following components in part by weight: adding 6g of microcrystalline cellulose and 8g of starch into the composition obtained in example 1, uniformly mixing, granulating, tabletting and coating.
Example 20
The embodiment of the invention provides a tablet with anti-fatigue and immunoregulation functions, which comprises the following components in part by weight: 27g of microcrystalline cellulose and 34g of starch are added into the composition obtained in the example 2, and the mixture is uniformly mixed, granulated, tabletted and coated to obtain the composition.
Example 21
The embodiment of the invention provides a tablet with anti-fatigue and immunoregulation functions, which comprises the following components in part by weight: adding 10g of microcrystalline cellulose and 15g of starch into the composition obtained in example 3, uniformly mixing, granulating, tabletting and coating.
Example 22
The embodiment of the invention provides a tablet with anti-fatigue and immunoregulation functions, which comprises the following components in part by weight: adding 15g of microcrystalline cellulose and 20g of starch into the composition obtained in the example 4, uniformly mixing, granulating, tabletting and coating.
Example 23
The example of the present invention provides a toxicological experiment of the tablet obtained in the above example 19:
1. acute toxicity test of mice: 40 clean Kunming mice with the weight of 18-22 g and half male and female are selected and tested according to the Horn method. The highest dose was 21500mg/kg.bw (distilled water as solvent, the same below), and the gavage was performed twice within 24 hours by the equal-volume gavage method (0.2ml/10g.bw), and continuously observed for two weeks. The signs of intoxication and death were recorded.
2. Acute toxicity test in rats: 40 Wistar rats with the weight of 180-. The highest dose was 21500mg/kg.bw, with four gavages within 24 hours using an equal-volume gavage (1ml/100g.bw) and observed for two weeks continuously. The signs of intoxication and death were recorded.
3. And (3) genetic toxicity test:
(1) ames test: the experiment adopts a flat plate doping method, and the mixed liquid is added or not added with S9. The dosages are 0.005, 0.025, 0.25, 1.0 and 5.0 mg/dish respectively, and a blank control group and a positive control group are additionally arranged. The positive control group used 2-AF20 ug/dish, 2,4, 7-TNFane 1 ug/dish, NaN32.5 ug/dish, mitomycin C4.0 ug/dish, 1, 8-dihydroxy anthraquinone 50 ug/dish, 3 plates per dose group, repeated twice. The strains are T A97(a), T A98, T A100 and TA102 strains.
(2) Mouse marrow pleochromocyte micronucleus test: selecting 50 clean Kunming mice with weight of 25-30 g, randomly dividing the mice into five groups, each group comprises 10 mice and each half of the mice is male and female; three dose groups were set: 2500. 5000 and 10000mg/kg of body weight, setting a negative control group (distilled water) and a positive control group (CP 40mg/kg) at the same time, taking samples twice at an interval of 24 hours for each group, taking sternum bone marrow slices 6 hours after the second sample taking, counting 1000 pleochromocyte of each animal, and calculating the micronucleus occurrence rate.
(3) Mouse teratospermia test: selecting 25 clean-grade Kunming male mice with the weight of 30-35 g, and randomly dividing the mice into five groups, wherein each group comprises five mice; three dose groups were set: 2500. 5000, 10000mg/kg body weight, negative control group (distilled water) and positive control group (CP 40mg/kg), and oral administration for 5 days. Animals were sacrificed on day 35 from the first sample presentation, bilateral epididymis were sectioned, 1000 sperm were counted per animal, and the teratogenicity rate was calculated.
