CN103689599A - Health-care food capable of enhancing immunity and preparation method thereof - Google Patents
Health-care food capable of enhancing immunity and preparation method thereof Download PDFInfo
- Publication number
- CN103689599A CN103689599A CN201310729930.6A CN201310729930A CN103689599A CN 103689599 A CN103689599 A CN 103689599A CN 201310729930 A CN201310729930 A CN 201310729930A CN 103689599 A CN103689599 A CN 103689599A
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- Prior art keywords
- preparation
- coix seed
- immunity
- add
- health food
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
- A23L7/198—Dry unshaped finely divided cereal products, not provided for in groups A23L7/117 - A23L7/196 and A23L29/00, e.g. meal, flour, powder, dried cereal creams or extracts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Botany (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a health-care food capable of enhancing the immunity and a preparation method thereof. The health-care food is prepared from the following raw materials of dendrobium nobile, semen coicis and gen-seng at a certain ratio. The health-care food has the functions of enhancing the physique and the immunity of the organism, has a certain promote effect on the cell-mediated immunity function, the antibody-mediated immunity function, the function of phagocytosis of peritoneal macrophage, and NK cell viability, is a good item for nourishing, bodybuilding and life lengthening, is safe, and has no side effects.
Description
Technical field
The present invention relates to a kind of health food that strengthens immunity and preparation method thereof, belong to the technical field of health food.
Background technology
Immunity refers to that human immune system resists the ability of pathogenic infection.Immune system is comprised of thymus gland (i.e. atrophy gradually, degeneration after puberty), spleen and lymphoid tissue etc.A sound human immune system is the main system of defense of body, can protect body to be without prejudice, and helps body to eliminate external bacterium, virus, repairs Organ and tissue in damaged condition and makes it recover original function etc.And everyone immunity is different, the people that immunity is strong is difficult for sick, and the weak people of immunity is easily brought out various illnesss by pathogenic bacterial infection.Immunoglobulin (Ig) is to be produced by immune system, and the power of immunity depends on quantity and the vigor of its globulin, and immunoglobulin (Ig) is that domestic and international scientific circles generally acknowledge and is the material of the raising human body resistance against diseases that is widely used.Immunization programs for children allelotaxis imperfection before 0～12 year old, has the immunoglobulin secretion amount of antibody activity few in body, immunologic function is unsound, and child is at this moment as easy as rolling off a log sick.During 13～45 years old, be immunologic function relatively stable period, immunologic function comparatively perfect.But because the people of this one-phase is easily subject to the pressure influence of life, study, work, antibody and growth factor secretory volume in body are fluctuated widely, thisly will cause the immunoglobulin content in body unstable, cause immunity degradation and sick.After 46 years old, the immune organ function of human body falls into a decline, and has the immunoglobulin (Ig) of antibody activity and growth factor secretion to decline, immunity degradation in body; Start to bring out geriatric disease.According to World Health Organization investigation, approximately there is more than 35% people in the whole world in fatigue state, and especially middle-aged male crowd fatigue state person is up to 60-75%.Expert points out, long-term poor sleep, continue overtired easy initiation pharyngo-laryngitis chronica, neck or the illness such as axillary gland swells and ache, DOMS and multiple joint property arthralgia, finally cause immunity degradation, and after immunity degradation, make again " burnout syndrome " more give prominence to and worsen, thereby form a vicious circle.Along with social development, the progress of science and technology, the quickening of rhythm of life, the enriching of culture and material life, the fatigue state of a middle-aged person in inferior health not only, also there is overtired phenomenon in each big-and-middle student of universities and colleges.So researching and developing a kind of Chinese medicine health-care series products that strengthens body autoimmunity has extensively and profound significance.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of health food that strengthens immunity and preparation method thereof, and these health products have builds up health, and improves the effect of immunity of organisms, and has no side effect.
For solving the problems of the technologies described above, the present invention realizes by the following technical solutions:
Strengthen a health food for immunity, by active drug composition and auxiliary material, made, according to listed as parts by weight, the active drug composition of use is: HERBA DENDROBII 500-800 part, coix seed 800-1200 part, American Ginseng 200-500 part.
Specifically, the health food of described enhancing immunity is according to listed as parts by weight, and the active drug composition of use is: 670 parts of HERBA DENDROBIIs, 1000 parts of coix seeds, 330 parts of American Ginsengs.
The preparation method of the health food of described enhancing immunity is: get HERBA DENDROBII, coix seed and American Ginseng, combine with pharmaceutic adjuvant, and make corresponding preparation according to conventional preparation method.
