CN104415214A - Application of black garlic extract in adjusting animal intestinal flora - Google Patents
Application of black garlic extract in adjusting animal intestinal flora Download PDFInfo
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- CN104415214A CN104415214A CN201310390623.XA CN201310390623A CN104415214A CN 104415214 A CN104415214 A CN 104415214A CN 201310390623 A CN201310390623 A CN 201310390623A CN 104415214 A CN104415214 A CN 104415214A
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- Prior art keywords
- bulbus allii
- black bulbus
- allii extract
- animal
- medicine
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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Abstract
The invention discloses an application of a black garlic extract in adjusting animal intestinal flora, an application of the black garlic extract in preparation of medicines, the medicines, foods, applications of the medicines and the foods in adjusting the animal intestinal flora, and a method for adjusting the animal intestinal flora. Inventors discover that by adopting the black garlic extract, numbers of probiotics such as bifidobacterium, ruminococcus and the like in animal intestines can be significantly improved, so that the animal intestinal flora can be significantly adjusted, and thus the immunity of the animal intestines can be enhanced by virtue of the action of the intestinal flora, and diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumors and the like can be prevented or treated.
Description
Technical field
The present invention relates to the novelty teabag of black Bulbus Allii extract, particularly, relate to black Bulbus Allii extract and regulating the purposes in microbial population of animal intestinal tract.More specifically, the present invention relates to black Bulbus Allii extract and regulating the purposes in microbial population of animal intestinal tract, black Bulbus Allii extract is preparing the purposes in medicine, medicine and food and regulating the purposes in microbial population of animal intestinal tract, and regulates the method for microbial population of animal intestinal tract.
Background technology
A large amount of symbiosis floras is there is in normal human, these antibacterial major parts colonize in the intestinal of people, and quantity, more than 1,000 trillion, is equivalent to 10 times of human body cell sum, its microbial gene quantity is about 3,000,000, is approximately more than 100 times of human genome gene dosage.The gene of magnanimity like this can help microorganism to adapt to changeable environment, defines the mutualism relation inseparable with human body simultaneously.Completing of the Human Genome Project, make the mankind more deep to the understanding of oneself, but the genome understanding the mankind can not hold the health of the mankind completely, the micropopulation of forever settling down in human body intestinal canal is extremely far-reaching on the impact of human health, little of digestive tract environment, digestive problems, large to fat or thin problem, hypertension, diabetes, hyperlipidemia, cancer etc., the life and health of the mankind is closely bound up with them, and therefore human body intestinal canal microbial genome is also described as the second cover genome of the mankind.
At present, research confirms that the various diseases of human body is as all closely related with intestinal microbial population in diabetes, hypertension, hyperlipidemia, cancer etc.Intestinal microbial population fermentation polysaccharides metabolite mainly contains short key fatty acid (acetic acid, propanoic acid, butanoic acid etc.), gas (hydrogen, carbon dioxide, hydrogen sulfide and methane) and ammonia, and wherein hydrogen and carbon dioxide are the main components of gas in colon.The in vitro study of people's intestinal shows, butanoic acid can promote intestinal mucosa reparation and functional rehabilitation thereof, and the formation of inflammation-inhibiting cytokine, and the secretion of the tumor necrosis factor of intestinal epithelial cell can be reduced, play antiinflammatory and antitumor action.In addition, separately studies have reported that the change of Bacteroides and (or) Clostridia population in intestinal microbial population and the appearance of antibacterial Uncultured bacterium clonenbw1009b01c1, the hypertensive generation of Kazak ethnic population and development may be affected.Different intestinal microbial population spectrums can be caused a disease, can also diseases prevention.
But, regulate the research of microbial population of animal intestinal tract aspect still to need deeply at present.
Summary of the invention
The present invention is intended at least to solve one of technical problem existed in prior art.For this reason, one object of the present invention is to propose a kind of means that effectively can regulate microbial population of animal intestinal tract.
Inventor studies discovery, black Bulbus Allii extract can significantly improve the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in intestinal, regulate the intestinal microbial population of animal, and then play the effects such as enhancing immunity, fat-reducing, blood sugar lowering, blood pressure lowering, blood fat reducing, antitumor by the effect of intestinal microbial population, and without any side effect, can long-term taking.Wherein, it should be noted that, the main component of black Bulbus Allii extract is diallyl trisulfide (DATS) and diallyl disulfide (DADS), wherein topmost effective active composition is diallyl trisulfide (DATS) i.e. garlicin (allicin), it is slightly soluble in water, be dissolved in the organic solvents such as ethanol, benzene, ether, present stage also has no the research in regulating intestinal canal flora of black Bulbus Allii extract and garlicin and report.
Thus, according to an aspect of the present invention, the invention provides black Bulbus Allii extract and regulate the purposes in microbial population of animal intestinal tract.According to embodiments of the invention, black Bulbus Allii extract can significantly improve the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in animal intestinal, thus significantly can regulate the intestinal microbial population of animal, and then played the effects such as enhancing immunity, fat-reducing, blood sugar lowering, blood pressure lowering, blood fat reducing, antitumor by the effect of intestinal microbial population.Thus, can effectively regulate its intestinal microbial population to animals administer black Bulbus Allii extract, thus strengthen its immunity, the diseases such as prevention or treatment obesity, diabetes, hypertension, hyperlipidemia or tumor.
According to embodiments of the invention, black Bulbus Allii extract of the present invention can be prepared by following steps: select fresh black Bulbus Allii of uniform size, peeling, cleans the garlic clove of peeling, and clothing film is gone to the greatest extent, with rustless steel crusher in crushing 5min.To squeeze the juice 10min with juice extractor (8000rpm), then to filter after 5min through centrifuge with 60 order screen cloths and namely obtain black Bulbus Allii extract.
