CN105147838B - With the immune composition and its preparation method and application with antifatigue effect of enhancing - Google Patents
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Abstract
Have enhancing immune and the composition and its preparation method and application of antifatigue effect the present invention relates to a kind of, belongs to pharmaceutical technology field.The active ingredient of the composition is prepared by the medicinal material of following parts by weight：3 30 parts of Schisandra chinensis, 5 45 parts of ginseng, 2 18 parts of pilose antler, 10 90 parts of raspberry, 8 60 parts of Radix Astragali.Wherein Schisandra chinensis invigorating the lung and benefiting vital QI, to help a surname of lung to drop；Ginseng and Radix Astragali invigorating the spleen are supporing yang, with the transporting of row spleen；Pilose antler and raspberry Zishen fill fine, with the Yuanyang of kidney tonifying；Gas, blood, smart three's collaboration are mended altogether, are used for the health therapy of Modern human populations immunologic hypofunction and fatigue state, have extraordinary effect.
Description
Technical field
The present invention relates to pharmaceutical technology fields, have enhancing immune and the composition of antifatigue effect more particularly to a kind of
And its preparation method and application.
Background technology
With the change of today's social life mode, people's pressure gradually increases, and physical sport is opposite to be reduced, simultaneously because
The disorder of diet, rule of working and resting, causes immune function to be greatly reduced, and body is for a long time in fatigue, inferior health and non resistance shape
State.
Therefore, body resisting fatigue ability is improved, adjusting immune function becomes academic circles at present and focus of people's attention.
But product antifatigue, that adjusting is immune conventional at present is not only expensive, but also it is confined to short-term body shape
The adjusting of state, clinical effectiveness are not satisfactory.
Therefore, using China's Traditional Chinese Medicine tcm theory, developmental function is clear, and cheap enhancing is immune and antifatigue
Product meet public demand and have particularly important realistic meaning and social effect.
Invention content
Based on this, it is an object of the invention to overcome the deficiencies of existing technologies, provide a kind of immune and anti-tired with enhancing
The active constituent of the composition of work, the composition is made of natural Chinese medicinal herb of raw material, and manufactured composition has anti-tired
Labor and immune enhancing function, and take for a long time, it is without side-effects.
A kind of to have enhancing immune and the composition of antifatigue effect, the active ingredient of the composition is by following parts by weight
Medicinal material be prepared：
3-30 parts of Schisandra chinensis, 5-45 parts of ginseng, 2-18 parts of pilose antler, 10-90 parts of raspberry, 8-60 parts of Radix Astragali.
Traditional Chinese medicine has human body the systematology macro-view cognition of " harmony between man and nature ", when body is by certain inside and outside pathogenic factors
Invasion, cause the physiological function activity of internal organs, channels and collaterals etc. to be abnormal, negative and positive of qi and blood balance coordination relationship is destroyed
When existing various clinical symptoms, human body is just in morbid state.The internal cause of morbidity has：Deficiency of vial QI, zang-fu disharmony, qi and blood it is indisposed and
Inernal injuries caused by seven emotions etc., external cause refer mainly to the heresy and pestilential pathogen of the six external factors which cause diseases, and the wherein deficiency of vital energy is most important.As it can be seen that healthy tendency is to maintain people
Body normal activities, the base substance for resisting exopathogen and disease preventing and treating.Its prosperity or not it is related to the vital movement of human body
And disease lapses to.In this sense, the healthy tendency of traditional Chinese medicine includes the immune function of human body of modern medicine.
Chinese medicine thinks mainly insufficient with the native endowment occurrence and development of immunological diseases, the heresy of the diseases caused by external factors six external factors which cause diseases, ying, wei, qi, xue
It is imbalance, the dirty dysfunction of internal organs, raw etc. closely related because being known as in phlegm-turbidity and blood stasis blood.The heresy of the diseases caused by external factors six external factors which cause diseases is the transient cause of disease,
Native endowment deficiency, ying, wei, qi, xue imbalance, the dirty dysfunction of internal organs etc. are immanent causes.Chinese medicine thinks that the kidney being the origin of congenital constitution, thus
Emphasize that kidney has the important function for adjusting and maintaining immunologic balance and its stabilization.Kidney not only shows whole to immune adjustment effect
Adjusting in terms of body, at the same it is also related with intracellular adjusting.The virtual loss of kidney, it may appear that endocrine disorder.Such as thyroid gland
Hyperfunction disease is the performance of kidney-yin deficiency, and hypothyroidism is the performance of kidney-yang deficiency.Chinese medicine thinks that spleen is the foundation of acquired constitution.
Spleen is the main place of living away from home of the maximum glandular organ of human body and immunocyte.The virtual loss of spleen, cellular immunity and body fluid
Immune function is lower than normal person.Many immunological diseases such as lupus erythematosus, dry syndrome are all that taste are deficient, and body fluid is not
Caused by foot.Chinese medicine thinks that kidney, spleen and RBC acceptor garland rate and hematid immunity function are in close relations, and the hematopoiesis function of marrow mainly comes
From kidney, spleen.Kidney storing essence, main bone, raw marrow；Spleen, middle coke, which is bullied, takes juice, variation and it is red be meaning blood.Transporting and transforming function of the spleen and stomach essence of water and grain, it is necessary to
It, could metaplasia qi and blood under kidney yang impetus.Therefore, Chinese medicine improves the work of Erythrocyte complement receptor type I with drugs for nourishing qi and activating blood
Property, to improve red blood cell adhesion immune complex function, play erythrocyte immune effect.Treatment anaphylaxis red blood cell is reduced
Disease has good curative effect.Thus Chinese medicine immunology thinks that kidney, spleen, lung dialogue cell immune system and erythrocyte immune system are all
There is performance adjustment effect.