Specifically, the preparation method of the health food of described enhancing immunity is: coix seed is got part and pulverized, and crosses 80-200 mesh sieve, and gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, American Ginseng and HERBA DENDROBII were soaked after 0.3-1 hour, decoct 1-3 time, add 4-8 times of water gaging to decoct 0.5-3 hour at every turn, filter, merging filtrate, reduced pressure concentration becomes thick paste, add coix seed fine powder and pharmaceutic adjuvant to mix, drying under reduced pressure, pulverizes, then and makes corresponding preparation according to conventional preparation method.
More particularly, the preparation method of the health food of described enhancing immunity is: coix seed is got 22% of recipe quantity and pulverized, and crosses 120 mesh sieves, and gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, American Ginseng and HERBA DENDROBII add suitable quantity of water and soak after 0.5 hour, decoct twice, add 6 times of water gagings decocts 1 hour at every turn, filter merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38, add coix seed fine powder and pharmaceutic adjuvant to mix,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, then and make corresponding preparation according to conventional preparation method.
Described preparation is capsule.
Described capsule is so prepared: coix seed is got 22% of recipe quantity and pulverized, and crosses 120 mesh sieves, and gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, American Ginseng and HERBA DENDROBII add suitable quantity of water and soak after 0.5 hour, decoct twice, add 6 times of water gagings at every turn and decoct 1 hour, filter, merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38, adds coix seed fine powder to mix ,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, cross 40 mesh sieves, incapsulate, obtain.
The asthenic symptoms of the traditional Chinese medical science are mainly seen in the immunity disease patient of modern medicine, especially the patient of immunologic hypofunction.Most tonics can strengthen the immunologic function of body, in order to treatment, because of the disease that immunologic hypofunction causes, play the effect of immunopotentiator; In < < medical science main story > >, also point out: " empty person, the positive deficiency of vital energy is also; For look miserable shape is thin, it is god's fright due to decicincy of qi that declines ... all working as of asthenic symptoms, mends also.
The health food of enhancing immunity of the present invention is prepared from by HERBA DENDROBII, coix seed, American Ginseng and auxiliary material.HERBA DENDROBII cold nature wherein, taste is sweet, returns stomach, kidney channel, and have and grow smart effect except gas, tonifying five zang organs consumptive disease, reinforcing yin essence under spleen, for pyreticosis Tianjin wound, dry polydipsia, deficiency of stomach-Yin, food is wanted to vomit less; Abnormal heat is not moved back after being ill, fire excess from yin deficiency, hectic fever due to yin consumptive disease heat, and order is secretly not clear, the illnesss such as flaccidity of extremities.Coix seed gas is micro-, taste is micro-sweet, cool in nature.Return spleen, stomach, lung channel, there is clearing damp and promoting diuresis, invigorating the spleen to arrest diarrhea, except the effect of numbness, apocenosis, detoxicating and resolving a mass.For oedema, beriberi, difficult urination, splenasthenic diarrhea, arthritis with fixed pain caused by dampness contraction, lung carbuncle, acute appendicitis, wart, cancerous swelling.American Ginseng is cool in nature, sweet, the micro-hardship of taste; The thoughts of returning home, lung, kidney, have boosting qi and nourishing yin, and the effect of clearing heat and promoting fluid is usually used in that the deficiency of vital energy is cloudy loses, and abnormal heat is tired tired, coughs to breathe heavily heat in phlegm blood and quench one's thirst the illnesss such as dryness of the mouth and throat.In this compound, take HERBA DENDROBII reinforcing stomach reg fluid, nourishing Yin and clearing heat be monarch drug in a prescription; Joining coix seed clearing damp and promoting diuresis, invigorating the spleen to arrest diarrhea, can help stem of noble dendrobium invigorating spleen and reinforcing stomach, is ministerial drug; American Ginseng boosting qi and nourishing yin, clearing heat and promoting fluid, help stem of noble dendrobium clearing heat and promoting fluid, is adjutant.The same use of three medicines, plays reinforcing stomach reg fluid, nourishing Yin and clearing heat merit altogether.Full side's compatibility is reasonable, and giving consideration to both the incidental and fundamental can effectively play enhancing immunity, is the health food of good enhancing immunity.
experimental example 1: technical study
1. total saponin content assay method
The preparation of reference substance solution: precision takes ginsenoside Re's standard items 20.0mg, dissolves and is settled to 10ml with methyl alcohol, obtains ginsenoside Re's standard liquid of 2.0mg/ml.
The preparation of need testing solution: precision takes capsule 's content 2g of the present invention and puts in tool plug conical flask, adds 40ml water, ultrasonic 30min, then be settled to 100ml, and shake up, place, standby.
