CN109486700A - Lactobacillus paracasei N1115 prevents application and the corresponding probiotic powder, application of colitis - Google Patents
Lactobacillus paracasei N1115 prevents application and the corresponding probiotic powder, application of colitis Download PDFInfo
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- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/148—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with compounds of unknown constitution, e.g. material from plants or animals
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- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
Abstract
The present invention provides a kind of application of lactobacillus paracasei N1115 prevention colitis and corresponding probiotic powders, application, for early intervention or prevention colitis.The present invention can effectively prevent colitis using lactobacillus paracasei N1115；And the present invention can not only effectively prevent colitis using the probiotic powder that can prevent colitis of lactobacillus paracasei N1115 preparation, while in good taste.The present invention is suitable for all groups.
Description
Technical field
The invention belongs to bioengineering fields, are related to a kind of application for preventing colitis bacterial strain, specifically a kind of pair
The application of Lactobacillus casei N1115 prevention colitis；The present invention also provides utilize lactobacillus paracasei N1115 preparation
A kind of probiotic powder and its application.
Background technique
Inflammatory bowel disease (IBD) is one group of agnogenic chronic, non-specific inflammatory bowel disease, mainly includes ulcer
Property colitis and Crohn disease, if patient suffers from the disease, and symptom is unable to get effective control, and inflammation substance can be with blood
Whole body is flowed to, the systemic immunities diseases such as iritis, arthritis, erythema nodosum, therefore IBD patients' large intestine will be probably caused
The probability of cancer is also compared with ordinary person's height.
And the initial symptoms due to IBD patient and the pressure diarrhea that office worker is common are quite similar, along with most people
Crowd is not enough for IBD understanding, therefore usually obscures the initial stage sign of IBD with other gastrointestinal diseases, in turn results in and misses
Early detection, early treatment prime time.The disease past is common in western countries, and developing country's disease incidence is relatively
It is low, however in the past 20 years with the process of industrialization, urbanization, the disease incidence of Chinese IBD shows an increasing trend year by year.
Currently, the cause of disease of IBD is unclear, it is generally contemplated that hereditary, immune and intestinal flora takes part in IBD's jointly
Occur.But since IBD genesis mechanism is indefinite, so for the prevention of IBD, there is presently no clearly effective in the prior art
Ways and means, the domestic method for using Chinese medicine or therapy of combing traditional Chinese and Western medicine general at present, but it is ineffective.
Summary of the invention
The technical problem to be solved in the present invention is to provide for a kind of application of lactobacillus paracasei N1115 prevention colitis,
It is capable of the generation of early intervention prevention colitis.Lactobacillus paracasei N1115 of the invention is the tradition hair from China Inner Mongol
It is separated in kefir milk product, has been stored in Institute of Microorganism, Academia Sinica's Culture Collection Center, deposit number is
CGMCC No.4691；
The present invention also provides a kind of probiotic powder, lactobacillus paracasei N1115 joined in raw material, which can not only
Enough effectively prevention colitis, while can guarantee good mouthfeel, it is suitable for edible；
Present invention provides a kind of applications of above-mentioned probiotic powder, it is for early intervention or prevention colitis.
In order to solve the above technical problems, the technical solution used in the present invention is:
A kind of application of lactobacillus paracasei N1115 prevention colitis, for early intervention or prevention colitis.
The present invention also provides a kind of probiotic powders that can prevent colitis, and the raw material of the probiotic powder is made with parts by weight
Number meter includes: 10-15 parts of curcumin, 60-70 parts of oligofructose, cocoa power 10-15 parts high in fat；
Also contain lactobacillus paracasei N1115 0.8 × 10 in the probiotic powder12-1.2×1012cfu/g。
Present invention provides a kind of applications of above-mentioned probiotic powder, it is for early intervention or prevention colitis.
It is suitable for any crowd to take, is especially suitable for the irregular person of diet, the raw and cold vinegar-pepper person of diet hobby or functions of intestines and stomach
Weak has old age and the children for suffering from colitis risk, 1-2 times daily, each 2g.
Due to the adoption of the above technical solution, compared with prior art, the present invention acquired technological progress is:
(1) taking lactobacillus paracasei N1115 of the invention in early days can effectively prevent with IBD, and then prevent human body from suffering from
The diseases such as colitis；
(2) probiotic powder of the invention is using Chinese medical extract --- benefit is made in conjunction with lactobacillus paracasei N1115 in curcumin
Raw bacterium powder has the function of preferably preventing colitis；And the cocoa power and oligofructose being added in probiotic powder improve ginger
The not high problem of the solubility of flavine, even curcumin is more soluble；The anaerobism Conservation environment of probiotic powder solves ginger simultaneously
The problem of flavine stability difference, enables curcumin more easy to maintain；In addition curcumin has bitter taste, special pungent can be added
Cocoa power with chocolate flavor and chocolate bitter taste is covered, and avoids probiotic powder with Chinese medicine taste, to make this
The probiotic powder taste of invention is easier to the person's of being eaten receiving.
