CN104783182B - A kind of composition for improving immunity and preparation method thereof - Google Patents
A kind of composition for improving immunity and preparation method thereof Download PDFInfo
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Classifications
-
- 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
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicinal Preparation (AREA)
Abstract
The present invention relates to health product technology field, more particularly to a kind of composition for improving immunity and preparation method thereof.The present invention can significantly improve the immunity of human body after being compounded apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder.Experiment shows, after composition provided by the invention is to mouse stomach, the phagocytic index of mouse monokaryon macrophage can be improved to 1.43 ± 0.23 from 1.19 ± 0.17, there is significant difference, and by the activity of NK cells in mice from 26.84% ± 13.52% raising to 28.96% ± 13.65%.And the survey showed that for taking through 300 hypoimmunity personages, it is believed that effective people accounts for 99%, is significantly higher than control group.
Description
Technical field
The present invention relates to health product technology field, more particularly to a kind of composition for improving immunity and preparation method thereof.
Background technology
Immunity is the defense mechanism of human body itself, is that human bioequivalence and any foreign matter of the external intrusion of elimination are (viral, thin
Bacterium etc.), processing aging, damage, death, the own cells of denaturation, and identification and processing vivo mutations cell and virus are infected
The ability of cell.Immunity can be divided into nospecific immunity and specific immunity：1st, congenital immunity, it is that people has once being born
's；2nd, acquired immunity, obtained naturally in life process after people is born, or the method manually aided in is passive
Obtain.
Some special cells of body immune system can be by the thin of the bacterium in invasion body, virus and internal aging death
Born of the same parents, the cell being mutated and the material for causing allergy, are wholly swallowed and are eliminated, so as to maintain vivo environment
It is stable, keep body health.But body's immunity began to subtract late at 30 years old or so, and this change is quietly, slowly, persistently
Ground is carried out.When immune function of human body is lacked of proper care, or immune system is unsound, immune system can not normally play protective effect.
In the case, the infection such as bacterium, virus, fungi is easily caused, therefore most directly performance is exactly easy to be raw to hypoimmunity
Disease.It is often ill, the consumption of body is aggravated, so typically having a delicate constitution, being malnutritive, be One's spirits are drooping, fatigue and weak, food
It is intended to reduce, the performance such as sleep-disorder；Furthermore, it is necessary to be lot more time to recover, and usually recurrent exerbation.It can if things go on like this lead
Dedicate one's life to body and intelligence development is bad, also easily induce major disease.The underlying causes for causing above-mentioned phenomenon to occur are hypoimmunities
Or immunity is unsound.
Modern society, environmental problem is more and more prominent, and people's operating pressure is big, does not get enough athletic exercise, and then causes being immunized for people
Power worse and worse, improves the hot issue that immunity is referred to as modern's care, substantial amounts of to have the health care for improving immunity function
Product arise at the historic moment.At present, common health products species mainly has：Nutrition sexual health using vitamin and mineral as main component
Product, using natural or rare species as raw material extract the health products of effective nutritional ingredient, medical value is crossed with rare Chinese medicine
Animals and plants are the tonic type health products of primary raw material, the health products made of effective component extracting or with the beginning of animal from marine organisms
Breast is health products made of raw material.But it is existing improve in immune health products, can not have mostly clearly improve it is immune
The effect of power, majority of populations think to take DeGrain after the health products for improving immunity.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of composition for improving immunity and its preparation
Method, the composition provided by the invention for improving immunity can have the function that significantly clearly to improve immunity.
The invention provides a kind of composition for improving immunity, including apple stem cell freeze-dried powder and Paecilomyces hepiali chen
Bacterium powder.
Wherein, apple stem cell not only can quickly activate the stem cell of dormancy in body, but also can active remediation by
The hyperplasia of the stem cell of damage, stimulating human horn cell and fibroblast, increase skin elasticity, there is anti-aging and beauty to give birth to
The special effect of hair.Apple stem cell has the characteristic of plant stem cell：Powerful anti-oxidant and activity of fighting against senium, can significantly improve people
Body immunity and anti-cancer ability.
