CN108991505A - A kind of blueberry local tyrant and its application in antitumor - Google Patents
A kind of blueberry local tyrant and its application in antitumor Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses a kind of blueberry local tyrants, using blueberry as major ingredient, add local tyrant's compound bacteria, sugar, lemon, blueberry yeast cream, obtain after repeatedly fermenting.The present invention is fermented using different proportion flora, sufficiently discharges blueberry nutrient ingredient, and keep unique activity.Using mtt assay detection blueberry local tyrant to the fragmentation effect of human lung carcinoma cell, experiments have shown that blueberry local tyrant significantly inhibits the cell activity of non-small cell lung cancer cell H1299, lung adenocarcinoma cell H1975, to illustrate blueberry local tyrant with effect of anti-lung cancer.
Description
Technical field
The present invention relates to health product technology field, in particular to a kind of blueberry local tyrant and its application in antitumor.
Background technique
Blueberry is a kind of very high fruit pulp of nutritive value rich in the battalion such as vitamin abundant, protein and minerals
Element is supported, wherein the anthocyanin pigments of unique preciousness specific gravity in many fruit is unusual height in blueberry, and there are also rich in blueberry
Containing nutrients such as protein abundant, dietary fiber, fat, vitamins, the nutrient that these in blueberry are rich in all is higher than
The nutrition that other fruit are rich in.It also include the trace quantity mineral substance elements such as calcium abundant, iron, phosphorus, potassium, zinc, these mineral in blueberry
Element there are ratios to be apparently higher than other fruit.In addition vitamin C abundant has pre- anti-cancer in blueberry, resists heart disease
The effect of.Blueberry pectin is abundant, can dilute body fat, protects the health of human body cardiovascular and cerebrovascular.In addition, there are also rich in blueberry
Rich antioxidant, can be with the aging of delaying human body.
" local tyrant " is also known as " meat ganoderma lucidum ", is the 4th kind of life form of non-plant, non-animal and non-mushroom in nature.
" local tyrant " being commonly called as is the slime bacteria between protozoa and fungi, is moved in soil, and vitality is extremely strong, is that nature is non-
Often rare large-scale slime bacteria complex.Belong to " super-huge rare Acarasiales complex ", the characteristics of existing protist, also has true
The characteristics of bacterium, be organism living, and the world is rare.Local tyrant's this " Acarasiales complex " is the most ancient extinct plants and animal found so far
Living specimen.Local tyrant has the function of breathing, repairs itself, and fructification and bacterium solution are rich in organic germanium, nucleic acid, polysaccharide, dimension
The nutritional ingredients such as raw element and amino acid, lecithin, coenzyme.Its effective bioactive ingredients is in anticancer, antibacterial, cytotoxicity, life
The effect of reason regulation etc. is also gradually exploited.
Currently, the preparation method of traditional blueberry ferment is that beneficial bacterium is not added, spontaneous fermentation.But time-consuming for this method,
Protein content is low and lacks beneficial microbe.The purpose of the present invention is increasing local tyrant's compound bacteria on the basis of original, make blueberry
In albumen preferably resolve into peptide fragment, the absorption of human body not only can be promoted, increased beneficial to bacterial content and hair can be shortened
The ferment time.Since local tyrant is Acarasiales complex, although comprising mushroom do not study completely also clear, we are from local tyrant's body
Interior separation has cultivated one group of good microbial flora and has carried out repeatedly recombination screening using modern biotechnology, cultivates
One group of beneficial composite flora.Therefore combine the composite flora isolated with blueberry fermentation, a kind of new method is provided, and visit
Effect of the blueberry fermentation liquid to lung carcinoma cell is studied carefully.
Summary of the invention
In view of above-mentioned shortcoming, the purpose of the present invention is to provide a kind of blueberry local tyrants and preparation method thereof, pass through use
Blueberry local tyrant prepared by the present invention can inhibit lung carcinoma cell activity.
