CN105147838A - Composition with immunity-enhancing and fatigue-resisting effects and preparation method and application thereof - Google Patents
Composition with immunity-enhancing and fatigue-resisting effects and preparation method and application thereof Download PDFInfo
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- CN105147838A CN105147838A CN201510638938.0A CN201510638938A CN105147838A CN 105147838 A CN105147838 A CN 105147838A CN 201510638938 A CN201510638938 A CN 201510638938A CN 105147838 A CN105147838 A CN 105147838A
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Abstract
The invention relates to a composition with immunity-enhancing and fatigue-resisting effects and a preparation method and application thereof, and belongs to the technical field of medicine. The composition is prepared from, by weight, 3-30 parts of schisandra chinensis, 5-45 parts of ginseng, 2-18 parts of cornua cervi pantotrichum, 10-90 parts of raspberries and 8-60 parts of astragalus membranaceus. The schisandra chinensis reinforces the lung and replenishes qi to disperse lung qi, the ginseng and the astragalus membranaceus tonify the spleen and reinforce yang to achieve spleen movement and transformation, and the cornua cervi pantotrichum and the raspberries nourish the kidneys and replenish essence to reinforce kidney yang; with qi, blood and essence synergically reinforced, the composition has an excellent effect when applied to healthcare treatment of immunodeficiency and fatigue status of modern people.
Description
Technical field
The present invention relates to medical art, particularly relate to a kind of composition and method of making the same and the application with enhancing immunity and antifatigue effect.
Background technology
Along with society living-pattern preservation, people's pressure increases gradually, and physical sport reduces relatively, and simultaneously due to the disorder of diet, work and rest rule, cause immunologic function greatly to decline, body is in fatigue, subhealth state and non resistance state for a long time.
Therefore, improve body resisting fatigue ability, immunity moderation function becomes academic circles at present and the focus of people's concern.
But resisting fatigue conventional at present, the product of immunity moderation are not only expensive, and are confined to the adjustment of short-term condition, and clinical effectiveness is not satisfactory.
Therefore, utilize China's Traditional Chinese Medicine tcm theory, developmental function is clear and definite, and the low-cost product strengthening immunity and resisting fatigue meets public demand and has very important realistic meaning and social meaning.
Summary of the invention
Based on this, the object of the invention is to the defect overcoming prior art, there is provided a kind of and there is the compositions strengthening immunity and antifatigue effect, the active component of said composition is that raw material is made by natural Chinese medicinal herb, the compositions made has resisting fatigue and immune enhancing function, and long-term taking, have no side effect.
Have the compositions strengthening immunity and antifatigue effect, the effective ingredient of described compositions is prepared from by the medical material of following weight portion:
Fructus Schisandrae Chinensis 3-30 part, Radix Ginseng 5-45 part, Cornu Cervi Pantotrichum 2-18 part, Fructus Rubi 10-90 part, Radix Astragali 8-60 part.
Chinese medicine has the systematology macro-view cognition of " harmony between man and nature " to human body, when body is subject to the invasion and attack of paathogenic factor inside and outside some, the physiological function activity of internal organs, meridians etc. is caused to occur abnormal, when negative and positive of qi and blood balance coordination relation is damaged and occurs various clinical symptoms, human body is just in morbid state.The endogenous cause of ill of morbidity has: the indisposed and internal injury caused by excess of seven emotions of deficiency of vital QI, zang-fu disharmony, QI and blood etc., exopathogenic factor mainly refers to heresy and the pestilential pathogen of six climate exopathogens, and wherein the deficiency of vital energy is most important.Visible, healthy energy is the base substance maintaining human normal vital movement, resist exopathogen and disease preventing and treating.Whether its prosperity is related to the vital movement of human body and lapsing to of disease.In this sense, the healthy energy of Chinese medicine includes the immune function of human body of modern medicine.
The traditional Chinese medical science think the generation of immunological diseases and main, the diseases caused by exogenous pathogenic factor six climate exopathogens not enough with natural endowment of development evil, the four stages in the course of an epidemic febrile disease lack of proper care, the dirty dysfunction of internal organs, the interior life of phlegm-turbidity and blood stasis blood etc. are closely related because have.The heresy of diseases caused by exogenous pathogenic factor six climate exopathogens is the transient cause of disease, and natural endowment deficiency, the four stages in the course of an epidemic febrile disease imbalance, the dirty dysfunction of internal organs etc. are immanent causes.The traditional Chinese medical science thinks that the kidney being the origin of congenital constitution, thus emphasizes that kidney has adjustment and maintains immunologic balance and stable important function thereof.Kidney not only shows the adjustment of overall aspect to the regulating action of immunity, simultaneously also relevant with intracellular adjustment.The virtual loss of kidney, there will be endocrine disorder.As the performance that hyperthyroidism is deficiency of the kidney yin, hypothyroidism is the performance of insufficiency of kidney-YANG.The traditional Chinese medical science thinks that spleen is the foundation of acquired constitution.Spleen is the maximum glandular organ of human body, is also the place of mainly living away from home of immunocyte.The virtual loss of spleen, its cellular immunization and humoral immune function are all low than normal person.Many immunological diseases such as lupus erythematosus, dry syndrome are all that taste are deficient, caused by deficiency of body fluid.The traditional Chinese medical science think kidney, spleen and erythropoiesis and hematid immunity function in close relations, the hemopoietic function of bone marrow is mainly from kidney, spleen.Kidney storing essence, main bone, raw marrow; Spleen, middle Jiao is bullied extracting juice, change and red be meaning blood.Transporting and transforming function of the spleen and stomach essence of water and grain, must under kidney yang impetus, the angry blood of abilityization.Therefore, the traditional Chinese medical science uses drugs for nourishing qi and activating blood to improve the activity of Erythrocyte complement receptor type I, thus improves red blood cell adhesion immune complex function, plays erythrocyte immune effect.Good curative effect is had to treatment anaphylaxis aglobuliosis.Thus traditional Chinese medical science immunology is thought, kidney, spleen, lung dialogue cell immune system and erythrocyte immune system have performance regulating action.