It should be noted that, the animal species of term " animal " indication in expression way " black Bulbus Allii extract is regulating the purposes in microbial population of animal intestinal tract " used in this article is not particularly limited.According to embodiments of the invention, this animal can be any animal with intestinal organ, and preferred mammal, more preferably rat, mice, people, optimum is chosen.In addition, used in this article expression way " prevention or treatment " mainly refers to the diseases such as prevention or auxiliary treatment obesity, diabetes, hypertension, hyperlipidemia and tumor.
According to embodiments of the invention, take black Bulbus Allii extract without any side effect, thus the dosage of black Bulbus Allii extract is not particularly limited, as long as can improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal and the intestinal microbial population effectively regulating animal.According to concrete examples more of the present invention, described black Bulbus Allii extract is carried out administration with the dosage of 0.3-1.0mg garlicin/kg to described animal.According to other embodiments of the present invention, described black Bulbus Allii extract is carried out administration with the dosage of 0.5mg garlicin/kg to described animal.Thus, increased obviously by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, namely black Bulbus Allii extract regulates the Be very effective of microbial population of animal intestinal tract.
According to another aspect of the invention, regulating the effect in microbial population of animal intestinal tract based on above-mentioned black Bulbus Allii extract, present invention also offers black Bulbus Allii extract and preparing the purposes in medicine, described medicine is for regulating microbial population of animal intestinal tract.
Said medicine can be used in regulating microbial population of animal intestinal tract, and then can play the effects such as enhancing immunity, fat-reducing, blood sugar lowering, blood pressure lowering, blood fat reducing, antitumor by the effect of intestinal microbial population.Thus, according to embodiments of the invention, described medicine is used for enhancing immunity, prevention or treatment obesity, diabetes, hypertension, hyperlipidemia or tumor.
According to embodiments of the invention, every day is to the black Bulbus Allii extract of described animals administer 0.3-1.0mg garlicin/kg.Thus, can significantly improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, regulate the intestinal microbial population of animal, and then effectively prevent or treat the diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumor by the effect of intestinal microbial population.
According to a further aspect in the invention, present invention also offers a kind of medicine.According to embodiments of the invention, this pharmaceutical pack contains: black Bulbus Allii extract; And pharmaceutically acceptable excipient.Inventor is surprised to find, medicine of the present invention can significantly improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, regulate the intestinal microbial population of animal, and then effectively prevent or treat the diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumor by the effect of intestinal microbial population.
According to embodiments of the invention, the kind of described excipient is not particularly limited, as long as medicine can be made to form the dosage form of easily carrying out administration.According to concrete examples more of the present invention, described excipient is at least one being selected from binding agent, filler, film-coating polymer, plasticizer, fluidizer, disintegrating agent and lubricant.
According to embodiments of the invention, the dosage form of medicine of the present invention is not particularly limited, as long as can be convenient to carry out administration.According to concrete examples more of the present invention, described medicine is in the form of at least one being selected from capsule, pill, tablet, granule, liquid oral, oral pastes, aerosol and spray.According to preferred embodiments more of the present invention, described medicine is the form of capsule.Thus, be easy to carry out administration.
According to embodiments of the invention, the dosage of medicine of the present invention is not particularly limited, and in practical application, can select flexibly according to the health status of administration object.According to some embodiments of the present invention, the dosage of described medicine is 0.3-1.0mg garlicin/kg.Thus, significantly increased by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, regulate the Be very effective of microbial population of animal intestinal tract.
According to embodiments of the invention, described medicine is for regulating microbial population of animal intestinal tract.And then, the effects such as enhancing immunity, fat-reducing, blood sugar lowering, blood pressure lowering, blood fat reducing, antitumor can be played by the effect of intestinal microbial population.Thus, according to other embodiments of the present invention, described medicine is used for enhancing immunity, prevention or treatment obesity, diabetes, hypertension, hyperlipidemia or tumor.
In accordance with a further aspect of the present invention, present invention also offers a kind of food.According to embodiments of the invention, this food product packets contains: black Bulbus Allii extract; And acceptable additive in bromatology.According to embodiments of the invention, food of the present invention can significantly improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, the intestinal microbial population of effective adjustment animal, and then effectively prevent or treat the diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumor by the effect of intestinal microbial population.Meanwhile, food of the present invention also has all raw materials and comes from natural, and safe and reliable, raw material sources are extensively easy to get, and preparation technology is simple, storing and application safety, low production cost, steady quality, advantage with long preservation period.
It should be noted that, term " food " used in this article should make broad understanding, its can be any can by the form eaten, namely except the food form of routine, food of the present invention can also be health product (being also sometimes referred to as " functional food "), beverage etc.According to concrete examples more of the present invention, food of the present invention is at least one being selected from cookies, fruit jam, confection, cake.
According to a further aspect in the invention, present invention also offers foregoing medicine or food, regulate the purposes in microbial population of animal intestinal tract.Inventor finds, to the foregoing medicine of the present invention of animals administer or food, can significantly improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, the intestinal microbial population of effective adjustment animal, and then effectively prevent or treat the diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumor by the effect of intestinal microbial population.
It should be noted that, term " administration " used in this article should make broad understanding, except the drug administration of routine, to feed for can be understood as during food or to provide this food animal.
According to another aspect of the invention, present invention also offers a kind of method regulating microbial population of animal intestinal tract.According to embodiments of the invention, the method is to described animals administer black Bulbus Allii extract, foregoing medicine or food.According to embodiments of the invention, utilize method of the present invention can significantly improve by the quantity of the probiotic bacteria such as bacillus bifidus, Ruminococcus in administration animal intestinal, the intestinal microbial population of effective adjustment animal, and then effectively prevent or treat the diseases such as obesity, diabetes, hypertension, hyperlipidemia or tumor by the effect of intestinal microbial population.
