CN107177535A - Applications of the Lactobacillus plantarum YS2 in the food or medicine of prevention ulcerative colitis is prepared - Google Patents
Applications of the Lactobacillus plantarum YS2 in the food or medicine of prevention ulcerative colitis is prepared Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
-
- 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
- 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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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
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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
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Mycology (AREA)
- Nutrition Science (AREA)
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- Polymers & Plastics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract
It is CCTCC NO the invention discloses deposit number：Applications of the M2016748 Lactobacillus plantarum YS2 (Lactobacillus plantarum YS2) in the food or medicine of prevention ulcerative colitis is prepared, not only expand Lactobacillus plantarum YS2 application, its application value is improved, and new hope is brought to the treatment or health care of ulcerative colitis.
Description
Technical field
The invention belongs to technical field of microbe application, it is related to a kind of Lactobacillus plantarum answering in food or medicine is prepared
With.
Background technology
Yak yogurt is a kind of spontaneous fermentation dairy products in Qinghai-Tibet Tibetan area, contains abundant nutritional ingredient, tool
There are anti-oxidant, reduction cholesterol, regulation immunity.Due to special spontaneous fermentation condition, including fermentation raw material breast, hair
Ferment temperature, fermentation time, fermentation vessel, particularly special fermentative microorganism composition, cause yak yogurt to have flavour
And quality.
Human ulcerative colitis is a kind of IBD of 2 type helper T lymphocyte (Th2) mediation, its cause of disease and hair
Interpretation of the cause, onset and process of an illness system is unclear, and therapeutic advance is slow and lacks specificity.The colitis of azolactone induction is a kind of Th2 of IL-4 mediations
Type inflammation, its histologic characteristics and inflammation are distributed similar human ulcerative colitis, the mouse Colon induced by Ci , azolactones
Scorching model can study and assess curative effect of medication or functional food physiological activity as Ulcerative Colitis
Beneficial instrument.
The content of the invention
It is an object of the invention to the mouse colitis model of Tong Guo azolactones induction, investigation is separated from yak yogurt
The Lactobacillus plantarum YS2 arrived is to the preventive effect of ulcerative colitis, to research and develop the food that can prevent ulcerative colitis
Or medicine.
Through research, the present invention provides following technical scheme：
Deposit number is CCTCC NO：M2016748 Lactobacillus plantarum YS2 (Lactobacillus plantarum
YS2) the application in the food or medicine of prevention ulcerative colitis is prepared.
Lactobacillus plantarum YS2 (Lactobacillus plantarum YS2) is the yak from Yushu district, Qinghai Tibetan autonomous prefecture
It is isolated in ox yogurt, it is preserved in China typical culture collection center (abbreviation CCTCC, address on December 14th, 2016：
No. 299 Wuhan Universitys of Wuhan City, Hubei Province Wuchang District Bayi Road), deposit number is CCTCC NO：M2016748.
After Shi azolactones induction BALB/c mouse colitis of the present invention, by determining, serum is related in colon to be referred to
Mark detection Lactobacillus plantarum YS2 colitis preventive effect.Experimental result shows that Lactobacillus plantarum YS2 can significantly reduce colon
The disease activity index (Disease activity index, DAI) of scorching mouse, suppresses the colon lengths contracting that colitis is caused
It is short, improve the ratio of colon weight/colon lengths；Colitis mice colon myeloperoxidase can be significantly reduced
(MPO), NO, MDA content and raising GSH contents；IL-2 levels and reduction IL- in colitis mice serum can also be significantly improved
10 levels.RT-PCR experimental results show that Lactobacillus plantarum YS2 can significantly raise neural in colitis mice colon
Type NOS (nNOS), endothelium in type NOS (eNOS), c-Kit, SCF mRNA expression, downward induce type NOS (iNOS), IL-8,
CXCR2mRNA is expressed.As can be seen here, the mouse colitis that Lactobacillus plantarum YS2 is induced azolactone has good pre- preventive effect
Really, available for the food or medicine for preparing prevention ulcerative colitis.