Column chromatography: with Amberlite-xad2 macroporous absorbent resin dress post 3cm, then with medium-sized aluminium oxide dress post 1cm, use 25ml70% ethanol elution, with 25ml water, carry out wash-out again, discard eluent, the standby need testing solution 1m upper prop of accurate absorption carries out chromatography, with 25ml, washes post, with water-solubility impurities such as sweetening offs, discard eluent, then use the total saponin(e of 25ml 70% ethanol elution, collect eluent in evaporating dish, 60 ℃ of water bath methods, do colour developing with this.
Determination method: accurately add 0.2ml 5% vanillic aldehyde glacial acetic acid solution in the above-mentioned evaporating dish having volatilized, rotate evaporating dish, residue is dissolved, add again 0.8ml perchloric acid, mix in rear immigration 10ml clogged tube the jam-pack lid 15min that heats, the cooling rear glacial acetic acid 5.0ml that accurately adds in water-bath below 60 ℃, after shaking up, take corresponding reagent as blank, in 560nm place, measure absorbance.
2. Methods in Determination of Polysaccaride Content
The preparation of reference substance solution: get glucose reference substance appropriate, accurately weighed, adding distil water is made every 1ml containing the solution of glucose 0.1mg, obtains.
The preparation of need testing solution: precision takes capsule 's content 2g of the present invention, evaporate to dryness, residue is put in tool plug conical flask, add 95% ethanol 50ml water-bath backflow 1h, filter, residue washs with 95% ethanol 20ml, filter, discard filtrate, residue adding distil water 50ml water-bath backflow 1h, filter, distillation 20ml washing for residue, filters, merging filtrate, is settled to 100ml, pipettes 5ml filtrate and puts in 50ml measuring bottle, be diluted to scale, obtain.
Determination method: pipette respectively each 3ml of reference substance solution and need testing solution and put in 10ml tool plug conical flask, add 5% phenol solution 1ml, shake up, add rapidly concentrated sulfuric acid 5ml, shake up, room temperature is placed 1h, measures in 480nm place, obtains.
3. extract the selection of solvent
Fang Zhongsan taste medicine, coix seed is the dry mature kernal of grass Job's tears, about result of study shows, coixan has the function of the immunity of organisms of raising.HERBA DENDROBII is the dry stem of orchid HERBA DENDROBII, other materials such as its chemical composition main alkaloid class, flavonoids and polysaccharide, and wherein polysaccharide has many-sided immunological enhancement.American Ginseng is the dry rhizome of Araliaceae American Ginseng, and main active ingredient has Radix Panacis Quinquefolii polysaccharide, American ginseng saponin and other trace elements.
In sum, in side, the main total polysaccharide of active ingredient, total saponin(e are water soluble ingredient, available water boiling and extraction, and the medication of three medicine traditional Chinese medical science tradition is decoction, therefore extract solvent, selects water.
4. extraction process Orthogonal Experiment and Design: from medicinal material extract principle, extraction efficiency is except outside the Pass the character with medicinal material has, and also, with solvent species, solvent load, the factors such as extraction time, extraction time are relevant.Owing to determining that the party extracts solvent, be water, therefore investigate extraction process by water.
Take respectively amount of water, extraction time, extraction time is investigation factor, and each factor is established 3 levels, in Table 1.Select L9(3
4) orthogonal table experiment arrangement, take medicinal extract amount, total polyoses content, total saponin content as investigating index, and calculate the preferred extraction process of overall score, in Table 2.
Overall score=(total determination of polysaccharide value/total polyoses content maximum) * 50+(total saponin content measured value/total saponin content maximum) * 50
Table 1 factor level table
Table 2 orthogonal experiments
Analyze conclusion: with general comment, be divided into index, by table 2 intuitive analysis, can obtain optimum extraction process is A
1b
2c
2, extreme difference size is R
c>R
b>R
a, extraction time > extraction time > amount of water, determines that extraction process is: extract 2 times, add 6 times of water gagings at every turn and decoct 1 hour.
5. the research of concentration technology
Because of the extraction process by water adopting, for preventing that active ingredient is subject to heat damage for a long time, so the basic methods that adopts the technique of reduced pressure concentration (vacuum is conventional-0.08Mpa) to carry out concentration technology research, take the content concentration technology parameter best as assessment indicator optimizes of thick polysaccharide, total saponin(e.Get 300ml water extraction liquid, by the technological parameter of table 3, be concentrated into 30ml, the concentrated required time of record is also measured thick polysaccharide, total saponin content, the results are shown in Table 3.Result shows to adopt vacuum-concentrcted equipment in vacuum to be-0.08Mpa, while concentrating under 75 ℃ of conditions of thickening temperature, the consumption time is shorter, and the content of active ingredient is the highest, considers thick polysaccharide, total saponin(e retention rate and production efficiency and is defined as concentrated technological parameter.According to the character of medicine, equipment performance and operating condition, determine that clear cream relative density is 1.25～1.35 (80 ℃), the dry operation of more convenient so clear cream simultaneously.