The traditional medicine of China has different sayings to fatigue, as " tired ", " unable ", " asthenia ", " de-power " etc., think the appearance of fatigue phenomenon, close relationship is had with the imbalance of the five internal organs, as many in myasthenia of limbs relevant with taste, waist soreness is how relevant with kidney, and short wind is how relevant with lungs, insobriety is how relevant with heart, and endurance is how not relevant with liver.The whole vital movement of people, entirely rely in " vigour ", vigour natural endowment is nourished in the day after tomorrow (taste) in congenital (kidney), it is the source of body energy, vigour produces different functions from the combination of different organs, therefore, vigour weakness can cause each function inhibitory state at a low ebb of people, and this is also the tired basic reason produced.Therefore, tonification the five internal organs are not only wanted in the treatment for fatigue, more can not ignore supplementing " gas ".
Based on above-mentioned theory, the present invention is on the basis of summing up numerous scholar's viewpoint, in conjunction with the constitutive character of Modern human populations, raising body's immunity and resisting fatigue are ascribed in lung, spleen, kidney three main organs, i.e. lung governing qi, the spleen governs the blood, kidney spermatogenesis, theoretical for this, select Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum, Fructus Rubi and the Radix Astragali side of composition altogether, wherein Fructus Schisandrae Chinensis invigorating the lung and benefiting vital QI, falls to help a surname of lung; Radix Ginseng and Radix Astragali spleen invigorating supporing yang, with the transporting of row spleen; Cornu Cervi Pantotrichum and Fructus Rubi Zishen fill fine, with the Yuanyang of the kidney invigorating; Gas, blood, smart three is collaborative mends altogether, for the health therapy of Modern human populations immunologic hypofunction and fatigue state, has extraordinary effect.
Fang Zhong: Fructus Schisandrae Chinensis, sour in the mouth, sweet, warm in nature, return lung, the heart, kidney channel.There is convergence astringent or styptic treatment for spontaneous sweating, supplementing QI for promoting the production of body fluid, effect of kidney calming.For chronic cough dyspnea due to deficiency, emission, enuresis frequent micturition, incessant chronic diarrhea, spontaneous perspiration, night sweat, Tianjin wound is thirsty, and deficient pulse of losing heart, interior-heat is quenched one's thirst, palpitation and insomnia.Fructus Schisandrae Chinensis main component is lignanoid and polysaccharide; modern pharmacology research shows; Fructus Schisandrae Chinensis crude polysaccharides can the leukopenic immunosuppressive action of dose-dependant sex resistance caused by cyclophosphamide; to bone marrow hematogenesis function, there is protective effect; improve the immunity of body; enhancing body, to the resistance of destructive stimulus, alleviates body injury.Fructus Schisandrae Chinensis obviously can strengthen the phagocytic function of mice reticuloendothelial system macrophage.Fructus Schisandrae Chinensis can also make mice serum hemolysin level obviously raise, and mice secretory antibody function is strengthened, and namely Fructus Schisandrae Chinensis has the effect of enhancing non-specific immunity function and humoral immune function.Fructus Schisandrae Polysaccharide can also the atrophy of antagonism mouse aging lymphatic organ Thymus and spleen, has the effect promoting lymphocyte DNA synthesis simultaneously, increases lymphoblast and generate, promote Splenic immunologic.Fructus Schisandrae Chinensis can also promote IgG plasma cell showed increased in submaxillary gland, the being proportionate property of content of immunocyte quantity and immunoglobulin in salivary gland, and immunoglobulin increases and namely effectively prevents cause of disease from invading in saliva.Above research all reflects that Fructus Schisandrae Chinensis is significantly improved to immunologic function tool, and mainly shows immune organ weight and the increase of immunoglobulin content and the enhancing of phagocyte and antibiotic substance ability secreted by it.
