CN109266567A - The microbial fermenters and its preparation method and application of acquisition are cultivated under music environment - Google Patents
The microbial fermenters and its preparation method and application of acquisition are cultivated under music environment Download PDFInfo
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Abstract
The present invention relates to a kind of microbial fermenters and its preparation method and application, the microbial fermenters are that vegetable juice and certain kind of berries fruit juice mixed liquor ferment via streptococcus thermophilus, saccharomycete and acetic acid bacteria in the environment of playing music and obtained;Microbial fermenters of the invention have the purposes in terms of being used to prepare the drug aspect for promoting fat cell metabolism or preparing liver-protective drug.
Description
Technical field
The present invention provides a kind of microbial fermenters that acquisition is cultivated under music environment, and preparation method thereof, and with
Purposes in terms of the drug that preparation promotes fat cell metabolism or in terms of preparing liver-protective drug.
Background technique
For a long time, obesity is defined as and most of health problem and disease symptom related with the reduction service life.
The modern times of health are laid siege in people, gradually appear the complementary goods of miscellaneous nutrition, weight reducing on the market to meet
The demand of people;However, these nutrition, weight reducing complementary goods are mostly single by chemical synthesis or with what is be not present in natural food
Molecular structure is combined into, raw material sources be all Petroleum refining object, coal pitch derived object, chemical process treated sugar or place
Industrialization product after reason, but nutrient made of these chemistry refinements is to be difficult to be recognized, receive and utilize by cell, because
This will actively be excreted with urine or sweat more than nutrient.Due to the low absorptivity of those nutriments, it is low in vivo
Residence time, low bioavailability, chemical synthesis nutrient not up to declare that effect, content may be daily suggestion intakes
Even thousand times or more of hundred times, high dose would be possible to bring the risk for generating toxic reaction for human body;Although body can be actively
Extra nutrient is discharged, but also illustrates that unabsorbed chemotrophy element is that liver kidney is needed to be metabolized, will increase human body additional negative
Load;And chemical synthesis nutrient, which is eaten for a long time, cumulative toxicity and to generate side effect, or even has carcinogenic risk.
Therefore, how by the nutritional ingredient that produces from natural food materials, the change for having too big in life style is not needed
And toxin will not be caused or have undesirable influence to health, while health of body health can be reached again and reduce fat dual function
Effect is the target that people's long-term endeavour is pursued.
Summary of the invention
In order to solve the above technical problems, one aspect of the present invention provides a kind of microbial fermenters, the microbial fermentation
Object is that the mixed liquor of vegetable juice and certain kind of berries fruit juice mixes in the environment of playing music with streptococcus thermophilus, saccharomycete and acetic acid bacteria
Post-fermentation obtains.
Preferably, the music environment is the environment for playing classic melody, and the classic melody sound intermediate frequency is lower than 10,
The part of 000Hz accounts for 99% or more of whole first song, and the allusion melody duration is 1 minute to 3 minutes;Alternatively, to broadcast
The environment of pop is put, and the pop sound intermediate frequency accounts for 95% or more of whole first song lower than the part of 10,000Hz,
The pop duration lasts time is 1 minute to 3 minutes;Alternatively, to play the environment of rock and roll melody, and described shake
Rolling melody sound intermediate frequency is higher than 50% or more that 10,000Hz accounts for whole first song, and the rock and roll melody duration is 1 minute to 3
Minute.
Another aspect of the present invention provides the preparation method of mentioned microorganism fermentation material,
The preparation method includes the following steps:
Step 1, the empty training liquid of preparation: the vegetable juice of fresh squeezing and the certain kind of berries fruit juice of fresh squeezing are mixed with the volume ratio of 1:1~2:1 first
It closes, then with after 3~6 times of dilution of distilled water (volumes), carries out sterilization treatment, obtain the empty training liquid;
Step 2, it is inoculated with strain: by streptococcus thermophilus (Streptococcus thermophilus), S. cervisiae
(Saccharomyces cerevisiae) and acetic acid bacteria (Acetobacter aceti) are inoculated into described empty with liquid, three kinds of bacterium
The weight ratio of kind is 1:1:1;The total volume of the streptococcus thermophilus, saccharomycete and acetic acid bacteria and the vegetable juice and certain kind of berries fruit juice
The ratio of the volume of mixed liquor is 2:100~7:100;
Step 3, ferment: in the environment of playing music, temperature condition is ferment at constant temperature 150~260 at 25 DEG C~35 DEG C
Hour is to obtain microbial fermenters of the invention.