The beneficial effects of the present invention are：Prevention ulcerative colitis is being prepared the invention provides Lactobacillus plantarum YS2
Food or medicine in application, not only expand Lactobacillus plantarum YS2 application, improve its application value, and
Treatment or health care to ulcerative colitis bring new hope.
Brief description of the drawings
Fig. 1 is the agarose gel electrophoresis figure of Lactobacillus plantarum YS2 genomic DNA, and wherein M is λ DNA/Hind III
Marker, 1 is Lactobacillus plantarum YS2 genomic DNA.
Fig. 2 is the PCR amplifications of Lactobacillus plantarum YS2 16S rDNA genetic fragments, and wherein M is DNA Marker,
C is negative control, and 1 is the pcr amplification product of Lactobacillus plantarum YS2 16S rDNA genetic fragments.
Fig. 3 is Lactobacillus plantarum YS2 Dui azolactone inducing colitis mouse Colon tissue nNOS, eNOS and iNOS mRNA
The influence of expression.
Fig. 4 is what Lactobacillus plantarum YS2 Dui azolactone inducing colitis mouse Colon tissue c-Kit and SCF mRNA were expressed
Influence.
Fig. 5 is what Lactobacillus plantarum YS2 Dui azolactone inducing colitis mouse Colon tissues IL-8 and CXCR2mRNA were expressed
Influence.
Fig. 3 is into Fig. 5, and LB represents lactobacillus bulgaricus, and LP-YS2 represents Lactobacillus plantarum YS2.
Embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with accompanying drawing to the excellent of the present invention
Embodiment is selected to be described in detail.
The separation and identification of embodiment 1, Lactobacillus plantarum YS2
First, lactic acid bacteria isolates and purifies
Gather 10 parts of the spontaneous fermentation yak yogurt sample of Yushu district, Qinghai Tibetan autonomous prefecture herdsman family, every part of sample nothing
After bacterium glass rod is fully stirred evenly, draw 50mL and be put into sterile centrifugation tube, be placed in food sampling box and take back laboratory, 4 DEG C of refrigerators
Preserve.
Every part of yak yogurt sample takes 1mL, and 10 times of gradient dilutions are made to 10 with sterile saline-7.Take 10-5、10-6、
10-7Each 1mL dilutions of three dilution factors respectively at equipped with 15mL MRS culture mediums (containing mass ratio be 5% CaCO3) plate
In, mix, put 30 DEG C of 72 ± 3h of insulating box culture.Each three, dilution factor work is parallel.After bacterium colony is formed, its form is observed special
Levy, the different bacterium colonies that picking has molten calcium circle are inoculated in degreasing milk medium respectively, put 30 DEG C of 24~48h of culture, treat that bacterial strain is given birth to
After length is good, streak inoculation puts 30 DEG C of culture 48h in MRS agar mediums；Above step is repeated at least three times, until obtaining
Pure bacterium colony.
Pure bacterial strain is rule on MRS solid mediums, 30 DEG C of culture 48h are put, with 10 times of amplification sem observation bacterium colony sizes,
The appearance features such as shape, carry out Gram's staining, and the typical bacterial strain of picking form carries out catalase test.Gram is contaminated
Color is positive, catalase test is negative meningitidis strains and bacillus strain, are fixed tentatively as lactic acid bacteria.
2nd, the identification of Bacillus acidi lactici
By the lactic acid bacillus mycopremna that preliminary screening goes out carry out respectively growth temperature experiment (10 DEG C, 15 DEG C, 45 DEG C, 60 DEG C,
30min), pH value gradient test, mobility test, catalase test, oxidase test, hydrogen sulfide production test, gelatin liquefaction are real
Test, nitrate reduction test, indole test, the experiment of glucose aerogenesis, benzidine test, litmus milk tests, arginine production ammonia
Experiment, casein decomposition run and various sugar alcohol fermentation tests.