In addition, according to embodiments of the invention, black Bulbus Allii extract and comprise food of the present invention or the medicine of black Bulbus Allii extract, the effect strengthening animal immunizing power is better than the action effect of the Radix Astragali of single component, oligofructose, oligochitosan or yeast dextran.
Also it should be noted that, the novelty teabag of black Bulbus Allii extract of the present invention---regulating the purposes in microbial population of animal intestinal tract, the present inventor just surprisingly finds through arduous creative work and a large amount of experimental works just.
Additional aspect of the present invention and advantage will part provide in the following description, and part will become obvious from the following description, or be recognized by practice of the present invention.
Detailed description of the invention
Embodiments of the invention are described below in detail.Embodiment described below is exemplary, only for explaining the present invention, and can not be interpreted as limitation of the present invention.Unreceipted concrete technology or condition in embodiment, (such as show with reference to J. Pehanorm Brooker etc. according to the technology described by the document in this area or condition, " Molecular Cloning: A Laboratory guide " that Huang Peitang etc. translate, the third edition, Science Press) or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, be and by the conventional products of commercial acquisition, such as, can be able to purchase from Illumina company.
Black Bulbus Allii extract of the present invention can be buied from market, or prepares according to existing routine techniques extraction on demand, does not here tire out one by one and states.Describe the preparation method of black Bulbus Allii extract below in embodiment 1, but be not limitation of the present invention.
Embodiment 1: the preparation of black Bulbus Allii extract
Raw material: fresh black Bulbus Allii
Preparation method: select fresh black Bulbus Allii of uniform size, peeling, cleans the garlic clove of peeling, and clothing film is gone to the greatest extent, with rustless steel crusher in crushing 5min.To squeeze the juice 10min with juice extractor (8000rpm), then to filter after 5min through centrifuge with 60 order screen cloths and namely obtain black Bulbus Allii extract.
Embodiment 2: the preparation of black Bulbus Allii extract capsule
Raw material: black Bulbus Allii extract, granular starch
Preparation method: take black Bulbus Allii extract 8 weight portion, granular starch 6 weight portion, research of super-pine crush equipment processing is adopted to pulverize, obtain black Bulbus Allii extract, granular starch fine powder that grain diameter is 100 microns, homogenizing becomes mixing fine powders, then content material is placed in soft capsule pellet press, fills according to existing soft capsule production technology and be pressed into soft capsule.
Embodiment 3: the preparation of black Bulbus Allii extract cookies
Raw material: flour, black Bulbus Allii extract, sweeting agent, phospholipid, flavoring agent, sodium bicarbonate, vegetable oil, milk powder, fresh hen egg, water
Preparation method: by 5 weight portion black Bulbus Allii extracts, 100 weight portion flour mix homogeneously, add 4 weight portion sugar, 0.5 weight portion phospholipid, 1.0 weight portion refined salt, 0.2 weight portion sodium bicarbonate, 8 weight portion vegetable oil, 8 weight portion milk powder, 2 weight portion fresh hen eggs and 20 weight portion drinking waters, after being mixed by above-mentioned raw materials, make black Bulbus Allii extract cookies through adjusting powder, shaping, baking process.
Embodiment 4: the preparation of black Bulbus Allii extract fruit jam
Raw material: fresh fruit, black Bulbus Allii extract, Mel, essence
Preparation method: by 10 weight portion black Bulbus Allii extracts, 50 weight portion fresh fruits, 40 weight portion Mel, 2 weight portion essence mix homogeneously, drying technology of the package makes black Bulbus Allii extract fruit jam.
Embodiment 5: the preparation of black Bulbus Allii extract confection
According to the molten sugar of the processing technology of confection, filtration, sugar cook, be in harmonious proportion other additives time add black Bulbus Allii extract, then mix, cool, molding, packaging, make the confection containing black Bulbus Allii extract.This technique is except interpolation black Bulbus Allii extract operation, and other operation way is identical with general confection (as sesame seed candy) technique.
Embodiment 6: the preparation of black Bulbus Allii extract cake
Raw material: wheat flour, vegetable oil, wetting agent, emulsifying agent, saccharide, black Bulbus Allii extract
Preparation method: by 30 weight portion vegetable oil, 1.0 weight portion wetting agents, 5 parts by weight Emulsifier, 30 parts by weight of saccharide, insufflation gas after 25 weight portion black Bulbus Allii extracts mixing, is modulated into butyrous material, then mixes with 100 weight portion wheat flours, is shaped, toasts and obtain black Bulbus Allii extract cake.
Embodiment 7: rat acute toxicity test
Specific-pathogen free (SPF) SD rat 15 in 6 week age is divided into 3 groups at random, often organizes 5.The food prepared by embodiment of the present invention 2-6 is suspended in the methocel solution of 0.5%, respectively with the dosage of 1g/kg body weight, 5g/kg body weight and 10g/kg body weight once by oral administration to 5 the SD rats often organized, observe the death of rat, clinical symptoms and body weight change, and carry out blood test and hematochemical test, when postmortem, any abnormal phenomena of macroscopy thorax abdomen gastrointestinal tissue.
Experimental result shows: black Bulbus Allii extract product of the present invention does not cause any specific clinical symptoms, body weight change or death, and blood test, hematochemical test and postmortem also do not change.Thus, due to until the dosage of 10g/kg body weight can not cause any poisoning change of rat, can judge that its median lethal dose(LD 50) (LD50) is far longer than 10g/kg body weight, black Bulbus Allii extract product of the present invention is rated as safety non-toxic material.