The research of table 3 concentration technology
6. the research of drying process
For preventing that active ingredient is subject to heat damage for a long time, so the basic methods that adopts the technique of vacuum decompression dry (vacuum is conventional-0.08Mpa) to carry out drying process research, take the content Drying Technology Parameter best as assessment indicator optimizes of thick polysaccharide, total saponin(e.Get 300ml concentrate, by the technological parameter of table, be dried, the dry required time of record is also measured thick polysaccharide, total saponin content, the results are shown in Table 4.Consider thick polysaccharide, total saponin(e retention rate and production efficiency, final definite drying parameter is the vacuum of employing-0.08Mpa, 75 ℃ of drying under reduced pressure.
The research of table 4 drying process
experimental example 2: function assessment research
1 experimental animal and modeling
Kunming kind SPF level mouse, female, by Chongqing City's Experimental Animal Center, provided, weight is 16～18 g, puts into Animal House and adapts to after 3d, weight is that 18～22g starts modeling, intraperitoneal injection 100 mg/kg endoxan, 1 time/d, for three days on end.
2 grouping and dosages
Each experiment is divided into four groups, i.e. negative control group, high dose group, middle dosage group, low dose group (medicine is prepared according to the method for the embodiment of the present invention 1), 10 every group.High dose group is 20 times of one day doses of adult, and middle dosage group is 10 times, and low dose group is 5 times.By per day for adults 6 (2.4g), calculate, adding distil water is deployed into mouse dose 0.2ml/20g, control group administered physiological saline 0.2ml/20g.
3 test methods
3.1 body weight, internal organs/weight ratio are respectively organized mouse dosage administration in accordance with regulations, after gavage 30d, weigh continuously, and cervical vertebra dislocation is put to death, and dissects animal, takes out Thymus and spleen, weigh after blotting bloodstain with filter paper, calculate the ratio between internal organs and weight.
3.2 SPL transformation experiments adopt isotope to mix method.Each organizes mouse dosage administration in accordance with regulations, and after gavage 30 d, mouse boosting cell is respectively organized in preparation continuously, as reacting cells.The preparation of irritation cell: aseptic preparation C
57bL/ 6 mouse boosting cell suspensions, are that the mitomycin C of 25mg/ L is processed after 30min under 37 degrees Celsius by concentration, the centrifugal supernatant that goes, then wash after 2 times, with the RPMI1640 containing 10% calf serum, return to 5 * 10
9/ L, and join in 96 orifice plates.Cell is cultivated after 6d, and every hole adds
3it is 18.5*10 that H-TdR makes final concentration
46h is continued to cultivate in Bq/ hole.Collecting cell is in glass fiber filter paper, and liquid scintillation instrument is counted umber of pulse per minute.
3.3 delayed allergy experiments adopt ear swelling method.Each organizes mouse dosage administration in accordance with regulations, continuously after gavage 30 d, in every mouse hind leg intracutaneous injection in the 1st, the 5th day, 7% DNFB acetone soln 0. 02mL carried out sensitization, in the 8th day of the 1st sensitization, with 1% DNFB acetone soln 0. 02mL, coat auris dextra and attack, left ear coating physiological saline in contrast.Put to death animal next day, with the card punch of diameter 8mm, at the same position of left and right ear, lay round auricle respectively, weigh and the difference of calculating left and right ear weight is carried out one-way analysis of variance.
3.4 antibody producing cells detect and adopt Jerne improvement slide method.Each organizes mouse dosage administration in accordance with regulations, uses SRBC sensitization continuously after gavage 30 d, and after 5d, cervical vertebra dislocation is put to death, and take out liver and make cell suspension, technology cell in 5mlRPMI1640 nutrient solution, and cell concentration is adjusted into 5*10
6individual/ml.By after the culture medium heating for dissolving of top layer, put 50 ℃ of water bath heat preservations, with the Hank's liquid of equivalent pH7.2-7.4,2 times of concentration depending on closing, packing small test tube, every pipe 0.5ml, in pipe, add 50 μ l10%SRBC again, 20 μ l splenocyte suspensions, mix rapidly, be poured on the slide of oneself brush agarose thin layer, do parallel plate, after agar solidifies, slide level is buckled and is placed on horse. put into CO2gas incubator and hatch 1-1.5h and then with the complement of SA buffer solution dilution, join slide frame groove, continue after incubation 1-1.5h counting hemolysis plaque number.