In conclusion lactobacillus paracasei N1115 of the invention can effectively prevent colitis；And the present invention is done using secondary
Prepared by Lactobacillus paracasei N1115, which can prevent colitis probiotic powder, can not only effectively prevent colitis, while in good taste.
The present invention is suitable for all groups.
The present invention is described in further detail below in conjunction with specific embodiment.
Detailed description of the invention
Fig. 1 a be the embodiment of the present invention 1 in not using lactobacillus paracasei N1115 to mouse progress early intervention, and not into
The intestinal tube structure chart of row DSS induction；
Fig. 1 b is to carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and do not carry out DSS
The intestinal tube structure chart of induction；
Fig. 1 c is not carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and carry out
The intestinal tube structure chart of DSS induction；
Fig. 1 d is to carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and carried out DSS
The intestinal tube structure chart of induction；
Fig. 2 is the intestinal tissue inflammatory scoring figure of four groups of mouse intestinal tissues in the embodiment of the present invention 1；
Fig. 3 a is not carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and carry out
The colon cell apoptosis figure of DSS induction；
Fig. 3 b is to carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and carried out DSS
The colon cell apoptosis figure of induction；
Fig. 3 c is not carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and do not carry out
The colon cell apoptosis figure of DSS induction；
Fig. 3 d is to carry out early intervention to mouse using lactobacillus paracasei N1115 in the embodiment of the present invention 1, and do not carry out DSS
The colon cell apoptosis figure of induction;
Fig. 4 is the TNF expression figure for the four groups of mouse tested in the embodiment of the present invention 1；
Fig. 5 is the KC expression figure for the four groups of mouse tested in the embodiment of the present invention 1.
Specific embodiment
The application of 1 lactobacillus paracasei N1115 of embodiment prevention colitis
The present embodiment is tested using new born C57BL/6 mouse as subjects, and mouse used is U.S.'s vanderburg university
Offer female and male mice each 12, weight 1.5g, comprising:
Step 1: intervening: 24 new born C57BL/6 mouse being divided into two groups at random, every group of male and female each 6, one group is
Lactobacillus paracasei N1115 intervention group, one group is control group.Wherein the mouse of lactobacillus paracasei N1115 intervention group is from birth
Start within the 1st day to after being born the 7th day afterwards, daily stomach-filling lactobacillus paracasei N1115 bacterium number 107Contain bacterium number in CFU(20 μ l water
107CFU)；Mouse is the 8th day to the 14th day after birth, daily stomach-filling lactobacillus paracasei N1115 bacterium number 108CFU(20 μ l water
In contain bacterium number 108CFU).And control group mice, the daily same amount of pure water of stomach-filling.Mouse, which is born the 15th day, to be started, secondary cheese
Lactobacillus N1115 intervention group stops intervening using lactobacillus paracasei N1115, and control group is constant, and two groups of mouse are later with identical
The mode of stomach-filling pure water is fed to 6 week old, carries out DSS intervention study subsequently into step 2.
Step 2: colitis model is established
The adult mice of 6 week old all in step 1 is divided into 4 groups, wherein carried out lactobacillus paracasei N1115 intervene and
The mouse for not carrying out lactobacillus paracasei N1115 intervention is divided into two groups at random, then carries out the research of DSS Intervention trial.
When test, carried out lactobacillus paracasei N1115 intervention two groups of mouse in one group of carry out DSS intervention, another group without
DSS intervenes.One group of carry out DSS intervention in two groups of mouse of lactobacillus paracasei N1115 intervention is not carried out equally, another group is not
Carry out DSS intervention.By above-mentioned test, mouse has been divided into four groups, has been respectively as follows: no lactobacillus paracasei N1115 intervention+nothing
DSS intervenes；No lactobacillus paracasei N1115 intervenes+carry out DSS intervention；It is dry without DSS lactobacillus paracasei N1115+ has been carried out
In advance；Carried out lactobacillus paracasei N1115 intervene+carry out DSS intervention.