Preferably, the vegetable juice of the fresh squeezing is isometric proportioning concentration of broccoli, celery and asparagus;It is described fresh
The certain kind of berries fruit juice of squeezing be blueberry, Cranberry, red grape, white grape, red bayberry, mulberries, apple, carrot, sugarcane, passion fruit, pineapple and
Isometric proportioning concentration of lemon.
The microorganism that further aspect of the present invention provides mentioned microorganism fermentation material or above-mentioned preparation method prepares
Fermentation material is used to prepare the purposes in terms of the drug for promoting fat cell metabolism.
Preferably, the promotion fat cell metabolism is to promote lipolysis or promotion brown fat cell disintegration.
Preferably, the promotion lipolysis includes to increase Adipose trigtyceride lipase (adipose
Triglyceride lipase, ATGL) and hormone-sensitive lipase (hormone-sensitive lipase, LIPE)
Gene expression amount.
Preferably, the acceleration brown fat cell disintegration includes to increase fatty uncoupling proteins 1 (uncoupling
Protein1, UCP1) and fatty uncoupling protein-3 (UCP2) gene expression amount.
The microorganism that further aspect of the present invention provides mentioned microorganism fermentation material or above-mentioned preparation method prepares
Fermentation material is used to prepare the purposes in terms of liver-protective drug.
Preferably, the protection liver is to improve liver cell oxidation resistance or DNA repair ability.
Preferably, the DNA repair ability system of improving refers to the repair ability for improving DNA structure damage or improves DNA
The repair ability of bifilar fracture.
Preferably, the raising liver cell oxidation resistance includes to increase by 1 (superoxide of superoxide dismutase
Dismutase1, SOD1) and superoxide dismutase 2 (SOD2) gene expression amount, and improve the DNA structure damage repair
Reactivation power includes to increase excision to repair (the excision repair cross-complementing gene of complementary chiasma gene 1
1, ERCC1) and DNA damage cognition and reparative factor (DNA damage recognition and repair factor,
XPA gene expression amount), and the repair ability for improving DNA double stock fracture includes to increase x-ray to repair 1 (X- of Cross-complementing Gene
Ray repair cross complementing 1, XRCC1) and x-ray repair Cross-complementing Gene 2 (XRCC2) gene
Expression quantity.
Further aspect of the present invention provides a kind of pharmaceutical composition, which includes above-mentioned microbial fermenters
Or the microbial fermenters that such as above-mentioned preparation method prepares.
Detailed description of the invention
Fig. 1 is the content of the Like superoxide dismutase SOD-like of the microbial fermenters of the embodiment of the present invention 1;
Fig. 2 is superoxide dismutase 2 (SOD2) of the microbial fermenters to gastric epithelial cell of the embodiment of the present invention 1
The datagram of gene relative expression quantity;
Fig. 3 is superoxide dismutase 1 of the microbial fermenters to oxidative damage liver cell of the embodiment of the present invention 1
(SOD1) and the datagram of superoxide dismutase 2 (SOD2) gene relative expression quantity;
Fig. 4 is that the microbial fermenters of the embodiment of the present invention 1 repair complementary chiasma base to the excision of oxidative damage liver cell
Because of 1 (ERCC1) and DNA damage cognition and the datagram of reparative factor (XPA) gene relative expression quantity;
Fig. 5 is that the microbial fermenters of the embodiment of the present invention 1 repair Cross-complementing Gene to the x-ray of oxidative damage liver cell
1 (XRCC1) and x-ray repair the datagram of Cross-complementing Gene 2 (XRCC2) gene relative expression quantity;
Fig. 6 is oil red O stain (oil-red O of the microbial fermenters to fat metabolism of the embodiment of the present invention 1
Stain) the datagram analyzed;
Fig. 7 is the microbial fermenters of the embodiment of the present invention 1 to Adipose trigtyceride lipase (ATGL) and hormone-sensitive
The datagram of property lipase (LIPE) gene relative expression quantity;
Fig. 8 is the microbial fermenters of the embodiment of the present invention 1 to fatty uncoupling proteins 1 (UCP1) and fatty uncoupling
The datagram of albumen 2 (UCP2) gene relative expression quantity.