Radix Ginseng, sweet in the mouth, micro-hardship, property is flat, tepor, returns spleen, lung, heart channel.Effect is strongly invigorating primordial QI, invigorating the spleen to benefit the lung, promoting the production of body fluid to quench thirst, Fructus Alpiniae Oxyphyllae of calming the nerves, and mends all void.The main component of Radix Ginseng is ginsenoside, polysaccharide and volatile oil, and modern study confirms, Radix Ginseng has multiple efficacies, as slow down aging and antioxidation, improves the adaptive capacity to environment of human body, antitumor and enhancing immunity etc.Ginsenoside Rg1 obviously can extend the time that mice power exhausts swimming, and ginsenoside Rb2 can strengthen the function of liver carbohydrate metabolism system and energy production system.
Cornu Cervi Pantotrichum, sweet in the mouth, salty, warm in nature, return kidney, Liver Channel.There is invigorating kidney-YANG, mend marrow, bone and muscle strengthening, adjust punching to appoint, the effects such as holder sore.Cornu Cervi Pantotrichum contains abundant aminoacid, lecithin, vitamin and trace element etc.Cornu Cervi Pantotrichum is warm in nature and not dry, has and rouses oneself and improve body function, to patient after asthenia universalis, prolonged illness, have good health-care effect.Cornu Cervi Pantotrichum can improve cellular immunization and the humoral immune function of body, promotes lymphocytic conversion, has the effect of immunopotentiating agent.It can increase body defence capability to external world, the immunologic balance in control agent and avoid disease to occur and promote wound healing, sick body rehabilitation, thus plays strengthening body, to antidotal effect.
The Radix Astragali, sweet in the mouth, warm in nature, return lung, spleen channel.There is invigorating QI to consolidate the body surface resistance, diuresis poison holding, evacuation of pus, effect of expelling pus and promoting granulation.Radix Astragali main component is polysaccharide and Flavonoid substances.Modern pharmacology research shows, the Radix Astragali has enhancing human body immunity function, blood fat reducing, diuresis, the effects such as blood pressure lowering.Modern study finds, the Radix Astragali significantly can increase the total white blood cells in blood, promotes phagocytic function and the sterilizing ability of neutrophilic granulocyte and macrophage.The Radix Astragali obviously can strengthen cellular immunization, promotes the lymphocyte transformation that PHA, COnA, PWM (phytolacca american) cause.
Fructus Rubi, sweet in the mouth, acid, warm in nature.Return liver, kidney channel.There is tonifying the kidney to consolidate the essence, effect of supporing yang improving eyesight.Fructus Rubi main component is terpenoid, steroidal, organic acid, polysaccharide and vitamin C.Modern pharmacology research shows, Fructus Rubi has the effect of antitumor, defying age, fat-reducing liver-protecting and enhancing immunologic function.4 kinds of extraction components of Fructus Rubi show the experimental result of spleen lymphocyte synthesis in vitro, 4 kinds of extract components of Fructus Rubi: aqueous extract, alcohol extract, crude polysaccharides and n-butyl alcohol component have obvious Proliferation of lymphocytes, to have or under effect that no mitogen ConA is auxiliary, Fructus Rubi all has the effect of obvious activated lymphocyte.The crude polysaccharides extracted from Fructus Rubi and n-butyl alcohol component have cAMP effect in elevate cellular to the lymphocyte of In vitro culture in early days in activation, and do not affect cGMP level.
Wherein in an embodiment, the effective ingredient of described compositions is prepared from by the medical material of following weight portion: Fructus Schisandrae Chinensis 8-12 part, Radix Ginseng 12-17 part, Cornu Cervi Pantotrichum 5-7 part, Fructus Rubi 25-35 part, Radix Astragali 16-24 part.
Wherein in an embodiment, the effective ingredient of described compositions is prepared from by the medical material of following weight portion: Fructus Schisandrae Chinensis 10 parts, Radix Ginseng 15 parts, 6 parts, Cornu Cervi Pantotrichum, Fructus Rubi 30 parts, the Radix Astragali 20 parts.
The invention also discloses the above-mentioned preparation method with the compositions strengthening immunity and antifatigue effect, comprise the following steps:
Prepare the first component: take Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum by weight, pulverize, add reflux, extract, after water, filter, when filtrate is concentrated into 40-60 DEG C, relative density is 1.10-1.15, adds by every gram of medical material the ethanol that 2-4ml volumn concentration is 90-98%, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component;
Prepare second component: take Fructus Rubi, the Radix Astragali by weight, pulverize, add reflux, extract, after water, filter, dry after filtrate recycling design, obtain second component;
Mixing: merge the first component and second component, obtain compositions.
The preparation method with the compositions strengthening immunity and antifatigue effect of the present invention, is extracted by the effective ingredient in medical material, improve the convenience that patient takes, and this preparation method is simple for process, be easy to carry out commercial production.