The traditional medicine in China has different sayings, such as " tired ", " inability ", " burnout ", " de- power " to fatigue, recognizes
For the appearance of fatigue phenomenon, the imbalance with the five internal organs has close relationship, if weakness of limbs is mostly related with taste, waist soreness mostly with
Kidney is related, and short wind is mostly related with lungs, and insobriety is mostly related with heart, and endurance is not mostly related with liver.People's
Entire vital movement relies entirely in " vigour ", and vigour natural endowment is nourished in congenital (kidney) in the day after tomorrow (taste), is the source of body energy
The combination of spring, vigour and different organs generates different functions, and therefore, vigour weakness can cause each function of people at a low ebb
Holddown, this is also the basic reason that fatigue generates.Therefore, help the five internal organs are not only wanted for the treatment of fatigue, can not neglects
Depending on the supplement to " gas ".
Based on above-mentioned theory, the present invention is on the basis of summarizing numerous scholar's viewpoints, in conjunction with the constitutive character of Modern human populations,
To improve body's immunity and it is antifatigue be attributed to lung, spleen, in three main organs of kidney, i.e. lung governing qi, The spleen governs the blood, kidney life
Essence selects Schisandra chinensis, ginseng, pilose antler, raspberry and the Radix Astragali side of composition, wherein Schisandra chinensis invigorating the lung and benefiting vital QI altogether for this theory,
To help a surname of lung to drop；Ginseng and Radix Astragali invigorating the spleen are supporing yang, with the transporting of row spleen；Pilose antler and raspberry Zishen fill fine, with the member of kidney tonifying
Sun；Gas, blood, smart three's collaboration are mended altogether, are used for the health therapy of Modern human populations immunologic hypofunction and fatigue state, have very
Good effect.
Fang Zhong：Schisandra chinensis, sour, sweet, warm-natured, return lung, the heart, kidney channel.With convergence astringent method, supplementing qi and promoting the production of body fluid, tonifying kidney and calming nerves
The effect of.For long-last cough and virtual asthma, emission, enuresis frequent micturition, endless diarrhea, spontaneous perspiration, night sweat, injury thirst, feeble pulse of losing heart is interior
Heat is quenched one's thirst, palpitation and insomnia.Schisandra chinensis main component is lignanoid and polysaccharide, and modern pharmacology research shows Schisandra chinensis Thick many candies
Can the leukopenic immunosuppressive action of dose-dependant sex resistance caused by cyclophosphamide, to marrow hemopoiesis function have protection make
With improving the immunity of body, enhancing body mitigates body injury to the resistance of destructive stimulus.Schisandra chinensis can be remarkably reinforced
The phagocytic function of mouse reticuloendothelial system macrophage.Schisandra chinensis can also make mice serum hemolysin level significantly raised, make
Mouse secretory antibody function enhances, i.e., Schisandra chinensis has the function of enhancing non-specific immune function and humoral immune function.Five
Taste polysaccharide can also antagonism mouse aging lymphoid organ thymus gland and spleen atrophy, while have promote lymphocyte DNA synthesis
Effect, increase lymphoblast generate, promote Splenic immunologic.Schisandra chinensis can also promote IgG thick liquid cells in salivary gland apparent
Increase, the being proportionate property of content of immunocyte quantity and immunoglobulin, immunoglobulin increase in saliva in salivary gland
Cause of disease is effectively prevent to invade.Above research all reflects that Schisandra chinensis is significantly improved to immune function tool, and mainly
Show the increase of immune organ weight and its secreted immunoglobulin content and the increasing of phagocyte and antibacterial material ability
By force.
Ginseng, sweet in flavor, slight bitter, mild-natured, tepor, returns spleen, lung, the heart channel of Hang-Shaoyin.Effect is to reinforce vital energy, and reinforces the spleen to benefit the lung, promotes the production of body fluid only
Yearningly, tranquilize the mind and promote the intelligence mends all void.The main component of ginseng is ginsenoside, polysaccharide and volatile oil, and modern study confirms, ginseng
With multiple efficacies, such as slows down aging and anti-oxidant, improve the adaptive capacity to environment of human body, antitumor and strengthen immunity etc..
Ginsenoside Rg1 can be obviously prolonged the time that mouse power exhausts swimming, and ginsenoside Rb2 can enhance liver glycometabolism system and energy
Measure the function of generation system.
Pilose antler, it is sweet in flavor, salty, it is warm-natured, return kidney, Liver Channel.With invigorating kidney yang, marrow is mended, strengthening the bones and muscles adjusts punching to appoint, torr sore poison etc.
Effect.Pilose antler contains abundant amino acid, lecithin, vitamin and trace element etc..Pilose antler is warm but not dry, have rouse oneself and
Improve body function has preferable health-care effect to patient after asthenia universalis, prolonged illness.Pilose antler can improve the cell of body
Immune and humoral immune function, promotes the conversion of lymphocyte, has the function of immunopotentiating agent.It can increase body to the external world
Defence capability, adjust internal immunologic balance and disease avoided to occur and promote wound healing, sick body rehabilitation, it is strong to play
Strong body, the effect for resisting aging.
