CN110354148A - Bifidobacterium adolescentis CCFM1061 is preparing the application in functional microbial inoculum, food and/or drug - Google Patents
Bifidobacterium adolescentis CCFM1061 is preparing the application in functional microbial inoculum, food and/or drug Download PDFInfo
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- CN110354148A CN110354148A CN201910765973.7A CN201910765973A CN110354148A CN 110354148 A CN110354148 A CN 110354148A CN 201910765973 A CN201910765973 A CN 201910765973A CN 110354148 A CN110354148 A CN 110354148A
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- 231100000283 hepatitis Toxicity 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 1
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- 229940074383 interleukin-11 Drugs 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
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- 231100000915 pathological change Toxicity 0.000 description 1
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- 229940111202 pepsin Drugs 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
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- 229940107700 pyruvic acid Drugs 0.000 description 1
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- 230000002588 toxic effect Effects 0.000 description 1
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
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- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
Classifications
-
- 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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- 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
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses bifidobacterium adolescentis CCFM1061 to prepare the application in functional microbial inoculum, food and/or drug, bifidobacterium adolescentis CCFM1061 can in enteron aisle rapid field planting, significantly improve fasting blood-glucose, the oral glucose tolerance of the type-II diabetes by STZ and high fat diet induction, area under the curve when reducing glucose tolerance；Significantly improving type-II diabetes, total cholesterol increases in the serum as caused by STZ and high fat diet, high-density lipoprotein cholesterol declines；The insulin resistance condition of type-II diabetes can be significantly improved；The level of inflammation in type-II diabetes liver organization can be significantly improved；Significantly improve the pathology damage of the tissues such as type-II diabetes pancreas, liver.In addition, bifidobacterium adolescentis CCFM1061 has stronger adsorption capacity to perfluoro caprylic acid (PFOA), there is the ability for alleviating PFOA toxicity.
Description
Technical field
The invention belongs to technical fields, and in particular to bifidobacterium adolescentis CCFM1061 is preparing functional microbial inoculum, food
And/or the application in drug.
Background technique
Non-alcohol fatty liver (non-alcoholic fatty liver disease, NAFLD) is liver cell
Steatosis caused by middle fat excess accumulation, is a kind of symptom of liver metabolism disorder, not plus intervenes and can gradually be evolved into
Non-alcoholic fatty type hepatitis (NASH) and then deterioration are the high liver fibrosis of the death rate, cirrhosis, liver cancer.In recent years, with
Improvement of living standard, more and more high-fat, high-energy diet lay out dining table, and the amount of exercise of people is not therewith
Increase, so that being increased year by year by the incidence of the metabolic syndrome of representative of non-alcohol fatty liver.Due to its disease
Reason mechanism is not perfect, and the drug having no specifically for NAFLD is sold in the market, and therapeutic modality can only be from diet and life style
On improved, while being aided with the drug for reducing blood lipid or blood glucose to reach better effect.
In recent years, expanding economy causes our people's living-pattern preservation, activity to reduce, and fat ratio is obvious
Increase, diabetes obtain illness rate and significantly increase.International Diabetes Federation (IDF) indicates that 2017, the whole world had 4.25 hundred million years
Age, the people greater than 19 years old suffered from diabetes, if maintaining this trend, after about 30 years, diabetic's number will be up to 6.93 hundred million
People.Therefore, control diabetes have become extremely urgent thing.
Type-II diabetes are with fasting blood-glucose raising, high-density lipoprotein cholesterol reduction and triglycerides raising, pancreas islet
The performances such as element resistance exist simultaneously, and are extremely a variety of danger on pathological change basis with glycometabolism, lipid metaboli and protein metabolism
Factor aggregation, inspires the clinical syndrome of the various cardiovascular and cerebrovascular disease occurrence and development such as atherosclerosis.Due to two type glycosurias
Disease is the pathological state of a variety of Metabolite abnormal aggregations, and cluster generation is related with insulin resistance, has become painstaking effort at present
The hot spot that pipe disease and liver diseases research field are paid close attention to jointly.In addition, type-II diabetes are along with the disorderly of intestinal microecology
Disorderly, while being also closely related with psychotic disorders such as depression and anxiety.In addition some researches show that occupational exposure is in the worker of PFOA
Type II diabetes mortality risk increase, therefore in daily life PFOA exposure, be the mankind's type II diabetes morbidity it is potential
Risk.
It is mostly at present to surround to alleviate hypoinsulinism and insulin resistance two for the drug therapy of type-II diabetes
Aspect, melbine, thiazolidinediones including mitigating insulin resistance；Sulfonylureas, the Rosiglitazone of good control blood glucose
Deng.These drugs have certain therapeutic effect, but with aggravation, dosage increase, drug drug interaction, drug poison
Side effect etc. also dramatically increases, and alimentary canal is caused adverse reaction occur, and shows certain liver renal toxicity, such as takes two for a long time
First biguanides can cause the gastrointestinal tract for stimulating some patients to cause discomfort and may will affect absorption of the patient to vitamin B12,
Rosiglitazone can cause liver dysfunction and oedema etc..
Summary of the invention
The purpose of this section is to summarize some aspects of the embodiment of the present invention and briefly introduce some preferable implementations
Example.It may do a little simplified or be omitted to avoid our department is made in this section and the description of the application and the title of the invention
Point, the purpose of abstract of description and denomination of invention it is fuzzy, and this simplification or omit and cannot be used for limiting the scope of the invention.
In view of above-mentioned technological deficiency, the present invention is proposed.
Therefore, as one aspect of the present invention, the present invention overcomes the deficiencies in the prior art, and it is double to provide the youth
Discrimination bacillus CCFM1061 is preparing the application in functional microbial inoculum, food and/or drug.
In order to solve the above technical problems, the present invention provides the following technical scheme that bifidobacterium adolescentis CCFM1061 is making
Application in standby functionality microbial inoculum, food and/or drug, in which: the bifidobacterium adolescentis CCFM1061 can be used in preparation and change
Microbial inoculum, food and/or the drug of fasting blood-glucose and impaired glucose tolerance caused by kind type-II diabetes.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, to be improved caused by type-II diabetes
Microbial inoculum, food and/or the drug that total cholesterol increases in serum, high-density lipoprotein cholesterol declines.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, to be improved caused by type-II diabetes
The microbial inoculum, food and/or drug of inflammation in liver organization.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and significantly improve type-II diabetes and make
At pancreas, liver organization pathology damage microbial inoculum, food and/or drug.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061 can also be used to preparation absorption perfluoro caprylic acid, alleviate entirely
Microbial inoculum, food and/or the drug of fluorine octanoic acid toxicity.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and alleviate type-II diabetes mouse
Microbial inoculum, food and/or the drug of constipation.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061, which can also be used to prepare non-alcohol fatty liver, to be made
At hepatic injury microbial inoculum, food and/or drug.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and improve non-alcoholic fatty liver
Microbial inoculum, food and/or the drug of the oxidation resistance of sick liver.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves blood caused by high fat diet
Microbial inoculum, food and/or the medicine that clear total cholesterol, blood glucose, liver tg and low density lipoprotein cholesterol content rise
Object.