Specific embodiment
Below with reference to specific embodiment shown in the drawings, the present invention will be described in detail.But these embodiments are simultaneously
The present invention is not limited, structure that those skilled in the art are made according to these embodiments, method or functionally
Transformation is included within the scope of protection of the present invention.
Example embodiment is described more fully with reference to the drawings.However, example embodiment can be with a variety of shapes
Formula is implemented, and is not understood as limited to embodiment set forth herein;On the contrary, thesing embodiments are provided so that the present invention will
Fully and completely, and by the design of example embodiment comprehensively it is communicated to those skilled in the art.
The present invention uses fluid as the medium of sound transmission, (about with the microorganism speed of sound audible in fermentation
For 1463m/s), accelerate intercellular reaction.After sound stimulation, microbial activities power is significantly improved, so that be metabolized, synthesize,
The effect for the acceleration that oxidation and reducing power all obtain, this is because making microorganism after sound wave person plants alternate stress field action
Change slightly, the mobility of intracellular nutriment and the enhancing (Ca of permeability occur for structure2+、Mg2+Etc.), thus
The variation for leading to ion concentration inside and outside film makes cellular endogenous auxin and its substance in relation to influencing growth that directional migration occur,
The ability of substance is absorbed and operated to change cell.Therefore, the present invention was fermented by the mixed liquor of vegetable juice and certain kind of berries fruit juice
Music is played in journey, fermentation material can be made to receive the energy of frequency and wavelength different in melody, generates microbial fermenters.Together
When, the present invention is in the Like superoxide dismutase (superoxide for measuring the microbial fermenters after fermentation
Dismutase-like, SOD-like) content, gastric cells resistance to oxidation, liver cell resistance to oxidation, liver cell DNA repair ability
And promote fat cell metabolism ability.
Definition
Herein described vegetables include leaf vegetables, bean sprouts class, melon and fruit class, solanaceous vegetables, potato class, root vegetables, flower vegetables etc.
Vegetables, but not limited to this;It also include the fruit such as kernel approaches, pomaceous fruit, melon, but not limited to this.
Certain kind of berries fruit described herein include blueberry, Cranberry, red grape, white grape, red bayberry, mulberries, apple, carrot, sugarcane,
Passion fruit, pineapple and lemon etc., but not limited to this.
It is for statistical analysis using Excel software.Data indicate with average value ± standard deviation (SD), a difference between this
It is different that (student's t-test) analysis is examined with student t.
As in this article the numerical value that uses be approximation, all experimental datas all indicate in the range of 20%, preferably
In the range of 10%, most preferably in the range of 5%.
The method of the microbial fermenters of the invention cultivated under music environment of embodiment 1
In a preferred embodiment of the present invention, by vegetable juice (fresh juicing) and certain kind of berries fruit juice (fresh juicing) with 3 to 2
After ratio mixing, then with 5 times of distilled water dilutions, as empty training liquid;Wherein certain kind of berries fruit juice includes: blueberry, Cranberry, red grape, white
Grape, red bayberry, mulberries, apple, carrot, sugarcane, passion fruit, pineapple and lemon, their equal proportion mixed liquor;Vegetable juice
Include: broccoli, celery and asparagus, their equal proportion mixed liquor.Sky training liquid is handled with pasteurism, then
Access streptococcus thermophilus (Streptococcus thermophilus), S. cervisiae (Saccharomyces
Cerevisiae) and acetic acid bacteria (Acetobacter aceti) is into empty training liquid, carries out primary fermentation, condition is 30 DEG C ± 1 DEG C
To obtain microbial fermenters of the invention, wherein the total volume of those bacterial strains is the 2 of empty training liquid product within ferment at constant temperature 192 hours
To 7%;It is divided into three groups in fermentation process, dials put three kinds of different genres of musics respectively, is divided into classical music (Mozart:
Four No. ten symphony-Opus Ones), pop music (Maroon 5:sugar) and rock music (Maroon 5:Maps), wherein classic
Pleasure is that account for 99% or more of whole first song, pop music lower than 10,000Hz in song 1 to 3 minute be low in song 1 to 3 minute
Accounting for 95% or more of whole first song, rock music in 10,000Hz is to be higher than 10,000Hz in song 1 to 3 minute to account for whole first song
50% or more.