Radix Astragali, sweet in flavor, warm-natured, return lung, the spleen channel.With invigorating qi for strengthening superficies, the effect of diuretic and maintain drug, apocenosis, expelling pus and promoting granulation.It is yellow
Stilbene main component is polysaccharide and Flavonoid substances.Modern pharmacology research shows that Radix Astragali has enhancing body's immunity, drops blood
The effects that fat, diuresis, decompression.Modern study finds that Radix Astragali can dramatically increase the total white blood cells in blood, promotes neutral grain thin
The phagocytic function and sterilizing ability of born of the same parents and macrophage.Cellular immunity can be remarkably reinforced in Radix Astragali, promote (the America PHA, COnA, PWM
Phytolacca acinosa) caused by lymphocyte transformation.
Raspberry, it is sweet in flavor, sour, it is warm-natured.Return liver and kidney channel.With invigorating the kidney and stopping nocturnal emission, the effect of supporing yang improving eyesight.Raspberry is main
Ingredient is terpenoid, steroidal, organic acid, polysaccharide and vitamin C.It is anti-swollen that modern pharmacology research shows that raspberry has
Tumor, anti-aging, fat-reducing liver-protecting and the effect for enhancing immune function.4 kinds of extraction components of raspberry are thin to spleen lymph in vitro
Born of the same parents synthesis the experimental results showed that, 4 kinds of extract components of raspberry：Aqueous extract, alcohol extract, Thick many candies and n-butanol group
Dividing has apparent Proliferation of lymphocytes, and under the action of being assisted with or without mitogen ConA, raspberry all has obviously
The effect of activated lymphocyte.The Thick many candies and n-butanol component extracted from raspberry are swashing the lymphocyte of in vitro culture
There are the intracellular cAMP effects that increase in early stage living, horizontal without influencing cGMP.
The active ingredient of the composition is prepared by the medicinal material of following parts by weight in one of the embodiments,：Five
8-12 parts of taste, 12-17 parts of ginseng, 5-7 parts of pilose antler, 25-35 parts of raspberry, 16-24 parts of Radix Astragali.
The active ingredient of the composition is prepared by the medicinal material of following parts by weight in one of the embodiments,：Five
10 parts of taste, 15 parts of ginseng, 6 parts of pilose antler, 30 parts of raspberry, 20 parts of Radix Astragali.
The invention also discloses above-mentioned to have enhancing immune and the preparation method of the composition of antifatigue effect, including with
Lower step：
Prepare the first component：Schisandra chinensis, ginseng, pilose antler are weighed by weight, is crushed, refluxing extraction after water is added, and are filtered,
Relative density is 1.10-1.15 when filtrate is concentrated into 40-60 DEG C, and it is 90- that 2-4ml volumn concentrations, which are added, by every gram of medicinal material
98% ethyl alcohol, alcohol precipitation 8-12h are filtered, and are dried to get the first component after filtrate recycling design；
Prepare the second component：Raspberry, Radix Astragali are weighed by weight, is crushed, and refluxing extraction after water, filtration, filtrate is added
It is dried to get the second component after recycling design；
Mixing：Merge the first component and the second component to get composition.
The present invention's has enhancing immune and the preparation method of the composition of antifatigue effect, by the active ingredient in medicinal material
It extracts, improves the convenience that patient takes, and the preparation method is simple for process, be easy to carry out industrial production.
The actual conditions for preparing the first component are in one of the embodiments,：Schisandra chinensis, people are weighed by weight
Ginseng, pilose antler crush, and water is added according to the amount of every gram of medicinal material 8-12ml, impregnates 25-35min, refluxing extraction 50-70min, filters,
Water, refluxing extraction 25-35min, filtration, merging filtrate, filtrate concentration is added according still further to the amount of every gram of medicinal material 8-12ml in the dregs of a decoction
It is 1.10-1.15 to relative density at 40-60 DEG C, it is 90-98%'s that volumn concentration, which is added, by the amount of every gram of medicinal material 2-4ml
Ethyl alcohol, alcohol precipitation 8-12h are filtered, and are dried to get the first component after filtrate recycling design；
It is described prepare the second component actual conditions be：Raspberry, Radix Astragali are weighed by weight, are crushed, according to every gram of medicine
Water is added in the amount of material 8-12ml, impregnates 25-35min, refluxing extraction 50-70min, filters, according still further to every gram of medicinal material 8- in the dregs of a decoction
Water is added in the amount of 12ml, and refluxing extraction 25-35min is filtered, merging filtrate, is dried to get second group after filtrate recycling design
Point.
The actual conditions for preparing the first component are in one of the embodiments,：Schisandra chinensis, people are weighed by weight
Ginseng, pilose antler crush, and water is added according to the amount of every gram of medicinal material 10ml, impregnates 30min, refluxing extraction 60min, filters, in the dregs of a decoction again
Water is added according to the amount of every gram of medicinal material 10ml, refluxing extraction 30min, filtration, merging filtrate, filtrate is concentrated into relatively close at 50 DEG C
Degree is 1.10-1.15, the ethyl alcohol that volumn concentration is 95% is added by the amount of every gram of medicinal material 3ml, alcohol precipitation 8-12h is filtered, filter
It is dried to get the first component after liquid recycling design；
It is described prepare the second component actual conditions be：Raspberry, Radix Astragali are weighed by weight, are crushed, according to every gram of medicine
Water is added in the amount of material 10ml, impregnates 30min, refluxing extraction 60min, filters, and the amount in the dregs of a decoction according still further to every gram of medicinal material 10ml adds
Enter water, refluxing extraction 30min is filtered, merging filtrate, is dried to get the second component after filtrate recycling design.