3.5 serum hemolysins are measured the HD50 value method of measuring that adopts.Each organizes mouse dosage administration in accordance with regulations, uses SRBC sensitization continuously after gavage 30 d, after 5 days, extracts eyeball and gets blood in centrifuge tube, places about 1h serum is fully separated out, and the centrifugal 10min of 2000r/min, collects serum.With SA buffer solution, dilute.Serum 1ml after dilution is put in vitro. add successively 10% SRBC 0.5ml, complement 1ml.Separately establish the not control tube of increase serum (replacing with SA buffer solution).Put in 37 ℃ of waters bath with thermostatic control and be incubated after skilful 20min. ice bath cessation reaction, the centrifugal 10min of 2000r/min, get feelings liquid 1ml and add Dou Shi reagent 3ml, get 10% SRBC 0.25ml simultaneously and add Dou Shi reagent to 4mL, fully mix, place after 10min, in 540nm, sentence control tube and do blank, measure and respectively manage OD value respectively, be half hemolytic dose (serum HC
50).
3.6 carbon are cleaned up experiment and are respectively organized mouse dosage administration in accordance with regulations, after continuously after gavage 30 d, the tail vein of mouse is 25% prepared Chinese ink by every 10g weight 0.1ml implantation concentration, injection prepared Chinese ink interval 2,10min, get blood 20 μ L from adjoining veniplex in mouse respectively, joins the Na of 2 ml
2cO
3in (w=0.1%) solution.With Na
2cO
3solution is made blank, with 722 visible spectrophotometers, under 600 nm, measures absorbance.Then mouse is put to death, get liver and spleen, with filter paper, blot organ surface blood stains, weigh, calculate phagocytic index α.
Mouse dosage administration in accordance with regulations is respectively organized in 3.7 macrophage phagocytic chicken red blood cell experiments, continuously after gavage 30 d, to the chicken erythrocyte suspension lml of every mouse peritoneal injection 20%, after 30min, animal is put to death in cervical vertebra dislocation, dissect mouse peritoneal, inject 2ml physiological saline, draw peritoneal fluid and make smear, with Jim Sa working solution dyeing 15～30 min, again with distilled water rinsing, dry, under microscope, count, calculate phagocytic index and phagocytic percentage.
3.8 NK cytoactive detection mtt assay: getting in the Yac-1 of exponential phase cell is target cell.Each is effector cell after organizing mouse spleen lymphocyte and putting in blake bottle adherent 2h and remove adherent macrophage, is calculated as follows kill rate:
Kill rate (%)=1-(OD
(E+ T)-OD
e)/OD
t
OD
(E+ T): refer to NK
+target cell hole OD value.OD
e: refer to same group of effector cell's control wells OD value.OD
t: refer to respective target cell control well OD value.
4 results
4.1 pairs of Mice Body quality, spleen, thymus index impact and negative control group comparison, each administration group Mouse Weight increases difference not remarkable (P > 0. 05); In spleen/weight and thymus gland/weight, high dose group all can significantly improve mouse spleen or thymus index, and difference has conspicuousness (P < 0. 05).(in Table 5).
The impact (n=10, x ± s) of table 5 on Mice Body quality and spleen, thymus gland quality
Note: compare * P < 0.05 with control group
4.2 transform impact and negative control group comparison to mouse spleen lymphocyte, and each dosage group significantly suppresses the mice spleen lymphocytes proliferation of Con A induction, and difference has statistical significance.(?P＜0.?05）。(in Table 6).
Table 6 transforms impact (n=10, x ± s) to mouse spleen lymphocyte
| Group | n | Con A （cpm） ? |
| Negative control group | 10 | 13568±2186 |
| Low dose group | 10 | 9054±1856* |
| Middle dosage group | 10 | 7695±1547* |
| High dose group | 10 | 5385±1225* |
Note: compare * P < 0.05 with control group
4.3 on mouse delayed allergy impact and negative control group comparison, each dosage group mouse right ear is obvious tumefaction after 1% DNFB acetone soln is attacked, weight is higher than left ear, and wherein, the difference of high dose group left and right ear weight is apparently higher than control group, and difference has statistical significance.(?P＜0.?05）。(in Table 7).
Table 7 is on mouse delayed allergy impact (n=10, x ± s)
| Group | n | Left and right ear weight difference (* 10 -3） |
| Negative control group | 10 | 8.76±3.19 |
| Low dose group | 10 | 10.86±2.38 |
| Middle dosage group | 10 | 12.52±2.68* |
| High dose group | 10 | 14.76±3.23* |
Note: compare * P < 0.05 with control group
4.4 on the impact of mouse antibodies cellulation and negative control group comparison, and middle and high dosage group mice spleen antibody forming cell obviously strengthens, and difference has statistical significance.(?P＜0.?05）。(in Table 8).