Embodiment 8: black Bulbus Allii extract regulates human body intestinal canal flora impact experiment
The black Bulbus Allii extract product prepared by embodiment 2-6 is evaluated according to the regulating intestinal canal flora function human experiment experimental evaluation way of Ministry of Public Health " health food inspection and assessment technical specification (2003) ".Specific as follows:
1, the grouping of test-meal experiment:
The grouping situation of test-meal experiment sees the following form 1:
Table 1 test-meal experiment grouped table
2, human experiment experimental technique
Experimenter is divided at random 5 groups, often organize 10 people, group result is as follows: group 1 is normal population, and group 2 is type ii diabetes patient, and group 3 is hyperlipemic patients, and group 4 is hyperglycemic patients, and group 5 is tumor patient.Before experimenter's test-meal, all asepticly take experimenter's feces, 16S rDNA checks order inspection intestinal microbial population, as the reference of background level.
By the grouping situation of table 1, group 1 takes black Bulbus Allii extract capsule, group 2 takes black Bulbus Allii extract cookies, group 3 takes black Bulbus Allii extract fruit jam, group 4 takes black Bulbus Allii extract confection, group 5 takes black Bulbus Allii extract cake, after 4 weeks, take experimenter's feces, 16S rDNA checks order and checks intestinal microbial population, contrast with background level, contrast the multiple that its relative abundance increases.
3, intestinal microbial population detection method
First fecal sample carries out DNA extraction, V3 ~ V5 region of pcr amplification 16SrDNA, then with 454 platform order-checkings, and order-checking direction V5->V3.Average 7795 tag of each sample initial data, read long about 400bp.The final tag number average out to 3235 for analyzing, reads long about 265bp.
16S analyzes main mothur software (http://www.mothur.org/wiki/Mothur_manual) that adopts and comprises Quality Control, OTU cluster, the analyses such as annotation, and on the basis completing data species taxonomy, analyzes the relative abundance of each species.
4, experimental result
The result of the test of the present embodiment is as shown in table 2 below.
The sequencing data abundance contrast (improving multiple compared with the background level before test-meal) of each Pseudomonas of table 2 people intestinal microbial population
The experimental result of table 2 shows, after test-meal black Bulbus Allii extract product of the present invention, the intestinal microbial population of normal population, diabetes, hyperglycemia, hyperlipidemia, tumor patient all obtains improvement in various degree, black Bulbus Allii extract product regulating intestinal canal flora Be very effective of the present invention.
Embodiment 9: the experiment of black Bulbus Allii extract enhancing immunity function
According to following steps, the research embodiment of the present invention 3 gained black Bulbus Allii extract cookies is on the impact of animal immunizing power:
1, test material
Laboratory animal: BALB/C1 monthly age secondary mouse, male and female half and half, body weight 20 ± 2g, is in a good state of health, and is provided, production licence number by Wuhan University Experimental Animal Center: SCXK(Hubei Province) 2008-0004.Often criticize animal random packet.
The recommended dose of the black Bulbus Allii extract cookies of embodiment 3 gained is mice 250mg/kgbw every day (body weight), recommends 0.5 times, 1 times and 2 times as low dose group (125mg/kgbw), middle dosage group (250mg/kgbw), high dose group (500mg/kgbw) respectively using mice.
If 4 positive controls,
Positive control 1 group is astragalus polysaccharides capsule (primary raw material: the Radix Astragali, oligomeric isomaltose, Nei Monggol Shuangqi Pharmaceutical Co., Ltd., the strong word G20040082 of state's food),
Positive control 2 groups is oligofructose oral liquid (primary raw material: oligofructose, Zhuhai Shengyuan Biotechnology Co., Ltd., the strong word G20050336 of state's food),
Positive control 3 groups is chitosan oligosaccharide capsule (primary raw material: oligochitosan, Xiamen Blue Bay Science and Technology Co., Ltd., the strong word G20110158 of state's food),
Positive control 4 groups is yeast dextran capsule (primary raw material: yeast dextran, Nanjing Potomac Beauty&Health Care Co., Ltd., the strong word G20110445 of state's food).
Other main reagent reagent is: sheep red blood cell (SRBC) (SRBC), Hank ' s liquid, guinea pig serum, prepared Chinese ink.
The wherein compound method of Hank ' s liquid:
(1) stock solution:
NaCl:80.00g, KCl:4.00g, Na
2hPO
412H
2o:1.52g, KH
2pO
4: 0.60g, glucose: 4.0g, 0.4% phenol red liquid: 50.00mL, two water that heats up in a steamer adds and is settled to 100mL, 115 DEG C of autoclaving 15min, stored frozen.
(2) working solution:
Add the two of 9 times of volumes to stock solution and heat up in a steamer water, 115 DEG C of autoclaving 15min.Face the 5.6%NaHCO of used time sterilizing
3solution adjust pH is to 7.2-7.4 (solution is orange red).
The preparation of normal saline: 9.00g NaCl is dissolved in 991g pair and heats up in a steamer in water the NaCl solution namely obtaining 0.9%.
2, test method
Administration by gavage is taked in test.Every day gavage once, blank group fills with equal volume distilled water.Each group of mice gives tested material after 30 days continuously, measures indices.
2.1 impacts on Mouse Weight, thymus/body weight, spleen/body weight
Mouse stomach de-neck execution afterwards in 30 days, weighs.Get Thymus and spleen, weigh Thymus and spleen weight respectively, measure thymus/body weight value, and spleen/body weight value.
2.2 on the impact of mouse antibodies cellulation (PFC)
The mensuration of antibody-producting cell, hemolytic plaque test is the method for the single antibody forming cell of a kind of vitro detection (plasma cell).