Wherein in an embodiment, the actual conditions of described preparation first component is: take Fructus Schisandrae Chinensis by weight, Radix Ginseng, Cornu Cervi Pantotrichum, pulverize, water is added according to the amount of every gram of medical material 8-12ml, soak 25-35min, reflux, extract, 50-70min, filter, water is added according to the amount of every gram of medical material 8-12ml again in medicinal residues, reflux, extract, 25-35min, filter, merging filtrate, when filtrate is concentrated into 40-60 DEG C, relative density is 1.10-1.15, the ethanol that volumn concentration is 90-98% is added by the amount of every gram of medical material 2-4ml, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component,
The described actual conditions preparing second component is: take Fructus Rubi, the Radix Astragali by weight, pulverize, add water according to the amount of every gram of medical material 8-12ml, soak 25-35min, reflux, extract, 50-70min, filter, add water according to the amount of every gram of medical material 8-12ml again in medicinal residues, reflux, extract, 25-35min, filter, merging filtrate, dry after filtrate recycling design, obtain second component.
Wherein in an embodiment, the actual conditions of described preparation first component is: take Fructus Schisandrae Chinensis by weight, Radix Ginseng, Cornu Cervi Pantotrichum, pulverize, water is added according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter, water is added according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, when filtrate is concentrated into 50 DEG C, relative density is 1.10-1.15, the ethanol that volumn concentration is 95% is added by the amount of every gram of medical material 3ml, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component,
The described actual conditions preparing second component is: take Fructus Rubi, the Radix Astragali by weight, pulverize, add water according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter, add water according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, dry after filtrate recycling design, obtain second component.
The invention also discloses above-mentioned compositions and prepare the application in the medicine or health product having and strengthen immunity and antifatigue effect.
Wherein in an embodiment, the dosage form of described medicine or health product is tablet, granule, hard capsule, soft capsule, oral liquid, powder, mixture, pill or drop pill.
Above-mentioned dosage form all can adopt customary adjuvant and preparation process to prepare.
Compared with prior art, the present invention has following beneficial effect:
Of the present invention have the compositions strengthening immunity and antifatigue effect, selects Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum, Fructus Rubi and the Radix Astragali side of composition altogether, wherein Fructus Schisandrae Chinensis invigorating the lung and benefiting vital QI, fall to help a surname of lung; Radix Ginseng and Radix Astragali spleen invigorating supporing yang, with the transporting of row spleen; Cornu Cervi Pantotrichum and Fructus Rubi Zishen fill fine, with the Yuanyang of the kidney invigorating; Gas, blood, smart three is collaborative mends altogether, for the health therapy of Modern human populations immunologic hypofunction and fatigue state, has extraordinary effect.
Further, after being coordinated by each raw material, have extraordinary effect with specific ratio, the oral said composition that gives, after 30 days, has significant difference by macrophage phagocytic experiment and the display of lymphocyte proliferation assay result; Tested by mice burden swimming, the display of hepatic glycogen determination experiment, Serum lactic acid content determination experiment and serum urea nitrogen analysis experimental result has significant difference.
Detailed description of the invention
Explain explanation the present invention in detail below in conjunction with specific embodiment, but any restriction is not caused to the present invention.
The crude drug adopted in following examples and adjuvant are common commercially available.
Embodiment 1
There is the compositions strengthening immunity and antifatigue effect, be prepared from by following medical material:
Fructus Schisandrae Chinensis 100 grams, Radix Ginseng 150 grams, 60 grams, Cornu Cervi Pantotrichum, Fructus Rubi 300 grams, the Radix Astragali 200 grams.
Above-mentioned raw materials is followed these steps to preparation:
1, the first component is prepared.
Get above-mentioned Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum medical material, pulverize, obtain coarse powder, insert in multi-function extractor, add distilled water according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter; Add distilled water according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, when filtrate is concentrated into 50 DEG C, measure relative density 1.10-1.15; In filtrate, the ethanol that volumn concentration is 95% is added by the amount of every gram of medical material 3ml, alcohol is sink to about 70% of total liquid quality and (namely adds water-soluble substances precipitation after alcohol, liquid quality after removing precipitation declines, for about 70% of liquid quality total before precipitate with ethanol), precipitate with ethanol spends the night (about 10h), relative density 1.20-1.30 is measured when reclaiming ethanol and be concentrated into 50 DEG C, dry, obtain the first component.
2, second component is prepared.
Get above-mentioned Fructus Rubi, Milkvetch Root, pulverize, obtain coarse powder, insert in multi-function extractor, add distilled water according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter; Add distilled water according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, when filtrate is concentrated into 50 DEG C, measure relative density 1.05-1.10, dry, obtain second component.
3, mix.
Merge the first component and second component, obtain compositions A.
Embodiment 2
There is the compositions strengthening immunity and antifatigue effect, be prepared from by following medical material:
Fructus Schisandrae Chinensis 30 grams, Radix Ginseng 450 grams, 20 grams, Cornu Cervi Pantotrichum, Fructus Rubi 90 grams, the Radix Astragali 80 grams.
Method with reference to embodiment 1 prepares compositions B.
Embodiment 3
There is the compositions strengthening immunity and antifatigue effect, be prepared from by following medical material:
Fructus Schisandrae Chinensis 300 grams, Radix Ginseng 50 grams, 180 grams, Cornu Cervi Pantotrichum, Fructus Rubi 100 grams, the Radix Astragali 600 grams.
Method with reference to embodiment 1 prepares compositions C.
Embodiment 4
There is the oral liquid strengthening immunity and antifatigue effect, can take as health product.
