CN106387854B - A kind of Halth-care composition of strengthen immunity - Google Patents
A kind of Halth-care composition of strengthen immunity Download PDFInfo
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- CN106387854B CN106387854B CN201610726850.9A CN201610726850A CN106387854B CN 106387854 B CN106387854 B CN 106387854B CN 201610726850 A CN201610726850 A CN 201610726850A CN 106387854 B CN106387854 B CN 106387854B
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- 125000003729 nucleotide group Chemical group 0.000 description 1
- 235000003715 nutritional status Nutrition 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000009965 odorless effect Effects 0.000 description 1
- 238000003305 oral gavage Methods 0.000 description 1
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- 230000008506 pathogenesis Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
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- 230000002062 proliferating effect Effects 0.000 description 1
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- 238000000926 separation method Methods 0.000 description 1
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- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011655 sodium selenate Substances 0.000 description 1
- 229960001881 sodium selenate Drugs 0.000 description 1
- 235000018716 sodium selenate Nutrition 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
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- 238000007619 statistical method Methods 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
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- 235000003702 sterols Nutrition 0.000 description 1
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- 239000002344 surface layer Substances 0.000 description 1
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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 Material From Animals Or Micro-Organisms (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses a kind of Halth-care compositions of strengthen immunity, the composition includes that Ganodenna Lucidum P.E 100-150 parts by weight, peacilomyce hepiahi bacterium powder 100-150 parts by weight, sodium selenite 0.01-0.1 parts by weight and L-Glutamine 50-150 parts by weight, the composition have the effect of significant strengthen immunity.The invention also discloses the preparation methods of the preparation containing the composition.
Description
Technical field
The invention belongs to field of health care products, and in particular to a kind of Halth-care composition of strengthen immunity.
Background technique
It is later that word appearance is immunized in traditional Chinese medicine, sees 19th century " immune class side " earliest.From the point of view of its original idea, it is immunized just
It is disease harm to be exempted, ensures the health of human body.From this, motherland's medicine is early to immune Connotation Definition and modern medicine
Phase is almost the same to immune being defined on Spirit Essence.This function embodiment is in immune defense, immune stable, immune prison
Depending on three aspects.
In terms of immunoprophylaxis, the coordinating and unifying of the Overall View and body that are mainly reflected in theory of traditional Chinese medical science.Overall View is recognized
An entirety for human body, human body be naturally also an entirety, naturally day alternates with night, the change of Four seasons change and environment,
The physiological function activity that will influence human body, by the self-control of human body itself, with the variation of reform of nature circle, realize people with
The unity of opposites of nature, therefore advocate and acting in accordance with the changes of YIN YANG in four seasons is wanted to change in terms of health care, actively maintain putting down for human body and external environment
Weighing apparatus, to keep the inherent coordinated balance of human body itself.If external environment variation is fierce, the adaptability or suitable of human body is surmounted
Should be able to power decline, then will affect the dysequilibrium of inside of human body and fall ill.Modern medicine also thinks that human body is complete with one
Immune system, there are many immunocompetent cells with its a variety of lymphokine and antibody, ties up in the case where nerve is adjusted with endocrine hormone
Hold the physiological equilibrium of people.Neuroendocrine-immunoregulatory network system is formed, defence is played and resists the effect of disease.
In terms of immune stabilizations, the equilibrium between yin and yang theory of Chinese medicine, the five internal organs correlation theory, relationship of promotion and restriction of five elements theory etc. all be immunized
Stablize closely related.The dynamic equilibrium that yin-yang under normal circumstances is under certain " degree " will lead to disease once " degree " is unbalance
The generation of disease.Just so-called " An excess of yang leads to disorder of yin ", " An excess of yin leads to disorder of yang ", " predominant yang generating heat ", " An excess of yin brings abort cold syndrome ".The marrow of yin-yang theory
It is " permit hold both ends and with wherein " ", pursue " yin and yang in equilibrium " and " put down by a definite date "；Traditional Chinese medicine regards human body as one with five
The organic whole that each internal organs centered on dirty influence each other with regulation is shown in the do not feel like eating of stomach not receiving such as Yangming fu-viscera excess, or
See the cough of Lung Qi upward reversal, wheeze；Internal heat is assigned, kidney Ji waterborne, then coordinating water and fire, on the contrary then breakdown of the normal physiological coordination between the heart and the kidney；" the five internal organs all can again
Make us cough, non-only lung "；" seeing the disease of liver, knowing the liver disease will be transmitted into the spleen, in the ban real spleen " etc..It can be seen that motherland's medicine from etiology and pathogenesis, diagnosis
And the everyways such as treatment show the dynamic stability of the five internal organs correlation and generation-inhibition in five elements inhibition and generation." Kang Ze evil, holds and is between the five-element
System ".Immunologic homeostasis can remove the cell of damage, aging, death, and immunoregulation effect is effectively performed, if stablized
Functional disturbance is then easy to appear autoimmune disease.