The process of intervention modeling is carried out in the above process using DSS are as follows: 6 week old mouse, using DSS(3%, molecular weight 36-
50 kDa) intervene 4 days, induce damage of intestines and acute colitis；And sterile water conduct is then used without the DSS 6 week old mouse intervened
Control.After the completion of DSS intervenes, all mouse are put to death, detection inflammation damnification, Epithelial Cell Apoptosis, proinflammatory cytokine etc. refer to
Mark.
Three, evaluation index:
Colonic tissue damage and Injury score: the colonic tissue slice of all test mices is passed through after hematoxylin eosin staining
The detection of Application Optics microscope.Then, by the sample of the entire colonic tissue from same mouse, reference literature points-scoring system,
Intestinal tract injury is assessed.
The detection of colonic tissue Apoptosis: ApopTagTMIn SituOligo Ligation(situ oli-gonucleotide is applied
Connection, ISOL) kit handles sample using T4 DNA ligase according to guidance method, it is sliced in differential interference
(DIC) microscopically observation.
Cytokine gene expression detection: the colonic tissue of each detection sample is broken into homogenate, is separated and is tried using RNA
Agent box separates total serum IgE in sample, with the DNA enzymatic removal DNA interference of no RNA enzyme；Then, using large capacity cDNA Reverse Transcription
Box and 7300RT-PCR system carry out reverse transcription；As a result it is analyzed using Sequence Detection System software.
Lactobacillus paracasei N1115 early intervention carries out the influence that DSS intervenes mouse epithelial damage to the later period
Mouse intestinal tissue figure that Fig. 1 a is Non-LP N1115/Water, Fig. 1 b are the small of LP N1115 treatment/Water
It is the normal intestinal tissue for not carrying out DSS intervention shown in mouse intestinal tissue figure, i.e. Fig. 1 a and Fig. 1 b, just by Fig. 1 a, Fig. 1 b
Normal histology shows that the arrangement of mouse Colon epithelial cell is complete, and clear in structure, lamina propria has a small amount of lymphocyte, goblet cell
Abundant, glandular tube structure is normal, and the inflammatory damage scoring of epithelial cell is illustrated in figure 20.
Fig. 1 c is the mouse intestinal tissue figure of Non-LP N1115/DSS, i.e., not using lactobacillus paracasei N1115 to mouse
Early intervention is carried out, and directlys adopt DSS induction, then occurs serious Traumatic Colon and acute colitis symptom in Fig. 1 c,
With ulcer, crypts destroys and serious inflammatory, therefore, the inflammatory damage scoring such as Fig. 2 of the epithelial cell of Fig. 1 c
It show 9.63 ± 0.83(* P < 0.01), which prompts the colon models success of DSS induction.
However, the mouse that early utilization lactobacillus paracasei N1115 intervenes, then after carrying out DSS induction, as shown in Figure 1 d,
The damage of epithelial cell significantly reduces, and the inflammatory damage scoring of epithelial cell is illustrated in figure 2 6.11 ± 0.83(P <
0.05).Result prompt, the early intervention of lactobacillus paracasei N1115 significantly reduce DSS to the damage of intestinal epithelial cell
Wound has the function of certain prevention colitis.
Influence of the lactobacillus paracasei N1115 early intervention to suckling mouse adult hindgut Epithelial Cell Apoptosis
Gut integrity destruction is DSS induction one of intestinal tract injury and the important feature of colitis, and the present embodiment passes through Apop
Taq, ISOL detection kit, the influence to lactobacillus paracasei N1115 early intervention to intestinal epithelial cell apoptosis are commented
Valence, as a result as Fig. 3 a to Fig. 3 d is shown.Wherein the Non-LP N1115/DSS of Fig. 3 a indicates not utilize lactobacillus paracasei
N1115 carries out early intervention to mouse and directly DSS induces intestines mouse enteritis, colon epithelial cell application Apop Taq, ISOL
The fluorescence signal of stronger red can be detected in reagent, illustrates that DSS results in mouse intestinal integrity violations, Apoptosis is bright
It is aobvious.Non-LP N1115/Water and Fig. 3 d LP N1115 treatment/Water of control group Fig. 3 c is respectively not to small
Mouse carries out DSS induction, therefore fluorescence signal is weaker, illustrates that apoptotic cell is less, gut integrity is preferable.
However, LP N1115 treatment/ DSS indicates to carry out mouse using lactobacillus paracasei N1115 in Fig. 3 b
Early intervention, after DSS induction, compared with positive controls, fluorescence signal is significantly reduced, and colon cell apoptosis obviously subtracts
It is weak.The result further illustrates that lactobacillus paracasei N1115 early intervention there is protection to make the apoptosis of intestinal epithelial cell
With, have certain prevention colitis, weaken the effect of Epithelial Cell Apoptosis.