This oral liquid obtains by the following method:
The compositions A that Example 1 prepares, adds dextrin in said composition A or Mel is in harmonious proportion, and filling bottle, makes oral liquid, to obtain final product.
Embodiment 5
There is the capsule strengthening immunity and antifatigue effect, can take as health product.
This capsule obtains by the following method:
The compositions A that Example 1 prepares, adds in the dry powder of above-mentioned composition A by pregelatinized Starch, mixing, granulates, and dries, adds magnesium stearate and Pulvis Talci, filled hard capsules, to obtain final product.
Embodiment 6
There is the tablet strengthening immunity and antifatigue effect, can take as health product.
This tablet obtains by the following method:
The compositions A that Example 1 prepares, adds in the dry powder of above-mentioned composition A by microcrystalline Cellulose, pregelatinized Starch, carboxymethyl starch sodium, mixing, granulates, and dry, granulate, adds magnesium stearate, and tabletting to obtain final product.
Comparative example 1
This comparative example prepares a kind of compositions, with embodiment 1 have strengthen immunity substantially identical with the compositions of antifatigue effect, difference is, the compositions of this comparative example is prepared from by following medical material:
Radix Ginseng 150 grams, 60 grams, Cornu Cervi Pantotrichum, Fructus Rubi 300 grams, the Radix Astragali 200 grams.
Method with reference to embodiment 1 prepares compositions D, and wherein Radix Ginseng, Cornu Cervi Pantotrichum are prepared according to the preparation method of the first component, and Fructus Rubi, the Radix Astragali are prepared according to the preparation method of second component.
Comparative example 2
This comparative example prepares a kind of compositions, with embodiment 1 have strengthen immunity substantially identical with the compositions of antifatigue effect, difference is, the compositions of this comparative example is prepared from by following medical material:
Fructus Schisandrae Chinensis 100 grams, Radix Ginseng 150 grams, 60 grams, Cornu Cervi Pantotrichum, the Radix Astragali 200 grams.
Method with reference to embodiment 1 prepares compositions E, and wherein Fructus Schisandrae Chinensis, Radix Ginseng and Cornu Cervi Pantotrichum are prepared according to the preparation method of the first component, and the Radix Astragali is prepared according to the preparation method of second component.
Comparative example 3
This comparative example prepares a kind of compositions, with embodiment 1 have strengthen immunity substantially identical with the compositions of antifatigue effect, difference is, the compositions of this comparative example is prepared from by following medical material:
Fructus Schisandrae Chinensis 100 grams, Radix Ginseng 150 grams, 60 grams, Cornu Cervi Pantotrichum, Fructus Rubi 300 grams, the Radix Astragali 200 grams, Fructus Lycii, 200 grams.
Method with reference to embodiment 1 prepares composition F, and wherein Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum are prepared according to the preparation method of the first component, and Fructus Rubi, the Radix Astragali and Fructus Lycii are prepared according to the preparation method of second component.
Comparative example 4
This comparative example prepares a kind of compositions, with embodiment 1 have strengthen immunity substantially identical with the compositions of antifatigue effect, difference is, the compositions of this comparative example is prepared from by following medical material:
Fructus Schisandrae Chinensis 100 grams, Radix Ginseng 150 grams, 60 grams, Cornu Cervi Pantotrichum, Fructus Rubi 300 grams, Fructus Lycii 200 grams.
Method with reference to embodiment 1 prepares compositions G, and wherein Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum are prepared according to the preparation method of the first component, and Fructus Rubi, Fructus Lycii are prepared according to the preparation method of second component.
Experimental example 1
Compositions above-described embodiment and comparative example prepared carries out pharmacodynamic study.
1, immunostimulant experiment.
1.1 macrophage phagocytic experiments.
1.1.1 laboratory animal: kunming mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
1.1.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL; India ink, Rui Yong bio tech ltd, Shanghai provides.
1.1.3 method and result: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.After last administration, inject the india ink of dilution from mouse tail vein, calculate by every 10g body weight 0.1mL.Treat that prepared Chinese ink injects, immediately timing.To inject after prepared Chinese ink 2,10min, get blood 20 μ l from angular vein clump respectively, existing side by side is added to 2mL0.1%Na by it
2cO
3in solution.By spectrophotometer or full-automatic microplate reader at 600nm wavelength place densitometric value (OD), with Na
2cO
3solution makes blank.By sacrifice, get liver and spleen, blot organ surface blood stains with filter paper, weigh.The ability of mice carbonic clearance is represented with phagocytic index.Be calculated as follows phagocytic index a:
Table 1 macrophage phagocytic experimental result
As can be seen from the above table, the macrophage phagocytic index of the mice of compositions A-G group is all higher than blank group, the effect all having the phagocytic activity strengthening mouse macrophage is described, but, wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has best enhancing immune effect.
1.2 lymphproliferation response experiments.