It in terms of immunosurveillance, is mainly reflected in tumorigenic understanding, traditional Chinese medicine thinks, the formation of tumour is
Body " healthy energy is insufficient ", then caused by " perverse trend crouches it ".When body is influenced or the invasion of exopathogen by certain factors, cause dirty
The physiological activities such as internal organs, channels and collaterals are abnormal, and when negative and positive of qi and blood balance coordination relationship is destroyed, human body will illness.Perfect
Immune surveillance function is built upon on sound function of immune system (including non-specific and specific immunity) basis, is such as exempted from
Epidemic disease monitors mistake department, then tumour cell is escaped from monitoring, such as immune deficient patients or largely using immunosuppressor person, because exempting from
Epidemic disease monitoring is impaired, and tumour often easily occurs.
The immune defense of immune system, immune stabilization and immune surveillance function are also among certain dynamic equilibrium, are fitted
Degree is advisable.And the immune function of body is limited to the influence of inside and outside many factors, maintains the dynamic of body's immunity flat
Weighing apparatus, is just able to maintain the health of human body.
Modern society, due to dog-eat-dog, pressure increases, and rhythm of life is getting faster, and for diet, people are often cared for not
It is upper reasonably combined, it is balanced nutritious, cause immunity degradation.In in the period of facing extraneous viral prevalence, such crowd most vulnerable to
Infection.The big feature of the one of weak and sickly crowd is exactly that immunity is low, and the big office worker of operating pressure is especially apparent.This life shape
State is mainly related with study, the work excessively inducements such as fatigue, stress, professional stress.Usually go in for the study the mental of work
Labourer, businessman etc. most easily enter this state.Modern society's keen competition, fast pace, high pressure have aggravated the body and mind of people
Burden；Environmental pollution, undesirable living habit and alcohol drinking patterns etc., such as overeating, the excessive drinking of thermophilic cigarette, the high food of edible tri,
Cause to be deficient in vitamin with microelement etc., the physical fitness of people is reduced from another point of view, these are all to lead to inferior health
The inducement that state occurs.
Immunopotentiator is also referred to as immunologic adjuvant, is a kind of special by non-specific approach raising body fight original or microorganism
The substance of opposite sex reaction.In recent years, at home and abroad scholar is to find determining, efficient, stable, the nontoxic ideal Immune-enhancing effect of characteristic
Under the effort of agent, the research field of immunopotentiator is constantly widened, and newcomer, novel form constantly discover, and Exploration of Mechanism is constantly deep
Enter, clinical application range is more and more wider.
Immunopotentiator is classified as follows by raw material:
A minerals class, selenium, zinc etc.；
B Chinese herbal medicine class, Radix Angelicae Sinensis, ginseng, ganoderma lucidum, cordyceps sinensis, Poria cocos, fructus lycii etc. and its polysaccharide, glucoside and volatile ingredient；
C immunologic adjuvants, propolis etc.；
D microorganism formulation class, BCG vaccine, choleratoxin B subunit etc.；
E vitamins, VitAVitE, vitamin D, vitamin C etc.；
F amino acids, arginine, leucine etc.；
G nucleic acid preparation class, more (poly-) nucleotide, immune ribonucleic acid etc..
One of an important factor for selenium is influence body's immunity, can influence the immune function of body from many aspects.It is right
The understanding of effect of the selenium in immune function is mainly set about by positive and negative two aspects: observation selenium deficiency animal immune
The variation of function；Selenium deficiency animal selenium-supply is given, the variation of Immune Function In Animals is observed.The study found that shadow of the selenium to immune function
Very extensive complexity is rung, although some report results are still variant, comprehensive relevant information, effect packet of the selenium to immune system
Include: selenium participates in cellular immune processes: selenium can influence the proliferation and differentiation of lymphocyte, can promote lymphocytic emiocytosis lymph
The factor (such as interferon, interleukin etc.): selenium can enhance the cytotoxicity of lymphocyte；Selenium can also promote specificity humoral to exempt from
Epidemic disease function increases internal antibody level: selenium improves nospecific immunity effect: animal experiment study and human body epidemiology tune
Looking into also found, selenium can improve body to the resistivity of helminth and infectious disease pathogens to a certain extent.