As bifidobacterium adolescentis CCFM1061 of the present invention in preparing functional microbial inoculum, food and/or drug
A kind of preferred embodiment of application: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves microbial inoculum, the food of enteritis
And/or drug.
Beneficial effects of the present invention: bifidobacterium adolescentis CCFM1061 can in enteron aisle rapid field planting, significantly improve by
Fasting blood-glucose, the oral glucose tolerance of STZ and the type-II diabetes of high fat diet induction, when reducing glucose tolerance below curve
Product；Significantly improve type-II diabetes total cholesterol raising, high-density lipoprotein gallbladder in the serum as caused by STZ and high fat diet
Sterol decline；The insulin resistance condition of type-II diabetes can be significantly improved；It can significantly improve in type-II diabetes liver organization
The level of inflammation；Significantly improve the pathology damage of the tissues such as type-II diabetes pancreas, liver.In addition, bifidobacterium adolescentis
CCFM1061 has stronger adsorption capacity to perfluoro caprylic acid (PFOA), has the ability for alleviating PFOA toxicity；Bifidobacterium adolescentis
CCFM1061 is remarkably improved high sugar and acts on the lower proliferation of INS-1 cell and the expression of MafA gene；Two type glycosurias can be alleviated
The constipation situation of disease.The bifidobacterium adolescentis CCFM1061 can be resistant to simulation gastro-intestinal Fluid well, can be used for preparing alleviation
Type-II diabetes, constipation and PFOA toxicity pharmaceutical composition and fermented food, have very extensive application prospect.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, required use in being described below to embodiment
Attached drawing be briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for this
For the those of ordinary skill of field, without any creative labor, it can also be obtained according to these attached drawings other
Attached drawing.Wherein:
Fig. 1 is the colonial morphology of bifidobacterium adolescentis CCFM1061；
Fig. 2 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mouse fasting blood-glucose；
Fig. 3 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes Mouse oral glucose tolerance；
Fig. 4 is bifidobacterium adolescentis CCFM1061 to type-II diabetes Mouse oral glucose tolerance area under the drug-time curve
(AUCglucose) influence；
Fig. 5 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mice serum total cholesterol (TC) level；
Fig. 6 is bifidobacterium adolescentis CCFM1061 to type-II diabetes mice serum high-density lipoprotein cholesterol (HDL-
C) horizontal influence；
Fig. 7 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mouse islets element sensibility；
Fig. 8 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mouse liver inflammation；
Fig. 9 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mice pancreatic histopathology；
Figure 10 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes mouse liver histopathology；
Figure 11 is absorption situation of the bifidobacterium adolescentis CCFM1061 to PFOA；
Figure 12 is the influence that bifidobacterium adolescentis CCFM1061 acts on high sugar lower INS-1 cell proliferative conditions；
Figure 13 is the influence that bifidobacterium adolescentis CCFM1061 acts on high sugar lower INS-1 cell MafA gene expression；
Figure 14 is the influence that bifidobacterium adolescentis CCFM1061 arranges type-II diabetes mouse the first grain melena time；
Figure 15 is influence of the bifidobacterium adolescentis CCFM1061 to type-II diabetes stool in mice water content；
Figure 16 is bifidobacterium adolescentis CCFM1061 to characteristics bacterium such as Akkermansia Pseudomonas in NAFLD mouse intestinal
The influence of group；
Figure 17 is influence of the bifidobacterium adolescentis CCFM1061 to total cholesterol (TC) in NAFLD mouse liver；
Figure 18 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mice serum glutamic-pyruvic transaminase (ALT)；
Figure 19 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mice serum glutamic-oxalacetic transaminease (AST) level；
Figure 20 is bifidobacterium adolescentis CCFM1061 to NAFLD mice serum low density lipoprotein cholesterol (LDL-C) water
Flat influence；
Figure 21 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse fasting blood-glucose；
Figure 22 is influence of the bifidobacterium adolescentis CCFM1061 to the insulin resistance of NAFLD mouse
Figure 23 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse liver triglycerides (TG)
Figure 24 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse liver superoxide dismutase (SOD)
Figure 25 is bifidobacterium adolescentis CCFM1061 to NAFLD mouse liver glutathione peroxidase (GSH-Px)
It influences
Figure 26 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse liver inflammation；
Figure 27 is influence of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse liver histopathology；
Figure 28 is influence of the bifidobacterium adolescentis CCFM1061 to fatty liver cell Nrf2 gene expression.
Note: a, b, c indicate that group representated by different letters all has significant difference (P < 0.05).
Specific embodiment
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, right combined with specific embodiments below
A specific embodiment of the invention is described in detail.
In the following description, numerous specific details are set forth in order to facilitate a full understanding of the present invention, but the present invention can be with
Implemented using other than the one described here other way, those skilled in the art can be without prejudice to intension of the present invention
In the case of do similar popularization, therefore the present invention is not limited by the specific embodiments disclosed below.
Secondly, " one embodiment " or " embodiment " referred to herein, which refers to, may be included at least one realization side of the invention
A particular feature, structure, or characteristic in formula." in one embodiment " that different places occur in the present specification not refers both to
The same embodiment, nor the individual or selective embodiment mutually exclusive with other embodiments.
Bifidobacterium adolescentis CCFM1061 (Bifidobacterium adolescentis) was protected on 06 28th, 2019
It is hidden in Guangdong Province's Culture Collection, address is the compound the 59th of Xianlie Middle Road, Guangzhou City 100 5 building, building, and Guangdong Province is micro-
Biological study institute, deposit number are GDMCC No:60706.
The acquisition methods of bacterial strain are as follows:
(1) separation screening:
(1) take 1g from the fresh excreta of one girl baby of Jinan, Shandong Province servicemen norm.MMRS is coated on after gradient dilution
Solid medium is placed in lower 37 DEG C of anaerobic environment and cultivates 72 hours.