The invention also discloses combinations of the above objects to prepare with the immune drug or health care with antifatigue effect of enhancing
Application in product.
In one of the embodiments, the dosage form of the drug or health products be tablet, granule, hard capsule, soft capsule,
Oral solution, pulvis, mixture, pill or dripping pill.
Customary adjuvant can be used in above-mentioned dosage form and preparation process is prepared.
Compared with prior art, the invention has the advantages that：
The present invention's has enhancing immune and the composition of antifatigue effect, selects Schisandra chinensis, ginseng, pilose antler, raspberry
With the Radix Astragali side of composition, wherein Schisandra chinensis invigorating the lung and benefiting vital QI altogether, to help a surname of lung to drop；Ginseng and Radix Astragali invigorating the spleen are supporing yang, with the fortune of row spleen
Change；Pilose antler and raspberry Zishen fill fine, with the Yuanyang of kidney tonifying；Gas, blood, smart three's collaboration are mended altogether, and work(is immunized for Modern human populations
It can lowly and the health therapy of fatigue state, with extraordinary effect.
Also, after coordinating each raw material at a specific ratio, there is extraordinary effect, it is oral to give the composition 30 days
Afterwards, it is shown with significant difference by macrophage phagocytosis experiment and lymphocyte proliferation assay result；It is born a heavy burden by mouse
Swimming test, hepatic glycogen determination experiment, Serum lactic acid content determination experiment and serum urea nitrogen analysis experimental result display tool
Significant difference.
Specific implementation mode
Illustrate the present invention to explain in detail below in conjunction with specific embodiment, but any restrictions is not caused to the present invention.
Bulk pharmaceutical chemicals and auxiliary material employed in following embodiment are common commercially available.
Embodiment 1
It is a kind of to have enhancing immune and the composition of antifatigue effect, it is prepared by following medicinal material：
100 grams of Schisandra chinensis, 150 grams of ginseng, 60 grams of pilose antler, 300 grams of raspberry, 200 grams of Radix Astragali.
Above-mentioned raw materials are followed these steps to prepare：
1, the first component is prepared.
Above-mentioned Schisandra chinensis, ginseng, pilose antler medicinal material are taken, is crushed, coarse powder is obtained, is placed in multi-function extractor, according to every gram of medicine
Distilled water is added in the amount of material 10ml, impregnates 30min, refluxing extraction 60min, filtration；According still further to every gram of medicinal material 10ml's in the dregs of a decoction
Distilled water, refluxing extraction 30min, filtration, merging filtrate, measurement relative density 1.10- when filtrate is concentrated into 50 DEG C is added in amount
1.15；The ethyl alcohol that volumn concentration is 95% is added into filtrate by the amount of every gram of medicinal material 3ml, alcohol is sink to total liquid quality
About 70% (water-soluble substances precipitate i.e. plus after alcohol, remove the liquid quality after precipitation and decline, and are total liquid quality before alcohol precipitation
70% or so), alcohol precipitation is overnight (about 10h), recycles ethyl alcohol and measures relative density 1.20-1.30 when being concentrated into 50 DEG C, dry, obtains
First component.
2, the second component is prepared.
Above-mentioned raspberry, Milkvetch Root are taken, is crushed, coarse powder is obtained, is placed in multi-function extractor, according to every gram of medicinal material 10ml
Amount be added distilled water, impregnate 30min, refluxing extraction 60min, filtration；Amount in the dregs of a decoction according still further to every gram of medicinal material 10ml is added
Distilled water, refluxing extraction 30min, filtration, merging filtrate, filtrate measure relative density 1.05-1.10 when being concentrated into 50 DEG C, do
It is dry, obtain the second component.
3, it mixes.
Merge the first component and the second component to get composition A.
Embodiment 2
It is a kind of to have enhancing immune and the composition of antifatigue effect, it is prepared by following medicinal material：
30 grams of Schisandra chinensis, 450 grams of ginseng, 20 grams of pilose antler, 90 grams of raspberry, 80 grams of Radix Astragali.
Composition B is prepared with reference to the method for embodiment 1.
Embodiment 3
It is a kind of to have enhancing immune and the composition of antifatigue effect, it is prepared by following medicinal material：
300 grams of Schisandra chinensis, 50 grams of ginseng, 180 grams of pilose antler, 100 grams of raspberry, 600 grams of Radix Astragali.
Composition C is prepared with reference to the method for embodiment 1.
Embodiment 4
It is a kind of to have enhancing immune and the oral solution of antifatigue effect, it can be taken as health products.
The oral solution is made by the following method：
The composition A that Example 1 is prepared, dextrin is added into the composition A or honey reconciles, and filling bottle is made
Oral solution to get.
Embodiment 5
It is a kind of to have enhancing immune and the capsule of antifatigue effect, it can be taken as health products.
The capsule is made by the following method：
The composition A that Example 1 is prepared pregelatinized starch is added in the dry powder of above-mentioned composition A, mixing,
Granulation, drying are added magnesium stearate and talcum powder, filled hard capsules to get.
Embodiment 6
It is a kind of to have enhancing immune and the tablet of antifatigue effect, it can be taken as health products.