Therefore, the raw material rich in selenium, such as organic selenium: Se-enriched yeast, selenizing card can all be added in more health care product
Glue, selenoprotein, selenium-enriched edible mushroom powder and methylselenocysteine etc. are drawn, there are also part inorganic seleniums to be also used as selenium source replenishers,
Such as sodium selenate and sodium selenite.Sodium selenite is colourless crystallization, soluble easily in water, is the widest selenium hardening agent of current purposes,
Human body selenium is supplemented frequently as selenium source.Although sodium selenite is as selenium source replenishers, easy preparation at low cost, to the toxic pair of human body
Reaction and gastrointestinal irritation, and body is low to the degree of being absorbed and utilized of selenium.It is particularly preferably rich to have compared with multi-product using organic selenium
Selenium yeast, which obviate the toxicity of sodium selenite and gastrointestinal irritations, but are same as other biological raw material, and Se-enriched yeast is same
There is a problem of at high cost.
Summary of the invention
In order to solve the above technical problems, the purpose of the present invention is to provide a kind of Halth-care composition of strengthen immunity,
The composition is at low cost using sodium selenite as selenium source, and se complementary effect is good, improves immunity definite effect.
Above-mentioned purpose of the invention is achieved through the following technical solutions:
A kind of Halth-care composition of strengthen immunity, the composition include Ganodenna Lucidum P.E 100-150 parts by weight, bat
Moth paecilomycerol powder 100-150 parts by weight, sodium selenite 0.01-0.1 parts by weight, the composition further includes L-Glutamine
50-150 parts by weight.
Ganodenna Lucidum P.E is ganoderma lucidum hot water extract, and ganoderma lucidum (Ganoderma lucidum) is a kind of macro fungi, is belonged to
Basidiomycetes, Polyporus, Ganodermataceae.Recent studies indicate that the main chemical compositions of ganoderma lucidum have polysaccharide, ucleosides,
Furosemide feeding class, sterol, alkaloids and amino acids.
Peacilomyce hepiahi bacterium powder: peacilomyce hepiahi bacterium powder is had shown that through the units research such as institute of materia medica, China Medical institute
Chemical component is similar to natural cordyceps, has the effect of cordyceps sinensis, and there are also vasodilator, reduce blood pressure, slow down
Heart rate increases cAMP, stimulation adrenal gland generation steroid hormone, inhibits platelet aggregation, relaxes blood vessels smooth muscle, calmness and resist frightened
It a variety of physiological activity such as faints.
Sodium selenite: Na2SeO3, molecular weight 172.94, No. CAS: 10102-18-8, white crystals or crystalline powder are molten
Yu Shui does not dissolve in ethyl alcohol.
L-Glutamine: C5H10N2O3, molecular weight 146.15, No. CAS: 56-85-9, white crystals or crystalline substance powder, energy are molten
Yu Shui, it is odorless, it is nontoxic, nutritional supplement is often done, blending enriching substance is the most abundant free amino acid in muscle, accounts for about human body
The 60% of free amino acid total amount.Fasting plasma glutamine concentration is 500-750umol/L.Glutamine is not required ammonia
Base acid, it can be synthesized in human body by glutamic acid, valine, isoleucine.In disease, nutritional status is bad or high-intensity exercise
Etc. under stress situations, body increases the demand of glutamine.
In the present compositions, sodium selenite is selenium source, by influencing body from many aspects to body Selenium Supplement
Immune function, such as cellular immune processes: selenium can influence the proliferation and differentiation of lymphocyte, lymphocytic emiocytosis can be promoted to drench
Ba Yinzi (such as interferon, interleukin etc.)；Selenium can enhance the cytotoxicity of lymphocyte；Selenium can also promote specificity humoral
Immune function increases internal antibody level；Selenium can also improve nospecific immunity effect etc..
However body to inorganic selenium be absorbed and utilized and it is bad, if simply by increase sodium selenite content improve inhale
Receive availability, and toxic side effect by sodium selenite and gastrointestinal irritation are limited, especially inorganic selenium be not easy internal organs such as
It accumulates and utilizes in liver kidney, even if Blood Se concentration is higher, the content of human body Selenonic protein still cannot be promoted to increase, select organic selenium
Cost is again excessively high.The present inventor has found in the formulation procedures of research health care product, when the suitable L-Glutamine of addition
When, the degree of being absorbed and utilized of sodium selenite greatly improves, and not only increases blood selenium level, while the internal organs egg containing selenium such as increase liver kidney
White contents level.
And health care product group of the present invention containing Ganodenna Lucidum P.E, peacilomyce hepiahi bacterium powder, sodium selenite and L-Glutamine
It closes object and also shows excellent immunity humidification under synergistic effect.
Preferably, the composition includes Ganodenna Lucidum P.E 120-150 parts by weight, peacilomyce hepiahi bacterium powder 100-
130 parts by weight, sodium selenite 0.01-0.05 parts by weight, L-Glutamine 80-120 parts by weight.
As a kind of perferred technical scheme, the composition includes 130 parts by weight of Ganodenna Lucidum P.E, Paecilomyces hepiali chen
120 parts by weight of bacterium powder, 0.03 parts by weight of sodium selenite, 100 parts by weight of L-Glutamine.