Peacilomyce hepiahi bacterium powder is cordyceps, tool physiologically active moral nucleoside compound, peptides, PEARLITOL 25C, more
Sugar, Sudismase, ergosterol and abundant amino acid and vitamin needed by human, trace element, such as B1, B2, B12,
E, K, phosphorus, iron etc..Its main component and core effect include：1. polysaccharide：More than or equal to 8.0%, start macrophage, enhancing is exempted from
Epidemic disease and hematopoietic potential；2. adenosine：More than or equal to 0.3%, pressure of releiving and intense strain, improving sleep, enhancing is physical, endurance,
It is antifatigue.Man is not easy tired；Ms can be invigorated blood circulation, and lift qi and blood, and antenatal postpartum recuperating more preferably, is had rosy cheeks, the warm foot temperature of hand,
Beauty is healthy and strong.And it can effectively maintain healthy cholesterol；3. cordycepic acid (PEARLITOL 25C)：More than or equal to 14.0%, happy organ, only
Cloperastine is breathed heavily, and reduces respiratory problems, strengthens adrenaline；4. ergosterol：More than or equal to 0.65%, serum cholesterol can be reduced
And content of triglyceride, strengthen heart and cardiovascular function, suppress harmful cellular growths；5. Sudismase (SOD)：On a small quantity,
It is anti-oxidant, strengthen brain oxygen supply amount, lift mental and memory, pressure of releiving.Body or function aging can be resisted, make the heart, lung,
Kidney and brain are kept fit；SOD contents increase for removing oxygen radical caused by organism metabolism, or entered in vivo by the external world
Free radical caused by some drugses or toxicant metabolism and free radical can play a positive role as caused by radiation.6. albumen
Matter：31.0%~35.0%, it is to form human body cell and the basic substance of vital movement.
The present invention can significantly improve human body after being compounded apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder
Immunity.Experiment shows, being capable of gulping down mouse monokaryon-macrophage after composition provided by the invention is to mouse stomach
Bite index to improve to 1.43 ± 0.23 from 1.19 ± 0.17, there is significant difference, and by the activity of NK cells in mice oneself
26.84% ± 13.52% improves to 28.96% ± 13.65%.And take investigation result through 300 hypoimmunity personages
Show, it is believed that effective people accounts for 99%, is significantly higher than control group.
In an embodiment of the present invention, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder for (2~
5)：(4~8).
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is 3.5:6.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is 1:2.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is 1:4.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is 5:4.
In certain embodiments, the mass ratio of apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is 5:8.
The apple stem cell freeze-dried powder that the present invention uses can be bought for market also to be prepared using method provided by the invention,
It is implemented all within protection scope of the present invention.Preferably, apple stem cell freeze-dried powder is with method system provided by the invention
, this method using apple new life branch as raw material, after induced synthesis callus Multiplying culture and expand culture obtain apple
Stem cell, then be made after freezing.The preparation that the present invention provides composition, the freeze-dried powder are directly used in after the apple stem cell is lyophilized
Middle flavones and polyphenol content enrich.
The preparation method of apple stem cell freeze-dried powder comprises the following steps：
Step 1：The sterilization of apple new life branch, after removing xylem and marrow, inducing culture is inoculated in, Fiber differentiation obtains
Cambial cell；
Step 2：The cambial cell accesses proliferated culture medium, shaking table culture obtains unicellular through squamous subculture；
Step 3：The unicellular proliferated culture medium that is inoculated in is cultivated through expanding, obtains apple stem cell；
Step 4：It will freeze, crush after the apple stem cell multigelation, obtain apple stem cell freeze-dried powder；
The inducing culture is the MS solid mediums containing NAA, BA and caseinhydrolysate；
The proliferated culture medium is the MS fluid nutrient mediums containing 2,4-D, BA, caseinhydrolysate and activated carbon.
In an embodiment of the present invention, sterilization comprises the following steps：
Step 1：After apple new life branch is rinsed with water, the L-AA solution using concentration as 10~1000mg/L soaks
Bubble；
Step 2：With aseptic water washing after using volume fraction as 60%~95% ethanol water immersion；
Step 3：With aseptic water washing after using volume fraction as 10%~100% aqueous hydrogen peroxide solution immersion.
Preferably, the length of apple new life branch is 10cm.
Preferably, the concentration of L-AA is 120mg/L.
Preferably, the time soaked described in step 1 is 1min.
Preferably, the volume fraction of ethanol is 75% in ethanol water.