As a result isolated from 10 parts of yak yogurt samples and be purified into 20 plants of Bacillus acidi lacticis, be Gram-positive,
Catalase test is negative.By morphological observation and physiological and biochemical test, this 20 plants of bacterial strains are initially identified as 7 kinds of lactic acid
Bacillus, number consecutively is Lb1~Lb7, wherein, numbering is Lb5 Bacillus acidi lactici is initially identified as Lactobacillus plantarum
(Lactobacillus plantarum) (table 1).
The numbering of table 1 is LbThe morphological feature and physiological and biochemical test result of 5 Bacillus acidi lactici
Note：+, it is positive；-, it is negative；+ w, weakly positive.
3rd, the identification of Lactobacillus plantarum
It is L to take numberingbSupernatant is abandoned in the bacteria suspension of 5 Bacillus acidi lactici Liquid Culture, centrifugation, thalline is collected, according to bacterium base
Because a group DNA extraction kit (Beijing Suo Laibao scientific ＆ technical corporation) methods described extracts genomic DNA, Ago-Gel electricity is carried out
Swim (gel mass concentration ratio 0.8%, voltage 80V, electrophoresis 80min), after GelRed nucleic acid staining dyes, in gel imaging
(Fig. 1) is observed in system.The genomic DNA of extraction is diluted 200 times, concentration purity testing is carried out with ultraviolet specrophotometer.
Using primer 2 7F：5 '-AGAGTTTGATCCTGGCTCAG-3 ' (SEQ ID No.1) and 1492R： 5’-
GGCTACCTTGTTACGACTT-3 ' (SEQ ID No.2) enters performing PCR amplification (Fig. 2) to 16S rDNA genetic fragments.Amplification production
Thing is sequenced by Sangon Biotech (Shanghai) Co., Ltd. and Beijing Liuhe Huada Genomics Technology Co., Ltd, and will
Sequencing result (SEQ ID No.3) is compared with correlated series in Genbank.As a result show, numbering is Lb5 lactic acid bar
The similitude of the 16S rDNA gene fragment orders of bacterium and Lactobacillus plantarum strain CIP 103151 reaches
To 99%.
Therefore, numbering is Lb5 Bacillus acidi lactici is accredited as Lactobacillus plantarum (Lactobacillus plantarum), will
It is named as YS2, and is preserved in China typical culture collection center (abbreviation CCTCC, address on December 14th, 2016：Lake
No. 299 Wuhan Universitys of Bayi Road of Bei Sheng wuchang, wuhan area), deposit number is CCTCC NO：M2016748.
The prevention effect of embodiment 2, Lactobacillus plantarum YS2 Dui azolactone inducing mouse colitis
Main material, instrument and the experimental animal that the present embodiment is used are as follows：Lactobacillus plantarum YS2 (preserving numbers：CCTCC
NO：M2016748), lactobacillus bulgaricus (Chinese industrial Microbiological Culture Collection administrative center)；Azolactone (U.S. Sigma
Company)；MPO, NO, GSH, MDA, SOD determine kit (Bioengineering Research Institute is built up in Nanjing)；IL-2, IL-10 serum are thin
Intracellular cytokine determines kit (BioLegend companies of the U.S.)；Trizol reagents, OligodT18, RNase, dNTP, MLV are reversed
Record enzyme (Invitrogen companies of the U.S.)；ROX reference Dye and SYBR Premix Ex Taq II (Japanese TAKARA
Company)；RT-PCR primer (Beijing Tiangeng biochemical technology Co., Ltd).Biomate3S ultraviolet-visibles spectrophotometer,
The multi-functional ELIASA (Thermo Fisher Scientific companies of the U.S.) of A200PCR instrument, LUX；Tancon2500PCR
Gel imager (Shanghai Tian Neng Science and Technology Ltd.s)；(the limited public affairs of instrument are contained to ICEN-24R high speed freezing centrifuges by Hangzhou Austria
Department).SPF grades of BALB/c (body weight 25-30g) mouse of male 7 week old purchased from Medical University Of Chongqing's animal experimental center, (permit by animal
Card number：SCXK (Chongqing) 2012-0001).