Table 8 is on mouse antibodies cellulation impact (n=10, x ± s)
| Group | n | Antibody forming cell |
| Negative control group | 10 | 5.02±0.13 |
| Low dose group | 10 | 5.21±0.18 |
| Middle dosage group | 10 | 5.95±0.21* |
| High dose group | 10 | 6.28±0.19* |
Note: compare * P < 0.05 with control group
4.5 on mice serum hemolysin impact and negative control group comparison, each administration group serum HC that all can raise
50, significant difference (P < 0. 05).(in Table 9).
Table 9 is on mice serum hemolysin impact (n=10, x ± s)
| Group | n | Serum HC 50 |
| Negative control group | 10 | 123 |
| Low dose group | 10 | 228* |
| Middle dosage group | 10 | 276* |
| High dose group | 10 | 305* |
Note: compare * P < 0.05 with control group
4.6 clean up and macrophage phagocytic function impact and negative control group comparison mouse carbon, can obviously improve the carbon of mouse and clean up ability during with high dose gavage, and difference has statistical significance (P < 0. 05).Middle high dose group can obviously improve the ability that mouse macrophage is engulfed chicken red blood cell, significant difference (P < 0. 05).(in Table 10).
Table 10 is cleaned up mouse carbon and macrophage phagocytic function impact (n=10, x ± s)
| Group | n | Carbon is cleaned up phagocytic index | Phagocytic percentage/% | Phagocytic index |
| Negative control group | 10 | 4.43±0.76 | 34.33±10.83 | 0.45±0.14 |
| Low dose group | 10 | 5.02±0.86 | 35.28±14.20 | 0.47±0.12 |
| Middle dosage group | 10 | 4.89±0.73 | 53.58±15.36* | 0.61±0.17* |
| High dose group | 10 | 5.71±0.58* | 57.89±16.22* | 0.66±0.15* |
Note: compare * P < 0.05 with control group
The 4.7 pairs of NK cells in mice activity influences and negative control group comparison, each administration group all can improve NK cells in mice activity, significant difference (P < 0. 05).(in Table 11).
Table 11 pair NK cells in mice activity influence (n=10, x ± s)
| Group | n | Kill rate (%) |
| Negative control group | 10 | 43.2±4.5 |
| Low dose group | 10 | 61.7±6.5* |
| Middle dosage group | 10 | 69.5±3.3* |
| High dose group | 10 | 71.4±7.8* |
Note: compare * P < 0.05 with control group
5 conclusions
It is all positive that capsule various functions of the present invention is learned experimental result, proves that patent of the present invention capsule of the present invention has enhancing immunity function.
experimental example 3.Acute toxicity test
(1) experiment material and condition
1, animal: Kunming mouse, body weight 20 ± 2g, male and female half and half, are provided by Military Medical Univ No.3, P.L.A's Animal Experimental Study center.
2, health products: make capsule according to method described in the embodiment of the present invention 1, the used time is poured out content, are made into required confession reagent liquid with distilled water.
3, solvent: distilled water.
4, condition: temperature 22-25 ℃, humidity is 50%-60%.
(2) experimental technique
1, animal grouping: establish four dosage groups, every group of 10 mouse, male and female half and half.
2, dosage and give mode: experiment establishes 1000,2150,4640,4 dosage groups of 10000mg/ kg/d.Adopt the administration of per os gavage mode, every day, gavage was 1 time, Continuous Observation 7 days,
(3) experimental result: experimental result is in Table 12.
Table 12 capsule of the present invention is to chmice acute toxicity test result
As shown in Table 4, in the observation period, have no experiment mice and occur poisoning symptom and death condition, the LD of capsule of the present invention to mouse
50value is greater than 10000mg/kg, and according to acute toxicity grading criteria, its acute toxicity belongs to actual non-toxic type.
experimental example 4: Synergistic experiment
Applicant has carried out Synergistic experimental study to the present invention, by experiment data declaration beneficial effect of the present invention.
1 experimental animal and modeling
Kunming kind SPF level mouse, female, by Chongqing City's Experimental Animal Center, provided, weight is 16～18 g, puts into Animal House and adapts to after 3d, weight is that 18～22g starts modeling, intraperitoneal injection 100 mg/kg endoxan, 1 time/d, for three days on end.
2 medicines, grouping and dosage
2.1 medicine
A extract group:
Prescription: HERBA DENDROBII 670g
Technique: HERBA DENDROBII adds suitable quantity of water and soaks after 0.5 hour, decoct twice, add 6 times of water gagings decocts 1 hour at every turn, filter merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, in the ratio of the extract obtained 5000ml of being dissolved in water, make solution, obtain.