The tablet is made by the following method：
Microcrystalline cellulose, pregelatinized starch, sodium carboxymethyl starch are added the composition A that Example 1 is prepared
In the dry powder for stating composition A, mix, pelletize, drying, whole grain, be added magnesium stearate, tabletting to get.
Comparative example 1
This comparative example prepares a kind of composition, with embodiment 1 with the composition base for enhancing immune and antifatigue effect
This is identical, the difference is that, the composition of this comparative example is prepared by following medicinal material：
150 grams of ginseng, 60 grams of pilose antler, 300 grams of raspberry, 200 grams of Radix Astragali.
Be prepared composition D with reference to the method for embodiment 1, wherein ginseng, pilose antler according to the first component preparation method
It prepares, raspberry, Radix Astragali are prepared according to the preparation method of the second component.
Comparative example 2
This comparative example prepares a kind of composition, with embodiment 1 with the composition base for enhancing immune and antifatigue effect
This is identical, the difference is that, the composition of this comparative example is prepared by following medicinal material：
100 grams of Schisandra chinensis, 150 grams of ginseng, 60 grams of pilose antler, 200 grams of Radix Astragali.
Composition E is prepared with reference to the method for embodiment 1, wherein Schisandra chinensis, ginseng and pilose antler are according to the first component
Prepared by preparation method, Radix Astragali is prepared according to the preparation method of the second component.
Comparative example 3
This comparative example prepares a kind of composition, with embodiment 1 with the composition base for enhancing immune and antifatigue effect
This is identical, the difference is that, the composition of this comparative example is prepared by following medicinal material：
100 grams of Schisandra chinensis, 150 grams of ginseng, 60 grams of pilose antler, 300 grams of raspberry, 200 grams of Radix Astragali, the fruit of Chinese wolfberry, 200 grams.
Be prepared composition F with reference to the method for embodiment 1, wherein Schisandra chinensis, ginseng, pilose antler according to the first component system
Prepared by Preparation Method, raspberry, Radix Astragali and the fruit of Chinese wolfberry are prepared according to the preparation method of the second component.
Comparative example 4
This comparative example prepares a kind of composition, with embodiment 1 with the composition base for enhancing immune and antifatigue effect
This is identical, the difference is that, the composition of this comparative example is prepared by following medicinal material：
100 grams of Schisandra chinensis, 150 grams of ginseng, 60 grams of pilose antler, 300 grams of raspberry, 200 grams of the fruit of Chinese wolfberry.
Be prepared composition G with reference to the method for embodiment 1, wherein Schisandra chinensis, ginseng, pilose antler according to the first component system
Prepared by Preparation Method, raspberry, the fruit of Chinese wolfberry are prepared according to the preparation method of the second component.
Experimental example 1
The composition that above-described embodiment and comparative example are prepared carries out pharmacodynamic study.
1, Immune-enhancing effect is tested.
The phagocytosis experiment of 1.1 macrophages.
1.1.1 experimental animal：Kunming mice, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
1.1.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing；India ink, the Shanghai bio tech ltd Rui Yong carry
For.
1.1.3 method and result：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, group
Close object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.It is given in last
Diluted india ink is injected from mouse tail vein after medicine, is calculated by per 10g weight 0.1mL.Wait for that prepared Chinese ink injects, timing immediately.
2,10min, takes 20 μ l of blood from angular vein clump respectively, exists side by side and be added into 2mL 0.1%Na after injection prepared Chinese ink2CO3Solution
In.With spectrophotometer or full-automatic microplate reader at 600nm wavelength densitometric value (OD), with Na2CO3Solution makees blank pair
According to.Mouse is put to death, liver and spleen are taken, organ surface blood stains is blotted with filter paper, weighs.Indicate that mouse carbon is wide with phagocytic index
Clear ability.Phagocytic index a is calculated as follows：
1 macrophage of table swallows experimental result
As can be seen from the above table, the macrophage phagocytic index of the mouse of composition A-G groups is above blank group, explanation
Play the role of enhancing the phagocytic activity of mouse macrophage, still, wherein composition A-C groups are with obvious effects is better than D-G groups, spy
It is not composition A groups, there is best enhancing immune effect.
1.2 lymphproliferation responses are tested.
1.2.1 experimental animal：C57 mouse, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
1.2.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing；Platform expects blue (Trypan Blue) that Beijing ancient cooking vessel state prosperity is given birth to
Object Science and Technology Ltd. provides；Canavaline, sigma companies provide；Hanks liquid, GENVIEW companies provide；RPMI1640 is trained
Nutrient solution is matched silent your biochemistry product (Beijing) Co., Ltd of winged generation and is provided；Calf serum, the Zhejiang day limited public affairs of Hangzhoupro biotechnology
Department provides；2 mercapto ethanol, Beijing prosperity bio tech ltd of ancient cooking vessel state provide；Isopropanol, Tianjin great Mao chemical reagent works carry
For.
1.2.3 reagent pre-treatment.
Complete culture solution：RPMI1640 culture solutions are with 0.22 μm of membrane filtration degerming, with 10% calf serum of preceding addition,
1% glutamine (200mmol/L), penicillin (100U/mL), streptomysin (100ug/L) and 5 × 10- 5The 2- sulfydryls of mol/L
Ethyl alcohol,
ConA (canavaline) liquid：The solution of 100 μ g/mL, filtration sterilization, in low temperature refrigerator (- 20 are configured to distilled water
DEG C) preserve.
Sterile Hank's liquid：With preceding with 3.5% sterile NaHCO3Adjust pH7.2-7.4.