Lactobacillus paracasei N1115 intervenes the influence to TNF and KC expression after suckling mouse adult
Proinflammatory cytokine (such as TNF and KC) generation increases, this is a feature of the colitis of DSS induction.And the present embodiment
In all Tumor necrosis factor TNF (Mm00443259) and primer KC(Mm00433859), be purchased from Applied
Biosystems company.The quantitative phase with glyceraldehyde 3-phosphate dehydro-genase (GAPDH) mRNA of target gene mRNA in all samples
To abundance as reference standard, all cDNA samples are that 3 parts of a formula is analyzed.
The influence of inflammatory cytokine, is shown in the colitis model that lactobacillus paracasei N1115 early intervention induces DSS
Fig. 4 and Fig. 5, from Fig. 4 with the intestines it is found that compared with control group (not carrying out the mouse of DSS induction), induced by DSS in Fig. 5
The mRNA level in-site of the TNF and KC of inflammation group mouse dramatically increase (* P < 0.01, * P < 0.01).And by two groups by DSS induction
Enteritis group is compared, and DSS induction does not utilize secondary cheese using what lactobacillus paracasei N1115 early intervention group compared DSS induction
Lactobacillus N1115 early intervention group is compared, and the mRNA of the TNF and KC of lactobacillus paracasei N1115 early intervention group mouse are utilized
It is horizontal then significantly reduce (#P < 0.05, #P < 0.05).The lactobacillus paracasei N1115 of result prompt, early stage intervenes to intestines
Road colitis has prevention effect.
Embodiment 2-7 utilizes the probiotic powder of lactobacillus paracasei N1115 preparation
Embodiment 2-7 is respectively a kind of probiotic powder prepared using lactobacillus paracasei N1115, they are by curcumin, low
Fructooligosaccharides, cocoa power high in fat are uniformly mixed with lactobacillus paracasei and are made.The raw material proportioning of these embodiments see the table below 1:
1 embodiment 2-7 raw material proportioning of table
Embodiment 2-7 in conjunction with lactobacillus paracasei N1115, can preferably prevent colitis using Turmeric element, but by
Low in the solubility of curcumin, when reconciliation, needs to improve the solubility of curcumin, to enable the more effective fruit of the present embodiment, therefore this reality
It applies example and has carried out dissolubility test for curcumin, test the material of selection are as follows: curcumin, common flavoring agent and prebiotics, this
The flavoring agent and prebiotics that embodiment uses use cocoa power in the prior art, Fragaia ananassa Duchesne extract, inulin, oligofructose.
Experimental method:
By curcumin 5g with mixed respectively with cocoa power, Fragaia ananassa Duchesne extract, inulin, oligofructose according to mass ratio 1:1 after be added
In 40 DEG C of warm water of 100mL, glass bar stands 20min after stirring 30 seconds；Simultaneously respectively by 5g cocoa power, Fragaia ananassa Duchesne extract, chrysanthemum
Four kinds of powder, oligofructose samples to be tested are individually added into 40 DEG C of warm water of 100mL, and glass bar stands 20min after stirring 30 seconds；
Supernatant is abandoned after all 100mL solution is centrifuged, precipitating is placed in 50 DEG C of baking oven 2h in glass dish, weighs after drying,
Middle mixing example weight is denoted as A, and sample to be tested weight is denoted as a, curcumin meltage Y=5-(A-a).
Test result such as table 2:
Table 2
By table 2 it is found that dissolving preferable flavoring agent for curcumin is cocoa power and oligofructose, therefore the present embodiment again will
Preferable flavoring agent cocoa power is dissolved to curcumin and oligofructose carries out ratio adjustment, and carries out promotion turmeric in the same way
Plain solubility test, test procedure are as follows: by cocoa power and oligofructose with mass ratio be respectively 1:1,1:2,1:3,1:4,1:5,1:
6, respectively with 1 × 10 after the ratio mixing of 1:712Cfu/kg lactobacillus paracasei N1115 is mixed, and using using above-mentioned
Dissolution weight testing method detects it and promotes curcumin solvability.
Experimental group design and result such as table 3:
Table 3
By table 3 it is found that the adjustment of the two ratio does not have an apparent correlation for the solvability of curcumin, therefore from cost control
System considers to select cocoa power: oligofructose is added with mass ratio for the ratio of 1:7.