4. Feeding test for 30 days
80 initial weaning Wistar rats with the weight of 60-80g are selected and randomly divided into four groups, and each group comprises 20 rats. Breeding in single cage with half male and female. The test article has three test doses of 1.6g/kg body weight, 3.1g/kg body weight, and 6.2g/kg body weight (corresponding to 100 times of the recommended amount), and a negative control group is provided. The test substances were incorporated into the feed separately and fed to each group of animals for 30 consecutive days. The negative control group was given basal diet only. During the feeding period, animals in each group had free access to food and water, the mice were weighed weekly and the feed consumption was recorded once, and tail blood was collected at the end of the experiment for hematology and biochemistry examinations. The observation indexes include growth, food utilization, hematology examination (measured by a blood cell counter), and biochemistry examination (after serum is separated, a corresponding kit and a biochemical analyzer are adopted, and a 756MC ultraviolet-visible spectrophotometer). At the end of the experiment, the rats were sacrificed, the liver, spleen, kidney, and testis (ovary) were dissected and weighed, organ coefficients were calculated, and then histopathological examination (paraffin section) was performed on the liver, kidney, stomach, and intestine.
The experiment result shows that the composition of the invention can treat LD of big mice and mice of two sexes50Bw are all more than 21500mg/kg, which indicates that the product is nontoxic; the results of the three mutagenesis tests are negative; all indexes of the rat feeding test for 30 days have no obvious abnormality.
Example 24
The present example provides a study (animal test) of the anti-fatigue pharmacodynamics of the tablet obtained in the above example 19.
1. Experimental animals: the clean-grade male Kunming mice are 18-22 g in weight and are randomly grouped according to the weight.
2. Experimental samples: example 19 the recommended dose of the pulverized fine powder for human body is 3.72 g/day/60 kg body weight.
3. Selecting the dosage: the equivalent dose of the mouse is 10 times of the recommended dose of the human body, namely, 0.62g/kg body weight is ingested per day to be low dose, and then 2 times and 3 times of the low dose are respectively set into a dose group, namely 1.24 g/day/kg body weight (medium dose) and 1.86 g/day/kg body weight (high dose); distilled water was given to the control group, and the test substance was administered once a day in an equal volume to the stomach, and after 30 days, each index was measured.
4. The experimental method comprises the following steps:
(1) swimming test: 30 minutes after the last administration of the test substance, the tail root of the mouse was loaded with a lead skin of 5% of the weight of the mouse, and the mouse was placed in a swimming box at 25 ℃ and 35cm in water depth. The temperature of the water was adjusted to 25 ℃ before each batch of mice was launched into the water and the room temperature was kept constant at 25 ℃.5 mice were placed in each water tank at a time. The time from the start of swimming to exhaustion death was recorded with a stopwatch, and this time was the swimming time of the mouse.
(2) Serum urea nitrogen determination (diacetyl-oxime method): after 30 minutes from the last administration of the test substance, the mice were placed in water at 30 ℃ for 90 minutes, and blood was taken from the eyes and serum was measured. Using the kit method, the procedure is as shown in Table 1.
TABLE 1
The solutions to be tested were prepared according to table 1, mixed thoroughly, boiled in a boiling water bath for 10 minutes and then cooled in cold water. After zeroing with distilled water at a wavelength of 520nm, absorbance values of the standard, sample and blank were determined.
(3) Liver glycogen assay (anthraquinone method): after 30 minutes from the last administration of the test substance, the mice were placed in water at 30 ℃ for 90 minutes and sacrificed immediately. Taking the liver, rinsing with normal saline, then sucking the liver with filter paper, accurately weighing 200mg of the liver, adding 4ml of TCA, homogenizing for 1 minute in each tube, pouring the homogenate liquid into a centrifuge tube, centrifuging for 15 minutes at 3000 rpm, and transferring the supernatant into another test tube. Adding 4ml of TCA into the precipitate, homogenizing for 1 minute, centrifuging again for 15 minutes, taking supernatant, combining with the supernatant obtained by the first centrifugation, mixing fully, taking 1ml of supernatant, putting into a10 ml centrifuge tube, adding 4ml of 95% ethanol into each tube, and mixing fully until no interface is left between the two liquids. The mixture was stoppered with a clean stopper and allowed to stand overnight at room temperature. After the precipitation was complete, the tube was centrifuged at 3000 rpm for 15 minutes. Carefully pour off the supernatant and place the tube upside down for 10 minutes. The glycogen was dissolved in 2ml of distilled water, and the glycogen on the tube wall was washed off with water to completely dissolve it. Reagent blank: suck 2ml of distilled water into a clean centrifuge tube. Standard tubes: 0.5ml of a glucose standard (containing 100mg/dl glucose) and 1.5ml of distilled water were aspirated into the same centrifuge tube. And (3) determination: 10ml of anthrone reagent was added to each tube vigorously, and the flow was directed into the center of the tube, ensuring thorough mixing. When the anthrone reagent is injected into the tube, the tube is put into cold water, and after the temperature of the cold water is reached, the tube is immersed in a boiling water bath for 15 minutes, and then the tube is moved to a cold water bath and cooled to room temperature. The liquid in the tube was transferred to a cuvette and the absorbance of the sample tube and the standard tube was measured after zeroing with a reagent blank at a wavelength of 620 nm. Liver glycogen content (expressed in mg/g liver) was converted from liver weight.