50 BALB/c mouses are randomly divided into 5 groups, are normal group, model control group respectively, at lactobacillus bulgaricus
Reason group, Lactobacillus plantarum YS2 low concentrations treatment group and high concentration treatment group, every group 10.Normal group and model control group are normal
Freely take in diet and drink water 2 weeks；During this is 2 weeks except it is normal freely take in diet and drinks water in addition to, at lactobacillus bulgaricus
The 1.0 × 10 of the daily every gavage 2mL of reason group mouse9At CFU/mL lactobacillus bulgaricus, Lactobacillus plantarum YS2 low concentrations
The 1 × 10 of reason group and the daily every difference gavage 2mL of high concentration treatment group mouse8CFU/mL Lactobacillus plantarum YS2 and 1 ×
109CFU/mL Lactobacillus plantarum YS2.The belly of the 1st day all mouse carries out shaving, normal group with 2 × 2cm areas after 2 weeks
Mouse web portion smears 0.2mL ethanol；It is 3% azolactone solution (ethanol that other each group mouse web portions, which smear 0.2mL mass ratioes,
For solvent).Implement fasting to mouse, but allow mouse freely to take in drinking-water within 5th day.Mouse is implemented after fasting 24h to anaesthetize
(0.1mL/10g chloraldurate), then carries out bowel lavage：With silicone tube it is blunt nosed from mouse anus insert enteron aisle depth 3.5cm, normally
The ethanol solution that group mouse injection 0.15mL volume ratios are 50%, other each group mouse injection 0.15mL mass ratioes are 1% Evil
Conduit is extracted after oxazolone solution (ethanol solution that volume ratio is 50% is solvent), 20s, while mouse is inverted into 30s.Bowel lavage 7
The neck that breaks after it puts to death whole mouse, collects mice plasma and colon is standby.
1st, influences of the Lactobacillus plantarum YS2 to colitis symptoms
(the 20th day), 3 days (the 22nd day), 6 days (the 25th day) the DAI standards of grading calculating as described in table 2 respectively 1 day after bowel lavage
DAI values, DAI=(Body weight loss fraction+stool fraction+fraction of having blood in stool)/3.Colitis can cause body weight loss simultaneously
There is diarrhoea and the symptom such as bleeding, can be as measurement colitis using body weight, stool and the DAI having blood in stool as score standard
The standard of degree.
Table 2DAI standards of grading
The DAI value result of calculations of each group mouse are shown in Table 3, and DAI value of the normal group mouse in whole experimental period is remained
0.00 ± 0.00, DAI values increase and keep highest in each group always always after model control group mouse azolactone bowel lavage, protect
Plus the DAI values of Leah lactobacillus and Lactobacillus plantarum YS2 gavage Hou azolactone inducing colitis mouse are aobvious compared with model control group
Write and decline (p<0.05), wherein Lactobacillus plantarum YS2 high concentrations treatment group is reduced by up to.
The DAI values of each group mouse of table 3
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
Mouse Colon tissue is taken, the weight and length of colon is determined, the ratio of colon weight/colon lengths is calculated.Colon
The ratio of length and colon weight/colon lengths also weighs the standard of colitis degree.Compared with normal mouse, colon
The colon lengths of scorching mouse are shorter, colon weight/colon lengths ratios are lower.The colon lengths and colon weight of each group mouse
The ratio of amount/colon lengths is shown in Table 4, and the colon lengths of model control group mouse and the ratio of colon weight/colon lengths are most
It is low, normal group mouse highest, and the colon lengths and colon weight/colon of Lactobacillus plantarum YS2 high concentration treatment group mouse are long
The ratio of degree is closest to normal group.
The colon lengths of each group mouse of table 4 and the ratio of colon weight/colon lengths
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
The mouse colitis symptom of low azolactone induction can drop in above-mentioned experimental result explanation, Lactobacillus plantarum YS2, and with
The increase effect for concentration becomes apparent, and its effect is better than the lactobacillus bulgaricus with concentration.