As another preferred technical solution, which includes 120 parts by weight of Ganodenna Lucidum P.E, bat moth
130 parts by weight of paecilomycerol powder, 0.05 parts by weight of sodium selenite, 120 parts by weight of L-Glutamine.
A kind of preparation of the Halth-care composition containing strengthen immunity of the present invention, the preparation is by above-mentioned composition and auxiliary material
It is made, said preparation can be tablet, pill, capsule, powder or granule.The auxiliary material select often according to dosage form
Advise auxiliary material.
Preferably, the preparation of the Halth-care composition of strengthen immunity of the present invention is capsule.
The present invention also provides a kind of methods of the preparation of Halth-care composition for preparing strengthen immunity, including walk as follows
It is rapid:
Stock: Ganodenna Lucidum P.E, peacilomyce hepiahi bacterium powder, Co-60 irradiation sterilization, 4kGy are weighed；Powder is distinguished after irradiation
Broken 80 mesh of mistake, it is spare；Sodium selenite, L-Glutamine, microcrystalline cellulose, starch, silica material crushed 80 respectively
Mesh, it is spare；
Mixing: sodium selenite being uniformly mixed with starch equal increments method, obtains mixed powder, and mixed powder and ganoderma lucidum are extracted
The materials such as object, peacilomyce hepiahi bacterium powder, L-Glutamine, microcrystalline cellulose, silica and magnesium stearate are put into mixed together
40min is mixed in conjunction machine, is uniformly mixed material, is obtained total mixed powder；
Capsule filling: mixed material is filled, operating condition on automatic capsule filling machine: 18~26 DEG C, relatively
Humidity: 45%~60%.
The beneficial effects of the present invention are: Halth-care composition of the invention selects Ganodenna Lucidum P.E, Paecilomyces hepiali chen
Bacterium powder, sodium selenite and L-Glutamine compatibility, it is unexpected sodium selenite toxic side effect and gastrointestinal irritation do not occurring
In the case of, it greatly increases body and selenium is absorbed and utilized, not only improve blood selenium content, moreover it is possible to improve body internal organs Selenonic protein
Level, and excellent strengthen immunity effect, the health care product that the present invention enhances human immunity are shown in functional experiment
Composition has at low cost, the good advantage of effect.
Specific embodiment
Below by way of specific embodiment, above content of the invention is described in further detail.But it should not incite somebody to action
This range for being interpreted as the above-mentioned theme of the present invention is only limitted to example below.All technologies realized based on above content of the present invention
It all belongs to the scope of the present invention.
Ganodenna Lucidum P.E is purchased from Sanjiang Biologica Engineering Co., Ltd., Xi-an City；Peacilomyce hepiahi bacterium powder is looked forward to purchased from Zhejiang Wan Feng
Pharmaceutical Co. Ltd, industry group.
Embodiment 1
Table 1 is formulated
Ganodenna Lucidum P.E, peacilomyce hepiahi bacterium powder, Co-60 irradiation sterilization, 4kGy are weighed by formula ratio；After irradiation respectively
It crushes, crosses 80 meshes, it is spare.Sodium selenite, L-Glutamine, microcrystalline cellulose, starch, silica material crushed respectively
80 meshes, it is spare.
Mixing: sodium selenite is uniformly mixed with starch equal increments method, obtains mixed powder I.Mixed powder I and ganoderma lucidum are extracted
The materials such as object, peacilomyce hepiahi bacterium powder, L-Glutamine, microcrystalline cellulose, silica and magnesium stearate are put into mixed together
40min is mixed in conjunction machine, is uniformly mixed material, is obtained mixed powder II.
Capsule filling: mixed material being filled on automatic capsule filling machine, every 0.4g containing content, is operated
Condition: 18~26 DEG C, relative humidity: 45%~60%.
Capsule polishing: canned capsule is polished on capsule polisher, eliminates surface medicinal powder.
Embodiment 2
Table 2 is formulated
It is formulated according to table 2 and prepares product according to embodiment 1.
Embodiment 3
Table 3 is formulated
It is formulated according to table 3 and prepares product according to embodiment 1.
Embodiment 4
Table 4 is formulated
It is formulated according to table 4 and prepares product according to embodiment 1.
Comparative example 1
Table 5 is formulated
It is formulated according to table 5 and prepares product, every 0.4g containing content according to embodiment 1.
5 strengthen immunity function test of embodiment
1.1 samples: function is carried out according to sample (being referred to as sample 1 and sample 2) prepared by embodiment 1 and comparative example 1
Learn test.Sample is set shady and cool dry and comfortable ventilation and is saved.Population clothes recommend dosage for everyone (adult weight is calculated by 60kg) daily 6
Grain, i.e. 0.04g/kg BW are daily.Capsule 's content is taken to be tested.