Like superoxide dismutase (SOD-like) content detection of the microbial fermenters of the invention of embodiment 2
Superoxide dismutase (superoxide dismutase, SOD) is a kind of catalysis superoxide radical disproportionation
As oxygen and the ferment of hydrogen peroxide, those ferment are made in the cell for being exposed to oxygen with important oxidation resistant defence
With.Therefore, the present invention utilizes SOD active agent group (SOD activity kit, Enzo Life Science company, model
CAT#ADI-900-157 cellular oxidation Pressure Analysis) is carried out.
As a result as shown in Figure 1, compared to control group (ferment only not listened to music), no matter under any music environment
The microbial fermenters of (classical music, pop music and rock music) culture can all effectively improve Like superoxide dismutase (SOD-
Like) 1 times of content or more, it can deduce that it can be than the microbial fermenters do not cultivated under music environment more in oxidation resistance
It is promoted.Microbial fermenters of the invention contain the activity of higher superoxide dismutase as the result is shown for this, can have relatively plus
By force to the defence of oxidative pressure.
The gastric cells oxidation resistance of the microbial fermenters of the invention of embodiment 3
The present invention carries out the test of oxidation resistance with cell platform, utilizes quantitative real time aggregation enzyme chain reaction
The AGS mankind Weishang that (Quantitative real time PCR, qPCR) detection is handled with microbial fermenters of the invention
The gene expression amount of the superoxide dismutase 2 (SOD2) of chrotoplast (gastric epithelial cells), analyzes this hair
The gastric cells oxidation resistance of bright microbial fermenters.
The present invention is purchased from Unite States Standard biology product collecting center (American Type using human gastric epithelium
Culture Collection, ATCC), number CRL-1739.By the cell culture in 10% fetal calf serum (fetal of addition
Bovine serum, FBS) (GIBCO company, number 10438-026, the U.S.) and 1% penicillin/streptomycin
(penicillin/streptomycin) 1640 basal medium of RPMI of (GIBCO company, number 15140-122, the U.S.)
In (Roswell Park Memorial Institute) (GIBCO company, the U.S.).
Each hole inoculation 1.5 × 10 in 6 hole culture plates (GeneDireX company, TaiWan, China)5A mankind Weishang skin
Cell contains 5%CO in 37 DEG C with above-mentioned culture medium2Incubator culture 24 hours;Training is attached to not interfering with this
The culture medium is removed in the case where supporting the cell on disk, the culture medium more renewed later is separately added into three kinds of differences of 2% concentration
Microbial fermenters of the invention under music environment are handled 6 hours, and the cell to contain only culture medium is as control group, on
It states same experimental conditions and is all carried out in triplicate with every group.Human gastric epithelium is collected later.Use RNA extraction agent group
The RNA of the above-mentioned cell of (Geneaid company, TaiWan, China) extraction, recycling reverse transcriptase ( III
Reverse Transcriptase) (Invitrogen company, the U.S.) by RNA reverse transcription be cDNA, then use KAPAFAST qPCR reagent set (KAPA Biosystems company, the U.S.) is with real time aggregation enzyme chain reaction instrument (ABI
StepOne PlusTMSystem (Applied Biosystems company, the U.S.) carries out quantitative real time aggregation enzyme chain reaction
(qPCR), target gene (SOD2) and the expression with reference to gene are detected, with reference to gene (reference gene)
Expression is standardized the expression of target gene.