(2) colonial morphology, picking colony scribing line purifying are observed and recorded.
(3) in mMRS fluid nutrient medium, 37 DEG C are cultivated 48 hours, and gained bacterium colony carries out Gram's staining, record bacterium colony
Form.
(4) Gram negative bacterial strain and gram-positive cocci in reject bacterium colony is selected to obtain Gram-positive bar
Bacterium.After catalase analysis, reject catalase-positive organism strain retains catalase negative strain.
Embodiment 1: bifidobacterium adolescentis CCFM1061 has good tolerance to simulation gastro-intestinal Fluid
The bifidobacterium adolescentis CCFM1061 of freezen protective is inoculated in mMRS culture medium (+0.05% half Guang of MRS culture medium
Propylhomoserin hydrochloride) in, in 37 DEG C of Anaerobic culturel 48h of temperature, then after mMRS culture solution secondary culture 2~3 times, take the 1mL youth
The culture solution of Bifidobacterium CCFM1061, with 9.0mL pH 2.5 artificial simulation gastric juices (containing 1% pepsin, pH=2.5
MMRS culture medium) mixing, and the Anaerobic culturel at 37 DEG C, it is sampled respectively in 0h, 0.5h, 1h and 2h, with mMRS agar culture
Base casting culture carries out plate count, measures viable count and calculates its survival rate.
Survival rate be viable count logarithm in the culture solution in sampling in 0h when the ratio between viable count logarithm,
It is indicated with %.Take the culture solution of 1mL bifidobacterium adolescentis CCFM1061 that 9mL artificial simulation intestinal juice is added (containing 0.3% N of gallbladder
Salt, 1% trypsase, pH=8.0 mMRS culture medium) in, the Anaerobic culturel at 37 DEG C, respectively in 0h, 0.5h, 1h, 2h, 3h
It is sampled when with 4h, carries out plate count with the casting culture of mMRS agar medium, measure viable count and calculate its survival rate.
Survival rate be viable count logarithm in the culture solution in sampling in 0h when the ratio between viable count logarithm, with % table
Show.Experimental result is as shown in Table 1 and Table 2.The result shows that bifidobacterium adolescentis CCFM1061 has preferably artificial gastro-intestinal Fluid
Tolerance.
Tolerance of the 1 bifidobacterium adolescentis CCFM1061 of table in artificial simulation gastric juices
Tolerance of the 2 bifidobacterium adolescentis CCFM1061 of table in artificial simulation intestinal juice
Embodiment 2: bifidobacterium adolescentis CCFM1061 has no toxic side effect to C57BL/6J mouse
Bifidobacterium adolescentis CCFM1061 thallus is resuspended in 3% sucrose solution, be made concentration be 3.0 ×
109The bacteria suspension of CFU/mL.Healthy male C 57 BL/6 J mouse 8 of weight 16-20g or so are taken, adapt to environment after a week, often
It is primary to give the concentration bacteria suspension stomach-filling day, observes one week, record death and body weights.
These test results are listed in Table 3 below.These results indicate that feeding concentration 3.0 × 109The youth bifid bar of CFU/mL
Bacterium CCFM1061 does not cause to significantly affect to mouse, and weight is generated without significant changes, the no phenomena of mortality.Mouse appearance is without obvious disease
Manage symptom.
The variation and death condition of 3 mouse weight of table
Note :-: mouse is without death
Embodiment 3: bifidobacterium adolescentis CCFM1061 can reduce type-II diabetes mouse (empty stomach) blood glucose level
It takes healthy male C 57 BL/6 J mouse 40 of weight 16-20g, adapts to environment 1 week, be randomly divided into 5 groups: blank pair
According to group (NC), model control group (M), Rosiglitazone control group (RH), bifidobacterium adolescentis CCFM1061 intervention group
(CCFM1061), containing mouse 8 for every group of bifidobacterium adolescentis BA1 control group (BA1), the dosage of stomach-filling bacteria suspension is 3.0 ×
109CFU/mL is resuspended in 3% sucrose solution.Experimental animal grouping and processing method are shown in Table 4:
The grouping of 4 experimental animal of table
The 2-7 weeks: normal group mouse feeding normal diet, remaining mouse feeding high lipid food.
In the 11st week 1d, all mouse were deprived of food but not water 12h, normal group injection 50mmol/L citric acid-citric acid
Sodium buffer (pH 4.5), remaining group injection according to 100mg/ (kg weight) dosage injection 50mmol/L STZ (be protected from light on ice,
It is ready-to-use), wherein the preparation of STZ is dissolved with 50mmol/L citric acid-sodium citrate buffer solution.
Test latter stage collects mouse fresh excreta and freezes in -80 DEG C, extracts the macro genome in excrement, and survey using two generations
Sequence instrument analyzes intestinal microflora.When off-test, mouse is deprived of food but not water 12h, and 0.5mL/10g1% is injected intraperitoneally
Nembutal sodium solution anesthesia after, Culling heart blood, be aided with cervical dislocation execution.3000 × g of blood sample, under the conditions of 4 DEG C
It is centrifuged 15min, takes supernatant, -80 DEG C freeze for measuring associated serum index.Partial liver is immediately placed in the life of pre-cooling after collecting
Blood is removed in rinsing in reason salt water, is put into paraformaldehyde and fixes, remainder liver is quick-frozen in liquid nitrogen and is transferred to -80 DEG C of jellies
It deposits, it is subsequent that liver homogenate is made to be used to measure index of correlation, specifically the preparation method is as follows: a certain amount of liver organization is weighed, by 1:9
Ratio is added physiological saline and carries out tissue grinder, and 3000r is centrifuged 10min, takes supernatant to freeze spare in -80 DEG C.
Experimental result is as shown in Figure 2.Model group mouse fasting blood-glucose significantly increases, stomach-filling bifidobacterium adolescentis CCFM1061
Significantly reduce the fasting blood glucose level of model mice and close to blank control group.Its energy for reducing mouse fasting blood glucose level
Power is similar to Rosiglitazone medicine group.
Embodiment 4: bifidobacterium adolescentis CCFM1061 can enhance type-II diabetes mouse glucose tolerance ability
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.Before mouse is put to death, it is deprived of food but not water 12h, is filled
Stomach glucose solution (2g/kg weight), measures 0,30,60,120min blood glucose situation respectively.