1.2 experimental animals and grouping: select the cleaning grade of Institute of Experimental Animals, Chinese Academy of Medical Sciences's breeding breeding strong
Health male Kunming strain mice 210, weight is 18-22 grams, credit number: the capital SCXK- 2004-001.It is divided into 3 groups, every group
70, I group is immunized, carries out the mouse spleen lymphocyte transformation experiment of ConA induction；Immune II group, carries out delayed allergy
Experiment；Immune III group carries out serum hemolysin measurement and the measurement of antibody-producting cell number.
1.3 experimental situation conditions: experimental animal room temperature: 22-25 DEG C, relative humidity: 55-70%.
The selection of 1.4 dosage gives mode with tested material: being taken according to population and recommends dosage, each sample sets basic, normal, high dosage
Group, respectively 0.20g, 0.40g, 1.20g/kg/ day (being respectively equivalent to 5,10,30 times that human body recommends dosage), if a yin
Property control group, every group of 10 animals.Tested material is prepared with water, once a day orally administration, and items are surveyed after continuous gavage 33 days and are exempted from
Epidemic disease index.Intragastric administration on mice volume is 0.1mL/10g mouse weight.Negative control group replaces tested material with water.
1.5 key instruments and reagent: animal platform balance, electronic analytical balance, centrifuge, clean bench, carbon dioxide training
Support case, constant water bath box, microscope, semi-automatic biochemical analyzer, microplate reader etc..
Sterile surgical instrument, micro syringe, cell counter, the flat tissue culture plate in 24 holes and 96 holes, 96 holes are U-shaped thin
Born of the same parents' culture plate, glass dish, gauze, glass slide, test tube, 200 mesh screens, timer hemoglobin pipet etc..
Sheep red blood cell (SRBC) (SRBC), physiological saline, Hank's liquid (pH 7.2-7.4), RPMI1640 culture solution, small ox blood
Clearly, penicillin, streptomysin, ConA, 1% glacial acetic acid, the HCL solution of 1mol/L, isopropanol, MTT, DNFB, PBS buffer solution
(pH7.2-7.4), complement (guinea pig serum), SA buffer, agarose, YAC-1 cell, lithium lactate, nitro tetrazolium chloride,
Phenazine Dimethyl Sulfate, oxidized coenzyme I, the Tris-HCL buffer of 0.2mol/L, 1% NP40, india ink,
Sodium carbonate, chicken red blood cell, methanol, Gimsa dye liquor of 0.l% etc..
1.6 experimental method
1.6.1 ConA induction mouse spleen lymphocyte transformation experiment (mtt assay):
It is sterile to take spleen, it is placed in the plate for filling appropriate sterile Hank's liquid, cell suspension is made, through 200 mesh screen mistakes
Filter.It is washed 2 times with Hank's liquid, is centrifuged l0min (1000r/min) every time.Then cell is suspended in 1mL complete culture solution,
Living cell counting number is 3*10 with RPMI1640 culture solution adjustment cell concentration6A/mL..It is divided to two holes to be added cell suspension again
In 24 well culture plates, every hole 1mL, 75uL ConA liquid (being equivalent to 7.5uL/mL) is added in a hole wherein, another hole conduct pair
According to setting 5%CO2, 72h is cultivated in 37 DEG C of carbon dioxide incubators.4h gently sucks supernatant 0.7mL before culture terminates, and is added
0.7mL is free of the RPMI1640 culture solution of calf serum, while the hole MTT (5mg/mL) 50uL/ is added, and continues to cultivate 4h.Culture
After, 1mL acid isopropyl alcohol is added in every hole, and piping and druming mixes, is completely dissolved purple crystal, is then dispensed into 96 well culture plates
In, 3 parallel holes are made in each hole, with microplate reader, measure OD value with 570nm wavelength.The proliferative capacity of lymphocyte, which is used, to be added
The OD value in the hole ConA subtracts the OD value expression that the hole ConA is not added.
1.6.2 delayed allergy (DTH) (vola pedis thickens method)
After sensitization 4 days, left back vola pedis thickness be measured, then to mouse peritoneal injection 2% hematocrit SRBC (the every mouse of 0.2mL/)
20% (v/v) SRBC (the every mouse of 20ul/) is subcutaneously injected in measuring point, measures left back vola pedis thickness, same position for 24 hours after injection
It sets measurement three times, is averaged, indicate the degree of DTH to attack front and back vola pedis thickness difference (pedal swelling degree), it is tested
The difference of sample sets is significantly higher than the difference of control group, can determine that this experimental result positive.