Preferably, the time soaked described in step 2 is 1min.
Preferably, the volume of sterilized water is 2L in step 2.
Preferably, the volume fraction of hydrogen peroxide is 30% in aqueous hydrogen peroxide solution.
Preferably, the time soaked described in step 3 is 20min.
Preferably, the time rinsed described in step 3 is 10min.
Carrying out sterilization to apple new life branch with method for disinfection provided by the invention can sufficiently kill except apple innovation
Entrained fungi, bacterium or virus on bar.To ensure that apple new life branch is not disturbed in incubation, so as to ensure
Cell on apple new life branch more quickly dedifferentes.And test and show, method provided by the invention can be effectively by apple
Fruit new life branch cell de-differentiation, through inducing and expanding culture, substantial amounts of apple stem cell can be obtained within 48 days or so.
In embodiments of the present invention, the mass ratio of NAA, BA and caseinhydrolysate is 0.5：2：1.
Preferably, NAA concentration is 0.5mg/L in inducing culture.
Preferably, BA concentration is 2.0mg/L in inducing culture.
Preferably, the concentration of caseinhydrolysate is 1.0mg/L in inducing culture.
Preferably, inducing culture is solid medium, wherein the mass fraction of agar is 1%.
Preferably, the pH value of inducing culture is 6.0.
In an embodiment of the present invention, the inoculation described in step 1 is that the apple innovation of xylem and marrow is removed per g
Bar is inoculated in 10cm2Inducing culture.
In certain embodiments, Fiber differentiation is light culture；Temperature is 25 DEG C；Time is 10 days.
After Fiber differentiation, Cambium tissue is the tabular tissue of homogeneous growth, and its hetero-organization is then to be irregular
Assemble growth-gen.
In certain embodiments, squamous subculture is light culture；Temperature is 25 DEG C；Time is 10 days.
In an embodiment of the present invention, squamous subculture is inoculated in 10cm using culture medium is run into per g cambial cells2's
Inducing culture.
Through the squamous subculture of 10 days, cambial cell formed callus, per 10cm2Inducing culture on there are about callus
Organize 20.8g.
In some embodiments, in proliferated culture medium, 2,4-D, BA, the mass ratio of caseinhydrolysate and activated carbon be 2:1:1:
1。
Preferably, 2,4-D concentration is 2.0mg/L in proliferated culture medium.
Preferably, BA concentration is 1.0mg/L in proliferated culture medium.
Preferably, the concentration of caseinhydrolysate is 1.0mg/L in proliferated culture medium.
Preferably, the concentration of activated carbon is 1.0mg/L in proliferated culture medium.
Preferably, the pH value of proliferated culture medium is 6.0.
In an embodiment of the present invention, per g callus access 20mL proliferated culture mediums.
Preferably, sterilized a diameter of 0.5cm bead is also added during shaking table culture, in proliferated culture medium.
Preferably, the addition of bead is 100g/L proliferated culture mediums.
In embodiment, culture uses triangular flask, and 200mL proliferated culture mediums are added in 1000mL triangular flasks.
In an embodiment of the present invention, the condition of shaking table culture is per hour with 20W incandescent lamp irradiation 10min.
In certain embodiments, the temperature of shaking table culture is 25 DEG C；Rotating speed is 120 revs/min；Time is 14 days.
Preferably, fresh proliferated culture medium is added after 7 days in shaking table culture.
Preferably, the volume for adding fresh proliferated culture medium is 2 times of former proliferated culture medium.
After adding fresh proliferated culture medium, wherein the density of cell is 1 × 105/ml。
Shaking table culture is carried out using method provided by the invention, callus turns into single cell suspension, cell proliferation rate
Comparatively fast, filtered with the stainless steel mesh of 100 mesh, removing cell mass, bead and residue, acquisition cell density about 0.5~1 ×
106/ml。
Preferably, the density being inoculated with step 3 is 1 × 104/ml。
In certain embodiments, expanding culture is specially：5L proliferated culture mediums, training are added in 20L bioreactor
It is 1 × 10 that proliferated culture medium to cell concentration is added after supporting 7 days5/ ml, it is further cultured for 7 days.
The preparation method of the composition provided by the invention for improving immunity is, by apple stem cell freeze-dried powder and bat moth
Paecilomyces varioti bacterium powder mixes, and the composition for improving immunity is made.