Lumbar injection 0.2mL2% (v/v) SRBC immunity every mice.Immunity is after 5 days, and getting mouse spleen, to make cell concentration be 5 × 10
6the splenocyte liquid of individual/mL.Dissolve the solution that agarose makes l%, in 48 DEG C of water bath heat preservations, mix with Hank ' the s liquid of equivalent 2 times of concentration, divide and be filled in small test tube, often pipe 0.5mL.Add 0.05mL10%SRBC and 0.01mL splenocyte suspension, mix rapidly, be poured on slide.Incubation 1h in 37 DEG C of calorstats, is added in complement (guinea pig serum) in slide groove, then incubation 1.5h.Counting plaque.
2.3 impacts (mensuration of serum hemolysin) on mice hemolytic antibody nucleus formation
Lumbar injection 0.2mL2% (v/v) SRBC immunity every mice.Immunity, after 5 days, is plucked eyeball and is got blood in centrifuge tube, places 1h, 3000rpm centrifugal 5 minutes, collects serum.Serum normal saline dilution 400 times, adds the mice serum 1.0mL of dilution, the SRBC0.5mL of 10% (v/v), complement (with normal saline 1:10 dilution) 1.00mL successively.Separately establish the control tube of the not increase serum replaced with normal saline.Put 37 DEG C of water-baths and set to 0 DEG C refrigerator 30 minutes stopped reactions after 20 minutes, the centrifugal 5min of 3000rpm.Get supernatant 1.0mL, measure optical density value in spectrophotometer 540nm wavelength place.
2.4 impacts on mice carbonic clearance
Every mouse tail vein injection is diluted to the prepared Chinese ink of 4 times, 0.1mL/10g body weight.Timing immediately after prepared Chinese ink injects.2.0mL0.1%Na is added to respectively within l minute, 10 minutes, getting blood 0.02mL after injection prepared Chinese ink
2cO
3in solution (i.e. 1g/L), measure optical density value in 600nm wavelength place, with Na
2cO
3solution compares.Separately get liver and spleen is weighed.The ability of mice carbonic clearance is represented with phagocytic index (α).
3, result of the test
The measurement result of 3.1 body weight, thymus/body weight, spleen/body weight
Each tested material is to Mouse Weight, thymus/body weight ratio, and the impact of spleen/body weight ratio, and measurement result is as shown in table 3,4 and 5.
The each tested material of table 3 is on the impact of Mouse Weight
As shown in Table 3, before per os gives the tested material of mice various dose, and after 30 days, basic, normal, high 3 dosage groups are compared with blank group and positive controls, body weight, all without significant difference (P>0.05), namely feeds the sample of 3 basic, normal, high dosage groups and 4 positive controls to the body weight of mice without impact to mice.
Table 4 tested material is on the impact of mouse thymus/body weight ratio
| Group | Dosage | Number of animals (only) | Thymus/body weight ratio (g/g) | P value |
| Blank group | 0 | 12 | 0.0017±0.0004 | - |
| Positive control 1 group | 1 | 12 | 0.0021±0.0003 | 0.0321 |
| Positive control 2 groups | 0.5mL | 12 | 0.0020±0.0002 | 0.0321 |
| Positive control 3 groups | 1 | 12 | 0.0021±0.0001 | 0.0352 |
| Positive control 4 groups | 1 | 12 | 0.0020±0.0006 | 0.0259 |
| Low dose group | 125mg/kg·bw | 12 | 0.0024±0.0004 | 0.0254 |
| Middle dosage group | 250mg/kg·bw | 12 | 0.0025±0.0002 | 0.0215 |
| High dose group | 500mg/kg·bw | 12 | 0.0028±0.0008 | 0.0289 |
As shown in Table 4, per os gave the tested material of mice various dose after 30 days, compare with blank group, each dosage group thymus/all there were significant differences for body weight ratio (P < 0.05), namely increases all to some extent to the feed thymus/body weight ratio of sample to mice of 3 dosage groups and 4 positive controls of mice.
Table 5 tested material is on the impact of mouse spleen/body weight ratio
| Group | Dosage | Number of animals (only) | Thymus/body weight ratio (g/g) | P value |
| Blank group | 0 | 12 | 0.0041±0.0001 | - |
| Positive control 1 group | 1 | 12 | 0.0048±0.0003 | 0.0386 |
| Positive control 2 groups | 0.5mL | 12 | 0.0049±0.0005 | 0.0425 |
| Positive control 3 groups | 1 | 12 | 0.0050±0.0005 | 0.0415 |
| Positive control 4 groups | 1 | 12 | 0.0054±0.0002 | 0.0423 |
| Low dose group | 125mg/kg·bw | 12 | 0.0055±0.0004 | 0.0302 |
| Middle dosage group | 250mg/kg·bw | 12 | 0.0059±0.0008 | 0.0332 |
| High dose group | 500mg/kg·bw | 12 | 0.0060±0.0003 | 0.0456 |
As shown in Table 5, per os gave the tested material of mice various dose after 30 days, compare with blank group, each dosage group spleen/all there were significant differences for body weight ratio (P < 0.05), namely increases all to some extent to the feed spleen/body weight ratio of sample to mice of 3 dosage groups and 4 positive controls of mice.
3.2 antibody-producting cells (PFC) measurement result
Antibody-producting cell (PFC) testing result is as shown in table 6.
Table 6 tested material is on the impact of mouse antibodies cellulation number
| Group | Dosage | Number of animals (only) | Antibody-producting cell number | P value |
| (Ⅹ±S) | ||||
| Blank group | 0 | 12 | 70±19 | - |
| Positive control 1 group | 1 | 12 | 87±13 | 0.0458 |
| Positive control 2 groups | 0.5mL | 12 | 85±18 | 0.3210 |
| Positive control 3 groups | 1 | 12 | 82±21 | 0.3321 |
| Positive control 4 groups | 1 | 12 | 85±73 | 0.2104 |
| Low dose group | 125mg/kg·bw | 12 | 96±18 | 0.0348 |
| Middle dosage group | 250mg/kg·bw | 12 | 99±13 | 0.0401 |
| High dose group | 500mg/kg·bw | 12 | 85±17 | 0.0647 |
As shown in Table 6, per os gave the tested material of mice various dose after 30 days, compare with blank group, there were significant differences (P < 0.05) for the PFC quantity of positive control 1 group and low, middle dosage group, high dose group and 3 positive controls are described on mice PFC without impact, and positive control 1 group, low dose group, middle dosage group there is appreciable impact to mice PFC.