In order to achieve the above object, the invention adopts the following technical scheme:
A kind of blueberry local tyrant is by being made following preparation method:
(1) it prepares the fresh slurry of blueberry: selecting full, fresh blueberry fresh fruit, drain away the water after cleaning, be placed in abundant powder in pulverizer
The fresh slurry of blueberry is obtained after broken；
(2) prepare blueberry yeast cream: by the fresh slurry of blueberry be placed in watt cylinder it is quiet put 12h after, by the fresh slurry of blueberry: reddle is sugared: lemon: too
Year compound bacteria mass ratio 1:0.5:0.01:0.3 ratio addition reddle sugar, lemon, local tyrant's compound bacteria, must mix after being sufficiently stirred
Object is stood in the dry environment in cool place, and then the stirring of the every two moon is primary, every time addition mixture gross mass 2% before stirring
Local tyrant's compound bacteria obtains blueberry yeast cream after 12 months full；
(3) it is mixed: blueberry fresh fruit, local tyrant's compound bacteria, sugar, lemon, the blueberry yeast for being 100:5:5:1:50 by mass ratio
Slurry is placed in Wa Gangzhong, stirs evenly, and covers, reserves gas vent, and stirring is primary every three days；
(4) repeatedly fermentation: after two weeks, then 1% local tyrant's compound bacteria of step (3) gains gross mass, 10% blueberry ferment are added
Mother's slurry, 10% the fresh slurry of blueberry stir evenly, cover, reserve gas vent, stirring is primary weekly；
(5) blueberry local tyrant is obtained after after continuing half a year according to step (4) method, sealing, displaying 1 year.
Further, local tyrant's compound bacteria is by local tyrant's beneficial flora, Bifidobacterium, bacillus, saccharomycete, acetic acid bacteria
It is constituted according to the mass ratio of 0.35:0.25:0.15:0.15:0.1.
Further, local tyrant's beneficial flora includes Acarasiales and the branch acremoniums such as wart spore calcium skin bacterium, the double skin bacterium of flat pad
Category, trichoderma, Penicillium, aspergillus, Nectria, Gibberella, saccharomyces, Cladosporium, Stachybotrys, stone seat Pseudomonas and
The fungies such as broom Thamnidium.
Further, the watt cylinder buries 1/3 in underground.
Further, the mixing time is 4 hours.
The invention has the following advantages:
1, blueberry local tyrant be as addition local tyrant's composite flora after brew gained, for for the first time utilize local tyrant's composite flora to blueberry into
Row fermentation.
2, blueberry local tyrant is to be repeated several times to ferment and obtain according to traditional ancient method fermentation technique, can sufficiently discharge blueberry and support
Point.
3, it first demonstration that produced by the present invention have the function of inhibiting proliferation of lung cancer cells, is opened for blueberry mixed health product
Hair provides new thinking.
Detailed description of the invention
Fig. 1 is the inhibitory effect that blueberry local tyrant of the present invention grows Non-small cell lung carcinoma cell-H1299 cell.
Fig. 2 is the inhibitory effect that blueberry local tyrant of the present invention grows human lung adenocarcinoma cell-H1975 cell.
Specific embodiment
Embodiment
The preparation of blueberry local tyrant:
(1) it prepares the fresh slurry of blueberry: selecting full, fresh blueberry fresh fruit, drain away the water after cleaning, be placed in abundant powder in pulverizer
The fresh slurry of blueberry is obtained after broken；
(2) it prepares blueberry yeast cream: preparing watt cylinder, watt cylinder is buried 1/3 in underground, the fresh slurry of blueberry is then placed in quiet in watt cylinder put
After 12h, in the fresh slurry of blueberry: reddle sugar: lemon: reddle sugar, lemon is added in local tyrant's compound bacteria mass ratio 1:0.5:0.01:0.3 ratio
Lemon, local tyrant's compound bacteria obtain mixture after being sufficiently stirred 4 hours, stand in the dry environment in cool place, then the every two moon stirs
Once, local tyrant's compound bacteria of mixture gross mass 2% is added before stirring every time, obtains blueberry yeast cream after 12 months full；Local tyrant is multiple
Combined bacteria is by local tyrant's beneficial flora, Bifidobacterium, bacillus, saccharomycete, acetic acid bacteria according to 0.35:0.25:0.15:0.15:0.1
Mass ratio constitute, local tyrant's beneficial flora includes wart spore calcium skin bacterium, flat pads Acarasiales and the acremoniums, trichoderma such as double skin bacterium
Category, Penicillium, aspergillus, Nectria, Gibberella, saccharomyces, Cladosporium, Stachybotrys, stone seat Pseudomonas and broom branch are mould
The fungies such as category.