1.6.3 antibody-producting cell detection (Jerne improves slide method)
Sheep blood is taken, with brine 3 times, l0min (2000r/min) is centrifuged every time, with physiological saline by hematocrit
SRBC is made into the cell suspension of 2% (v/v), and 0.2mL is injected intraperitoneally in every mouse.5 days mouse are put to death after will be immune, take spleen, gently
It grinds, cell suspension, 200 mesh net filtrations is made with Hank's liquid, cell is finally suspended in 5mL by washing, centrifugation 2 times
In Hank's liquid.After surface layer culture medium (lg agarose adds distilled water to l00mL) is dissolved by heating, puts 45~50 DEG C of water-baths and protect
Temperature is mixed with Hank ' the s liquid of equivalent pH7.2-74,2 times of concentration, dispenses small test tube, every pipe 0.5mL, then into pipe plus use SA
10%SRBC50uL (v/v), the 25uL splenocyte suspension of liquid preparation, are poured into the slide of brush agarose thin layer after mixing rapidly
On, after to be solidified, flat buckle of slide is placed in glass frame, is put into carbon dioxide incubator and is incubated for 1.5h, it is dilute with SA buffer
The complement (1:8) released is added in glass frame groove, continues to be incubated for 1.5h, counts hemolysis plaque number.With plaque number/full splenocyte table
Show antibody-producting cell number.The plaque number of test sample group is significantly higher than the plaque number of control group, can determine that this experimental result
It is positive.
1.6.4 the measurement (HC50) of half hemolytic value
Sheep blood is taken, with brine 3 times, l0min (2000r/min) is centrifuged every time, with physiological saline by hematocrit
SRBC is made into the cell suspension of 2% (v/v), and 0.2mL is injected intraperitoneally in every mouse.After immune 5 days, extract the eyeball of mouse, take blood in
In centrifuge tube, about 1h is placed, 2000r/min is centrifuged l0min, and serum is collected in separation.Serum is diluted to 300 with SA buffer
Times, it takes 1mL to set in test tube, sequentially adds 10%SRBC 0.25mL, complement 1mL.The control tube of not increase serum is separately set, it is slow with SA
Fliud flushing replaces.It is placed in 37 DEG C of waters bath with thermostatic control after keeping the temperature 15min, ice bath terminates reaction.2000r/min is centrifuged 10min, takes supernatant
1mL adds Dou Shi reagent 3mL.The SRBC 0.25mL for taking 10% simultaneously adds Dou Shi reagent to 4mL, sufficiently mixed in another test tube
Even, after placing 10min, control tube makees blank pipe at 540nm, measures each pipe OD value respectively.The amount of hemolysin is molten with half
Blood number (HC50) indicates, is calculated as follows:
OD value * extension rate when sample half hemolytic value=sample OD value/SRBC half hemolysis
The HC50 of test sample group is significantly higher than the HC50 of control group, can determine whether experimental result for the positive.
1.7 experimental datas statistics: variance analysis statistical disposition is carried out using SPSS statistical software.
2. result
Influence of 2.1 samples to mouse cell immune function
2.1.1 influence of the sample to mouse delayed allergy (DTH)
Influence of 6 sample of table to mouse delayed allergy (DTH)
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
As seen from the above table, the sample of orally administration mouse various dose is after 33 days, the swelling degree of the paw difference of each dosage group
It is all larger than negative control group, but only sample 1 reaches significant difference, and middle high dose group difference has very significant (P
<0.01), each dosage group difference of sample 2 does not have conspicuousness (P>0.05).
Influence of 2.12 samples to lymphocyte transformation test
Influence of 7 sample of table to lymphocyte transformation test
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
As seen from the above table, compared with negative control group, lymphopoiesis ability increases each dosage group of sample 1 and 2,
But 2 no significant difference of sample (P>0.05), only sample 1 show the significance difference opposite sex (P<0.05), and high dose group otherness
Highly significant illustrates that sample 1 has the function of promoting lymphopoiesis, the conversion capability of mouse.
Influence of 2.2 samples to mouse humoral immune function
2.2.1 influence of the sample to mouse antibodies cellulation
8 sample of table is to mouse antibodies cellulation test experience result
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
As seen from the above table, each dosage group of sample 1 and 2 is compared with negative control group, and hemolysis plaque number increased, but sample
Low dose group no significant difference (P > 0.05) in 2, and all dosage groups of sample 1 and 2 high dose group of sample show significant difference
Property (P < 0.05), especially 1 high dose group of sample, difference have very significant (P < 0.01).
2.2.2 influence of the sample to mouse half hemolytic value (HC50)
Influence of 8 sample of table to mouse half hemolytic value (HC50)
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
As seen from the above table, compared with negative control group, sample half hemolytic value increased each dosage group of sample 1 and 2, but
Each dosage group of sample 2 without significant difference (P>0.05), and all dosage groups of sample 1 show significance difference it is anisotropic (P<
0.05), especially 1 high dose group of sample, difference have very significant (P < 0.01).