(4) And (3) blood lactic acid determination: after the test object is given for 30 minutes at the last time, the root of the mouse bears a lead skin which is 4 percent of the weight of the mouse, the mouse is put into a swimming box with the water temperature of 30 ℃ and the water depth of 35cm for swimming for 10 minutes and then taken out, 20 mu l of tail blood is immediately collected and added into a test tube containing 40 mu l of hemolytic agent for even mixing, and the blood lactic acid value is measured by a biological sensing analyzer. In addition, the blood lactic acid level was measured before and after swimming for 30 minutes at rest in the same manner.
5. Results
(1) Effect of composition on mouse body weight.
Initial body weights of each group of mice are shown in table 2.
TABLE 2
The mid-term body weights of the mice in each group are shown in Table 3.
TABLE 3
End-stage body weights of the mice in each group are shown in Table 4.
TABLE 4
As can be seen from tables 2-4, when the composition was orally administered to mice at different doses for 30 days, there was no significant difference in the body weight change of mice in each dose group compared to the control group (P > 0.05).
(2) The effect of the composition on the swimming time of the mice is shown in table 5.
TABLE 5
As can be seen from Table 5, when the composition was orally administered to the mice at different doses for 30 days, the swimming time of the mice in the medium and high dose groups was longer than that in the control group, and there was a significant difference (P < 0.05).
(3) The effect of the composition on mouse serum urea nitrogen is shown in table 6.
TABLE 6
As can be seen from Table 6, the composition was orally administered to the mice at different doses for 30 days, and the serum urea nitrogen in exercise was significantly different in the middle-dose group than in the control group (P < 0.05).
(4) The effect of the composition on liver glycogen in mice is shown in table 7.
TABLE 7
As can be seen from Table 7, when the composition was orally administered to the mice at different doses for 30 days, the liver glycogen content of the mice in the medium and high dose groups was significantly higher than that in the control group, and there was a significant difference (P < 0.05).
(5) Effect of the composition on blood lactate in mice.
The effect of the composition on the amplitude of the increase in blood lactate (mmol/L) after exercise in mice is shown in Table 8.
TABLE 8
As can be seen from Table 8, the amplitude of the increase in blood lactic acid after exercise was significantly different (P < 0.05) in the low and high dose groups when the composition was orally administered to the mice at different doses for 30 days, compared with the control group.
The effect of the composition on the extent of blood lactate (mmol/L) elimination after exercise in mice is shown in Table 9.
TABLE 9
As can be seen from Table 9, the amplitude of blood lactate elimination after exercise was not significantly different in each dose group of mice when the composition was orally administered to the mice at different doses for 30 days (P > 0.05).
6. Small knot
The composition with different dosages is orally administered to a mouse for 30 days, so that the swimming time of the mouse can be obviously prolonged, the serum urea nitrogen of the mouse after exercise can be obviously reduced, the blood lactic acid rising amplitude of the mouse after exercise can be obviously reduced, and the liver glycogen content of the mouse can be obviously improved. Thus, the composition has an anti-fatigue effect.
Example 25
The present example provides a study (animal test) of the immunomodulatory pharmacodynamics of the tablets obtained in example 19 above.