As a result as shown in Fig. 2, compared to the cell of culture medium is contained only as control group, no matter in any music environment
Under (classical music, pop music and rock music) culture microbial fermenters processing gastric epithelial cell, anti-oxidant correlation SOD2
Gene expression amount can effectively improve 20% or more.
The liver cell oxidation resistance of the microbial fermenters of the invention of embodiment 4
The present invention is with hydrogen peroxide (Hydrogen peroxide, H2O2) inducing hepatocyte oxidative damage, due to known super
Superoxide dismutase (SOD) can be catalyzed superoxides, reduce free-radical contents, antioxidant important in vivo of behaving;This hair
The bright oxidative damage liver handled using quantitative real time aggregation enzyme chain reaction (qPCR) detection with microbial fermenters of the invention
The superoxide dismutase 1 (SOD1) of cell and the gene expression amount of superoxide dismutase 2 (SOD2), are analyzed of the invention
The liver cell oxidation resistance of microbial fermenters.
The present invention is received using HepG2 liver cell (liver hepatocellular cells) purchased from Unite States Standard biology product
Hiding center, number HB-8065.Hepatocyte culture conditions method as described in Example 1, and with 4mL H2O2Inducing cell oxidation
It damages, then behind microbial fermenters processing of the invention 6 hours be separately added under three kinds of different music environments of 2% concentration,
The oxidative damage liver cell handled with microbial fermenters of the invention is detected with quantitative real time aggregation enzyme chain reaction (qPCR)
SOD1 and SOD2 gene expression amount, detection mode is as described in Example 3, above-mentioned same experimental conditions all with every group in triplicate
It carries out.
As a result as shown in figure 3, compared to the cell of culture medium is contained only as control group and only with H2O2Induced oxidation damage
The cell of wound cultivates SOD1 and the SOD2 base of the liver cell of microbial fermenters processing under classical music and pop music music environment
Because expression quantity all obviously increases;Meanwhile with H2O2Induced oxidation damage and using the cell of the empty training liquid processing of embodiment 1 as pair
According to group, SOD1 and the SOD2 gene expression amount of oxidative damage liver cell can be also promoted, but to cultivate microorganism under classical music environment
SOD1 and the SOD2 gene expression amount of the liver cell of fermentation material processing are best.Show that microbial fermenters of the invention can promote liver
Cellular oxidation ability also has the effect of protection liver cell fairly good.
The liver cell DNA repair ability of the microbial fermenters of the invention of embodiment 5
Since the cell of known oxidative damage can generate free radicals, and free radical will lead to DNA and be undermined gene mutation.This
Invention is with hydrogen peroxide-induced HepG2 Oxidative Damage in Liver, then is separately added under three kinds of different music environments of 2% concentration
After microbial fermenters of the invention are handled 6 hours, detected with quantitative real time aggregation enzyme chain reaction (qPCR) with of the invention
The expression of the revision points of the DNA structure damage and distrand DNA fracture of the oxidative damage liver cell of microbial fermenters processing
Amount;Wherein DNA structural damage revision points include that (the excision repair cross- of complementary chiasma gene 1 is repaired in excision
Complementing gene 1, ERCC1) and DNA damage recognizes and reparative factor (DNA damage recognition
And repair factor, XPA);The revision points of distrand DNA fracture include that x-ray repairs 1 (X-ray of Cross-complementing Gene
Repair cross complementing 1, XRCC1) and x-ray reparation Cross-complementing Gene 2 (XRCC2), detection mode
As described in Example 3, above-mentioned same experimental conditions are all carried out with every group in triplicate.