Experimental result is as shown in Figure 3 and Figure 4, and model group mouse is poor to the tolerance of glucose, after stomach-filling glucose,
Blood glucose value is significantly raised, and declines slowly, and stomach-filling bifidobacterium adolescentis CCFM1061 significantly reduces AUCglucoseArea, and
With normal group without significant difference.This just illustrates that bifidobacterium adolescentis CCFM1061 can significantly improve oral glucose tolerance, and act on
Effect is better than bifidobacterium adolescentis BA1.These results are consistent with blood glucose target result, prompt bifidobacterium adolescentis CCFM1061
Blood glucose level can be further decreased by enhancing glucose tolerance.
Embodiment 5: bifidobacterium adolescentis CCFM1061 can reduce the water of type-II diabetes mice serum total cholesterol (TC)
It is flat
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.When off-test, mouse is deprived of food but not water
After the Nembutal sodium solution anesthesia of 12h, intraperitoneal injection 0.5mL/10g 1%, Culling heart blood.3000 × g of blood sample, 4 DEG C of conditions
Lower centrifugation 10min, takes supernatant, according to the content of total cholesterol (TC) in the detection method measurement blood of kit.
Experimental result is as shown in Fig. 5.As seen from Figure 5, model group mice serum total cholesterol level is significantly raised,
Stomach-filling bifidobacterium adolescentis CCFM1061 reduces the content of serum total cholesterol, is better than to the recovery capability of total cholesterol level
Control drug and bifidobacterium adolescentis BA1.
Embodiment 6: it is solid that bifidobacterium adolescentis CCFM1061 can increase type-II diabetes mice serum high-density lipoprotein gallbladder
The level of alcohol (HDL-C)
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.When off-test, mouse is deprived of food but not water
After the Nembutal sodium solution anesthesia of 12h, intraperitoneal injection 0.5mL/10g 1%, Culling heart blood, blood sample centrifuging and taking supernatant, according to
The content of detection method measurement serum middle-high density lipoprotein cholesterol (HDL-C) of kit.
Experimental result is as shown in Fig. 6.It can be seen from experimental result compared with Normal group, model group mouse blood
Aloof from politics and material pursuits density lipoprotein-cholesterol content significantly reduces, and serum high-density rouge egg can be improved in stomach-filling bifidobacterium adolescentis CCFM1061
The content of white cholesterol, and bifidobacterium adolescentis CCFM1061 is aobvious to the recovery capability of serum High Density Lipoprotein Cholesterol level
Work is better than bifidobacterium adolescentis BA1.
Embodiment 7: the insulin sensitivity of type-II diabetes mouse can be improved in bifidobacterium adolescentis CCFM1061
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.When off-test, mouse is deprived of food but not water
After the Nembutal sodium solution anesthesia of 12h, intraperitoneal injection 0.5mL/10g 1%, Culling heart blood, blood sample centrifuging and taking supernatant, according to
The content of detection method measurement serum insulin (INS) of kit, and combine fasting blood-glucose result to calculate insulin resistance and refer to
Number.
Experimental result is as shown in Fig. 7.It can be seen from experimental result compared with Normal group, model group mouse pancreas
Insulin resistance index significantly increases, and stomach-filling bifidobacterium adolescentis CCFM1061 can reduce the insulin resistance index of mouse, improves
The insulin sensitivity of mouse, and bifidobacterium adolescentis CCFM1061 is significantly stronger than the recovery capability of mouse islets element sensibility
Compare bacterium bifidobacterium adolescentis BA1.
Embodiment 8: bifidobacterium adolescentis CCFM1061 can improve the inflammatory conditions of type-II diabetes mouse liver
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.When off-test, mouse is deprived of food but not water
After the Nembutal sodium solution anesthesia of 12h, intraperitoneal injection 0.5mL/10g 1%, Culling heart blood, cervical dislocation is put to death.Take liver-
80 DEG C freeze, and when measurement weighs a certain amount of liver organization, and physiological saline is added in 1:9 ratio and carries out tissue grinder, 3000r centrifugation
10min takes supernatant, measures interleukin-11 β (IL-1 β) according to the detection method of kit, is corrected with liver protein concentration.
Experimental result is as shown in Fig. 8.It can be seen from experimental result compared with Normal group, model group Mouse Liver
Dirty IL-1 β is significantly increased, and stomach-filling bifidobacterium adolescentis CCFM1061 can be relieved the inflammatory conditions of mouse liver, and youth bifid bar
Bacterium CCFM1061 is significantly stronger than bifidobacterium adolescentis BA1 to the relief capabilities of mouse liver inflammation.
Embodiment 9: bifidobacterium adolescentis CCFM1061 can be relieved the tissue damage of type-II diabetes mice pancreatic and liver
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.When off-test, mouse is deprived of food but not water
After the Nembutal sodium solution anesthesia of 12h, intraperitoneal injection 0.5mL/10g 1%, Culling heart blood, cervical dislocation is put to death.Take pancreas,
The part such as liver production paraffin section, observes tissue morphology under light microscopic after HE is dyed and takes pictures, carry out pathological evaluation.Specifically
Steps are as follows:
(1) fixed: tissue sample is washed with physiology salt, is put into immediately solid in neutral paraformaldehyde fixer (4%)
Fixed, the general set time is within 72h.
(2) wash: flowing water rinses or impregnates a few hours or stays overnight.
(3) be dehydrated: sample is successively dehydrated through 70%, 80%, 90% ethanol solutions at different levels, and each 30min places into 95%1
Secondary 20min, 100%2 each 10min.
(4) transparent :+1/2 dimethylbenzene mixed liquor 10min of 1/2 absolute alcohol, I 10min of dimethylbenzene, II 10min (are to transparent
Only).
(5) sample waxdip: is put into paraffin (62 DEG C) wax 2h thoroughly.
(6) it embeds: with maximum face in bottom, making maximum shared by the covering weave face cut out.
(7) it is sliced: using hand microtome, wax stone is cut into the segment of 5 μ m thicks.
(8) it opens up piece and bonding die (fishing piece): opening water-bath, water temperature is made to maintain 42 DEG C, make to be sliced and smooth spread over the water surface
On.
(9) it bakes piece: glass slide is put into together with object slide stand to 55 DEG C of drying box, about 2h to wax melts.
(10) aquation: paraffin section dewaxes each 10min through dimethylbenzene I, II, be then placed in 100%, 95%, 90%,
80%, each 5min in 70% alcoholic solutions at different levels, places into 3min in distilled water.
(11) just contaminate: slice, which is put into hematoxylin, dyes about 20s.
