CN107022502A - Application of the clostridium butyricum in food hypersenstivity food or medicine is prevented and treated - Google Patents
Application of the clostridium butyricum in food hypersenstivity food or medicine is prevented and treated Download PDFInfo
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- CN107022502A CN107022502A CN201610280941.4A CN201610280941A CN107022502A CN 107022502 A CN107022502 A CN 107022502A CN 201610280941 A CN201610280941 A CN 201610280941A CN 107022502 A CN107022502 A CN 107022502A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/145—Clostridium
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- 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
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
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Abstract
The invention discloses a kind of clostridium butyricum, it is CGMCC NO.0313.1 in the deposit number of China General Microbiological culture presevation administrative center, applies in the food or medicine of food hypersenstivity is prevented.Relation of the invention by furtheing investigate anaphylactia and probiotics, by clostridium butyricum applied in the food or medicine that prevent food hypersenstivity, achieves unexpected technique effect, to prevent food hypersenstivity from providing new solution.
Description
Technical field
The invention belongs to microorganism new opplication field, more particularly, to a kind of clostridium butyricum in preventing and treating food mistake
Application in quick food or medicine.
Background technology
The incidence of anaphylactia is raised year by year, and according to World Health Organization, the population in the whole world 20% experienced
Quick property disease such as atopic dermatitis, food hypersenstivity, asthma, allergic rhinitis, allergic conjunctivitis, wherein with air flue, food mistake
It is quick more typical, and child's incidence within 1 years old is higher.The common original milk of allergy of induction allergic reaction generation,
Peanut, seafood, pollen, acarid, medicine etc., allergy can gently can be weighed, for serious allergic reaction, if uncontrolled cause
Death, along with allergic reaction, the decline of quality of life and the economic costs directly or indirectly that may occur at any time, allergy
The difficulty faced of crowd is unthinkable.
After baby due soon, a variety of floras are just occurred in that in intestines and stomach, about one week, gut flora field planting is complete
Into.Human body intestinal canal flora only has more than 200 kinds in cradle, and adult plants bacterium about more than the 400-500 and constituted, the micro- life of enteron aisle
Thing total amount is about 1013-1014, and about 1012 microorganisms are contained in every gram of excrement.
The pathogenesis of anaphylactia is complicated, it is generally recognized that Thl/Th2 immune imbalances, excessive Th2 immune responses
A series of allergic symptom is triggered.In all allergies, esophagus and human airway epithelial cells barrier function damage and allergic reaction
It is closely related, impaired esophagus and airway epithelia barrier, it is allowed to which intact antigen or other harmful substances pass through, into the anti-of body
Original chance contacts immunocyte, and as host, contact allergy original can trigger different degrees of allergic reaction again.Clinical main
Medicine for treatment, such as glucocorticoid, antihistamine, leukotriene receptor retarding agent, mainly mitigate allergic symptom, and specificity is exempted from
Epidemic disease treatment can induce body to anaphylactogen immune tolerance, but the treatment food hypersenstivity that all can not thoroughly cure.
The content of the invention
It is an object of the invention to by various probiotics are carried out with screening extensively and profoundly, finally filter out one plant of butyric acid
Clostridium, can apply to prevent and treat in food hypersenstivity food or medicine.
The clostridium butyricum, be in the deposit number of China General Microbiological culture presevation administrative center
CGMCC NO. 0313.1, the application in the food or medicine of food hypersenstivity is prevented.
Further, the clostridium butyricum is the CFU/ml of 3-5 × 1011.
Relation of the invention by furtheing investigate anaphylactia and probiotics, clostridium butyricum is applied to anti-
Only in the food or medicine of food hypersenstivity, unexpected technique effect is achieved, to prevent food hypersenstivity from providing newly
Solution.
Brief description of the drawings
Fig. 1 is 28 days Gain weight schematic diagrames of experimental example each group mouse of the present invention（N=8, ± s）；
Fig. 2 is experimental example optical microphotograph Microscopic observation mouse jejunum Pathologic changes result schematic diagram of the present invention（×100）；
Fig. 3 is experimental example serum total Ig E level testing result schematic diagram of the present invention（N=8, ± s）；
Fig. 4 is experimental example levels of IL-4 testing result schematic diagram of the present invention（N=8, ± s）；
Fig. 5 is the horizontal testing result schematic diagram of experimental example serum IL -13 of the present invention（N=8, ± s）；
Fig. 6 is serum IL-10 testing result schematic diagram（N=8, ± s）
Embodiment
Below in conjunction with the accompanying drawings and specific embodiment, the present invention is described in further details.
Experimental example
Cleaning grade BALB/c mouse, male, 6 week old, body weight 18-22g, purchased from The Fourth Military Medical University's Experimental Animal Center.Experiment
Group and control group are separately raised, and feeding environment temperature humidity is suitable, and water, feed, bedding and padding are totally sufficient.Bacterial strain is by Shandong Ke Xingsheng
Tetramune Co., Ltd provides.