The test of taste modification effect:
Since curcumin is Chinese medicine, with bitter taste and special pungent, directly curcumin and lactobacillus paracasei N1115 are tied
It closes, mouthfeel is very poor, it is therefore desirable to which flavoring agent is effectively covered and modified to curcumin taste.Since above-mentioned experiment discovery can
Can powder can promote the dissolution of curcumin, while cocoa power is with strong bitter taste, therefore considers relatively adjustment proper ratio, leads to
Hardship, the perfume (or spice) of the chocolate of cocoa power are crossed to modify curcumin taste.
Taste tests are carried out after curcumin is mixed with cocoa power with mass ratio 2:1,1:1,1:2,1:3, and respectively
Taste Blind Test has been carried out in cell, supermarket, university.
Object and requirement: 90 people of male is investigated altogether, 81 people of women, the age was at 10~69 years old.
1) test method
Test specimen appraises through comparison sample from chocolate flavoured, pungent and taste of traditional Chinese medicine and preference degree four, by each comparation and assessment
Score value appraises through comparison product by the method for weighting multiplied by the practical score of the sample is added as after scoring weight, three indexs
The weight that scores and illustrated in table 4:
4 product test evaluation table of table
2) test result
Sample taste test statistics the results are shown in Table 5, and most of consumer thinks to deserve the quality of cocoa power and curcumin in sample
When than being higher than 1:1, the taste of traditional Chinese medicine and pungent of test specimen are weaker, and taste modification effect is preferable.
Table 5
The application of 8 the provided probiotic powder of embodiment 2-7 of embodiment
Each probiotic powder provided by embodiment 2-7, is tried using new born C57BL/6 mouse as subjects
It tests, mouse used is the female that vanderburg university in the U.S.'s provides and male mice each 12, weight 1.5g, comprising:
Step 1: intervening: 24 new born C57BL/6 mouse are divided into two groups at random, one group is probiotic powder intervention group,
One group is control group.Wherein the mouse of intervention group after birth the 1st day to the 7th day after birth, daily stomach-filling probiotic powder
(stomach-filling amount is with contained lactobacillus paracasei N1115 bacterium number for 107CFU is calculated)；Mouse is the 8th day to the 14th day after birth, often
(stomach-filling amount is with contained lactobacillus paracasei N1115 bacterium number for 10 for its stomach-filling probiotic powder8CFU is calculated).And control group mice, often
The same amount of pure water of its stomach-filling.Mouse, which is born the 15th day, to be started, and intervention group stops intervening, and control group is constant, two groups small later
Mouse is fed in a manner of identical stomach-filling pure water to 6 week old, carries out DSS intervention study subsequently into step 2.
Step 2: colitis model is established
The adult mice of 6 week old all in step 1 is divided into 4 groups, wherein having carried out probiotic powder intervention and not carried out prebiotic
The pre- mouse of bacterium dried bean noodles is randomly divided into two groups, then carries out the research of DSS Intervention trial.When test, intervened two groups
One group of carry out DSS intervention in mouse, another group is intervened without DSS.One group in two groups of mouse that do not intervened equally
DSS intervention is carried out, another group is intervened without DSS.By above-mentioned test, mouse four groups are divided into, has been respectively as follows: without prebiotic
Bacterium dried bean noodles is pre-+intervenes without DSS；No probiotics dried bean noodles is pre-+carry out DSS intervention；Carried out probiotic powder intervention+without DSS intervene；Into
Probiotic powder intervention+carry out DSS intervention is gone.
The process of intervention modeling is carried out in the above process using DSS are as follows: 6 week old mouse, using DSS(3%, molecular weight 36-
50 kDa) intervene 4 days, induce damage of intestines and acute colitis；And sterile water conduct is then used without the DSS 6 week old mouse intervened
Control.After the completion of DSS intervenes, all mouse are put to death, detection inflammation damnification, Epithelial Cell Apoptosis, proinflammatory cytokine etc. refer to
Mark.
Three, evaluation index:
Colonic tissue damage and Injury score: the colonic tissue slice of all test mices is passed through after hematoxylin eosin staining
The detection of Application Optics microscope.Then, by the sample of the entire colonic tissue from same mouse, reference literature points-scoring system,
Intestinal tract injury is assessed.
The detection of colonic tissue Apoptosis: ApopTagTMIn SituOligo Ligation(situ oli-gonucleotide is applied
Connection, ISOL) kit handles sample using T4 DNA ligase according to guidance method, it is sliced in differential interference
(DIC) microscopically observation.
Cytokine gene expression detection: the colonic tissue of each detection sample is broken into homogenate, is separated and is tried using RNA
Agent box separates total serum IgE in sample, with the DNA enzymatic removal DNA interference of no RNA enzyme；Then, using large capacity cDNA Reverse Transcription
Box and 7300RT-PCR system carry out reverse transcription；As a result it is analyzed using Sequence Detection System software.