(12) it washes: rinsing about 15min with tap water flowing water.Make to be sliced color and become blue, but it is noted that flowing water cannot be excessive,
It falls off to prevent slice.
(13) break up: slice being put into 1% ethanol solution hydrochloride and is faded, 7s.See that slice reddens, color is shallower.
(14) rinse: slice, which places into flushing 15-20min in tap water flowing water, makes its restore blue.
(15) it redyes: immersing eosin stain, take out be dehydrated immediately.
(16) be dehydrated: will slice successively cross 95% ethyl alcohol I, 95% ethyl alcohol II, 70% ethyl alcohol, place into 80% ethyl alcohol 50s,
Dehydrated alcohol 2min.
(17) transparent: slice is put into 1/2 dehydrated alcohol, 1min in 1/2 dimethylbenzene, each 2min in dimethylbenzene I, II.
(18) mounting: slice uses neutral gum to hide agent as envelope after dimethylbenzene is transparent, and natural gum can be diluted with dimethylbenzene
To suitable consistency.
Experimental result is as shown in attached drawing 9 and 10.Model group mouse islets quantity tails off it can be seen from experimental result, out
There is the change of vesicle rouge in existing atrophy phenomenon, liver cell, there is a morphologic features of early stage fibrosis, and stomach-filling bifidobacterium adolescentis
CCFM1061 can obviously improve above-mentioned lesion, and effect is significantly better than bifidobacterium adolescentis BA1.
Embodiment 10: there is good PFOA adsorption capacity in vitro；
Thallus absorption carries out purifying and activation culture to bifidobacterium adolescentis CCFM1061, is inoculated with by 1% (v/v) inoculum concentration
In MRS fluid nutrient medium, 37 DEG C of culture 18h.Then thallus is collected in 8000r/min centrifugation 5min, takes precipitating physiology salt
Continue to be centrifuged 5min in 8000r/min after water cleaning, precipitating is gone to obtain viable bacteria body cell, i.e. wet thallus.Wet thallus is resuspended in
In 50mg/LPFOA solution, and so that final cell concentration is reached 1g dry mycelium/L and (wet thallus is resuspended in without the ultrapure of PFOA
Blank control is used as in water).The pH of the PFOA solution containing bacterium solution is adjusted rapidly to 3.0 using the NaOH or HCl solution of 0.1M,
The influence that PFOA is adsorbed can be ignored by adding a small amount of NaOH or HCl (less than 0.5ml) its ionic strength.It will then be equipped with
The 250ml conical flask of 100ml sample liquid is placed in 37 DEG C, 150rpm anaerobism shaking table culture, is measured by sampling after 6h, 2 parallel tests take
Average value.
The measurement of PFOA adsorbance: after adsorption experiment, sample liquid is centrifuged 5min in 8000r/min, and with 0.22 μm of moisture film
Filtering, PFOA concentration uses the UPLC-MS with Waters SYNAPT MS system to measure, using Acquity UPLC BEH c18
Column (2.1 × 100mm, 1.7 μm, Waters Co.), 35 DEG C of column temperature, 1 μ L of sample volume.Acetonitrile solution with 100% (v/v) is (molten
Liquid A) and 0.1% (v/v) aqueous formic acid (solution B) be used as eluent, carry out gradient cleaning, flow velocity is 0.3mL/min.
5 condition of gradient elution of table
| t/min | 0-0.5 | 0.5-5.0 | 5.0-7.0 | 7.0-7.5 |
| Solvent A ratio | 70% | 70-100% | 100% | 100-70% |
Mass Spectrometry Conditions: ionization source is the source ESI；MRM detection；MS+ detection；Capillary (capillary)；3.0kV；Conc
(centrum): 40.00V；Source Temperature (radiation source temperature): 120 DEG C；Desolvation (desolvation) temperature:
400℃；Conc Gas Flow:50L/h；Desolvation Gas Flow:700L/h. gas flow rate is 0.1ml/min；Matter
Son is than scanning range: 100-2000；Surface sweeping time 1s is spaced 0.061s.As a result with MassLynxV4.1 (Waters company) point
Analysis；Lactic acid bacteria is calculated to the adsorbance of PFOA according to the concentration difference of absorption front and back PFOA.Measurement result is listed in Figure 11, and the youth is double
Discrimination bacillus CCFM1061 is 69.48% ± 2.87% to the adsorption rate of the PFOA of 50mg/L.
Embodiment 11: bifidobacterium adolescentis CCFM1061 can promote the proliferation and Maf A of the INS-1 cell of high glucose induction
The expression of mRNA
Experiment is divided into 5 groups: normal group (the Nostoc commune Vanch liquid of the glucose containing 11.1mmol/L), and high sugar group (contains 22.2mmol/
The sugared culture solution of the height of L glucose), Rosiglitazone group (Rosiglitazones of+80 μm of ol/L of high sugar culture solution), (the high sugar culture of BA1 group
Liquid+contain 1*109CFU/mL BA1 bacterium solution) CCFM1061 group (high sugar culture solution+contain 1*109CFU/mL CCFM1061 bacterium solution).
By INS-1 cell (number: BH-AC0530) be incubated at RPMI-1640 culture solution (glucose containing 11.1mmol/L,
10%FBS, 50 μm of ol/L 2 mercapto ethanols, 1mmol/L pyruvic acid, 10mmol/L HEPES) in, and 37 DEG C are put into, 5%CO2
Incubator in.
CCK-8 method detects cell Proliferation: cell dissociation in good condition is centrifuged and is inoculated on 96 orifice plates, each hole about 5
×103A cell, the periphery hole of plate not inoculating cell, to prevent edge effect while being added PBS solution thereto.It is pasted to cell
The RPMI-1640 culture medium for containing 0.5% fetal calf serum is added in each hole for wall, and synchronization process is for 24 hours.Synchronization terminates, foundation point
Corresponding culture medium culture 48h is added to each hole in group, and every group sets three multiple holes, while zeroing hole is arranged.Pharmaceutical intervention terminates, and sucks
Old culture medium, PBS are cleaned 2 times, and 180 μ L serum free mediums and 20 μ L CCK-8 solution are added, and are incubated for 3-4h.Incubation terminates,
Each hole absorbance value is measured under 450nm using microplate reader.