Present invention also offers a kind of health products for improving immunity, including the combination provided by the invention for improving immunity
Thing.
Raising immunity health products provided by the invention also include acceptable auxiliary material in health products.
The formulation of the health products provided by the invention for improving immunity is tablet or capsule.
The capsule of immunity is improved present invention also offers a kind of, including composition provided by the invention, lubricant and is filled out
Fill agent.
Preferably, lubricant is magnesium stearate；Filler is starch.
In an embodiment of the present invention, composition, lubricant and filling provided by the invention in the capsule of immunity are improved
The mass ratio of agent is (60~130)：0.5：(0~30).
In certain embodiments, composition provided by the invention in the capsule of immunity, lubricant and filler are improved
Mass ratio is 95：15：0.5.
In certain embodiments, the preparation method of the capsule of immunity is improved, for composition provided by the invention is crushed
Afterwards, with filler and mix lubricant, capsule is filled, the capsule for improving immunity is made.
The present invention can significantly improve human body after being compounded apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder
Immunity.Experiment shows, being capable of gulping down mouse monokaryon-macrophage after composition provided by the invention is to mouse stomach
Bite index to improve to 1.43 ± 0.23 from 1.19 ± 0.17, there is significant difference, and by the activity of NK cells in mice oneself
26.84% ± 13.52% improves to 28.96% ± 13.65%.And take investigation result through 300 hypoimmunity personages
Show, it is believed that effective people accounts for 99%, is significantly higher than control group.
Embodiment
The invention provides a kind of composition for improving immunity and preparation method thereof, those skilled in the art can use for reference
Present disclosure, it is suitably modified technological parameter realization.In particular, all similar replacements and change are to this area skill
It is it will be apparent that they are considered as being included in the present invention for art personnel.The present invention method and application by compared with
Good embodiment is described, related personnel substantially can not depart from present invention, in spirit and scope to methods herein
It is modified or suitably changes with combining with application, realizes and using the technology of the present invention.
The instrument that the present invention uses is all common commercially available product, can all be bought in market.
With reference to embodiment, the present invention is expanded on further：
The preparation of the apple stem cell freeze-dried powder of embodiment 1
1st, the formula of culture medium：
1.1 inducing culture：MS solid medium+NAA0.5mg/L+BA 2mg/L+CH 1mg/L, pH value 6.0；
1.2 proliferated culture medium：MS fluid nutrient medium+2,4-D 2.0mg/L+BA 1.0mg/L+CH1.0mg/L+AC
1.0mg/L, pH value 6.0；
2nd, the sterilization of apple branch
The apple tree without fertilizer and pesticide pollution, green planting is selected, 10 10cm length are selected according to certain judgment criteria
Newborn branch；Newborn branch surface attachments are washed with sterilized water 2L；Newborn branch is put into 120mg/L L-AAs
Immersion 1 minute；Newborn branch is put into 75% ethanol solution and sterilized 1 minute, it is clean with 2L aseptic water washings；Place into
Sterilized 20 minutes in 30% hydrogenperoxide steam generator, and with aseptic water washing 10 minutes.
3rd, the induction of Cambium tissue
The apple new life branch of sterilizing is peeled off into the tissue such as forming layer, bast, cortex and epidermis with vaccinating lancet, abandons wood
Matter portion and marrow；The tissue of acquisition is seeded in fresh solid medium, inoculation 10cm is organized with every 1g2Solid medium
Density is inoculated with, and is continuously cultivated 10 days for 25 DEG C in controlled darkroom, is obtained Cambium tissue.
4th, squamous subculture Cambium tissue
The Cambium tissue of regeneration and its hetero-organization are separated；Inoculation 10cm is organized with every 1g2Solid medium it is close
Degree, the Cambium tissue of regeneration is seeded in fresh solid medium, is continuously cultivated 10 days for 25 DEG C in controlled darkroom,
Obtain callus.