2nd, influences of the Lactobacillus plantarum YS2 to mouse Colon tissue MPO, NO, GSH and MDA content
The physiological saline of 9 times of quality is added in clean mouse Colon tissue, it is using Ultrasonic Pulverization that colon is even
Slurry, is then measured by kit specification to MPO, NO, GSH and MDA content in mouse Colon tissue.MPO is in colon
Content improve and mean that stream enters colon in neutrophil leucocyte, be the table that neutrophil accumulation is reduced in Inflamed tissue
It is existing.INOS generates the damage that NO aggravates colon during colitis, while increasing in colon with NO content
Plus MPO activity also strengthens therewith, inflammation aggravation.Colitis may further result in the big volume productions of ROS (active oxygen) and RNS (active nitrogen)
It is raw, histocyte is occurred toxic reaction, colon is inflamed damage.Be inflamed reaction after ROS a large amount of generations
The content of GSH in body oxidation/anti-oxidant balance, reduction colon will be destroyed, substantial amounts of peroxidatic reaction of lipid aggravates fat
Matter peroxidating end-product MDA generation.
As shown in Table 5, MPO, NO, MDA content highest in model control group mouse Colon tissue, GSH contents are minimum；Just
Often the colon of group mouse shows opposite trend, and MPO, NO, MDA content are minimum, GSH content highests；Relative to model
Control group, lactobacillus bulgaricus and Lactobacillus plantarum YS2 can reduce MPO in colitis mice colon, NO,
MDA contents and raising GSH contents, but Lactobacillus plantarum YS2 effect is stronger.
MPO, NO, GSH and MDA content of each group mouse Colon tissue of table 5
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
3rd, influences of the Lactobacillus plantarum YS2 to mice serum cell factor IL-2 and IL-10 level
Mice serum is taken, IL-2 and IL-10 cell factors in mice serum are determined by kit specification using ELISA method
Content.The colitis of azolactone induction is a kind of cell-mediated inflammation of Th2.The mechanism that inflammation is produced is between Th2/Th1
Immunological network unbalance cause colitis.IL-10 is the cell factor that a kind of Th2 cells are produced, and IL-2 is that Th1 cells are produced
Cell factor, all directly related with colitis, too low IL-2 levels and too high IL-10 levels can mean that colitis degree
Deepen.
As shown in Table 6, normal group mice serum cell factor IL-2 levels are significantly higher than other group of mouse (p<0.05), and
IL-10 levels are substantially less than other group of mouse (p<0.05)；Compared with other each groups, Lactobacillus plantarum YS2 high concentration treatment groups
The closest normal group of mice serum cell factor IL-2 and IL-10 level.
Each group mice serum cell factor IL-2 and the IL-10 level of table 6
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
4th, the influence that Lactobacillus plantarum YS2 is expressed mouse Colon tissue correlation mRNA
The colon of mouse is taken, colon's total serum IgE is extracted with RNAzol after crushing, is diluted to 1 μ g/ μ L；Take 2 μ L dilute
Total RNAs extraction liquid after releasing, sequentially adds 1 μ L oligodT18, RNase, dNTP, MLV enzyme and 10 μ L 5 × buffer,
37 DEG C of 120min, 99 DEG C of 4min, 4 DEG C of 3min synthesis cDNA；Then use primer described in table 7, RT-PCR amplification nNOS, eNOS,
INOS, c-Kit, SCF, IL-8 and CXCR2 mRNA expression, internal reference is used as using housekeeping gene GAPDH；Finally with containing mass ratio
Agar electrophoresis for 1% ethidium bromide checks pcr amplification product, and carries out semi-quantitative analysis with Image1.44 softwares.
Table 7RT-PCR primer sequences
(1) influence that Lactobacillus plantarum YS2 is expressed mouse Colon tissue nNOS, eNOS and iNOS mRNA
NOS points are nNOS, eNOS and iNOS, and research confirms that the NO that eNOS is produced has crucial work to the damage of control colon
With the excessive NO that iNOS is produced then accelerates colitis to damage, and nNOS, which is lowered, also makes iNOS expression strengthen a large amount of release NO simultaneously.