Maf A mRNA expression measurement: Trizol method extract RNA, inhale abandon 6 orifice plates in original fluid, while be pre-chilled
PBS is cleaned 2 times, and 1.0mL Trizol lytic cell is separately added into each hole and celliferous lysate is gone to no enzyme EP and is managed,
Liquid-transfering gun is blown and beaten to no obvious sediment and stands 5min.0.2mL chloroform is added to each EP pipe, acutely shakes 15s, is placed at room temperature for
2-3min.4 DEG C, 12000rpm is centrifuged 15min, draws supernatant 0.4m L or so, is transferred in another no enzyme EP pipe, is added
The isopropanol of 0.5mL, is mixed by inversion, and is stored at room temperature 10min.4 DEG C, 12000rpm is centrifuged 10min, carefully discards supernatant, and is added
75% ethyl alcohol of 1.0mL is simultaneously mixed by inversion.4 DEG C, 12000rpm centrifugation 5min, abandoning supernatant, drying at room temperature 2-5 minutes.20 μ L are added
DEPC handles water dissolution, is stored in 80 DEG C for use.Measure RNA concentration and quality, and according to reverse transcription reagent box specification into
Row reverse transcription.The cDNA that reverse transcription obtains carries out q RT-PCR detection, wherein MafA specific primer: F:5'-
Atcactctgcccaccatcac-3', R:5'-atgacctcctccttgctgaa-3'.PCR system are as follows: F (10 μM), 0.50 μ
L；R(10μM),0.50μL；C DNA Template, 1.00 μ L；dd H2O, 3.00 μ L；Mix, 5.00 μ L.PCR program: 95 DEG C,
2min；
(95 DEG C, 30sec；60 DEG C, 30sec；72 DEG C, 20sec) * 35；72 DEG C, 5min；Target gene passes through Real-time
After PCR detection, using 2-△△CTMethod carries out Relative gene expression analysis.First each group rat INS- is analyzed with CFX Manager software
The expression quantity of 1 cell target gene, then to organize expression quantity normally as 1, other each groups in comparison, calculate each group gene expression
It is horizontal.
CCK-8 method testing result is as shown in figure 12, compared with normal group, high sugar effect group cell growth be substantially reduced (P <
0.05), Rosiglitazone cellular control unit proliferation higher sugar group is significantly increased (P < 0.05), and CCFM1061 group is compared with high sugared group
Cell proliferative condition also obviously increases (P < 0.05).
Maf A mRNA expression such as Figure 13 shows that the expression quantity of the MafA mRNA of high sugar effect group cell is obviously low
In normal group (P < 0.05), and the Maf A mrna expression amount higher sugar of Rosiglitazone positive controls and CCFM1061 group acts on
Group is obvious to rise (P < 0.05).
Embodiment 12: bifidobacterium adolescentis CCFM1061 can improve the constipation situation of type-II diabetes mouse
C57BL/6J mice group, modeling and processing method are the same as embodiment 3.Terminate the previous day in experiment, after stomach-filling,
Mouse is singly only put into the cage box for being lined with blotting paper, excrement is collected, weighing is weight in wet base, after freeze-drying, as dry weight, according to such as
Lower formula calculates excrement water content.
Excrement water content (%)=(excrement weight in wet base-excrement dry weight)/excrement weight in wet base
First day of the 15th week, blank group and model group stomach-filling prepared Chinese ink, filled bacterium group and medicine group stomach-filling contains each self-priming
The prepared Chinese ink of gastric content records the time of each small mouse's head grain row melena since stomach-filling prepared Chinese ink.
Excrement water content, the first grain melena time experimental result of row are as shown in FIG. 14 and 15, as seen from the figure, compared to model
Group, bifidobacterium adolescentis CCFM1061, which can significantly improve excrement water content, to be shortened and arranges the first grain melena time to normal level, and is imitated
Fruit is better than bifidobacterium adolescentis BA1.
Embodiment 13: adjustment effect of the bifidobacterium adolescentis CCFM1061 to NAFLD mouse intestinal flora
It takes healthy male C 57 BL/6 J mouse 48 of weight 16-20g, adapts to environment 1 week, be randomly divided into 6 groups: blank pair
According to group (NC), model control group (M), Rosiglitazone control group (RC), Simvastatin control group (SC), bifidobacterium adolescentis
CCFM1061 intervention group (CCFM1061), Lactobacillus rhamnosus L10 intervention group (LC), every group contains mouse 8.Experimental animal grouping
And processing method is shown in Table 6:
The grouping of 6 experimental animal of table
Test latter stage collects mouse fresh excreta and freezes in -80 DEG C, extracts the macro genome in excrement, and survey using two generations
Sequence instrument analyzes intestinal microflora.When off-test, mouse is deprived of food but not water 12h, and 0.5mL/10g 1% is injected intraperitoneally
Nembutal sodium solution anesthesia after, Culling heart blood, be aided with cervical dislocation execution.3000 × g of blood sample, under the conditions of 4 DEG C
It is centrifuged 15min, takes supernatant, -80 DEG C freeze for measuring associated serum index.Partial liver is immediately placed in the life of pre-cooling after collecting
Blood is removed in rinsing in reason salt water, is put into 4% neutral paraformaldehyde solution and fixes, remainder liver is quick-frozen in liquid nitrogen and shifts
It is frozen to -80 DEG C, it is subsequent that liver homogenate is made to be used to measure index of correlation, specifically the preparation method is as follows: weighing a certain amount of liver
Tissue is added physiological saline in 1:9 ratio and carries out tissue grinder, and 3000r is centrifuged 10min, takes supernatant to freeze spare in -80 DEG C.
Flora analysis experimental result is as shown in figure 16, significantly improves Akkermansia in NAFLD mouse intestinal and belongs to rich
Degree reduces the diseases such as obesity, diabetes, fatty liver and enteritis and occurs.
Compared with model group, NAFLD mouse is after bifidobacterium adolescentis CCFM1061 intervention, in enteron aisle
Akkermansia belong to abundance significantly improve, and a large number of studies show that, Akkermansia belong to obesity, diabetes, fatty liver with
And the diseases such as enteritis are negatively correlated, show that the present invention has the diseases such as reduction obesity, diabetes, fatty liver and enteritis pathogenetic
Function.
Embodiment 14: bifidobacterium adolescentis CCFM1061 significantly reduces the level of NAFLD mice serum total cholesterol (TC)
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.
Experimental result is as shown in figure 17.Model group mice serum total cholesterol level is significantly raised, stomach-filling youth bifid bar
The TC that bacterium CCFM1061 significantly reduces model mice is horizontal and close to blank control group.Its ability for reducing mice serum TC
It is similar to Simvastatin medicine group.