1.2.1 laboratory animal: C57 mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
1.2.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL; Platform expects orchid (TrypanBlue), and prosperous bio tech ltd of Beijing ancient cooking vessel state provides; Canavaline, sigma company provides; Hanks liquid, GENVIEW company provides; RPMI1640 culture fluid, Sai Mo flies your biochemistry goods (Beijing) company limited of generation to be provided; Calf serum, Tian Hang bio tech ltd, Zhejiang provides; 2 mercapto ethanol, prosperous bio tech ltd of Beijing ancient cooking vessel state provides; Isopropyl alcohol, Tianjin great Mao chemical reagent work provides.
1.2.3 reagent pre-treatment.
Complete culture solution: RPMI1640 culture fluid is degerming with 0.22 μm of membrane filtration, with front adding 10% calf serum, 1% glutamine (200mmol/L), penicillin (100U/mL), streptomycin (100ug/L) and 5 × 10
-5the 2 mercapto ethanol of mol/L,
ConA (canavaline) liquid: the solution being mixed with 100 μ g/mL with distilled water, filtration sterilization, preserves at cryogenic refrigerator (-20 DEG C).
Aseptic Hank's liquid: with front with 3.5% aseptic NaHCO
3adjust pH7.2-7.4.
MTT liquid: 5mgMTT is dissolved in the PBS of 1mLpH7.2, now with the current.
Acid isopropyl alcoholic solution: the HCl adding 4mL1mol/L in 96mL isopropyl alcohol, prepared before use.
1.2.4 medication: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.
1.2.5 splenocyte suspension preparation
Within second day after last administration, put to death each treated animal, asepticly get spleen, be placed in the plate filling appropriate (3-5ml) aseptic Hank's liquid, and place one piece of gauze on spleen, gently spleen is ground with large syringe inner core, make individual cells suspension.Filter through 200 eye mesh screens, wash 2 times with Hank's liquid, each centrifugal 10min (1000r/min).Then by cell suspension in the complete culture solution of 2mL, with the blue dyeing counting viable count of platform phenol (should more than 95%), adjustment cell concentration be 3 × 10
6individual/mL.
1.2.6 lymphopoiesis measures.
Add in 24 well culture plates by a cell suspension point holes, every hole 1mL, a hole adds 75 μ lConA liquid (being equivalent to 7.5 μ g/mL), and 5%CO in contrast, is put in another hole
2, 37 DEG C of CO
272h is cultivated in incubator.Cultivation terminates front 4h, and every hole sucks supernatant 0.7mL gently, adds 0.7mL not containing the RPMI1640 culture fluid of calf serum, adds MTT (5mg/mL) 50 μ l/ hole simultaneously, continues to cultivate 4h.After cultivation terminates, every hole adds 1mL acid isopropyl alcohol, ultrasonic vibration (2 seconds) or manually blow and beat mixing, and purple crystal is dissolved completely.Then be dispensed in 96 well culture plates, subpackage 3 hole, each hole, as Duplicate Samples, with enzyme-linked immunosorbent assay instrument, measures optical density value with 570nm wavelength.
1.2.7 experimental result.
Lymphocyte proliferation assay result is as shown in the table.
Table 2 lymphocyte proliferation assay result
As can be seen from the above table, the multiplication capacity of the mouse lymphocyte of compositions A-G group is all higher than blank group, the effect all having the multiplication capacity strengthening mouse lymphocyte is described, but, wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has best enhancing immune effect.
In sum, compositions of the present invention carries out prescription under the guidance of theory of Chinese medical science, with mouse macrophage phagocytic activity for index, evaluate the cellular immune function of mice, and with lymphopoiesis ability for observation index, evaluate the lymphocyte immunologic function of mice, result shows, compositions of the present invention can improve immune function of mice, has obvious immunological enhancement.
2, resisting fatigue experiment.
2.1 swimming with a load attached to the body experiments.
2.1.1 laboratory animal: kunming mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
2.1.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL.
2.1.3 method and result: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.
After last administration 30min, mouse tail load 5% body weight lead, is placed in swimming trunk went swimming, depth of water 30cm, water temperature 25 DEG C-28 DEG C, and record is swum the time started to dead mouse, and result is as shown in the table.
Table 3 mice burden swimming experimental result
As can be seen from the above table, the mice swimming time of compositions A-G group, all higher than blank group, illustrates the effect all having and extend mice swimming time, but, wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has best antifatigue effect.
2.2 hepatic glycogen content determination experiments.
2.2.1 laboratory animal: kunming mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
2.2.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL; Hepatic glycogen testing cassete, Nanjing is built up Bioengineering Research Institute and is provided.
2.2.3 method and result: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.
The continuous gavage of mice 30 days, after last administration 30min, cervical dislocation is put to death, and get liver immediately, after normal saline flushing, blot with filter paper, accurately take liver 100mg, homogenate, with kit measurement, result is as shown in the table.
Table 4 Mouse Liver glycogen content determination experiment result
As can be seen from the above table, the Mouse Liver glycogen content of compositions A-G group, all higher than blank group, illustrates the effect of the hepatic glycogen content that is improved, but, wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has best antifatigue effect.
The mensuration of 2.3 Serum lactic acid contents
2.3.1 laboratory animal: kunming mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
2.3.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL; Blood lactase acid testing cassete, Nanjing is built up Bioengineering Research Institute and is provided.