5th, unicellular culture
200ml fluid nutrient mediums, the 20g of sterilization treatment, diameter 0.5cm bead are added in 1000ml triangular flasks；
10g callus is transferred in triangular flask；Triangular flask is placed in 25 DEG C, cultivated on 120 revs/min of shaking table, per small
When irradiated 10 minutes with 20W incandescent lamp；7th day, it was wherein cell to add the fresh fluid nutrient mediums of 400ml to cell density
Density be 1 × 105/ ml, then proceed to culture 7 days；Filtered after culture with the stainless steel mesh of 100 mesh, remove cell mass, glass
Glass pearl and residue, obtain single cell suspension,
6th, amplification culture
Single cell suspension is transferred in 20L bioreactors and carries out large-scale culture, accommodates 5L's in bioreactor
Proliferated culture medium.When cultivating the 7th day, 10L fresh liquid culture medium is added, cell suspension is obtained after continuing culture.
7th, the preparation of freeze-dried powder
Apple stem cell suspension is filtered with 1um filters, removes culture medium, and the viscose shape cell for harvesting retention takes out；Will
Cell is put into pre-freeze 3 hours in -30 DEG C of refrigerator；It is put into super low temperature quick frozen 2 hours in liquid nitrogen container；Take out after thawing 30 minutes,
Place into it is quick-frozen in pre-freeze and liquid nitrogen container in -30 DEG C of refrigerators, 2 times successively；Cell is taken out, maintains 10 minutes, enters low at room temperature
In warm frozen vacuum dryer, after vacuum freeze drying, 60 mesh freeze-dried powders are ground at a high speed.
Embodiment 2 improves the preparation of the capsule of immunity
First 20g apple stem cells freeze-dried powder, 40g peacilomyce hepiahi bacterium powder are crushed and sieved, after by powder
Broken screened material is well mixed with 0.5g magnesium stearates in mixer, is filled, polished with Autocapsulefillingmachine；
Finally it is sub-packed in the hard shell capsules of No. 1, that is, is made with the capsule for improving immunity.
Embodiment 3 improves the preparation of the capsule of immunity
First 50g apple stem cells freeze-dried powder, 80g peacilomyce hepiahi bacterium powder are crushed and sieved, after by powder
Broken screened material is well mixed with 30g starch, 0.5g magnesium stearates in mixer, is filled out with Autocapsulefillingmachine
Fill, polish；Finally dispense in the hard shell capsules of No. 0, that is, be made with the capsule for improving immunity.
Embodiment 4 improves the preparation of the capsule of immunity
First 20g apple stem cells freeze-dried powder, 80g peacilomyce hepiahi bacterium powder are crushed and sieved, after by powder
Broken screened material is well mixed with 10g starch, 0.5g magnesium stearates in mixer, is filled out with Autocapsulefillingmachine
Fill, polish；Finally it is sub-packed in the hard shell capsules of No. 2, that is, is made with the capsule for improving immunity.
Embodiment 5 improves the preparation of the capsule of immunity
First 35g apple stem cells freeze-dried powder, 60g peacilomyce hepiahi bacterium powder are crushed and sieved, after by powder
Broken screened material is well mixed with 15g starch, 0.5g magnesium stearates in mixer, is filled out with Autocapsulefillingmachine
Fill, polish；Finally it is sub-packed in the hard shell capsules of No. 1, that is, is made with the capsule for improving immunity.
Embodiment 6 improves the preparation of the capsule of immunity
First 50g apple stem cells freeze-dried powder, 40g peacilomyce hepiahi bacterium powder are crushed and sieved, after by powder
Broken screened material is well mixed with 20g starch, 0.5g magnesium stearates in mixer, is filled out with Autocapsulefillingmachine
Fill, polish；Finally it is sub-packed in the hard shell capsules of No. 0, that is, is made with the capsule for improving immunity.
The preparation of the control sample 1 of comparative example 1
First 95g peacilomyce hepiahi bacterium powder is crushed and sieved, after the material to have pulverized and sieved and 15g are formed sediment
Powder, 0.5g magnesium stearates are well mixed in mixer, are filled, polished with Autocapsulefillingmachine；Finally it is sub-packed in 1
Number hard shell capsules in, that is, be made with improve immunity capsule.
The preparation of the control sample 12 of comparative example 2
First 95g apple stem cell freeze-dried powders are crushed and sieved, after the material to have pulverized and sieved and 15g are formed sediment
Powder, 0.5g magnesium stearates are well mixed in mixer, are filled, polished with Autocapsulefillingmachine；Finally it is sub-packed in 1
Number hard shell capsules in, that is, be made with improve immunity capsule.