B extract group:
Prescription: coix seed 1000g+ American Ginseng 330g
Technique: the 222g that coix seed is got recipe quantity pulverizes, and crosses 120 mesh sieves, gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, the West are participated in suitable quantity of water and are soaked after 0.5 hour, decoct twice, add 6 times of water gagings at every turn and decoct 1 hour, filter merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38, add coix seed fine powder to mix ,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, ratio in the extract obtained 5000ml of being dissolved in water is made solution, obtains.
C extract group:
Prescription: HERBA DENDROBII 670g+coix seed 1000g+ American Ginseng 330g
Technique: coix seed is got 220g and pulverized, crosses 120 mesh sieves, obtains 121 ℃ of fine powders, 0.1MPa moist heat sterilization 30 minutes, standby, all the other medicinal materials add the appropriate immersion of water 0.5 hour, decoct twice, add 6 times of water gagings at every turn and decoct 1 hour, filter merging filtrate,-0.08Mpa, 75 ℃ are concentrated into the thick paste that relative density is 1.35～1.38 (80 ℃), add coix seed fine powder to mix ,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, ratio in the extract obtained 5000ml of being dissolved in water is made solution, obtains.
2.2 grouping and dosage
Each experiment is divided into four groups, i.e. negative control group, A extract group (HERBA DENDROBII), B extract group (coix seed+American Ginseng), C extract group (HERBA DENDROBII+coix seed+American Ginseng), 10 every group.Pharmaceutical composition amount is 0.2ml/20g, control group administered physiological saline 0.2ml/20g.
3 test methods
3.1 SPL transformation experiments adopt isotope to mix method.Each organizes mouse dosage administration in accordance with regulations, and after gavage 30 d, mouse boosting cell is respectively organized in preparation continuously, as reacting cells.The preparation of irritation cell: aseptic preparation C
57bL/ 6 mouse boosting cell suspensions, are that the mitomycin C of 25mg/ L is processed after 30min under 37 degrees Celsius by concentration, the centrifugal supernatant that goes, then wash after 2 times, with the RPMI1640 containing 10% calf serum, return to 5 * 10
9/ L, and join in 96 orifice plates.Cell is cultivated after 6d, and every hole adds
3it is 18.5*10 that H-TdR makes final concentration
46h is continued to cultivate in Bq/ hole.Collecting cell is in glass fiber filter paper, and liquid scintillation instrument is counted umber of pulse per minute.
Mouse dosage administration in accordance with regulations is respectively organized in 3.2 macrophage phagocytic chicken red blood cell experiments, continuously after gavage 30d, to the chicken erythrocyte suspension lml of every mouse peritoneal injection 20%, after 30min, animal is put to death in cervical vertebra dislocation, dissect mouse peritoneal, inject 2ml physiological saline, draw peritoneal fluid and make smear, with Jim Sa working solution dyeing 15～30 min, again with distilled water rinsing, dry, under microscope, count, calculate phagocytic index and phagocytic percentage.
3.3 NK cytoactive detection mtt assay: getting in the Yac-1 of exponential phase cell is target cell.Each is effector cell after organizing mouse spleen lymphocyte and putting in blake bottle adherent 2h and remove adherent macrophage, is calculated as follows kill rate:
Kill rate (%)=1-(OD
(E+ T)-OD
e)/OD
t
OD
(E+ T): refer to NK
+target cell hole OD value.OD
e: refer to same group of effector cell's control wells OD value.OD
t: refer to respective target cell control well OD value.
4 results
4.1 transform impact and negative control group comparison to mouse spleen lymphocyte, and each dosage group significantly suppresses the mice spleen lymphocytes proliferation of Con A induction, and difference has statistical significance.(?P＜0.?05）。Adopt Jin's formula to evaluate drug combination to the impact of mice spleen lymphocytes proliferation inhibiting rate, result synergy index q value is that 1.26(is in Table 13).
Table 13 transforms impact (n=10, x ± s) to mouse spleen lymphocyte
Note: compare * P < 0.05 with control group
4.2 on macrophage phagocytosis of mice impact and negative control group comparison, and each group all can obviously improve the ability that mouse macrophage is engulfed chicken red blood cell, significant difference (P < 0. 05).Adopting Jin's formula to evaluate drug combination affects macrophage phagocytosis of mice, and result synergy index q value is that 1.21(is in Table 14).
Table 14 is on macrophage phagocytosis of mice impact (n=10, x ± s)
Note: compare * P < 0.05 with control group
The 4.3 pairs of NK cells in mice activity influences and negative control group comparison, each administration group all can improve NK cells in mice activity, significant difference (P < 0. 05).Adopt Jin's formula to evaluate drug combination to NK cells in mice activity influence, result synergy index q value is that 1.23(is in Table 15).