As a result as shown in Figures 4 and 5, compared to containing only the cell of culture medium as control group and only with H2O2Induction
The cell of oxidative damage, no matter under which kind of music environment (classical music, pop music and rock music) cultivate microbial fermenters
ERCC1, XPA, XRCC1 and XRCC2 gene expression amount of the oxidative damage liver cell of processing all obviously increase;Meanwhile with
H2O2Induced oxidation damages and not to listen to music the cell of ferment product control group processing, can promote oxidative damage liver cell
ERCC1 and XPA gene expression amount (such as Fig. 4);With H2O2Induced oxidation damage and the cell handled with the empty training liquid of embodiment 1,
Also XRCC1 and the XRCC2 gene expression amount (such as Fig. 5) of oxidative damage liver cell can be promoted, but micro- to cultivate under classical music environment
ERCC1, XPA, XRCC1 and XRCC2 gene average expression amount of the liver cell of biofermentation object processing are best.Show this hair
Bright microbial fermenters can promote the repair ability (ERCC1 and XPA) of liver cell DNA structure damage and in distrand DNA
(XRCC1 and XRCC5) also has the ability repaired in gene expression, also has the effect of for protection liver cell fairly good.
The promotion fat cell metabolism ability of the microbial fermenters of the invention of embodiment 6
The present invention is with dexamethasone (dexamethasone, DEXA) and isobutyl methylxanthine
(isobutylmethylxanthine, IBMX) induces mouse 3T3-L1 PECTORAL LIMB SKELETON (3T3-L1 preadipocytes) point
Mature fat cell is turned to, the microbial fermenters processing of the invention being separately added into again under three kinds of different music environments later
Afterwards, confirm that the promotion fat cell of microbial fermenters of the invention decomposes effect with oil red O stain (oil-red O stain)
The expression quantity for the fat metabolism gene that fruit and qPCR detection are handled with microbial fermenters of the invention.
6.1 3T3-L1 cell experiment analysis methods
Firstly, 3T3-L1 PECTORAL LIMB SKELETON is placed in each hole inoculation 1.5 × 10 of 6 hole cell cultures5A cell, to
Cell culture to densification is considered as the 0th day, replacement at this time added with 10% fetal calf serum, 10 μ g/ml insulin, 1 μM of DEXA,
Eagle culture medium (dulbecco's modified eagle medium, DMEM) is improved with the Du Shi of 0.5mMIBMX, culture
48 hours;Culture medium is replaced in third day, is cultivated 48 hours;Replacement culture medium is added with 10% fetal calf serum and 10 μ g/ within 5th day
The DMEM culture medium of mL insulin, every two days one subcultures of replacement, lasting differentiation was to the 6th day, when cell begins to differentiate into rouge
Microbial fermenters of the invention under three kinds of 1% or 2% concentration different music environments are separately added into after fat cell to various kinds
In this, continuous observation 8 days;Oil red dyeing is carried out, oil red stain is the stain for having specificity to neutral fat cell;It will culture
Base sucks, and washs secondary with PBS and fixes cell 30 minutes with 10% formaldehyde;It is secondary again with oil red O dye with PBS washing cell
Agent is dissolved in isopropanol and contaminates a hour, finally moves back dye with PBS, can observe under the microscope;It will be contaminated with 100% isopropanol again
The lipid solubilization of color, and light absorption value (OD) is measured under wavelength 510nm.
The present invention confirms the promotion rouge of microbial fermenters of the invention using oil red O stain (oil-red O stain)
Fat cell disintegration effect.As a result as shown in fig. 6, in fat metabolism experiment, make compared to the fat cell for containing only culture medium
It is handled for control group, with the fat cell of the 2% of embodiment 1 empty training liquid processing and with 1% or 2% taste pellet ferment fatty thin
Born of the same parents, no matter under 1% or 2% music environment (classical music, pop music and rock music) culture microbial fermenters processing
Fat cell all has the ability for remarkably promoting lipolysis, and the concentration of microbial fermenters more high effect is better.