MTT liquid：5mgMTT is dissolved in the PBS of 1mL pH7.2, it is now with the current.
Acid isopropyl alcoholic solution：The HCl of 4mL 1mol/L, prepared before use are added in 96mL isopropanols.
1.2.4 medication：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, combination
Object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.
1.2.5 prepared by splenocyte suspension
Each group animal was put to death in second day after the last administration, it is sterile to take spleen, it is placed in and fills appropriate (3-5ml) sterile Hank's
In the plate of liquid, and one piece of gauze is placed above in spleen, is gently ground spleen with large syringe inner core, it is outstanding that individual cells are made
Liquid.It is filtered through 200 mesh screens, is washed 2 times with Hank's liquid, centrifuge 10min (1000r/min) every time.Then cell is suspended in
In the complete culture solution of 2mL, with platform phenol orchid dyeing counting viable count (should be 95% or more), adjustment cell concentration is 3 × 106
A/mL.
1.2.6 lymphopoiesis measures.
Holes is divided to be added in 24 well culture plates cell suspension, per hole 1mL, a hole adds 75 μ l Con A liquid (to be equivalent to 7.5
μ g/mL), 5%CO as a contrast, is set in another hole2, 37 DEG C of CO272h is cultivated in incubator.Culture terminates preceding 4h, is gently sucked per hole
Supernatant 0.7mL is added 0.7mL and is free of the RPMI1640 culture solutions of calf serum, while 50 holes μ l/ MTT (5mg/mL) are added,
Continue to cultivate 4h.After culture, 1mL acid isopropyl alcohol is added per hole, ultrasonic vibration (2 seconds) or manually blows and beats mixing, makes purple
Color crystallization is completely dissolved.Then it is dispensed into 96 well culture plates, each hole dispenses 3 holes as Duplicate Samples, uses enzyme linked immunosorbent detection
Instrument measures OD value with 570nm wavelength.
1.2.7 experimental result.
The results are shown in table below for lymphocyte proliferation assay.
2 lymphocyte proliferation assay result of table
As can be seen from the above table, the proliferative capacity of the mouse lymphocyte of composition A-G groups is above blank group, explanation
Play the role of enhancing the proliferative capacity of mouse lymphocyte, still, wherein composition A-C groups are with obvious effects is better than D-G groups, spy
It is not composition A groups, there is best enhancing immune effect.
In conclusion the composition of the present invention carries out prescription under the guidance of theory of traditional Chinese medical science, swallowed with mouse macrophage
Ability is index, evaluates the cellular immune function of mouse, and using lymphopoiesis ability as observation index, evaluates the leaching of mouse
Bar cellular immune function the results show that immune function of mice can be improved in the composition of the present invention there is apparent Immune-enhancing effect to make
With.
2, antifatigue experiment.
2.1 swimming with a load attached to the body are tested.
2.1.1 experimental animal：Kunming mice, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
2.1.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing.
2.1.3 method and result：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, group
Close object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.
After last dose 30min, 5% weight lead of mouse tail load is placed in swimming trunk went swimming, depth of water 30cm,
25 DEG C -28 DEG C of water temperature, record swimming start the time to dead mouse, and the results are shown in table below.
3 mice burden swimming experimental result of table
As can be seen from the above table, the mouse swimming time of composition A-G groups is above blank group, illustrates have extension small
The effect of mouse swimming time, still, wherein composition A-C groups are with obvious effects is better than D-G groups, especially composition A groups, has most
Good antifatigue effect.
2.2 hepatic glycogen content determination experiments.
2.2.1 experimental animal：Kunming mice, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
2.2.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing；Bioengineering Research Institute is built up in hepatic glycogen testing cassete, Nanjing
It provides.
2.2.3 method and result：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, group
Close object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.
Mouse continuous gavage 30 days, after last dose 30min, cervical dislocation is put to death, and is taken liver immediately, is rushed with physiological saline
It after washing, is blotted with filter paper, accurately weighs liver 100mg, be homogenized, with kit measurement, the results are shown in table below.
4 mouse hepatic glycogen content determination experiment result of table
As can be seen from the above table, the mouse hepatic glycogen content of composition A-G groups is above blank group, and explanation is improved
The effect of hepatic glycogen content, still, wherein composition A-C groups are with obvious effects is better than D-G groups, especially composition A groups, has most
Good antifatigue effect.
The measurement of 2.3 Serum lactic acid contents
2.3.1 experimental animal：Kunming mice, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
2.3.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing；Bioengineering Research Institute is built up in blood lactase acid testing cassete, Nanjing
It provides.
2.3.3 method and result：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, group
Close object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.
Mouse continuous gavage 30 days after last dose 30min, uses capillary to take a blood sample 20 μ l from the mouse intraocular corner of the eyes immediately,
Then it does not bear a heavy burden and stops after temperature is 30 DEG C of water went swimming 10min, take a blood sample 20 μ l immediately, takes a blood sample again after rest 20min.
With kit measurement, the results are shown in table below for Serum lactic acid content.
5 mouse Serum lactic acid content determination experiment result (mg/g) of table
As can be seen from the above table, mouse blood lactase acid lift-off value is apparent after the composition of composition A-G groups can make movement
It reduces, and blood lactase acid level restores more apparent after rest, still, wherein composition A-C groups are with obvious effects is better than D-G groups,
Especially composition A groups have the function of best increase fatigue recovery speed.