(3) it is mixed: preparing watt cylinder, watt cylinder is buried 1/3 in underground, blueberry that mass ratio is 100:5:5:1:50 is fresh
Fruit, local tyrant's compound bacteria, sugar, lemon, blueberry yeast cream are placed in Wa Gangzhong, are sufficiently stirred 4 hours, and capping reserves gas vent, and every three
Its stirring is primary；
(4) repeatedly fermentation: after two weeks, then 1% local tyrant's compound bacteria of step (3) gains gross mass, 10% blueberry ferment are added
Mother's slurry, 10% the fresh slurry of blueberry stir evenly, cover, reserve gas vent, stirring is primary weekly；
(5) blueberry local tyrant is obtained after after continuing half a year according to step (4) method, sealing, displaying 1 year.
Growth, living environment according to microorganism, the environment that yeasting selects air clear, clean.
The watt cylinder that watt cylinder can choose tradition dress wine is fermented (its temperature change is small, is protected from light), buries cylinder body 1/3 in ground
Under.
Test example 1
The culture of lung carcinoma cell
1, material and method
1.1 material
Bacterial strain: Non-small cell lung carcinoma cell-H1299 cell, lung adenocarcinoma cell-H1975 cell (are ground purchased from Shanghai life science
Study carefully institute's cell bank)
Culture medium: Dulbecco ' s modified Eagle ' s medium (DMEM) culture solution adds the calf of 10% (v/v)
Serum
1.2 method
(1) cell suspension growth is in the DMEM culture solution containing 10% calf serum, in 37 DEG C, saturated humidity, 5% CO2Culture
It is cultivated in case.
(2) 2~3d change 1 culture solution.
(3) cell of logarithmic growth phase is collected for testing.
Test example 2
Inhibiting effect of the blueberry local tyrant to Non-small cell lung carcinoma cell-H1299 cell, lung adenocarcinoma cell-H1975 cell Proliferation
1, material and method
1.1 material
Non-small cell lung carcinoma cell-H1299 cell, lung adenocarcinoma cell-H1975 cell in logarithmic phase: embodiment 2 is used
The method obtains.
1.2 method
(1) with 2 × 104A/mL cell concentration, by tumor cell inoculation in 96 orifice plates, every 100 μ L cell suspension of hole is (every
Group sets 3 multiple holes), culture is for 24 hours.
(2) using not celliferous DEME culture solution as blank control group, the indigo plant of different volumes is added into the cell of culture
Certain kind of berries local tyrant, while cultivating respectively for 24 hours.
(3) every group of 10 μ L of addition, 5 mg/mL MTT continue after cultivating 4h behind every hole, absorb supernatant, 100 μ are added
The DMSO of L, gently concussion promotes the dissolution of crystallization.
(4) microplate reader (under 492nm wavelength) measurement absorbance value (A), replication 3 times.Observe result.
As shown in Figure 1, the inhibiting effect that blueberry local tyrant of the invention grows non-small cell lung cancer cell-H1299 cell
With the being positively correlated property of volume that blueberry local tyrant is added, wherein inhibiting rate reaches 100% when >=7 μ L are handled for 24 hours；
As shown in Fig. 2, inhibiting effect that blueberry local tyrant of the invention grows lung adenocarcinoma cell-H1975 cell and blueberry is added
The being positively correlated property of volume of local tyrant, wherein inhibiting rate reaches 90% or more when 2 μ L are handled for 24 hours.