2.3.3 method and result: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.
The continuous gavage of mice 30 days, after last administration 30min, to take a blood sample 20 μ l from the mice ophthalmic corner of the eyes with blood capillary immediately, does not then bear a heavy burden to stop after temperature is the water went swimming 10min of 30 DEG C, and take a blood sample 20 μ l immediately, again takes a blood sample after rest 20min.With kit measurement, its Serum lactic acid content result is as shown in the table.
Table 5 Mouse Blood lactic acid content determination experiment result (mg/g)
As can be seen from the above table, after the compositions of compositions A-G group can make motion, Mouse Blood lactic acid lift-off value obviously reduces, and blood lactase acid level recovers comparatively obvious after having a rest, but, wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has the effect of best increase fatigue recovery speed.
2.4 serum urea nitrogen analysis experiments
2.4.1 laboratory animal: kunming mice, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
2.4.2 medicine and reagent: get compositions A that above-described embodiment and comparative example prepare, B, C, D, E, F, G, during experiment, distilled water dissolved dilution to contained crude drug amount is 50mg/mL; Serum urea nitrogen testing cassete, Nanjing is built up Bioengineering Research Institute and is provided.
2.4.3 method and result: animal is divided into 8 groups, namely blank group, compositions A group, compositions B group, compositions C group, compositions D group, compositions E group, composition F group, compositions G group.
Blank group gives distilled water; Administration group is respectively according to the amount gavage of 10mg/10g (every 10g body weight mice administration 10mg), and the concentration of each group gavage solution used is configured to given low 0.2ml/10g respectively, and dosage period is 30 days.
The continuous gavage of mice 30 days, after last administration 30min, be not swimming with a load attached to the body 90min in the water of 30 DEG C in temperature, after rest 60min, pluck eyeball and get blood, the centrifugal 10min of 3000rpm, gets supernatant, and with its serum urea nitrogen of kit measurement, result is as shown in the table.
Table 6 mice serum urea nitrogen content determination experiment result
As can be seen from the above table, the compositions of compositions A-G group can obviously reduce mice serum urea nitrogen content, and enhancing body, to the adaptive capacity of strenuous exercise, makes body endurance increase.But wherein compositions A-C group successful is better than D-G group, particularly compositions A group, has the effect of best increase body endurance.
In sum, compositions of the present invention carries out prescription under the guidance of theory of Chinese medical science, with the mice burden swimming time for observation index, evaluates the anti-fatigue ability of mice; And with lactate level in blood after mouse movement for observation index, have rated mice aerobic metabolism and anti-fatigue ability; Result shows, and compositions of the present invention can improve aerobic metabolism level and antifatigue effect in Mice Body.
Experimental example 2
Compositions above-described embodiment and comparative example prepared carries out toxicologic study.
1, acute toxicity testing.
1.1 laboratory animals: kunming mice, 40, body weight 18-22g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
1.2 medicines and reagent: the compositions A prepared by Example 1, during experiment with distilled water dissolved dilution to the aqueous solution of 2g crude drug/ml.
1.3 methods and result: each gavage amount is 0.6ml/, and be total to gavage three times in 24 hours, total amount is 1.8ml, is equivalent to primary dose 180g crude drug/kg/d, be equivalent to the 600-800 of 70kg mankind's normal clinical consumption doubly.After mice administration, in 30 days there is not death and abnormal phenomena in conventional raising.Prove that the compositions A that obtains of the present invention has no side effect, clinically to use safely.
2, long term toxicity test.
2.1 laboratory animals: Wistar rat, 120, body weight 180-220g, male and female half and half, Changchun High-technology Medical Animal Experiment Research Center provides.
2.2 medicines and reagent: the compositions A prepared by Example 1, during experiment with distilled water dissolved dilution to the aqueous solution of 2g crude drug/ml.
2.3 grouping and medications: laboratory animal is divided into 4 groups at random by sex body weight, often organize 30.1 group of gavage gives distilled water, 0.2ml/100g; 2,3,4 groups of gavages give test medicine, and dosage is respectively: 50mg/100g, 100mg/100g, 200mg/100g, and namely every 100g body weight gives the crude drug amount of 50mg, 100mg or 200mg, and each group concentration is configured to the given low of 0.2ml/100g respectively.Take continuously after 6 months and all put to death for each group above.
2.4 Indexs measure
2.4.1 general status detects: body weight, diet, emotion, movable energy.
2.4.2 biochemical index: red blood cell count(RBC), numeration of leukocyte, platelet count, hemoglobin counting, serum albumin, cholesterol and blood glucose etc.
2.4.3 pathological examination: the heart, liver, spleen, lung, kidney, brain, stomach, thymus, adrenal gland etc.
2.5 experimental results show, after rat takes 6 months medicines of the present invention continuously, the general status such as body weight, diet, emotion, movable energy are good, each main organs and Serum Indexes no abnormality seen, prove compositions A that the present invention obtains to body without obvious long term toxicity effect.