- lactoglobulin (BLG, Sigma, Buchs, Switzerland), cholera toxin（CTX (Cholera
Toxin, List Biological Laboratories, Campbell, CA, USA), total IgE quantification kits（The U.S.
Chondrex companies）, IL-4, IL-10, IL-13, IFN-γ ELISA quantification kits（RD Systems, US）Deng.
Animal packet and model are set up：After weighing, according to the principle being grouped at random, BALB/c mouse is divided into four groups, respectively
For Normal group, food hypersenstivity group, clostridium butyricum prevention group and clostridium butyricum treatment group, every group 8.Normal group is modeled
Give physiological saline to compare, its excess-three group is the 7th, 14 and 21 days when carry out sensitization, gavage sensitization liquid 2ml/ only/time, 2ml sensitization
Liquid contains the μ g cholera toxins of 20mg-lactoglobulin (BLG, Sigma, Buchs, Switzerland)+adjuvant 10
（CTX, List Biological Laboratories, Campbell, CA, USA)；The 28th day of modeling, progress excites 1
Secondary, it is 3ml//times to excite liquid, and 3ml exciting liquids contain 100mg BLG.
Probiotics intervention experiment：While setting up model, prevention group and treatment group's gavage bacterium solution are intervened.Bacterium powder -20
DEG C preserve, bacterium powder normal saline dilution to 5 × 1011CFU/ml, 4 DEG C of preservations after packing.Prevention group was from the 1st to 21 day gavage
Bacterium solution 0.2ml/ days, the 22nd to 28 day gavage physiological saline 0.2ml/ days；Treatment group was from the 1st to 21 day gavage physiological saline
0.2ml/ days, the 22nd to 28 day gavage bacterium solution 0.2ml/ days；Normal group and the daily gavage sterile physiological salt of food hypersenstivity group
Water 0.2ml/ days, altogether gavage 28 days.After mouse fasting 12 hours, mouse experiment body weight is recorded, mouse Gain weight is calculated；Pluck
Eyeball takes and mouse jejunum is taken after blood, row HE dyeing；Total IgE, IFN-γ, IL-4, IL-5, IL-13 in ELISA method detection serum
With IL-10 level.
Observe mouse weight growth pattern：Mouse sensitization and after exciting, in fact it could happen that stool changes, diarrhoea number of times increases
It is many, vomiting, anorexia, weight loss, One's spirits are drooping depressed, or even occur and have blood in stool and dead, this experiment mainly passes through reality
Front and rear mouse weight change is tested to weigh the order of severity of mouse allergy symptom.
Mouse jejunum pathological change
Four groups of mouse completion eyeball, which takes, takes mouse jejunum 5cm after blood, with the enteric cavity of 4 DEG C of normal saline flushing mouse, rush repeatedly
Wash 3 times, intestinal tube is cut into tissue and is about 0.8cm intestinal segments, fixed after 48 hours, put after label with 10% neutral formalin
Enter in embedded box, by sample deliver to The Fourth Military Medical University's Pathological Staff Room be dehydrated, it is transparent, waxdip, embedding, section, de-
Wax, rehydration, HE dyeing, dehydration, mounting, the film making of light Microscopic observation.
EUSA（ELISA）：
After mouse fasting 12 hours, give after 3.5% chloraldurate intraperitoneal injection of anesthesia mouse, pluck eyeball and take blood, 2ml/, room temperature
After standing 2 hours, 2500rpm, room temperature is centrifuged 20 minutes, collects serum, -80 DEG C freeze after packing.In ELISA method detection serum
Total IgE, IL-4, IL-5, IL-10, IL-13 and IFN-γ level；The method of by specification is operated and determined OD values, is used
CurveExpert1.4 Software on Drawing standard curves, calculate each sample well concentration, multiplied by with the reality of extension rate, as sample
Border concentration.
Statistical analysis
Statistical analysis is carried out using SPSS19.0 softwares, measurement data data are represented with ± s, use one-way analysis of variance
（ANOVA）Difference compares between being organized,P<0.05 is that difference is statistically significant.
As a result
Substantially show and Gain weight：Each group weight ratio relatively draws, significant difference（F=149.6；P＜ 0.05）.Normal group
Mouse spirit is good, and appetite is good, and stool is normal, hair luster, and weightening is obvious.Food hypersenstivity group mouse weight substantially mitigates,
And occur stool change, have the fidgets, perpendicular hair, the allergic symptom such as be urinary and fecal inconvenience.Prevention group and treatment group's body weight have increase,
Other symptoms have mitigated compared with food hypersenstivity group（As a result Fig. 1, table 1 are seen）.