Influence of the antifatigue capsule of embodiment 7 to mouse physical signs
According to the Ministry of Public Health《Health food is examined and technology value disciplines》(2003), 120 kunming mices are pressed into body weight
It is randomly divided into 4 groups, every group 30, continuous 30 days.
1. experimental group：The capsule 's content of the offer of embodiment 5 is provided, is 1g/mL by contents melting to concentration, gavage is given
Mouse is given, gavage amount is 10mL/kg.
2. control group 1：The capsule 's content of the offer of comparative example 1 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
3. control group 2：The capsule 's content of the offer of comparative example 2 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
4. blank control group：Deionized water gavage gives mouse, and gavage amount is 10mL/kg.
10 mouse are randomly selected from every group, weigh body weight respectively, and spleen, thymus gland are taken out after carrying out dislocation execution,
Place's spleen, thymic weight are weighed respectively.Calculate internal organs/weight ratio (mg/10g expressions).As a result it is as shown in table 1：
The mouse physical signs of table 1
| Group | Number of animals (only) | Spleen/weight ratio (mg/10g) | Thymus gland/body mass ratio (mg/10g) |
| Experimental group | 10 | 41.45±2.87 | 18.79±1.51 |
| Control group 1 | 10 | 41.36±3.02 | 18.65±1.86 |
| Control group 2 | 10 | 42.09±2.44 | 19.03±1.22 |
| Blank control group | 10 | 42.27±2.63 | 18.21±2.01 |
* the P compared with control group is represented<0.05, there is significant difference
It was found from data, compared with control group, the capsule that embodiment 5 provides is to mice organs/equal nothing of weight ratio
Significantly affect, show that capsule provided by the invention has no toxic side effect to mouse.The capsule that other embodiments of the invention are provided
Testing result is similarly.
Influence of the antifatigue capsule of embodiment 8 to mouse monokaryon-macrophage phagocytic function
According to the Ministry of Public Health《Health food is examined and technology value disciplines》(2003), 120 kunming mices are pressed into body weight
It is randomly divided into 4 groups, every group 30, continuous 30 days.
1. experimental group：The capsule 's content of the offer of embodiment 5 is provided, is 1g/mL by contents melting to concentration, gavage is given
Mouse is given, gavage amount is 10mL/kg.
2. control group 1：The capsule 's content of the offer of comparative example 1 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
3. control group 2：The capsule 's content of the offer of comparative example 2 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
4. blank control group：Deionized water gavage gives mouse, and gavage amount is 10mL/kg.
10 mouse are randomly selected from every group, continuous gavage is after 30 days, every chicken red blood cell of mouse peritoneal injection 20%
Suspension 1mL, 30min；Afterwards, cervical dislocation is put to death, and is fixed on mouse plate, and abdominal skin is cut off in center, is given birth to through Intraperitoneal injection
Salt solution 2mL is managed, rotates mouse plate 1min, suctions out abdominal cavity washing lotion 1mL, mean droplet is on 2 slides, putting 37 DEG C of 30min of wet box；Take
Go out rinse, dry in physiological saline, be fixed, Giemsa PBS dyeing 3min, distilled water rinsing is dried, microscopy.Calculate phagocytosis
Percentage and phagocytic index.As a result such as table 2：
2 mouse monokaryons of table-macrophage phagocytic function
| Group | Number of animals (only) | Phagocytic percentage (%) | Phagocytic index |
| Blank control group | 10 | 32.75±4.20 | 1.19±0.17 |
| Control group 1 | 10 | 35.79±3.85△ | 1.23±0.13*△ |
| Control group 2 | 10 | 36.44±4.62△ | 1.25±0.18*△ |
| Test group | 10 | 41.67±6.83* | 1.43±0.23** |
* represent compared with control group, P<0.05, there is significant difference.
* expressions are compared with control group, P<0.01, there is pole significant difference.
△ represents test group compared with control group, P<0.05, there is significant difference.
As a result show, the phagocytic percentage come detected by blank control group is 32.75 ± 4.20, the phagocytosis of control group 1
Percentage is 35.79 ± 3.85,36.44 ± 4.62, and there was no significant difference compared with blank control group, and test group is 41.67
± 6.83, all there is significant difference P compared with control group<0.05.Compared with control group 1~2, phagocytic percentage is test group
P<0.05, there is significant difference.