Probiotic powder early intervention carries out the influence that DSS intervenes mouse epithelial damage to the later period
It is shown by normal histology, the arrangement of mouse Colon epithelial cell is complete, and clear in structure, lamina propria has a small amount of lymph thin
Born of the same parents, goblet cell is abundant, and glandular tube structure is normal, and the inflammatory damage scoring of epithelial cell is 0.
The result shows that: early intervention is not carried out to mouse using probiotic powder and directlys adopt DSS and induces, then is occurred tight
The Traumatic Colon and acute colitis symptom of weight, with ulcer, crypts destroys and serious inflammatory, epithelial cell
Inflammatory damage scoring be 9.62 ± 0.829(* P < 0.01), the result prompt DSS induction colon models success.
However, the mouse of early utilization probiotic powder intervention, then after carrying out DSS induction, the damage of epithelial cell is significantly dropped
Low, the inflammatory damage scoring of epithelial cell is 6.1 ± 0.83(P < 0.05).The early stage of result prompt, probiotic powder is dry
Damage of the DSS to intestinal epithelial cell is significantly reduced in advance, has the function of certain prevention colitis.
Influence of the probiotic powder early intervention to suckling mouse adult hindgut Epithelial Cell Apoptosis
Gut integrity destruction is DSS induction one of intestinal tract injury and the important feature of colitis, and the present embodiment passes through Apop
Taq, ISOL detection kit, the influence to probiotic powder early intervention to intestinal epithelial cell apoptosis are evaluated, as a result table
Show and early intervention and directly DSS induction intestines mouse enteritis, colon epithelial cell application are not carried out to mouse using probiotic powder
The fluorescence signal of stronger red can be detected in Apop Taq, ISOL reagent, and it is broken to illustrate that DSS results in mouse intestinal integrality
Bad, Apoptosis is obvious.The mouse of DSS induction is not carried out, and fluorescence signal is weaker, illustrates that apoptotic cell is less, gut integrity
Preferably.
However, early intervention is carried out to mouse using probiotic powder, and after DSS induction, compared with positive controls, fluorescence
Signal significantly reduces, and colon cell apoptosis obviously weakens.The result further illustrates that probiotic powder early intervention is on enteron aisle
The apoptosis of chrotoplast has protective effect, has certain prevention colitis, weakens the effect of Epithelial Cell Apoptosis.
Influence of the probiotic powder intervention to TNF and KC expression after suckling mouse adult
Proinflammatory cytokine (such as TNF and KC) generation increases, this is a feature of the colitis of DSS induction.And the present embodiment
In all Tumor necrosis factor TNF (Mm00443259) and primer KC(Mm00433859), be purchased from Applied
Biosystems company.The quantitative phase with glyceraldehyde 3-phosphate dehydro-genase (GAPDH) mRNA of target gene mRNA in all samples
To abundance as reference standard, all cDNA samples are that 3 parts of a formula is analyzed.
The influence of inflammatory cytokine in the colitis model that probiotic powder early intervention induces DSS, (i.e. with control group
The mouse of DSS induction is not carried out) it compares, the mRNA level in-site of the TNF and KC of the enteritis group mouse by DSS induction dramatically increase
(* P < 0.01, * P < 0.01).And compare two groups of enteritis groups by DSS induction, DSS induction utilizes probiotic powder early stage
Intervention group compares not comparing using probiotic powder early intervention group for DSS induction, utilizes probiotic powder early intervention group mouse
The mRNA level in-site of TNF and KC then significantly reduces (#P < 0.05, #P < 0.05).Result prompt, the probiotic powder intervention of early stage
There is prevention effect to enteric colonic inflammation.
Claims (3)
1. a kind of application of lactobacillus paracasei N1115 prevention colitis, it is characterised in that: the lactobacillus paracasei N1115
For early intervention or prevention colitis.