Table 15 pair NK cells in mice activity influence (n=10, x ± s)
Note: compare * P < 0.05 with control group
5 conclusions
Three groups of experiment synergy indexes are respectively 1.26,1.21,1.23, due to q>1.15, show drug combination after effect strengthen, illustrate that HERBA DENDROBII and coix seed, American Ginseng share and have obvious synergistic effect.
Compared with prior art, it is that primary raw material is made health food that the present invention selects HERBA DENDROBII, coix seed, American Ginseng, have and build up health, strengthen the function of immunity of organisms, cellular immunity, humoral immunity, macrophage phagocytic function and NK cytoactive are all had to certain promotion, strengthening by means of tonics, the good merchantable brand of health care.And safety is little, has no side effect; More amazing, can also to produce Synergistic after square Chinese traditional medicine coupling effect, has reached goal of the invention.
The specific embodiment
Embodiment 1:
Prescription: HERBA DENDROBII 670g, coix seed 1000g, American Ginseng 330g.
Preparation method: coix seed is got 220g and pulverized, crosses 120 mesh sieves, obtains 121 ℃ of fine powders, 0.1MPa moist heat sterilization 30 minutes, standby, all the other medicinal materials add the appropriate immersion of water 0.5 hour, decoct twice, add 6 times of water gagings at every turn and decoct 1 hour, filter merging filtrate,-0.08Mpa, 75 ℃ are concentrated into the thick paste that relative density is 1.35～1.38 (80 ℃), add coix seed fine powder to mix ,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, cross 40 mesh sieves, pack capsule into No. 1, make 1000, obtain.
Specification: 0.4g/ grain
Usage and dosage: oral, 2 times on the one, each 3.
Claims (7)
1. strengthen a health food for immunity, by active drug composition and auxiliary material, made, it is characterized in that: according to listed as parts by weight, the active drug composition of use is: HERBA DENDROBII 500-800 part, coix seed 800-1200 part, American Ginseng 200-500 part.
2. strengthen as claimed in claim 1 the health food of immunity, it is characterized in that: according to listed as parts by weight, the active drug composition of use is: 670 parts of HERBA DENDROBIIs, 1000 parts of coix seeds, 330 parts of American Ginsengs.
3. described in preparation claim 1 or 2, strengthen the method for the health food of immunity, it is characterized in that: get HERBA DENDROBII, coix seed and American Ginseng, combine with pharmaceutic adjuvant, and make corresponding preparation according to conventional preparation method.
4. strengthen according to claim 3 the preparation method of the health food of immunity, it is characterized in that: coix seed is got part and pulverized, cross 80-200 mesh sieve, gained fine powder moist heat sterilization 20-50 minute, standby; Residue coix seed, American Ginseng and HERBA DENDROBII were soaked after 0.3-1 hour, decoct 1-3 time, add 4-8 times of water gaging to decoct 0.5-3 hour at every turn, filter, merging filtrate, reduced pressure concentration becomes thick paste, add coix seed fine powder and pharmaceutic adjuvant to mix, drying under reduced pressure, pulverizes, then and makes corresponding preparation according to conventional preparation method.
5. strengthen according to claim 4 the preparation method of the health food of immunity, it is characterized in that: coix seed is got 22% of recipe quantity and pulverized, and crosses 120 mesh sieves, gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, American Ginseng and HERBA DENDROBII add suitable quantity of water and soak after 0.5 hour, decoct twice, add 6 times of water gagings decocts 1 hour at every turn, filter merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38, add coix seed fine powder and pharmaceutic adjuvant to mix,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, then and make corresponding preparation according to conventional preparation method.
6. according to the preparation method who strengthens the health food of immunity described in claim 3 or 4, it is characterized in that: described preparation is capsule.
7. strengthen according to claim 5 the preparation method of the health food of immunity, it is characterized in that: described capsule is so prepared: coix seed is got 22% of recipe quantity and pulverized, and crosses 120 mesh sieves, gained fine powder moist heat sterilization 20-50 minute is standby; Residue coix seed, American Ginseng and HERBA DENDROBII add suitable quantity of water and soak after 0.5 hour, decoct twice, add 6 times of water gagings at every turn and decoct 1 hour, filter, merging filtrate, the thick paste that when-0.08Mpa, 75 ℃ are concentrated into 80 ℃, relative density is 1.35～1.38, adds coix seed fine powder to mix ,-0.08Mpa, 75 ℃ of drying under reduced pressure, pulverize, cross 40 mesh sieves, incapsulate, obtain.
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