Embodiment 15: it is horizontal that bifidobacterium adolescentis CCFM1061 reduces NAFLD mice serum glutamic-pyruvic transaminase (ALT)
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.It is measured according to the detection method of ALT kit
The content of glutamic-pyruvic transaminase (ALT) in blood.
Experimental result is as shown in figure 18.Model group mouse empty stomach ALT is significantly increased, and bifidobacterium adolescentis CCFM1061's is dry
The ALT for significantly reducing NAFLD mouse in advance is horizontal, and the ability for reducing mouse fasting blood glucose level is similar to Simvastatin, and
The intake of Lactobacillus rhamnosus L10 does not reverse the raising of ALT, and the ALT level of noticeable Rosiglitazone group is aobvious
It writes and is higher than model group, prompt long-term use Rosiglitazone that can cause drug induced injury to liver.
Embodiment 16: bifidobacterium adolescentis CCFM1061 reduces the level of NAFLD mice serum glutamic-oxalacetic transaminease (AST)
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.Blood is measured according to the detection method of kit
The content of middle glutamic-oxalacetic transaminease (AST).
Experimental result is as shown in figure 19.As seen from Figure 19, model group mice serum AST content is significantly raised, stomach-filling
Bifidobacterium adolescentis CCFM1061 significantly reduces the content of serum AST, and its trend is consistent with ALT trend, prompts youth bifid
Bacillus CCFM1061 can alleviate hepar damnification.
Embodiment 17: bifidobacterium adolescentis CCFM1061 reduces NAFLD mice serum low density lipoprotein cholesterol (LDL-
C level)
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.It is low close according to the detection method measurement of kit
Spend the content of lipoprotein cholesterol (LDL-C).
Experimental result is as shown in Fig. 20.It can be seen from experimental result compared with Normal group, model group mouse blood
Clear low density lipoprotein cholesterol content significantly increases, and stomach-filling bifidobacterium adolescentis CCFM1061 can reduce serum low-density rouge egg
The content of white cholesterol, and bifidobacterium adolescentis CCFM1061 is bright to the readjustment ability of serum LDL cholesterol level
It is aobvious to be better than Lactobacillus rhamnosus L10.
Embodiment 18: the fasting blood glucose level of bifidobacterium adolescentis CCFM1061 reduction NAFLD mouse
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.
Experimental result is as shown in figure 21.Model group mouse fasting blood-glucose significantly increases, bifidobacterium adolescentis CCFM1061's
Intervention significantly reduces the fasting blood glucose level of NAFLD mouse, and fasting blood-glucose control ability is significantly stronger than Lactobacillus rhamnosus
The intervention of L10, and its ability for reducing mouse fasting blood glucose level is similar to Rosiglitazone.
Embodiment 19: the insulin resistance of bifidobacterium adolescentis CCFM1061 alleviation NAFLD mouse
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.Pancreas islet is measured according to the detection method of kit
The content of plain (INS), and fasting blood-glucose result is combined to calculate insulin resistance index.
Experimental result is as shown in figure 22.Compared to the blank group, High cholesterol diet high in fat is after 24 weeks, model group mouse islets
Element is resisted index and is significantly increased, and NAFLD mouse islets element resists index and decreases after Lactobacillus rhamnosus L10 intervenes, but its
Effect is not so good as bifidobacterium adolescentis CCFM1061, and bifidobacterium adolescentis CCFM1061 is prompted to can be improved the pancreas islet of NAFLD mouse
Plain sensibility may have certain remission effect to type-II diabetes.
Embodiment 20: bifidobacterium adolescentis CCFM1061 reduces the level of triglycerides (TG) in liver
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.Glycerol is measured according to the detection method of kit
The content of three esters (TG), is corrected with liver protein concentration.
Experimental result is as shown in figure 23.It can be seen from experimental result compared with Normal group, model group mouse liver
TG is significantly increased, and stomach-filling bifidobacterium adolescentis CCFM1061 reduces the level of TG in NAFLD mouse liver, and bifidobacterium adolescentis
CCFM1061 is suitable with Simvastatin to the regulating power of liver TG.
Embodiment 21: bifidobacterium adolescentis CCFM1061 improves the level of superoxide dismutase from liver (SOD), with
Liver protein concentration is corrected.
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.According to superoxide dismutase (SOD) reagent
The content of SOD in the specification measurement liver of box.
Experimental result is as shown in figure 24.The SOD level of blank control group is higher than model group it can be seen from experimental result,
But without significant difference, stomach-filling bifidobacterium adolescentis CCFM1061 significantly improves the level of SOD in NAFLD mouse liver and is higher than pungent
Statin intervention group and Rosiglitazone intervention group are cut down, and Lactobacillus rhamnosus L10 does not show similar result after intervening.
Embodiment 22: bifidobacterium adolescentis CCFM1061 improves liver Glutathione Peroxidase (GSH-Px)
Level is corrected with liver protein concentration.
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.According to superoxide dismutase (SOD) reagent
The content of GSH-Px in the specification measurement liver of box.
Experimental result is as shown in figure 25.The level of model group liver Glutathione Peroxidase (GSH-Px) is higher than sky
White group, but without significant difference, stomach-filling bifidobacterium adolescentis CCFM1061 significantly improves NAFLD Gsh In Mouse Liver peptide peroxide
The level of compound enzyme (GSH-Px) and Lactobacillus rhamnosus L10 do not significantly improve the level of GSH-Px after intervening.
Embodiment 23: bifidobacterium adolescentis CCFM1061 mitigates the level of inflammation in NAFLD mouse liver
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.According to saying for interleukin-6 (IL-6) kit
The concentration of IL-6, is corrected in bright book measurement liver with liver protein concentration.
Experimental result is as shown in figure 26.High cholesterol diet high in fat is after 24 weeks it can be seen from experimental result, model group
The horizontal significant raising of IL-6, stomach-filling bifidobacterium adolescentis CCFM1061 significantly reduce IL-6 level in NAFLD mouse liver, and its
Inflammation remission effect is better than Simvastatin and Rosiglitazone, and NAFLD mouse inflammation alleviates effect after Lactobacillus rhamnosus L10 intervenes
Fruit is general.
Embodiment 24: bifidobacterium adolescentis CCFM1061 alleviates the tissue damage of NAFLD mouse liver
C57BL/6J mice group, modeling and processing method are the same as embodiment 2.The neutral paraformaldehyde in part 4% is taken to fix
Liver make paraffin section, observe and tissue morphology and take pictures under light microscopic after HE is dyed, carry out pathological evaluation.