The gene expression amount of 6.2 fat cells is analyzed
3T3-L1 PECTORAL LIMB SKELETON is placed in each hole inoculation 2 × 10 of 6 hole cell cultures by 3T3L15A cell,
It is considered as the 0th day to cell culture to densification, replacement at this time is added with 10% fetal calf serum, 10 μ g/ml insulin, 1 μM of ground plug
Meter Song (DEXA), Du Shi improvement Eagle culture medium (DMEM) with 0.5mM isobutyl methylxanthine (IBMX), culture 48 are small
When;Culture medium is replaced in third day, is cultivated 48 hours;Replacement culture medium is added with 10% fetal calf serum and 10 μ g/mL pancreases within 5th day
The DMEM culture medium of island element, every two days one subcultures of replacement, lasting differentiation was to the 6th day, when cell begins to differentiate into fat carefully
It is separately added into the microbial fermenters of the invention under three kinds of different music environments after born of the same parents into each sample, it is small to carry out processing 24
When;Collect cell.The RNA of above-mentioned cell is extracted using RNA extraction agent group (Geneaid company, TaiWan, China), is recycled anti-
Transcriptase (III Reverse Transcriptase) (Invitrogen company, the U.S.) invert RNA
Record is cDNA, is then usedFAST qPCR reagent set (KAPA Biosystems company, the U.S.) is with reality
When Polymerase Chain Reaction instrument (ABI StepOne PlusTMSystem, Applied Biosystems company, the U.S.) determined
It measures the chain reaction of real time aggregation enzyme (qPCR), detects the expression of the fat metabolism gene of microbial fermenters processing of the invention
Amount;Wherein fat metabolism gene includes Adipose trigtyceride lipase (the adipose triglyceride for promoting lipolysis
Lipase, ATGL) and hormone-sensitive lipase (hormone-sensitive lipase, LIPE) gene;And accelerate palm fibre
Color fat cell is decomposed comprising promoting fatty uncoupling proteins 1 (uncoupling protein1, UCP1) and fatty uncoupling
Albumen 2 (UCP2) gene, detection mode is as described in Example 3, and above-mentioned same experimental conditions all to carry out in triplicate.
As a result as illustrated in the schematic views of figures 7 and 8, compared to containing only the fat cell of culture medium as control group and with embodiment 1
2% empty training liquid processing fat cell, no matter under which kind of music environment (classical music, pop music and rock music) culture is micro-
The expression quantity of ATGL and LIPE gene of the fat cell of biofermentation object processing all obviously increases, and indicates of the invention micro-
Biofermentation object has the ability for promoting lipolysis really;And the fat cell of pop music culture microbial fermenters processing
The expression quantity of UCP1 and UCP2 gene is best, indicates that microbial fermenters of the invention really have brown fat and promotees
Into the effect of decomposition.
In conclusion the present invention puts different types of sound by the mixed liquor fermentation process of vegetable juice and certain kind of berries fruit juice groups
It is happy, fermentation material can be made to receive the energy of frequency and wavelength different in melody, the microbial fermenters of generation.In fermentation process
In, which kind of music (classical music, pop music or rock music) no matter is given, microbial fermenters of the invention can all effectively improve class
1 times of component content of superoxide dismutase (SOD-like) or more can deduce that it can be than in music ring in oxidation resistance
The microbial fermenters cultivated under border are more promoted.The present invention further carries out the test of oxidation resistance using cell platform, with
Microbial fermenters of the invention under the different music environments of three kinds of 2% concentration handle gastric epithelial cell cell, then with quantitative
Real time aggregation enzyme chain reaction (qPCR) method detects SOD2 gene expression amount, as the result is shown microbial fermenters of the invention
Have the effect of promoting anti-oxidant SOD2 gene expression amount.
In addition, microbial fermenters of the invention are splendid for protecting liver also to have the effect of, under classical music environment
SOD1 and the SOD2 gene expression amount for cultivating the liver cell of microbial fermenters processing increase, and further go into seriously microorganism hair
Ferment object can also promote DNA repair ability, promote DNA structure injury repair gene (ERCC1 and XPA) and be broken in distrand DNA
Revision points (XRCC1 and XRCC5) expression quantity.
Furthermore in the experiment for promoting fat cell metabolism ability, with 1% and 2% concentration microbial fermentation of the invention
The fat cell of object processing has the ability for promoting lipolysis, and to cultivate microbial fermenters processing under pop music environment
The fat metabolism gene (ATGL and LIPE) of fat cell and accelerate brown fat cell disintegration gene (UCP1 and UCP2)
Expression quantity increases, and indicates that microbial fermenters of the invention have the effect of promoting to decompose for brown fat.