2. a kind of probiotic powder, it is characterised in that the raw material that the probiotic powder is made includes: curcumin 10-15 based on parts by weight
Part, 60-70 parts of oligofructose, cocoa power 10-15 parts high in fat；
Also contain lactobacillus paracasei N1115 0.8 × 10 in the probiotic powder12-1.2×1012cfu/kg。
3. a kind of application of probiotic powder according to claim 2, it is characterised in that: the probiotic powder is for early stage
Intervene or prevent colitis.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110892940A (en) * | 2019-11-20 | 2020-03-20 | 内蒙古伊利实业集团股份有限公司 | New application of lactobacillus paracasei K56 in relieving intestinal inflammation |
| CN110893195A (en) * | 2019-09-30 | 2020-03-20 | 内蒙古伊利实业集团股份有限公司 | Lactobacillus paracasei ET-22 with function of relieving intestinal inflammation |
| CN113396975A (en) * | 2021-07-01 | 2021-09-17 | 光明乳业股份有限公司 | Application of lactobacillus paracasei in preparing fermented dairy product in tumor |
| CN114159478A (en) * | 2021-11-22 | 2022-03-11 | 江南大学 | Bacterial powder composition for relieving inflammatory colitis and preparation method and application thereof |
Citations (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3017493B1 (en) * | 1999-02-25 | 2000-03-06 | 明治乳業株式会社 | Autoimmune disease prevention composition |
| CN1338940A (en) * | 1999-01-15 | 2002-03-06 | 爱尔兰企业(贸易上称作爱尔兰生物学研究公司) | Bidifobacterium in the treatment of inflammatory disease |
| WO2002053706A1 (en) * | 2000-12-29 | 2002-07-11 | Seong-Kun Cho | Novel lactobacillus paracasei subsp. paracasei strain antibacterial against helicobacter pylori and escherichia coli 157:h7 |
| EP1312667A1 (en) * | 2000-08-25 | 2003-05-21 | Wakamoto Pharmaceutical Co., Ltd. | Probiotics products containing lactic acid bacterium |
| CN1556853A (en) * | 2001-07-26 | 2004-12-22 | Probiotics lactobacillus casei bacterial strain | |
| CN1639317A (en) * | 2001-07-26 | 2005-07-13 | 营养健康有限公司 | Probiotic lactobacillus salivarius strains |
| CN101067119A (en) * | 2006-11-29 | 2007-11-07 | 康哲医药研究(深圳)有限公司 | Fermenting lactobacillus CMS II002 and its application |
| JP2008212006A (en) * | 2007-02-28 | 2008-09-18 | Meiji Milk Prod Co Ltd | Intestinal tract barrier function-recovering agent and intestinal tract barrier permeability exasperation inhibitor |
| KR20090116051A (en) * | 2008-05-06 | 2009-11-11 | 주식회사한국야쿠르트 | Lactobacillus brevis hy7401 having protective effect against colitis and products containing thereof as effective component |
| JP2011004620A (en) * | 2009-06-23 | 2011-01-13 | Tokai Igaku Kensa Kenkyusho:Kk | Genus lactobacillus strain and food and medicine containing the strain |
| CN102373173A (en) * | 2011-11-11 | 2012-03-14 | 石家庄君乐宝乳业有限公司 | Lactobacilluscasei N1115 and immune regulation function and application thereof |
| CN106497854A (en) * | 2017-01-19 | 2017-03-15 | 南京农业大学 | Lactobacilluss D8 and its application |
| CN106819106A (en) * | 2016-12-28 | 2017-06-13 | 石家庄君乐宝乳业有限公司 | Inactivation type leben with immunoloregulation function and preparation method thereof |
| CN107164263A (en) * | 2017-05-27 | 2017-09-15 | 西南大学 | A kind of adjustable function of intestinal canal, the lactobacillus fermenti HY01 for preventing colitis and application thereof |
| CN107619811A (en) * | 2017-11-08 | 2018-01-23 | 光明乳业股份有限公司 | Lactobacillus plantarum CCFM200 bacterial strains and application |
| CN108077925A (en) * | 2017-12-15 | 2018-05-29 | 石家庄君乐宝乳业有限公司 | The preparation method of synthetism rasp berry extract and the probiotic powder of tool enhancing immune function |
| CN108315285A (en) * | 2018-05-08 | 2018-07-24 | 西藏高原之宝牦牛乳业股份有限公司 | A kind of Lactobacillus paracasei and preparation method thereof |
-
2018
- 2018-08-31 CN CN201811008074.4A patent/CN109486700A/en not_active Withdrawn
Patent Citations (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338940A (en) * | 1999-01-15 | 2002-03-06 | 爱尔兰企业(贸易上称作爱尔兰生物学研究公司) | Bidifobacterium in the treatment of inflammatory disease |