Above-mentioned function test proves that Halth-care composition of the invention has the function of exact strengthen immunity, and because
All there is unexpected effect compared with the composition of comparative example or negative control group for the synergistic effect of L-Glutamine
Fruit.
Test is absorbed and utilized to sodium selenite in 6 rat of embodiment
Serum selenium can reflect the intake situation of the recent selenium of body, and selenium is main in body as microelement necessary to animal
It to play a role in the form of selenoprotein (as contained selenocystein), glutathione peroxidase (GSH-PX) is that research is more
One of selenoprotein, and selenoprotein expression and activity adjusted by Se nourishment level；GSH-PX be mammal content most
Selenoprotein abundant, activity are adjusted by the selenium in animal viscera is horizontal, can be added blood selenium using GSH-PX vigor as biology
With the index utilized.
1.1 materials and methods
1.1 experimental animals: the SPF grade healthy SD male of Institute of Experimental Animals, Chinese Academy of Medical Sciences's breeding breeding is selected
Rat 70, weight is 140-160 grams, credit number: the capital SCXK- 2009-002.
1.2 experimental materials: low selenium feed: materials of wheat, corn and soybean etc. are purchased from the low selenium in Henan Province's Sanmenxia City Lingbao City
Area is processed by experimental animal center of henan province, and through Co60 radiation sterilization, Se content≤0.03ug/g；Respectively by 1 He of embodiment
Sample 1 and sample 2 prepared by comparative example 1.
1.3 instruments and reagent
ADVIA1200 automatic clinical chemistry analyzer, KY-2000 semi-automatic biochemical analyzer, DS-671 type electronic balance,
DTS-3 low speed autobalancing centrifuge, DY89-1 type electric driven glass refiner, ASF-930 dual channel atomic fluorescence photometers；Paddy Guang
Sweet peptide peroxidase (GSH-Px) kit (lot number 20090522)；Rat thioredoxin reductase (TrxR), ELISA inspection
Test agent box (LOT:08-12).
1.4 experimental method
70 male SD rats are randomly divided into 7 groups by weight, raises and Se-low animal is established with low selenium feed.Weekly from
1 progress tail vein blood of every group of extraction, surveys its Blood Se concentration, after 7 weeks when blood selenium average value is reduced to 0.25mg/L or so, then
7 groups are randomly divided by weight, 1 group of control group, sample 1 and 2 each 3 groups of sample, every group 10, the daily stomach-filling pure water of control group, sample
2 groups of product 1 and sample distinguishes the corresponding sample of stomach-filling daily, and the dosage according to the daily recommended amounts rat oral gavage of sample is respectively
0.4,0.8,1.2g/kg (being equivalent to 10,20,30 times of dosage of people's recommendation) weight, continuous gavage 1 month, last stomach-filling was broken afterwards for 24 hours
Head takes blood, is centrifugated serum, and win hepatic and renal tissue, filter paper blots for use after being rinsed with ice-cold normal saline.
1.5 index determining
Blood selenium content: Hydride atomic fluorescent spectrum method for detecting.It weighs suitable liver and renal tissue prepares 10%
Tissue homogenate, 3000r/min are centrifuged 15min, take supernatant, illustrate to measure glutathione peroxidase respectively by kit
(GSH-Px) vigor content.The measurement biuret method of albumen in homogenate.
1.6 statistical analysis data useIt indicates, the t of two independent samples of comparison of average value is examined.
2. result
2.1 rat blood selenium levels
Influence of 9 sample of table to Blood Se concentration
Note: * is compared with negative control group, P < 0.05.
As seen from the above table, sample 1 and 2 can increase rat blood selenium content, and each dosage group of sample 1 and 2 middle and high doses of sample
For amount group compared with negative control group, difference has conspicuousness (P < 0.05).
2.2 rat livers and renal glutathione peroxidase (GSH-PX) vigour
Influence of 10 sample of table to liver GSH-PX vigor
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
As seen from the above table, sample 1 and each dosage group of sample 2 are compared with negative control group, liver glutathione peroxidase
Enzyme activity increases, and each dosage group of sample 1 and 2 high dose group vigor of sample increase difference and have conspicuousness P < 0.05, especially sample
The difference highly significant (P < 0.01) of each dosage group of product 1.
Influence of 11 sample of table to kidney GSH-PX vigor
Note: * is compared with negative control group, P < 0.05；* is compared with negative control group, P < 0.01.
By table 11 as it can be seen that sample 1 and each dosage group of sample 2 are compared with negative control group, kidney GSH-PX vigor has increasing
Add, still, difference is not significant compared with the control group for the low of sample 2, high dose group, and the middle and high dosage group of sample 1 and control group ratio
Compared with difference highly significant (P < 0.01).