From Fig. 3 and table 8, nNOS, eNOS mRNA expression highests of normal group mouse Colon tissue, and iNOS mRNA
Expression it is minimum；NNOS, eNOS mRNA of model control group mouse express minimum, iNOS mRNA expression highest；Relative to
Model control group, lactobacillus bulgaricus and Lactobacillus plantarum YS2 can be raised significantly in colitis mice colon
NNOS, eNOS mRNA expression and the expression (p for lowering iNOS mRNA<0.05), but Lactobacillus plantarum YS2 effect it is stronger.
Semi-quantitative analysis (the relative model control group table of table 8 mouse Colon tissue nNOS, eNOS and iNOS mRNA expression
Up to multiple)
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
(2) influence that Lactobacillus plantarum YS2 is expressed mouse Colon tissue c-Kit and SCF mRNA
Ulcerative colitis not only shows to suffer from diarrhoea and have blood in stool, while also there is colonic motor disorder.Research has shown that ICC
(Cajal interstitial cells) is relevant with colonic activity, directly participates in colitis process.When there is IBD, SCF is to maintaining
ICC quantity and function have direct effect, and SCF is c-Kit natural ligand, ICC increasing after SCF/Kit signal pathways sustain damage
Growing and breaking up can decline, so as to aggravate colitis.
From Fig. 4 and table 9, relative to model control group, lactobacillus bulgaricus and Lactobacillus plantarum YS2 can show
Write the expression (p of c-Kit and SCF mRNA in up-regulation colitis mice colon<0.05), but Lactobacillus plantarum YS2 effect
It is stronger.
Semi-quantitative analysis (the relative model control group expression times of table 9 mouse Colon tissue c-Kit and SCF mRNA expression
Number)
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
(3) influence that Lactobacillus plantarum YS2 is expressed mouse Colon tissue IL-8 and CXCR2mRNA
IL-8 has inflammatory activity and chemotaxis, and CXCR is IL-8 acceptors, and the morbidity of IL-8 and CXCR2 with colon cancer has
Close, document also reports the high expression that IL-8 and CXCR2 occurs under colon cancer state.
From Fig. 5 and table 10, relative to model control group, lactobacillus bulgaricus and Lactobacillus plantarum YS2 can
Significantly lower the expression (p of IL-8 and CXCR2mRNA in colitis mice colon<, but Lactobacillus plantarum YS2 0.05)
Effect is stronger.
Semi-quantitative analysis (the relative model control group expression times of table 10 mouse Colon tissue IL-8 and CXCR2mRNA expression
Number)
Note：Alphabetical different expression group differences are significantly (P ＜ 0.05).
Finally illustrate, the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although pass through ginseng
According to the preferred embodiments of the present invention, invention has been described, it should be appreciated by those of ordinary skill in the art that can
So that various changes are made to it in the form and details, the present invention limited without departing from appended claims
Spirit and scope.