| JP3017493B1 (en) * | 1999-02-25 | 2000-03-06 | 明治乳業株式会社 | Autoimmune disease prevention composition |
| EP1312667A1 (en) * | 2000-08-25 | 2003-05-21 | Wakamoto Pharmaceutical Co., Ltd. | Probiotics products containing lactic acid bacterium |
| WO2002053706A1 (en) * | 2000-12-29 | 2002-07-11 | Seong-Kun Cho | Novel lactobacillus paracasei subsp. paracasei strain antibacterial against helicobacter pylori and escherichia coli 157:h7 |
| CN1556853A (en) * | 2001-07-26 | 2004-12-22 | Probiotics lactobacillus casei bacterial strain | |
| CN1639317A (en) * | 2001-07-26 | 2005-07-13 | 营养健康有限公司 | Probiotic lactobacillus salivarius strains |
| CN101067119A (en) * | 2006-11-29 | 2007-11-07 | 康哲医药研究(深圳)有限公司 | Fermenting lactobacillus CMS II002 and its application |
| JP2008212006A (en) * | 2007-02-28 | 2008-09-18 | Meiji Milk Prod Co Ltd | Intestinal tract barrier function-recovering agent and intestinal tract barrier permeability exasperation inhibitor |
| KR20090116051A (en) * | 2008-05-06 | 2009-11-11 | 주식회사한국야쿠르트 | Lactobacillus brevis hy7401 having protective effect against colitis and products containing thereof as effective component |
| JP2011004620A (en) * | 2009-06-23 | 2011-01-13 | Tokai Igaku Kensa Kenkyusho:Kk | Genus lactobacillus strain and food and medicine containing the strain |
| CN102373173A (en) * | 2011-11-11 | 2012-03-14 | 石家庄君乐宝乳业有限公司 | Lactobacilluscasei N1115 and immune regulation function and application thereof |
| CN106819106A (en) * | 2016-12-28 | 2017-06-13 | 石家庄君乐宝乳业有限公司 | Inactivation type leben with immunoloregulation function and preparation method thereof |
| CN106497854A (en) * | 2017-01-19 | 2017-03-15 | 南京农业大学 | Lactobacilluss D8 and its application |
| CN107164263A (en) * | 2017-05-27 | 2017-09-15 | 西南大学 | A kind of adjustable function of intestinal canal, the lactobacillus fermenti HY01 for preventing colitis and application thereof |
| CN107619811A (en) * | 2017-11-08 | 2018-01-23 | 光明乳业股份有限公司 | Lactobacillus plantarum CCFM200 bacterial strains and application |
| CN108077925A (en) * | 2017-12-15 | 2018-05-29 | 石家庄君乐宝乳业有限公司 | The preparation method of synthetism rasp berry extract and the probiotic powder of tool enhancing immune function |
| CN108315285A (en) * | 2018-05-08 | 2018-07-24 | 西藏高原之宝牦牛乳业股份有限公司 | A kind of Lactobacillus paracasei and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| 孙贵范: "《助力癌症康复 1825+癌症的自然医学疗法策略》", 31 July 2018, 中国协和医科大学出版社 * |
| 寻晓爽等: "干酪乳杆菌N1115对人结肠癌细胞增殖活性影响", 《预防医学论坛》 * |
| 张婵等: "益生菌治疗炎症性肠病的研究进展 ", 《中国微生态学杂志》 * |
| 曹永强等: "副干酪乳杆菌N1115发酵乳对小鼠便秘的改善作用", 《食品科学》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110893195A (en) * | 2019-09-30 | 2020-03-20 | 内蒙古伊利实业集团股份有限公司 | Lactobacillus paracasei ET-22 with function of relieving intestinal inflammation |
| CN110893195B (en) * | 2019-09-30 | 2023-03-14 | 内蒙古伊利实业集团股份有限公司 | Lactobacillus paracasei ET-22 with function of relieving intestinal inflammation |
| CN110892940A (en) * | 2019-11-20 | 2020-03-20 | 内蒙古伊利实业集团股份有限公司 | New application of lactobacillus paracasei K56 in relieving intestinal inflammation |
| WO2021098764A1 (en) * | 2019-11-20 | 2021-05-27 | 内蒙古伊利实业集团股份有限公司 | New application of lactobacillus paracasei k56 in relieving intestinal inflammation |
| EP4062763A4 (en) * | 2019-11-20 | 2024-01-10 | Inner Mongolia Yili Ind Group | New application of lactobacillus paracasei k56 in relieving intestinal inflammation |
| CN113396975A (en) * | 2021-07-01 | 2021-09-17 | 光明乳业股份有限公司 | Application of lactobacillus paracasei in preparing fermented dairy product in tumor |
| CN114159478A (en) * | 2021-11-22 | 2022-03-11 | 江南大学 | Bacterial powder composition for relieving inflammatory colitis and preparation method and application thereof |
| CN114159478B (en) * | 2021-11-22 | 2022-11-15 | 江南大学 | Bacterial powder composition for relieving inflammatory colitis and preparation method and application thereof |
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