In conclusion Halth-care composition formula of the invention is rationally, each component reaches preferable effect, L- glutamy
The reasonably combined absorption and utilization for dramatically increasing body to selenium of amine, blood selenium content obviously increases, and promotes blood selenium in each internal organs
Make full use of, the increase of liver kidney GSH-PX vigor highly significant, and finally to mouse immunity promotion show excellent effect
Fruit.
The invention is not limited to specific embodiments above-mentioned.The present invention, which expands to, any in the present specification to be disclosed
New feature or any new combination, and disclose any new method or process the step of or any new combination.
Claims (7)
1. a kind of Halth-care composition of strengthen immunity, the composition includes Ganodenna Lucidum P.E 100-150 parts by weight, bat moth
Paecilomycerol powder 100-150 parts by weight, sodium selenite 0.01-0.1 parts by weight, which is characterized in that the composition further includes L-
Glutamine 50-150 parts by weight.
2. the Halth-care composition of strengthen immunity as described in claim 1, which is characterized in that the composition includes ganoderma lucidum
Extract 120-150 parts by weight, peacilomyce hepiahi bacterium powder 100-130 parts by weight, sodium selenite 0.01-0.05 parts by weight, L-
Glutamine 80-120 parts by weight.
3. the Halth-care composition of strengthen immunity as described in claim 1, which is characterized in that the composition includes ganoderma lucidum
130 parts by weight of extract, 120 parts by weight of peacilomyce hepiahi bacterium powder, 0.03 parts by weight of sodium selenite, 100 weight of L-Glutamine
Measure part.
4. the Halth-care composition of strengthen immunity as described in claim 1, which is characterized in that the composition includes ganoderma lucidum
120 parts by weight of extract, 130 parts by weight of peacilomyce hepiahi bacterium powder, 0.05 parts by weight of sodium selenite, 120 weight of L-Glutamine
Measure part.
5. a kind of preparation of the Halth-care composition of strengthen immunity described in any one of 1-4 containing claim, feature exist
In the preparation is tablet, pill, capsule, powder or granule.
6. the preparation of strengthen immunity as claimed in claim 5, which is characterized in that the preparation is capsule.
7. a kind of preparation method of capsule as claimed in claim 6, includes the following steps:
Stock: Ganodenna Lucidum P.E, peacilomyce hepiahi bacterium powder, Co-60 irradiation sterilization, 4kGy are weighed；It crushed respectively after irradiation
80 meshes, it is spare；Sodium selenite, L-Glutamine, microcrystalline cellulose, starch, silica material crushed 80 meshes respectively,
It is spare；
Mixing: sodium selenite being uniformly mixed with starch equal increments method, obtains mixed powder, by mixed powder and Ganodenna Lucidum P.E,
Peacilomyce hepiahi bacterium powder, L-Glutamine, microcrystalline cellulose, silica and magnesium stearate are put into mixing in mixing machine together
40min is uniformly mixed material, obtains total mixed powder；
Capsule filling: mixed material is filled, operating condition on automatic capsule filling machine: 18~26 DEG C, relatively wet
Degree: 45%~60%.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1762999A (en) * | 2005-09-01 | 2006-04-26 | 徐州工程学院 | Seleno compound amino acid and its preparation method |
| CN103127184A (en) * | 2013-03-25 | 2013-06-05 | 威海康博尔生物药业有限公司 | Preparation for improving organism immunity and preventing tumors |
| CN103461991A (en) * | 2013-08-05 | 2013-12-25 | 买世禄 | Production method for health care food adopting amino acids to combine with multiple trace elements to promote growth and development |
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| CN102860508A (en) * | 2012-09-28 | 2013-01-09 | 无锡市天赐康生物科技有限公司 | Immunity-enhancing and anti-fatigue health-care food and preparation method thereof |
| CN103907910A (en) * | 2013-01-09 | 2014-07-09 | 湖北创力药业有限公司 | Industrial production process of ganoderma Chinese caterpillar fungus capsule enhancing immunity function |
| CN103054909B (en) * | 2013-02-04 | 2014-09-10 | 福建仙芝楼生物科技有限公司 | Capsule for alleviating physical fatigue and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1762999A (en) * | 2005-09-01 | 2006-04-26 | 徐州工程学院 | Seleno compound amino acid and its preparation method |
| CN103127184A (en) * | 2013-03-25 | 2013-06-05 | 威海康博尔生物药业有限公司 | Preparation for improving organism immunity and preventing tumors |
| CN103461991A (en) * | 2013-08-05 | 2013-12-25 | 买世禄 | Production method for health care food adopting amino acids to combine with multiple trace elements to promote growth and development |
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