1 28 days each group mouse Gain weights of table（N=8,±s）
Note：A, is compared with normal groupP＜ 0.05；B, is compared with food hypersenstivity groupP＜ 0.05；C, is compared with clostridium butyricum prevention groupP＜ 0.05；D, treatment group is compared with clostridium butyricumP＜ 0.05
2.2 jejunum HE dye pathological change：Normal group mouse intestinal mucosa result is clear, intestinal villus marshalling, fine hair without
Substantially damage, no inflammation cellular infiltration, and its excess-three group has different degrees of epithelium of intestinal mucosa to come off is intestinal villus oedema, disconnected
Split, a large amount of inflammatory cell infiltrations, show that BLG allergy causes intestinal mucosa lesion under enteric epithelium.Clostridium butyricum prevention group and control
Treatment group, jejunum inflammatory cell infiltration degree mitigates（See Fig. 2A, B, C, D；A is Normal group；B is food hypersenstivity group；C is junket
Sour clostridium prevention group；D is clostridium butyricum treatment group）.
Serum total Ig E level：Serum total Ig E level has differences（F=97.02；P＜ 0.05）, food hypersenstivity group serum is total
IgE（809.1±87.65）Higher than Normal group（283.3±83.18）, significant difference（ P＜ 0.05）；Prevention group（284.5
±76.82）And treatment group（135.3±91.40）Significantly lower than food hypersenstivity group, the result of multiple comparisons shows that difference is equal in group
Write（Prevention group VS food hypersenstivity groups,P<0.05；Treatment group VS food hypersenstivity groupsP<0.05）.
The serum total Ig E level testing result of table 2（N=8,±s）
Note：A, is compared with normal groupP＜ 0.05；B, is compared with food hypersenstivity groupP＜ 0.05；C, is compared with clostridium butyricum prevention groupP＜ 0.05；D, treatment group is compared with clostridium butyricumP＜ 0.05
Cytokine of Serum level：The level of each group mice serum IL-5 and IFN-γ is extremely low after testing, can not measure
Imitate OD values, it is impossible to calculate valid density.Levels of IL-4, each group is compared, difference（F=41.12；P＜ 0.05）, its
Middle food hypersenstivity group（17.42±2.118）It is significantly raised, after oral clostridium butyricum, prevention group（4.48±2.085）And treatment group
（11.27±2.189）All substantially reductions, significant difference（Prevention group VS food hypersenstivity groups,P<0.05；Treatment group's VS food hypersenstivities
GroupP<0.05）（It is shown in Table 3, Fig. 4）；Had differences between IL-13, each group（F=23.08；P＜ 0.05）, wherein food hypersenstivity group
（156.2±11.65）It is significantly raised, after oral clostridium butyricum, prevention group（142.7±7.7）And treatment group（126.0±
6.502）Reduction is obvious, notable with food hypersenstivity group comparing difference（Prevention group VS food hypersenstivity groups,P<0.05；Treatment group VS is eaten
Thing allergy groupP<0.05）（It is shown in Table 3, Fig. 5）；Compare between serum IL-10, group and have differences（F=23.11；P＜ 0.05）,
Clostridium butyricum prevention group（30.14±5.553）And treatment group（37.53±7.810）, with food hypersenstivity group（25.65±6.185）
Compare, hence it is evident that rise, significant difference, with prevention control group（23.10±1.9）Compare, serum IL-10 is significantly raised（P<
0.05）（Prevention group VS food hypersenstivity groups,P<0.05；Treatment group VS food hypersenstivity groupsP<0.05）（It is shown in Table 3, Fig. 6）.
The Cytokine of Serum ELISA testing results of table 3（N=8,±s）
Note：A, is compared with normal groupP＜ 0.05；B, is compared with food hypersenstivity groupP＜ 0.05；C, is compared with clostridium butyricum prevention groupP＜ 0.05；D, treatment group is compared with clostridium butyricumP＜ 0.05.
Claims (2)
1. a kind of clostridium butyricum, is CGMCC in the deposit number of China General Microbiological culture presevation administrative center
NO. 0313.1, the application in the food or medicine of food hypersenstivity is prevented.
2. application of the clostridium butyricum as claimed in claim 1 in the food or medicine of food hypersenstivity is prevented, institute
It is the CFU/ml of 3-5 × 1011 to state clostridium butyricum.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610280941.4A CN107022502A (en) | 2016-05-03 | 2016-05-03 | Application of the clostridium butyricum in food hypersenstivity food or medicine is prevented and treated |
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| CN201610280941.4A CN107022502A (en) | 2016-05-03 | 2016-05-03 | Application of the clostridium butyricum in food hypersenstivity food or medicine is prevented and treated |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114437996A (en) * | 2022-03-25 | 2022-05-06 | 广西医科大学第一附属医院 | Heat-inactivated clostridium butyricum for preventing or treating asthma and preparation method thereof |
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| CN1989827A (en) * | 2005-12-30 | 2007-07-04 | 赵宏伟 | Application of butyric acid bacteria in the preparation of candy |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114437996A (en) * | 2022-03-25 | 2022-05-06 | 广西医科大学第一附属医院 | Heat-inactivated clostridium butyricum for preventing or treating asthma and preparation method thereof |
| CN114437996B (en) * | 2022-03-25 | 2024-03-15 | 广西医科大学第一附属医院 | Heat-inactivated clostridium butyricum for preventing or treating asthma and preparation method thereof |
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