The present invention is experimentally confirmed: blueberry local tyrant obtained through the invention has the activity for inhibiting lung carcinoma cell.It is real
Test the result shows that:
(1) inhibiting effect and addition blueberry local tyrant produced by the present invention to the growth of non-small cell lung cancer cell-H1299 cell
Being positively correlated property of volume, wherein >=7 μ L processing for 24 hours when inhibiting rate reach 100%；
(2) inhibiting effect produced by the present invention to the growth of lung adenocarcinoma cell-H1975 cell and the volume that blueberry local tyrant is added
Being positively correlated property, wherein inhibiting rate reaches 90% or more when 2 μ L are handled for 24 hours.
The above content is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be said that
Specific implementation of the invention is only limited to these instructions.For the technical field of the invention, present inventive concept is not being departed from
Under the premise of make several equivalent substitute or obvious modifications, and performance or use is same or like, all shall be regarded as belonging to this hair
Bright scope of patent protection determined by the appended claims.
Claims (9)
1. a kind of blueberry local tyrant, which is characterized in that blueberry local tyrant is by being made following preparation method:
(1) it prepares the fresh slurry of blueberry: selecting full, fresh blueberry fresh fruit, drain away the water after cleaning, be placed in abundant powder in pulverizer
The fresh slurry of blueberry is obtained after broken；
(2) prepare blueberry yeast cream: by the fresh slurry of blueberry be placed in watt cylinder it is quiet put 12h after, by the fresh slurry of blueberry: reddle is sugared: lemon: too
Year compound bacteria mass ratio 1:0.5:0.01:0.3 ratio addition reddle sugar, lemon, local tyrant's compound bacteria, must mix after being sufficiently stirred
Object is stood in the dry environment in cool place, and then the stirring of the every two moon is primary, every time addition mixture gross mass 2% before stirring
Local tyrant's compound bacteria obtains blueberry yeast cream after 12 months full；
(3) it is mixed: blueberry fresh fruit, local tyrant's compound bacteria, sugar, lemon, the blueberry yeast for being 100:5:5:1:50 by mass ratio
Slurry is placed in Wa Gangzhong, stirs evenly, and covers, reserves gas vent, and stirring is primary every three days；
(4) repeatedly fermentation: after two weeks, then 1% local tyrant's compound bacteria of step (3) gains gross mass, 10% blueberry ferment are added
Mother's slurry, 10% the fresh slurry of blueberry stir evenly, cover, reserve gas vent, stirring is primary weekly；
(5) blueberry local tyrant is obtained after after continuing half a year according to step (4) method, sealing, displaying 1 year.
2. blueberry local tyrant according to claim 1, it is characterised in that: local tyrant's compound bacteria is by local tyrant's beneficial flora, double
Discrimination bacillus, bacillus, saccharomycete, acetic acid bacteria are constituted according to the mass ratio of 0.35:0.25:0.15:0.15:0.1.
3. blueberry local tyrant according to claim 1, it is characterised in that: local tyrant's beneficial flora include wart spore calcium skin bacterium,
It is flat pad the Acarasiales such as double skin bacterium and acremonium, trichoderma, Penicillium, aspergillus, Nectria, Gibberella, saccharomyces,
The fungies such as Cladosporium, Stachybotrys, stone seat Pseudomonas and broom Thamnidium.
4. blueberry local tyrant according to claim 1, it is characterised in that: the watt cylinder buries 1/3 in underground.
5. blueberry local tyrant according to claim 1, it is characterised in that: the mixing time is 4 hours.
6. a kind of application of blueberry local tyrant as described in claim 1-5 any one in antitumor.
7. application according to claim 6, which is characterized in that the tumour is lung cancer.
8. application according to claim 7, which is characterized in that the lung cancer is non-small cell lung cancer.
9. application according to claim 8, which is characterized in that the non-small cell lung cancer is lung cancer H1299 cell and lung
Cancer H1975 cell.
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