The measurement result of 3.3 serum hemolysins
The measurement result of serum hemolysin is as shown in table 7.
Table 7 tested material is on the impact of mice serum hemolysin
| Group | Dosage | Number of animals (only) | HC 50 | P value |
| Blank group | 0 | 12 | 120±17 | - |
| Positive control 1 group | 1 | 12 | 135±24 | 0.1556 |
| Positive control 2 groups | 0.5mL | 12 | 137±20 | 0.2574 |
| Positive control 3 groups | 1 | 12 | 140±13 | 0.0325 |
| Positive control 4 groups | 1 | 12 | 130±15 | 0.2568 |
| Low dose group | 125mg/kg·bw | 12 | 162±21 | 0.0089 |
| Middle dosage group | 250mg/kg·bw | 12 | 167±18 | 0.0075 |
| High dose group | 500mg/kg·bw | 12 | 142±23 | 0.0698 |
As shown in Table 7, per os gave the tested material of mice various dose after 30 days, compared with blank group, the HC of low dose group, middle dosage group
50all there is pole significant difference (P < 0.01), the HC that positive control is 3 groups
50there were significant differences (P < 0.05), and low dose group of the present invention and middle dosage group are described, all can improve mice serum hemolysin HC significantly in pole
50, positive control 3 groups can significantly improve mice serum hemolysin HC
50, and positive control 1 group, positive control 2 groups, positive control 4 groups, high dose group are to mice serum hemolysin HC
50without impact.
3.4 mice carbonic clearance experimental results
Mice carbonic clearance experimental result is as shown in table 8.
Table 8 tested material is on the impact of mice carbonic clearance
| Group | Dosage | Number of animals (only) | Phagocytic index α (Ⅹ ± S) | P value |
| Blank group | 0 | 12 | 3.19±0.21 | - |
| Positive control 1 group | 1 | 12 | 4.17±0.39 | 0.0349 |
| Positive control 2 groups | 0.5mL | 12 | 4.19±0.41 | 0.0321 |
| Positive control 3 groups | 1 | 12 | 3.82±0.23 | 0.2100 |
| Positive control 4 groups | 1 | 12 | 3.98±0.45 | 0.5236 |
| Low dose group | 125mg/kg·bw | 12 | 4.00±0.21 | 0.0325 |
| Middle dosage group | 250mg/kg·bw | 12 | 4.53±0.21 | 0.0072 |
| High dose group | 500mg/kg·bw | 12 | 4.75±0.49 | 0.0086 |
As shown in Table 8, per os gave the tested material of mice various dose after 30 days, compare with blank group, middle dosage group of the present invention, high dose group have pole significant difference (P<0.01) to mouse macrophage phagocytic activity, low dose group, positive control 1 group, positive control 2 groups have significant difference (P<0.05) to mouse macrophage phagocytic activity, remaining group on mice carbonic clearance ability all without impact.This shows that black Bulbus Allii extract product of the present invention has the effect strengthening mononuclear-macrophage phagocytic function, can strengthen the non-specific immunity of mice.
Before and after the experiment being improved immunity function by the black Bulbus Allii extract product described in the invention described above, result shows, each batch of Mouse Weight, thymus/body weight ratio, spleen/body weight ratio are all without significant difference.In the experiment of mouse antibodies cellulation test experience, serum hemolysin determination experiment and mice carbonic clearance, its result has significant difference.Thus, can think that black Bulbus Allii extract product of the present invention has the function and efficacy of enhancing immunity, and the effect of black Bulbus Allii extract product of the present invention to enhancing immunity is better than the action effect of the Radix Astragali of single component, oligofructose, oligochitosan, yeast dextran.
Embodiment 10: the hypoglycemic zoopery of black Bulbus Allii extract
According to the auxiliary hyperglycemic function zoopery evaluation method of Ministry of Public Health " health food inspection and assessment technical specification (2003) ", embodiment 3-6 gained black Bulbus Allii extract product is evaluated.Specific as follows:
1, test method: get male SD rat 40, prepares low dose of streptozotocin according to standard method and causes diabetes rat model, be divided into 4 groups at random after three days according to rat blood sugar value, often organizes 10.Each group of continuous gastric infusion 8 days, every day 1 time, dosage is 4.5mg garlicin/kg, and last two hours after administration gets blood, measures blood glucose value.
2, subjects: tested SD rat is divided into 4 groups at random, often organize 10, tested SD rat primary diet control and movable constant, group 1 takes black Bulbus Allii extract cookies, group 2 takes black Bulbus Allii extract cake, group 3 takes black Bulbus Allii extract fruit jam, group 4 takes black Bulbus Allii extract confection.
3, experimental result
The hypoglycemic result of different experiments group is as shown in table 9.
Table 9 different experiments group blood sugar decreasing effect compares
Note: (1) compares * P<0.01 with blank group; (2) compare with blank group
△p<0.05
The animal test results of table 9 shows, black Bulbus Allii extract product of the present invention all has decline effect to the hyperglycemia of Experimental Diabetes of Mice, show black Bulbus Allii extract product of the present invention and there is hypoglycemic function, to diabetes, there is significant auxiliary therapeutic action.