The phagocytic index 1.19 ± 0.17 come detected by blank control group, the phagocytic index of control group 1 for 1.23 ±
0.13rd, 1.25 ± 0.18, there is significant difference compared with blank control group, and test group is 1.43 ± 0.23, is had extremely aobvious
Write sex differernce P<0.01.Test group is compared with control group 1~2, phagocytic index P<0.05, there is significant difference.
To the testing result of capsule made from other embodiments of the invention similarly.
Influence of the antifatigue capsule of embodiment 9 to NK cells in mice activity
According to the Ministry of Public Health《Health food is examined and technology value disciplines》(2003), 120 kunming mices are pressed into body weight
It is randomly divided into 4 groups, every group 30, continuous 30 days.
1. experimental group：The capsule 's content of the offer of embodiment 5 is provided, is 1g/mL by contents melting to concentration, gavage is given
Mouse is given, gavage amount is 10mL/kg.
2. control group 1：The capsule 's content of the offer of comparative example 1 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
3. control group 2：The capsule 's content of the offer of comparative example 2 is provided, is 1g/mL by contents melting to concentration, gavage
Mouse is given, gavage amount is 10mL/kg.
4. blank control group：Deionized water gavage gives mouse, and gavage amount is 10mL/kg.
10 mouse are randomly selected from every group, for continuous gavage after 30 days, cervical dislocation puts to death mouse, sterile to take spleen,
It is placed in the small plate for filling appropriate sterile Hank ' s liquid, gently spleen is torn up with tweezers, through 200 mesh sieve net filtrations, list is made
Cell suspension is standby.Cell suspension is washed with Hank ' s liquid 2 times, and each 1000r/min centrifuges 10min.Supernatant is abandoned, by cytoplasm
Upspring, add 0.5mL aqua sterilisa 20s, 2 times of Hank ' s liquid of 0.5mL and 8mL Hank ' s liquid are added after splitting erythrocyte,
1000r/min centrifuges 10min, is resuspended with 1mL containing 10% calf serum RPMI1640 complete culture solutions, expects blue dyeing counting with platform
Viable count (should be more than 95%), it is finally 2 × 10 with RPMI1640 complete culture solutions adjustment cell concentration7Individual/mL.Take target
(effect target is than 50 by cell and each 100 μ L of effector cell:1)), add in U-shaped 96 well culture plate；Target cell Spontaneous release hole adds target thin
Born of the same parents and each 100 μ L of nutrient solution, target cell maximum release aperture add target cell and each 100 μ L of 1%NP40；Above-mentioned items are all provided with three and put down
Row hole, in 37 DEG C, 5%CO24h is cultivated in incubator, 96 well culture plates are then centrifuged into 5min with 1500r/min, is drawn per hole
In the well culture plate of 100 μ L horizontalizations bottom of supernatant 96, while the μ L of LDH matrix liquids 100 are added, react 3min, add 1mol/L's per hole
The μ L of HCL 30, OD value (OD) is determined at ELIASA 490nm, calculate NK cytoactives, as a result such as table 3：
The NK cytoactives of table 3
* represent compared with control group, P<0.05, there is significant difference.
△ represents test group compared with control group, P<0.05, there is significant difference.
The NK cytoactives (%) come detected by blank control group are 26.84 ± 13.52, and the NK cells of control group 1 are lived
Property (%) be 26.17 ± 15.38, there was no significant difference with control group compared with, and experimental group be 28.96 ± 13.65, have significantly
Sex differernce P<0.05.Meanwhile test group has significant difference P compared with control group 1~2<0.05.To other realities of the invention
Apply the testing result of capsule made from example similarly.
Effective sex investigation of the antifatigue capsule of embodiment 10
Randomly select the volunteer of 300 hypoimmunities, the age is 28~55 years old, be have a delicate constitution, nutrition not
It is good, One's spirits are drooping, fatigue and weak, appetite reduction, sleep-disorder, easily sick and be not easy the crowd of the performances such as rehabilitation, repeated infection.
3 groups are randomly assigned into,
Control group 1 gives reference substance 1, and control group 2 gives reference substance 2, and experimental group gives the capsule of the preparation of embodiment 5.