2.4 serum urea nitrogen analysis are tested
2.4.1 experimental animal：Kunming mice, weight 18-22g, half male and half female, in the high-new medical animal experiment research in Changchun
The heart provides.
2.4.2 drug and reagent：Composition A, B, C, D, E, F, G that above-described embodiment and comparative example are prepared are taken, it is real
With distilled water dissolved dilution to contained crude drug amount it is 50mg/mL when testing；Serum urea nitrogen testing cassete, Nanjing are built up bioengineering and are ground
Study carefully and is provided.
2.4.3 method and result：Animal is divided into 8 groups, i.e. blank group, composition A groups, composition B groups, composition C groups, group
Close object D groups, composition E groups, composition F groups, composition G groups.
Blank group gives distilled water；Administration group is filled according to the amount of 10mg/10g (10mg is administered per 10g weight mouse) respectively
The concentration of stomach, solution used in each group gavage is each configured to given low 0.2ml/10g, and dosage period is 30 days.
Mouse continuous gavage 30 days, after last dose 30min, the not swimming with a load attached to the body 90min in the water that temperature is 30 DEG C, not
It after ceasing 60min, plucks eyeball and takes blood, 3000rpm centrifuges 10min, takes supernatant, with its serum urea nitrogen of kit measurement, as a result
As shown in the table.
6 mice serum urea nitrogen content determination experiment result of table
As can be seen from the above table, the composition of composition A-G groups can be substantially reduced mice serum urea nitrogen content, enhancing
Body makes body endurance increase the adaptability of strenuous exercise.But wherein composition A-C groups are with obvious effects is better than D-G
Group, especially composition A groups have the function of best increase body endurance.
In conclusion the composition of the present invention carries out prescription under the guidance of theory of traditional Chinese medical science, with the mice burden swimming time
For observation index, the anti-fatigue ability of mouse is evaluated；And it using lactate level in blood after mouse movement as observation index, has rated small
Mouse aerobic metabolism and anti-fatigue ability；The results show that aerobic metabolism can be improved in Mice Body is horizontal and anti-for the composition of the present invention
Fatigue effect.
Experimental example 2
The composition that above-described embodiment and comparative example are prepared carries out toxicologic study.
1, acute toxicity testing.
1.1 experimental animal：Kunming mice, 40, weight 18-22g, half male and half female, the high-new medical animal experiment in Changchun is ground
Center offer is provided.
1.2 drugs and reagent：Composition A prepared by Example 1 is given birth to distilled water dissolved dilution to 2g when experiment
The aqueous solution of medicine/ml.
1.3 methods and result：Each gavage amount is 0.6ml/, and gavage is total to three times in 24 hours, total amount 1.8ml, phase
When in primary dose 180g crude drugs/kg/d, being equivalent to 600-800 times of 70kg mankind's normal clinical dosage.30 after mouse administration
Dead and abnormal phenomenon does not occur for conventinal breeding in it.Prove that the composition A that the present invention obtains has no toxic side effect, clinic can be safe
It uses.
2, long term toxicity test.
2.1 experimental animal：Wistar rats, 120, weight 180-220g, half male and half female, the high-new medical faunae reality in Changchun
Research center offer is provided.
2.2 drugs and reagent：Composition A prepared by Example 1 is given birth to distilled water dissolved dilution to 2g when experiment
The aqueous solution of medicine/ml.
2.3 groupings and medication：Experimental animal is randomly divided into 4 groups, every group 30 by gender weight.1 group of gavage is given
Distilled water, 0.2ml/100g；2,3,4 groups of gavages give test medicine, and dosage is respectively：50mg/100g、100mg/100g、
200mg/100g, i.e., give the crude drug amount of 50mg, 100mg or 200mg per 100g weight, and each group concentration is each configured to 0.2ml/
The given low of 100g.The above each group is all put to death after continuously taking 6 months.
2.4 Indexs measure
2.4.1 general status detects：Weight, diet, mood, movable energy.
2.4.2 biochemical index：Red blood cell count(RBC), white blood cell count(WBC), platelet count, hemoglobin counting, serum
Albumen, cholesterol and blood glucose etc..
2.4.3 pathological examination：The heart, liver, spleen, lung, kidney, brain, stomach, thymus gland, adrenal gland etc..
2.5 the experimental results showed that, after rat continuously takes 6 months drugs of the present invention, weight, diet, mood, movable energy
Equal general status are good, each main organs and Serum Indexes no abnormality seen, it was demonstrated that the composition A that the present invention obtains is to body without bright
Aobvious long term toxicity effect.
In conclusion acute and long_term toxcity the experimental results showed that, patent health-care composition safety of the present invention, without obviously
Toxic side effect；It is oral to give patent drug of the present invention after 30 days, experiment and lymphocyte proliferation assay are swallowed by macrophage
As a result display has significant difference；It is tested by mice burden swimming, hepatic glycogen determination experiment, Serum lactic acid content determination experiment
It is shown with significant difference with serum urea nitrogen analysis experimental result.According to《Health food is examined advises with assessment technique
Model》(2003 editions), this zooscopy conclusion confirm that patent health-care composition of the present invention has enhancing immune function and antifatigue work
With.
Each technical characteristic of embodiment described above can be combined arbitrarily, to keep description succinct, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, it is all considered to be the range of this specification record.