Embodiment 11: the zoopery of black Bulbus Allii extract blood fat reducing
According to the auxiliary lipid-lowering function zoopery evaluation method of Ministry of Public Health " health food inspection and assessment technical specification (2003) ", embodiment 3-6 gained black Bulbus Allii extract product is evaluated.Specific as follows:
1, test material and method:
Wistar male rat 40, body weight 200 ± 20g, is provided by Wuhan University's Experimental Animal Center.Bring out rat hyperlipidemia pathological model formula (high lipid food): in normal feedstuff, add 1% cholesterol, 10% egg yolk, 10% Adeps Sus domestica, dry in bulk.
The preparation method of Hyperlipemia model rat: Wistar male rat is divided at random 4 groups (often organizing 10), i.e. hyperlipemia rat (hyperlipidemia model control group), to feed high lipid food (based on high lipid food formula feedstuff 93.8%, cholesterol 1.0%, Adeps Sus domestica 5.0%, cholate 0.2%), do not give tested material;
Medication:
Tested Hyperlipemia model rat is divided into 4 groups at random, often organize 10, the former diet control of tested Hyperlipemia model rat and movable constant, group 1 takes black Bulbus Allii extract cookies, group 2 takes black Bulbus Allii extract cake, group 3 takes black Bulbus Allii extract fruit jam, group 4 takes black Bulbus Allii extract confection.
All animals are all raised 20 days under equivalent environment, and dosage is 4.5mg garlicin/kg.After last gives the 4h of each tested material, get hematometry serum total cholesterol (TC mmol/L, iron chloride development process), serum triacylglycerol (TG mmol/L, acetylacetone method), HDL-C (HDL-C mmol/L, phosphotungstic acid method).
2, experimental result
The content of each group of rat blood serum total TG, TC and HDL-C is in table 10.
The effectiveness comparison of TC, TG, HDL-C falls in table 10 different experiments group
Note: (1) * represents P<0.05; (2) * * represents P<0.01
The experimental result of table 10 shows: test-meal is after 30 days, take the cholesterol of the rat of black Bulbus Allii extract product prepared by embodiment of the present invention 3-6, triglyceride and comparatively all have obvious reduction before test-meal, high density lipoprotein comparatively has obvious rising before test-meal, shows that black Bulbus Allii extract product of the present invention has significant auxiliary therapeutic action to hyperlipemia.
Embodiment 12: the human experiment experiment of black Bulbus Allii extract blood sugar lowering, blood fat reducing
According to auxiliary hyperglycemic, the hypolipemic function human experiment experimental evaluation way of Ministry of Public Health " health food inspection and assessment technical specification (2003) ", embodiment 2-6 gained black Bulbus Allii extract product is evaluated.Specific as follows:
1, human experiment and test method:
Select 50 routine type Ⅱdiabetes mellitus people, wherein male 35 people by the principle of voluntariness, women 15 people, the range of age 43-75 year, without complication such as severe cardiac Liver and kidney, contrast design after taking in before test-meal.
Experimenter is divided into 5 groups at random, often organize 10 people, experimenter's former medicine kind, diet control and activity are constant, group 1 takes black Bulbus Allii extract capsule, group 2 takes black Bulbus Allii extract cookies, group 3 takes black Bulbus Allii extract cake, group 4 takes black Bulbus Allii extract fruit jam, group 5 takes black Bulbus Allii extract confection, daily 1 time, in one after each meal, take 30 days continuously, dosage is 0.5mg garlicin/kg.
The change of monitoring patient blood pressure, defecation, body weight, and the blood glucose (with glucose oxidase method) measuring empty stomach and 2 hours after the meal; Measure blood fat (T-CHOL TC, triglyceride TG and high density lipoprotein HDL-C).
2, judgment criteria:
Effective: fundamental symptoms disappears, the blood glucose of empty stomach or after the meal 2h is comparatively treated front decline and is no more than 30.0%.
Effective: fundamental symptoms obviously improves, the blood glucose of empty stomach and after the meal 2h is comparatively treated front decline and is no more than 10.0%.
Invalid: fundamental symptoms is not improved, the blood glucose of empty stomach and after the meal 2h is comparatively treated front decline and is less than 10.0%.
3, human experiment result:
Test-meal, after 30 days, takes the patient of each black Bulbus Allii extract product, compared with before test-meal, and the blood pressure of patient, defecation, body weight there are no significant difference, but fasting glucose, 2h-plasma glucose all have obvious attenuating, and concrete outcome is in table 11.
Impact about on the blood fat before and after test-meal: the patient taking each black Bulbus Allii extract product, compared with before test-meal, the cholesterol of test-meal patient after 30 days, triglyceride comparatively all have obvious reduction before test-meal, and high density lipoprotein comparatively has obvious rising before test-meal, and concrete outcome is in table 12.
As can be seen here, black Bulbus Allii extract product of the present invention has the health care of obvious auxiliary hyperglycemic, blood fat reducing, and to the harmless effect of tested population health.
Table 11 foretastes crowd's blood glucose test results
Table 12 foretastes crowd's lipids detection result
Note: (1) * represents P<0.05; (2) * * represents P<0.01
Embodiment 13: black Bulbus Allii extract antitumor animal is tested
According to the adjunct antineoplastic function zoopery evaluation method of Ministry of Public Health " health food inspection and assessment technical specification (2003) ", embodiment 3-6 gained black Bulbus Allii extract product is evaluated.Specific as follows:
1, test material and method:
BALB/c mouse (4-6 age in week, male, body weight 20-24g) 50, is seeded in the right axil of BALB/c mouse by the hepatoma cells MM45T.Li of exponential phase subcutaneous, every only inoculation 5 × 10
7individual cell, sets up Transplanted tumor model.