In sum, acute and long_term toxcity experimental result shows, patent health-care composition safety of the present invention, without obvious toxic-side effects; Orally give patent drug of the present invention after 30 days, shown by macrophage phagocytic experiment and lymphocyte proliferation assay result and there is significant difference; Tested by mice burden swimming, the display of hepatic glycogen determination experiment, Serum lactic acid content determination experiment and serum urea nitrogen analysis experimental result has significant difference.According to " health food inspection and assessment technical specification " (2003 editions), this zooscopy conclusion proved patent of invention health-care composition has enhancing immunologic function and antifatigue effect.
Each technical characteristic of the above embodiment can combine arbitrarily, for making description succinct, the all possible combination of each technical characteristic in above-described embodiment is not all described, but, as long as the combination of these technical characteristics does not exist contradiction, be all considered to be the scope that this description is recorded.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be construed as limiting the scope of the patent.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (8)
1. have the compositions strengthening immunity and antifatigue effect, it is characterized in that, the effective ingredient of described compositions is prepared from by the medical material of following weight portion:
Fructus Schisandrae Chinensis 3-30 part, Radix Ginseng 5-45 part, Cornu Cervi Pantotrichum 2-18 part, Fructus Rubi 10-90 part, Radix Astragali 8-60 part.
2. according to claim 1 have the compositions strengthening immunity and antifatigue effect, and it is characterized in that, the effective ingredient of described compositions is prepared from by the medical material of following weight portion:
Fructus Schisandrae Chinensis 8-12 part, Radix Ginseng 12-17 part, Cornu Cervi Pantotrichum 5-7 part, Fructus Rubi 25-35 part, Radix Astragali 16-24 part.
3. according to claim 1 have the compositions strengthening immunity and antifatigue effect, and it is characterized in that, the effective ingredient of described compositions is prepared from by the medical material of following weight portion:
Fructus Schisandrae Chinensis 10 parts, Radix Ginseng 15 parts, 6 parts, Cornu Cervi Pantotrichum, Fructus Rubi 30 parts, the Radix Astragali 20 parts.
4. the preparation method with the compositions strengthening immunity and antifatigue effect described in any one of claim 1-3, is characterized in that, comprise the following steps:
Prepare the first component: take Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum by weight, pulverize, add reflux, extract, after water, filter, when filtrate is concentrated into 40-60 DEG C, relative density is 1.10-1.15, adds by every gram of medical material the ethanol that 2-4ml volumn concentration is 90-98%, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component;
Prepare second component: take Fructus Rubi, the Radix Astragali by weight, pulverize, add reflux, extract, after water, filter, dry after filtrate recycling design, obtain second component;
Mixing: merge the first component and second component, obtain compositions.
5. the preparation method with the compositions strengthening immunity and antifatigue effect according to claim 4, it is characterized in that, the actual conditions of described preparation first component is: take Fructus Schisandrae Chinensis by weight, Radix Ginseng, Cornu Cervi Pantotrichum, pulverize, water is added according to the amount of every gram of medical material 8-12ml, soak 25-35min, reflux, extract, 50-70min, filter, water is added according to the amount of every gram of medical material 8-12ml again in medicinal residues, reflux, extract, 25-35min, filter, merging filtrate, when filtrate is concentrated into 40-60 DEG C, relative density is 1.10-1.15, the ethanol that volumn concentration is 90-98% is added by the amount of every gram of medical material 2-4ml, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component,
The described actual conditions preparing second component is: take Fructus Rubi, the Radix Astragali by weight, pulverize, add water according to the amount of every gram of medical material 8-12ml, soak 25-35min, reflux, extract, 50-70min, filter, add water according to the amount of every gram of medical material 8-12ml again in medicinal residues, reflux, extract, 25-35min, filter, merging filtrate, dry after filtrate recycling design, obtain second component.
6. the preparation method with the compositions strengthening immunity and antifatigue effect according to claim 5, is characterized in that,
The actual conditions of described preparation first component is: take Fructus Schisandrae Chinensis, Radix Ginseng, Cornu Cervi Pantotrichum by weight, pulverize, water is added according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter, add water according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, when filtrate is concentrated into 50 DEG C, relative density is 1.10-1.15, adds by the amount of every gram of medical material 3ml the ethanol that volumn concentration is 95%, precipitate with ethanol 8-12h, filter, dry after filtrate recycling design, obtain the first component;
The described actual conditions preparing second component is: take Fructus Rubi, the Radix Astragali by weight, pulverize, add water according to the amount of every gram of medical material 10ml, soak 30min, reflux, extract, 60min, filter, add water according to the amount of every gram of medical material 10ml again in medicinal residues, reflux, extract, 30min, filter, merging filtrate, dry after filtrate recycling design, obtain second component.
7. the compositions described in any one of claim 1-3 has the application in the medicine or health product strengthening immunity and antifatigue effect in preparation.
8. compositions according to claim 7 has the application in the medicine or health product strengthening immunity and antifatigue effect in preparation, it is characterized in that, the dosage form of described medicine or health product is tablet, granule, hard capsule, soft capsule, oral liquid, powder, mixture, pill or drop pill.
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