Experimental result is as shown in figure 27.Model group mouse liver cell arranges sparse, liver fat drips it can be seen from experimental result
Quantity is more and not of uniform size, stick to each other between fat drips, lobuli hepatis have inflammatory cell infiltration, and balloon sample disease occurs for a small amount of liver cell
Become, and stomach-filling bifidobacterium adolescentis CCFM1061 can obviously improve above-mentioned lesion, and effect is substantially better than Lactobacillus rhamnosus L10
Intervention group.
Embodiment 25: bifidobacterium adolescentis CCFM1061 improves the level of Nrf2 in fatty liver cell
By L02 cell after containing the passage three times of 10% FBS continuous-stable, it is inoculated in 6 orifice plates, in 37 DEG C, 5%
CO2It is cultivated in environment for 24 hours, after cell is adherent, gives the blueness for individually accessing 1mL after 2mg/mL palmitinic acid is incubated for for 24 hours again
Spring Bifidobacterium CCFM1061 and Lactobacillus rhamnosus L10 (access PBS is as blank control) are incubated for for 24 hours.All incubations exist
37 DEG C, 5%CO2It is carried out in environment.Bifidobacterium adolescentis CCFM1061 stimulation test group, Lactobacillus rhamnosus L10 stimulation test
Group and each three holes of PBS control group, and in triplicate.
Culture solution is discarded, first cleans each hole with PBS buffer solution, each 1mL adds TRIZOL to crack after cleaning 3 times, into
Row cell RNA extracts.QPCR, which is carried out, after being cDNA by the RNA reverse transcription of extraction measures bifidobacterium adolescentis CCFM1061 and sandlwood
The expression of Nrf2 gene after sugared lactobacillus L10 and fatty liver cell are incubated for altogether.Nrf2 primer information is as shown in table 7.As a result
Using GAPDH as internal reference, it is expressed as
7 primer information of table
| Primer | It is positive | Reversely |
| NRF2 | CAACCCTTGTCACCATCTCA | GTGTTCTCACATTGGGCATC |
| GAPDH | AGGTCGGTGTGAACGGATTTG | TGTAGACCATGTAGTTGAGGTCA |
Experimental result is as shown in figure 28.Bifidobacterium adolescentis CCFM1061 stimulation significantly mentions it can be seen from experimental result
The high expression of the Nrf2 gene of fatty liver cell, the expression of the Nrf2 gene of Lactobacillus rhamnosus L10 stimulation group
Also there is raising but obvious not as good as bifidobacterium adolescentis CCFM1061 stimulation group, show that bifidobacterium adolescentis CCFM1061 may have
Certain oxidation resistance.
It should be noted that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to preferable
Embodiment describes the invention in detail, those skilled in the art should understand that, it can be to technology of the invention
Scheme is modified or replaced equivalently, and without departing from the spirit and scope of the technical solution of the present invention, should all be covered in this hair
In bright scope of the claims.
Claims (10)
1. bifidobacterium adolescentis CCFM1061 is preparing the application in functional microbial inoculum, food and/or drug, it is characterised in that: institute
Stating bifidobacterium adolescentis CCFM1061 can be used in preparing fasting blood-glucose and impaired glucose tolerance caused by improving type-II diabetes
Microbial inoculum, food and/or drug.
2. bifidobacterium adolescentis CCFM1061 as described in claim 1 is in preparing functional microbial inoculum, food and/or drug
Using, it is characterised in that: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves blood caused by type-II diabetes
Microbial inoculum, food and/or the drug that total cholesterol increases in clear, high-density lipoprotein cholesterol declines.
3. bifidobacterium adolescentis CCFM1061 as claimed in claim 2 is in preparing functional microbial inoculum, food and/or drug
Using, it is characterised in that: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves liver caused by type-II diabetes
The microbial inoculum, food and/or drug of inflammation in dirty tissue.
4. bifidobacterium adolescentis CCFM1061 according to any one of claims 1 to 3 prepare functional microbial inoculum, food and
Or the application in drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and significantly improve two types
Microbial inoculum, food and/or the drug of the pathology damage of pancreas, liver organization caused by diabetes.
5. bifidobacterium adolescentis CCFM1061 according to any one of claims 1 to 3 prepare functional microbial inoculum, food and
Or the application in drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061 can also be used to preparation absorption perfluoro caprylic acid,
Alleviate microbial inoculum, food and/or the drug of toxicity of perfluorooctanoic acid.
6. bifidobacterium adolescentis CCFM1061 as claimed in claim 5 is in preparing functional microbial inoculum, food and/or drug
Using, it is characterised in that: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and alleviate type-II diabetes mouse just
Secret microbial inoculum, food and/or drug.
7. the bifidobacterium adolescentis CCFM1061 as described in any one of claims 1 to 3,6 is preparing functional microbial inoculum, food
And/or the application in drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061 can also be used to prepare non-alcoholic rouge
Microbial inoculum, food and/or the drug of hepatic injury caused by fat hepatopathy.
8. the bifidobacterium adolescentis CCFM1061 as described in any one of claims 1 to 3,6 is preparing functional microbial inoculum, food
And/or the application in drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061 can also be used to preparation and improve non-alcohol
Microbial inoculum, food and/or the drug of the oxidation resistance of property fatty liver disease liver.
9. the bifidobacterium adolescentis CCFM1061 as described in any one of claims 1 to 3,6 is preparing functional microbial inoculum, food
And/or the application in drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves drink high in fat
Serum total cholesterol caused by eating, blood glucose, liver tg and the microbial inoculum of low density value protein cholesterol content rising are eaten
Product and/or drug.
10. the bifidobacterium adolescentis CCFM1061 as described in any one of claims 1 to 3,6 is preparing functional microbial inoculum, food
Application in product and/or drug, it is characterised in that: the bifidobacterium adolescentis CCFM1061, which can also be used to preparation, improves enteritis
Microbial inoculum, food and/or drug.
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| CN113005060A (en) * | 2021-03-15 | 2021-06-22 | 江南大学 | Application of bifidobacterium adolescentis CCFM1173 in preparation of functional microbial inoculum, food and/or medicament |
| WO2023029093A1 (en) * | 2021-08-31 | 2023-03-09 | 瑞微（深圳）生物科技有限公司 | Application of bifidobacterium adolescentis in preparation of drug for treating inflammation-related diseases |
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