1. Experimental animals: clean-grade female Kunming mice, the weight of which is 18-22 g.
2. Experimental samples: example 19 the recommended dose of the pulverized fine powder for human body is 3.72 g/day/60 kg body weight.
3. Selecting the dosage: the equivalent dose of the mouse is 10 times of the recommended dose of the human body, namely, 0.62g/kg body weight is ingested per day as a low dose, 2 times and 3 times of the low dose are respectively provided with a dose group, namely 1.24 g/day/kg body weight (medium dose) and 1.86 g/day/kg body weight (high dose), distilled water is given to a control group, and after 15 days of continuous equal-volume gastric perfusion of the test object, each index is measured.
4. The test method comprises the following steps:
(1) organ/body weight ratio was measured.
(2) Delayed allergy (DTH) (plantar thickening): the left hind paw plantar thickness was measured 4 days after intraperitoneal injection of 2% (v/v) Sheep Red Blood Cells (SRBC)0.2ml for mice after sensitization, followed by subcutaneous injection of 20% (v/v) SRBC (20. mu.l/mouse) at the site of measurement, and the thickness of the plantar part of the left paw was measured 24 hours after injection, and the same site was measured three times and averaged. The degree of DTH is expressed as the difference in anteroposterior plantar thickness (degree of swelling of the foot plantar surface).
(3) Spleen antibody-producing cell assay (Jeren modified slide method): sheep blood was taken, washed three times with physiological saline, and each mouse was intraperitoneally injected with 2% (v/v) SRBC0.2ml. Mice 5 days after SRBC immunization were sacrificed and spleens were removed and made into cell suspensions. Heating and dissolving surface layer culture medium, mixing with equivalent amount of double Hanks solution, subpackaging into small tubes with 0.5ml per tube, adding 50 μ l 10% SRBC (v/v, prepared by SA buffer solution) and 10 μ l spleen cell suspension into the tubes, rapidly mixing, pouring onto slide coated with agarose thin layer, after agar solidification, horizontally buckling the slide on a slide rack, placing into a carbon dioxide incubator for incubation for 1.5 hours, placing complement (1: 10) diluted by SA buffer solution into a groove of the slide rack, continuing incubation for 1.5 hours, and counting the number of hemolytic plaques.
(4) Mouse carbon particle clearance test: injecting 1:3 diluted India ink into mouse tail vein, and immediately timing when the ink is injected. 10 minutes after the injection of the ink, 20. mu.l of blood was collected from the angular venous plexus, and added to 2ml of Na2CO3The solution was used as a blank control. Mice were sacrificed and livers and spleens were weighed. The phagocytic index a was calculated.
5. Results
(1) Effect of composition on mouse body weight.
Initial body weights of each group of mice are shown in table 10.
Watch 10
The end body weights of each group of mice are shown in table 11.
TABLE 11
The results in tables 10 and 11 show that the weight of the mice given a dose of the composition orally is not different from that of the control group 15 days later (the reduction in the number of animals is due to unexpected death).
(2) The effect of the composition on the organ/body weight ratio of mice is shown in table 12.
TABLE 12
As can be seen from table 12, when the composition was orally administered to the mice at different doses for 15 days, there was no significant difference between the three dose groups of organ coefficients of spleen and thymus as compared with the control group.
(3) The effect of the composition on delayed allergy (DTH) in mice is shown in table 13.
Watch 13
As can be seen from table 13, when the composition was orally administered to mice at different doses for 15 days, the degree of swelling of the plantar surface of the foot was significantly different in the high dose group compared to the control group.
(4) The effect of the composition on the number of mouse spleen antibody-producing cells is shown in Table 14.
TABLE 14
As can be seen from Table 14, when the composition was orally administered to mice at different doses for 15 days, the number of antibody-producing cells was significantly different in the middle and high dose groups compared with the control group.
(5) The effect of the composition on mouse monocyte-macrophage carbon clearance function is shown in table 15.