It should be appreciated that although this specification is described in terms of embodiments, but not each embodiment only includes one
A independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should will say
As a whole, the technical solution in each embodiment may also be suitably combined to form those skilled in the art can for bright book
With the other embodiments of understanding.
The series of detailed descriptions listed above only for feasible embodiment of the invention specifically
Protection scope bright, that they are not intended to limit the invention, it is all without departing from equivalent implementations made by technical spirit of the present invention
Or change should all be included in the protection scope of the present invention.
Claims (9)
1. a kind of microbial fermenters, it is characterised in that: the microbial fermenters are the mixed liquor of vegetable juice and certain kind of berries fruit juice,
In the environment of playing music, post-fermentation acquisition is mixed with streptococcus thermophilus, saccharomycete and acetic acid bacteria.
2. microbial fermenters as described in claim 1, it is characterised in that:
The music environment is the environment for playing classic melody, and the classic part of the melody sound intermediate frequency lower than 10,000Hz accounts for
99% or more of whole head song, the allusion melody duration is 1 minute to 3 minutes;Alternatively, to play the ring of pop
Border, and the pop sound intermediate frequency accounts for 95% or more of whole first song lower than the part of 10,000Hz, the pop is held
Continuous duration time is 1 minute to 3 minutes;Alternatively, to play the environment of rock and roll melody, and the rock and roll melody sound intermediate frequency is high
50% or more of whole first song is accounted in 10,000Hz, the rock and roll melody duration is 1 minute to 3 minutes.
3. the preparation method of microbial fermenters as claimed in claim 1 or 2, it is characterised in that:
The preparation method includes the following steps:
Step 1, the empty training liquid of preparation: first being mixed the vegetable juice of fresh squeezing with the certain kind of berries fruit juice of fresh squeezing with the volume ratio of 1:1~2:1,
After diluting 3~6 times (volumes) with distilled water again, sterilization treatment is carried out, obtains the empty training liquid;
Step 2, it is inoculated with strain: by streptococcus thermophilus (Streptococcus thermophilus), S. cervisiae
(Saccharomyces cerevisiae) and acetic acid bacteria (Acetobacter aceti) are inoculated into described empty with liquid, three kinds of bacterium
The weight ratio of kind is 1:1:1;The total volume of the streptococcus thermophilus, saccharomycete and acetic acid bacteria and the vegetable juice and certain kind of berries fruit juice
The ratio of the volume of mixed liquor is 2:100~7:100;
Step 3, ferment: in the environment of playing music, temperature condition is ferment at constant temperature 150~260 hours at 25 DEG C~35 DEG C
To obtain microbial fermenters of the invention.
4. preparation method as claimed in claim 3, it is characterised in that:
The vegetable juice of the fresh squeezing is isometric proportioning concentration of broccoli, celery and asparagus;
The certain kind of berries fruit juice of the fresh squeezing is blueberry, Cranberry, red grape, white grape, red bayberry, mulberries, apple, carrot, sugarcane, hundred
Isometric proportioning concentration of fragrant fruit, pineapple and lemon.
What 5. microbial fermenters as claimed in claim 1 or 2 or as described in claim 3 or 4 preparation method prepared
The microbial fermenters are used to prepare the purposes in terms of the drug for promoting fat cell metabolism.
6. purposes as claimed in claim 5, it is characterised in that: the promotion fat cell metabolism is to promote lipolysis or promotion
Brown fat cell disintegration.
What 7. microbial fermenters as claimed in claim 1 or 2 or as described in claim 3 or 4 preparation method prepared
The microbial fermenters are used to prepare the purposes in terms of liver-protective drug.
8. purposes as claimed in claim 7, it is characterised in that: the protection liver be improve liver cell oxidation resistance or
DNA repair ability.
9. a kind of pharmaceutical composition, it is characterised in that: the pharmaceutical composition is sent out comprising microorganism as claimed in claim 1 or 2
The microbial fermenters that ferment object or as described in claim 3 or 4 preparation method prepare.
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