Medication:
Tested rat model is divided into 5 groups at random, often organize 10, former diet control and movable constant, group 1 takes black Bulbus Allii extract cookies, group 2 takes black Bulbus Allii extract cake, group 3 takes black Bulbus Allii extract fruit jam, group 4 takes black Bulbus Allii extract confection, group 5 takes normal saline, successive administration 30 days, dosage is 4.5mg garlicin/kg.Observe ordinary circumstance and tumor size change before and after mice treatment.
Become after tumor and within every 2 days, measure the most major diameter (a) of 1 tumor and most minor axis (b) with slide gauge, obtain tumor body approximate volumes (TV: volume) by formula TV=a × b2/2.Evaluate medicine tumor-inhibiting action by the following method: the animal of putting to death for the 2nd day in last administration takes tumor weight, and presses following formula:
Calculate medicine tumour inhibiting rate (IR): the average tumor of tumor growth tumour inhibiting rate=(matched group average tumor weight-black Bulbus Allii extract product group average tumor weight)/matched group heavy × 100%.Evaluate Antitumor Activity of Drugs effect with inhibition rate of tumor growth IR, the standard of curative effect evaluation: IR < 30 is invalid, IR >=30 are effective.
Result of the test sees the following form 13.
The effectiveness comparison of table 13 different experiments group Tumor growth inhibition
The result of the test of table 13 shows: compared with matched group, take the rat of black Bulbus Allii extract product of the present invention, test-meal after 30 days medicine tumour inhibiting rate all have obvious reduction, show that black Bulbus Allii extract product of the present invention has significant auxiliary therapeutic action to antitumor.
In the description of this description, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present invention or example.In this manual, identical embodiment or example are not necessarily referred to the schematic representation of above-mentioned term.And the specific features of description, structure, material or feature can combine in an appropriate manner in any one or more embodiment or example.
Although illustrate and describe embodiments of the invention, those having ordinary skill in the art will appreciate that: can carry out multiple change, amendment, replacement and modification to these embodiments when not departing from principle of the present invention and aim, scope of the present invention is by claim and equivalents thereof.
Claims (10)
1. black Bulbus Allii extract is regulating the purposes in microbial population of animal intestinal tract.
2. purposes according to claim 1, is characterized in that, described black Bulbus Allii extract is carried out administration with the dosage of 0.3-1.0mg garlicin/kg to described animal,
Optionally, described black Bulbus Allii extract is carried out administration with the dosage of 0.5mg garlicin/kg to described animal.
3. black Bulbus Allii extract is preparing the purposes in medicine, and described medicine is for regulating microbial population of animal intestinal tract.
4. purposes according to claim 3, is characterized in that, described medicine is used for enhancing immunity, prevention or treatment obesity, diabetes, hypertension, hyperlipidemia or tumor,
Optionally, every day is 0.3-1.0mg garlicin/kg to the dosage of described animal, preferred 0.5mg garlicin/kg.
5. a medicine, is characterized in that, comprises:
Black Bulbus Allii extract; And
Pharmaceutically acceptable excipient.
6. medicine according to claim 5, is characterized in that, described excipient is at least one being selected from binding agent, filler, film-coating polymer, plasticizer, fluidizer, disintegrating agent and lubricant,
Optionally, described medicine in the form of at least one being selected from capsule, pill, tablet, granule, liquid oral, oral pastes, aerosol and spray,
Optionally, the dosage of described medicine is 0.3-1.0mg garlicin/kg, preferred 0.5mg garlicin/kg,
Optionally, described medicine for regulating microbial population of animal intestinal tract,
Optionally, described medicine is used for enhancing immunity, prevention or treatment obesity, diabetes, hypertension, hyperlipidemia or tumor.
7. a food, is characterized in that, comprises:
Black Bulbus Allii extract; And
Acceptable additive in bromatology.
8. food according to claim 7, is characterized in that, described food is at least one being selected from cookies, fruit jam, confection and cake.
9. the medicine described in claim 5 or 6 or the food described in claim 7 or 8, regulating the purposes in microbial population of animal intestinal tract.
10. regulate a method for microbial population of animal intestinal tract, it is characterized in that, to the medicine described in described animals administer black Bulbus Allii extract, claim 5 or 6 or the food described in claim 7 or 8.
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| CN104886428A (en) * | 2015-06-17 | 2015-09-09 | 金乡县大蒜研究所 | Black garlic cake and preparation method thereof |
| CN108902970A (en) * | 2018-06-27 | 2018-11-30 | 安徽瑞康食品生物科技有限公司 | Edible composition containing pentosan, functional food and preparation method thereof, purposes |
| CN111789256A (en) * | 2020-07-30 | 2020-10-20 | 海南北纬十八度食品加工有限公司 | Pitaya flower polysaccharide composition with insulin balancing effect |
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| CN103169041A (en) * | 2011-12-21 | 2013-06-26 | 郑州固元堂医药科技有限公司 | Fermented black garlic powder production method |
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| CN103169041A (en) * | 2011-12-21 | 2013-06-26 | 郑州固元堂医药科技有限公司 | Fermented black garlic powder production method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104886428A (en) * | 2015-06-17 | 2015-09-09 | 金乡县大蒜研究所 | Black garlic cake and preparation method thereof |
| CN108902970A (en) * | 2018-06-27 | 2018-11-30 | 安徽瑞康食品生物科技有限公司 | Edible composition containing pentosan, functional food and preparation method thereof, purposes |
| CN111789256A (en) * | 2020-07-30 | 2020-10-20 | 海南北纬十八度食品加工有限公司 | Pitaya flower polysaccharide composition with insulin balancing effect |
| CN111789256B (en) * | 2020-07-30 | 2022-12-30 | 海南北纬十八度食品加工有限公司 | Pitaya flower polysaccharide composition with insulin balancing effect |
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