Watch 15
As can be seen from table 15, when the composition was orally administered to mice at different doses for 15 days, there was no significant difference in the mononuclear-phagocyte phagocytic index of each dose group as compared with the control group (the number of animals decreased due to accidental death).
6. Small knot
The composition with different dosages is orally administered to the mice for 15 days, so that the thickness of the plantar of the foot and the number of spleen antibody producing cells of the mice can be obviously improved. The composition can be judged to have an immunoregulatory effect according to the regulations of "evaluation procedure and test method of health food functionality".
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (8)
1. The composition with the functions of resisting fatigue and regulating immunity is characterized by comprising the following raw materials in parts by weight: 40-52 parts of epimedium, 5-23 parts of ginseng, 15-23 parts of astragalus and 14-22 parts of dogwood.
2. The composition with the functions of resisting fatigue and regulating immunity according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 52 parts of epimedium, 5 parts of ginseng, 15 parts of astragalus and 14 parts of dogwood.
3. The composition with the functions of resisting fatigue and regulating immunity according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 45 parts of epimedium, 23 parts of ginseng, 23 parts of astragalus and 22 parts of dogwood.
4. A preparation method of a composition with anti-fatigue and immunoregulation functions is characterized by comprising the following steps:
weighing the raw materials according to the proportion of the raw materials of the composition according to any one of claims 1 to 3, respectively extracting the astragalus, the dogwood, the epimedium and the ginseng with water, and mixing the extracts to obtain the composition; or extracting any two or more of radix astragali, Corni fructus, herba Epimedii and Ginseng radix with water, extracting the rest components with water separately, and mixing the extracts.
5. A preparation method of a composition with anti-fatigue and immunoregulation functions is characterized by comprising the following steps:
weighing the raw materials according to the proportion of the raw materials of the composition of any one of claims 1 to 3, and respectively extracting the astragalus, the dogwood and the epimedium with water to prepare mixed extracts; or extracting any two or more of radix astragali, Corni fructus and herba Epimedii with water, and extracting the rest with water separately to obtain mixed extract;
pulverizing Ginseng radix, and mixing with the mixed extract.
6. A preparation method of a composition with anti-fatigue and immunoregulation functions is characterized by comprising the following steps:
weighing the raw materials according to the proportion of the raw materials of the composition according to any one of claims 1 to 3, and respectively extracting the astragalus, the dogwood, the epimedium and a part of ginseng with water to prepare mixed extracts; or extracting any two or more of radix astragali, Corni fructus, herba Epimedii and part of Ginseng radix with water, and extracting the rest with water separately to obtain mixed extract;
pulverizing the rest part of Ginseng radix, and mixing with the mixed extract.
7. Use of the composition with anti-fatigue and immunoregulatory functions of any one of claims 1 to 3 in preparation of medicines or health foods.
8. An oral preparation having anti-fatigue and immunoregulatory functions, which comprises the composition prepared by the preparation method of claim 5, and an excipient; the oral preparation can be in the form of tablet, oral liquid, hard capsule, soft capsule, granule, powder, pill or teabag.
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Citations (2)
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| CN103520302A (en) * | 2013-10-30 | 2014-01-22 | 山西金科海生物科技有限公司 | Sea-buckthorn leaf anti-fatigue capsule and preparation method thereof |
| CN107875216A (en) * | 2017-11-20 | 2018-04-06 | 中国农业科学院特产研究所 | It is a kind of that there is composition for alleviating physical fatigue effect and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103520302A (en) * | 2013-10-30 | 2014-01-22 | 山西金科海生物科技有限公司 | Sea-buckthorn leaf anti-fatigue capsule and preparation method thereof |
| CN107875216A (en) * | 2017-11-20 | 2018-04-06 | 中国农业科学院特产研究所 | It is a kind of that there is composition for alleviating physical fatigue effect and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 神威牌精力源片 疲倦无力、腰酸腿疼、面色苍白怎么办？;神威健康网;《http://blog.sina.com.cn/s/blog_1488391a90102vs2b.html》;20150320;第1-3页，尤其是第1页第1段 * |
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