Each group volunteer's health products dosage is 30 days as one therapeutic course 1 tablet each time.After 30 days, investigation result such as table 4：
The effect research result of table 4
As a result show, the capsule effective percentage that the embodiment of the present invention 5 provides is significantly better than control group capsule 99%.To this
The testing result for the capsule that invention other embodiment provides is similarly.
It the above is only the preferred embodiment of the present invention, it is noted that come for those skilled in the art
Say, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as
Protection scope of the present invention.
Claims (7)
1. a kind of composition for improving immunity, it is characterised in that by apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder
Composition；The mass ratio of the apple stem cell freeze-dried powder and peacilomyce hepiahi bacterium powder is（2~5）：（4~8）；
The preparation method of wherein apple stem cell freeze-dried powder comprises the following steps：
Step 1：The sterilization of apple new life branch, after removing xylem and marrow, inducing culture is inoculated in, Fiber differentiation is formed
Confluent monolayer cells；
Step 2：The cambial cell accesses proliferated culture medium, shaking table culture obtains unicellular through squamous subculture；
Step 3：The unicellular proliferated culture medium that is inoculated in is cultivated through expanding, obtains apple stem cell；
Step 4：It will freeze, crush after the apple stem cell multigelation, obtain apple stem cell freeze-dried powder；
The inducing culture is the MS solid mediums containing NAA, BA and caseinhydrolysate；
The proliferated culture medium is the MS fluid nutrient mediums containing 2,4-D, BA, caseinhydrolysate and activated carbon.
2. the composition according to claim 1 for improving immunity, it is characterised in that apple stem cell freeze-dried powder and bat
The mass ratio of moth Paecilomyces varioti bacterium powder is 3.5:6.
3. a kind of health products for improving immunity, it is characterised in that including the composition described in any one of claim 1 ~ 2.
4. health products according to claim 3, it is characterised in that its formulation is tablet or capsule.
5. a kind of capsule for improving immunity, it is characterised in that including the composition described in any one of claim 1 ~ 2, lubrication
Agent and filler；
The mass ratio of composition, lubricant and filler described in any one of claim 1 ~ 2 is（60~130）：0.5：（0
~30）.
6. the capsule according to claim 5 for improving immunity, it is characterised in that the lubricant is magnesium stearate；Institute
It is starch to state filler.
7. the preparation method of the capsule of the raising immunity as described in any one of claim 5 ~ 6, it is characterised in that will by right
After asking the composition described in 1 ~ 2 any one to crush, with filler and mix lubricant, capsule is filled, is made and improves immunity
Capsule.
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| CN106820145A (en) * | 2017-01-13 | 2017-06-13 | 北京君合康医药科技有限公司 | A kind of health-care food composition of strengthen immunity and preparation method thereof |
| CN107349338A (en) * | 2017-07-17 | 2017-11-17 | 广东颜值科技有限公司 | A kind of stem of noble dendrobium stem cell composition and its oral formulations and application |
| CN109170905A (en) * | 2018-10-12 | 2019-01-11 | 江苏俊启生物科技股份有限公司 | The ingredient and process of peanut polysaccharide under low-temperature cold pressing |
| WO2020117309A1 (en) * | 2018-12-06 | 2020-06-11 | Rahimi Maryam | Plant stem cell product treatments |
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| WO2010095911A2 (en) * | 2009-02-23 | 2010-08-26 | 주식회사 운화 | Composition for enhancing immunity containing plant stem cell line derived from cambium of panax ginseng including wild ginseng or ginseng as an active ingredient |
| CN102459572B (en) * | 2009-05-26 | 2014-11-19 | 云火公司 | Plant stem cell derived from cambium of family gingkoaceae and method for isolation thereof |
| CN101856120A (en) * | 2010-03-23 | 2010-10-13 | 无锡市天赐康生物科技有限公司 | Health food for improving immunity of human body and preparation method thereof |
| CN102960718B (en) * | 2012-07-16 | 2013-10-30 | 赫康贸易(上海)有限公司 | Health products of plant stem cell |
| CN103989053A (en) * | 2014-04-29 | 2014-08-20 | 林树芳 | Selenium-rich chaff bamboo shoot tea combined ingredient containing plant stem cells and making method thereof |
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