Several embodiments of the invention above described embodiment only expresses, the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Range.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (8)
1. a kind of have enhancing immune and the composition of antifatigue effect, which is characterized in that the active ingredient of the composition by
The medicinal material of following parts by weight is prepared：
3-30 parts of Schisandra chinensis, 5-45 parts of ginseng, 2-18 parts of pilose antler, 10-90 parts of raspberry, 8-60 parts of Radix Astragali；
Described has enhancing immune and the preparation method of the composition of antifatigue effect, includes the following steps：
Prepare the first component：Schisandra chinensis, ginseng, pilose antler are weighed by weight, is crushed, and refluxing extraction after water, filtration, filtrate is added
Relative density is 1.10-1.15 when being concentrated into 40-60 DEG C, and it is 90-98%'s that 2-4ml volumn concentrations, which are added, by every gram of medicinal material
Ethyl alcohol, alcohol precipitation 8-12h are filtered, and are dried to get the first component after filtrate recycling design；
Prepare the second component：Raspberry, Radix Astragali are weighed by weight, is crushed, and refluxing extraction after water, filtration, filtrate recycling is added
It is dried to get the second component after solvent；
Mixing：Merge the first component and the second component to get composition.
2. according to claim 1 have enhancing immune and the composition of antifatigue effect, which is characterized in that the combination
The active ingredient of object is prepared by the medicinal material of following parts by weight：
8-12 parts of Schisandra chinensis, 12-17 parts of ginseng, 5-7 parts of pilose antler, 25-35 parts of raspberry, 16-24 parts of Radix Astragali.
3. according to claim 1 have enhancing immune and the composition of antifatigue effect, which is characterized in that the combination
The active ingredient of object is prepared by the medicinal material of following parts by weight：
10 parts of Schisandra chinensis, 15 parts of ginseng, 6 parts of pilose antler, 30 parts of raspberry, 20 parts of Radix Astragali.
4. claim 1-3 any one of them has enhancing immune and the preparation method of the composition of antifatigue effect, special
Sign is, includes the following steps：
Prepare the first component：Schisandra chinensis, ginseng, pilose antler are weighed by weight, is crushed, and refluxing extraction after water, filtration, filtrate is added
Relative density is 1.10-1.15 when being concentrated into 40-60 DEG C, and it is 90-98%'s that 2-4ml volumn concentrations, which are added, by every gram of medicinal material
Ethyl alcohol, alcohol precipitation 8-12h are filtered, and are dried to get the first component after filtrate recycling design；
Prepare the second component：Raspberry, Radix Astragali are weighed by weight, is crushed, and refluxing extraction after water, filtration, filtrate recycling is added
It is dried to get the second component after solvent；
Mixing：Merge the first component and the second component to get composition.
5. according to claim 4 there is the immune preparation method with the composition of antifatigue effect of enhancing, feature to exist
In the actual conditions for preparing the first component are：Schisandra chinensis, ginseng, pilose antler are weighed by weight, are crushed, according to every gram of medicine
Water is added in the amount of material 8-12ml, impregnates 25-35min, refluxing extraction 50-70min, filters, according still further to every gram of medicinal material 8- in the dregs of a decoction
Water is added in the amount of 12ml, and refluxing extraction 25-35min, filtration, merging filtrate, relative density is when filtrate is concentrated into 40-60 DEG C
The ethyl alcohol that volumn concentration is 90-98% is added by the amount of every gram of medicinal material 2-4ml in 1.10-1.15, and alcohol precipitation 8-12h is filtered,
It is dried to get the first component after filtrate recycling design；
It is described prepare the second component actual conditions be：Raspberry, Radix Astragali are weighed by weight, are crushed, according to every gram of medicinal material 8-
Water is added in the amount of 12ml, impregnates 25-35min, refluxing extraction 50-70min, filters, according still further to every gram of medicinal material 8-12ml in the dregs of a decoction
Amount be added water, refluxing extraction 25-35min, filtration, merging filtrate dries to get the second component after filtrate recycling design.
6. according to claim 5 there is the immune preparation method with the composition of antifatigue effect of enhancing, feature to exist
In,
It is described prepare the first component actual conditions be：Schisandra chinensis, ginseng, pilose antler are weighed by weight, are crushed, according to every gram of medicine
Water is added in the amount of material 10ml, impregnates 30min, refluxing extraction 60min, filters, and the amount in the dregs of a decoction according still further to every gram of medicinal material 10ml adds
Enter water, refluxing extraction 30min, filtration, merging filtrate, relative density is 1.10-1.15 when filtrate is concentrated into 50 DEG C, by every gram of medicine
The amount of material 3ml is added the ethyl alcohol that volumn concentration is 95%, alcohol precipitation 8-12h, filtering, dried after filtrate recycling design to get
First component；
It is described prepare the second component actual conditions be：Raspberry, Radix Astragali are weighed by weight, are crushed, according to every gram of medicinal material
Water is added in the amount of 10ml, impregnates 30min, refluxing extraction 60min, filters, and the amount in the dregs of a decoction according still further to every gram of medicinal material 10ml is added
Water, refluxing extraction 30min are filtered, merging filtrate, are dried to get the second component after filtrate recycling design.
7. claim 1-3 any one of them composition is being prepared with the immune drug or health care with antifatigue effect of enhancing
Application in product.
8. composition according to claim 7 is in preparing the drug or health products that have and enhance immune and antifatigue effect
Application, which is characterized in that the dosage form of the drug or health products be tablet, granule, hard capsule, soft capsule, oral solution, powder
Agent, mixture, pill or dripping pill.
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