<110>The college of education of Chongqing second
<120>Applications of the Lactobacillus plantarum YS2 in the food or medicine of prevention ulcerative colitis is prepared
<160>19
<210>1
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>Primer 2 7F
<400>1
agagtttgat cctggctcag 20
<210>2
<211>19
<212>DNA
<213>Artificial sequence
<220>
<223>Primer 1492R
<400>2
ggctaccttg ttacgactt 19
<210>3
<211>1469
<212>DNA
<213>Lactobacillus plantarum YS2（Lactobacillus plantarum YS2）
<220>
<223>16S rDNA genetic fragments
<400>3
gtgcaatggc ggcttgccta atacatgcaa gtcgaacgaa ctctggattg attggtgctt 60
gcatcatgat ttacatttga gggagtggcg aactggtgag taacacgtgg gaaacctgcc 120
cagaagcggg gaataacacc tggaaacaga tgctataccg cataacaact tggaccgcat 180
ggtccgagnt tgaaagatgg cttcggctat cacttttgga tggtcccgcg gcgtattagc 240
tagatggtgg ggtaacggct caccatggca atgatacgta gccgacctga gagggtaatc 300
ggccacattg ggactgactc acggccaaac tcctacggga ggcagcagta gggaatcttc 360
cacaatggac gaagtctgat ggagcaacgc cgcgtgagtg aaaagggttt cggctcgtaa 420
actctgttgt taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa 480
cagaaagcca cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg 540
tccggattta ttgggcgtaa agcgagcgca ggcggttttt taagtctgat gtgaaagcct 600
tcggctcaac cgaagaagtg catcggaaat gggaaacttg agtgcagaag aggacagtgg 660
aactccattg tagcggtgaa atgcgtagat atatggaaga acaccagtgg cgaaggcgct 720
gtctggtctg taactgacgc tgaggctcga aagtatggta gcaaacagga ttagataccc 780
tggtatccat accgtaaacg atgaatgcta agtgttggag ggtttccgcc cttcagtgct 840
gcagctaacg cattaagcat tccgcctggg gagtacggcc gcaaggctga aactcaaagg 900
aattacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagct acgcgaagaa 960
ccttaccagg tcttgacata ctatgcaaat ctaagagatt agacgttccc ttcggggaca 1020
tggatacagg tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg gttagtcccg 1080
cacgagcgca acccttatta tcagttgcca gcattaagtt gggcactctg gtgagactgc 1140
cggtgacaaa cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg 1200
ctacacacgt gctacaatgg atggtacaac agttgcgaac tcgcgagagt aagctaatct 1260
cttaaagcca ttctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg 1320
ctagtaatcc ggatcagcat gccgcggtga atacgttccc gggccttgac acaccgcccg 1380
tcacaccatg agagtttgta acacccaaag tcggtggggt aaccttttag gaaccagccg 1440
cctaaggggg acagatgatt agggtgggt 1469
<210>4
<211>23
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification nNOS mRNA expression
<400>4
gaataccagc ctgatccatg gaa 23
<210>5
<211>26
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification nNOS mRNA expression
<400>5
tcctccagga gggtgtccac cgcatg 26
<210>6
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification eNOS mRNA expression
<400>6
ggagaggctg catgacattg 20
<210>7
<211>22
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification eNOS mRNA expression
<400>7
ggtagagcca tagtggaatg ac 22
<210>8
<211>18
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification iNOS mRNA expression
<400>8
agagagatcg ggttcaca 18
<210>9
<211>18
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification iNOS mRNA expression
<400>9
cacagaactg agggtaca 18
<210>10
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification c-Kit mRNA expression
<400>10
agaccgaacg caactt 16
<210>11
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification c-Kit mRNA expression
<400>11
ggtgccatcc acttca 16
<210>12
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification SCF mRNA expression
<400>12
aaactggtgg cgaatc 16
<210>13
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification SCF mRNA expression
<400>13
cacgggtagc aagaac 16
<210>14
<211>21
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification IL-8 mRNA expression
<400>14
agaagcatgg cccagaaatc a 21
<210>15
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification IL-8 mRNA expression
<400>15
ggccttgtag acaccttggt 20
<210>16
<211>18
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification CXCR2 mRNA expression
<400>16
gaacaaaggc aaggctaa 18
<210>17
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification CXCR2 mRNA expression
<400>17
aacataacaa catctgggca 20
<210>18
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification GAPDH mRNA expression
<400>18
cggagtcaac ggatttggtc 20
<210>19
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification GAPDH mRNA expression
<400>19
agccttctcc atggtcgtga 20
Claims (1)
1. deposit number is CCTCC NO：M2016748 Lactobacillus plantarum YS2 (Lactobacillus plantarum YS2)
Application in the food or medicine of prevention ulcerative colitis is prepared.
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| CN110692726A (en) * | 2018-07-10 | 2020-01-17 | 重庆第二师范学院 | Lactobacillus plantarum CQPC01 and application thereof in preparation of food for improving constipation |
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