CN116492375B - Application of bifidobacterium longum NX-4 in preparation of medicines for preventing or treating intestinal inflammation - Google Patents
Application of bifidobacterium longum NX-4 in preparation of medicines for preventing or treating intestinal inflammation Download PDFInfo
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- 241001608472 Bifidobacterium longum Species 0.000 title claims abstract description 41
- 229940009291 bifidobacterium longum Drugs 0.000 title claims abstract description 41
- 206010061218 Inflammation Diseases 0.000 title claims abstract description 24
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- 230000000968 intestinal effect Effects 0.000 title claims abstract description 18
- 239000003814 drug Substances 0.000 title claims abstract description 15
- 238000002360 preparation method Methods 0.000 title claims description 7
- 229940079593 drug Drugs 0.000 title description 5
- 230000001580 bacterial effect Effects 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 6
- 238000004321 preservation Methods 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
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- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 21
- 230000002776 aggregation Effects 0.000 abstract description 9
- 238000004220 aggregation Methods 0.000 abstract description 9
- 244000005700 microbiome Species 0.000 abstract description 2
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 abstract 1
- 229920003045 dextran sodium sulfate Polymers 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 8
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical compound NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 description 7
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- 238000000034 method Methods 0.000 description 6
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- 239000006041 probiotic Substances 0.000 description 6
- 235000018291 probiotics Nutrition 0.000 description 6
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- 208000004232 Enteritis Diseases 0.000 description 4
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- 210000002540 macrophage Anatomy 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000000529 probiotic effect Effects 0.000 description 3
- 241000186000 Bifidobacterium Species 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 108010048367 enhanced green fluorescent protein Proteins 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 238000009630 liquid culture Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
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- 238000012360 testing method Methods 0.000 description 2
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010014418 Electrolyte imbalance Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108010043121 Green Fluorescent Proteins Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
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- 238000011534 incubation Methods 0.000 description 1
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- 230000005764 inhibitory process Effects 0.000 description 1
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- 208000037816 tissue injury Diseases 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
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- 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
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention discloses an application of bifidobacterium longum NX-4 in preparing a medicine for preventing or treating intestinal inflammation, and belongs to the technical field of microorganisms. The application of the bifidobacterium longum NX-4 in preparing the medicine for preventing or treating the intestinal inflammation disclosed by the invention can inhibit DSS-induced neutrophil aggregation to the intestinal tract of the zebra fish, so that theoretical reference and guiding basis are provided for developing the medicine for preventing or treating the intestinal inflammation by using the bifidobacterium longum NX-4.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of bifidobacterium longum NX-4 in preparation of medicines for preventing or treating intestinal inflammation.
Background
Enteritis is an inflammation of the intestinal tract caused by a series of factors such as bacteria, viruses, fungi or parasites, including inflammation of the small intestine and inflammation of the colon and rectum. The clinical manifestations mainly include fever, nausea, vomiting, abdominal pain, diarrhea, watery stool or mucopurulent bloody stool, and some patients may also have tenesmus. Dehydration and electrolyte disturbance can be caused when enteritis is serious, and even life is threatened.
Traditional methods of treating enteritis can be classified into modern medical treatment and traditional Chinese medicine treatment methods. The probiotics, serving as endogenous and immune defensive barriers of intestinal tracts, can antagonize pathogenic bacteria, has the characteristics of safety, controllability, effectiveness and small side effect, and is an ideal method for treating enteritis. In addition, the lack of powerful scientific research evidence of the function of probiotics seriously affects the popularization of the probiotics and products thereof. Based on the method, aiming at the function deep excavation of strain resources, the novel probiotic bacterial strain which has independent intellectual property, specific functional property and physiological property suitable for Chinese crowd is screened out, and the method is particularly important for improving the core competitiveness of Chinese probiotic production enterprises and promoting the development of Chinese probiotic products.
Therefore, providing the use of bifidobacterium longum NX-4 in the manufacture of a medicament for preventing or treating intestinal inflammation is a problem to be solved by the person skilled in the art.
Disclosure of Invention
In view of the above, the invention provides application of bifidobacterium longum NX-4 in preparing medicines for preventing or treating intestinal inflammation.
Inflammation is a reaction of the immune system to tissue injury and infection and is mainly characterized by the accumulation of leukocytes (granulocytes, macrophages) around the infected tissue. The immune system of zebra fish is very similar to that of mammal, and when wound happens, neutrophils and macrophages respond to traumatic inflammation almost simultaneously, the migration speed of neutrophils is high, the neutrophils are firstly recruited to the damaged part, and then the macrophages reach the damaged part. After several hours, the inflammation began to subside, and macrophages and neutrophils left the injury site. The inflammation of the intestinal tract of the zebra fish is induced by Dextran Sodium Sulfate (DSS) to promote the immune response of the zebra fish neutrophils.
The transgenic neutrophil fluorescent fish (green) is adopted, and the DSS-induced zebra fish intestinal neutrophil quantity can be obviously increased compared with the normal zebra fish under a fluorescent microscope.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the application of bifidobacterium longum NX-4 in preparing a medicament for preventing or treating intestinal inflammation is provided, wherein the preservation number of bifidobacterium longum NX-4 is CGMCC No.20112 (see patent number 202011181477.6).
Further, the bifidobacterium longum NX-4 is applied to preparation of medicines for inhibiting DSS-induced neutrophil aggregation to zebra fish intestinal tracts.
Further, the bifidobacterium longum NX-4 is a bacterial suspension.
Compared with the prior art, the application of the bifidobacterium longum NX-4 in preparing the medicine for preventing or treating the intestinal inflammation is disclosed, wherein the bifidobacterium longum NX-4 is obtained by separating and screening healthy infant feces, and the bifidobacterium longum NX-4 can inhibit DSS-induced neutrophil aggregation at the intestinal tract of the zebra fish, so that theoretical reference and guiding basis are provided for developing the medicine for preventing or treating the intestinal inflammation by using the bifidobacterium longum NX-4.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only embodiments of the present invention, and that other drawings can be obtained according to the provided drawings without inventive effort for a person skilled in the art.
FIG. 1 is a visual diagram showing that bifidobacterium longum NX-4 inhibits DSS-induced neutrophil aggregation to zebra fish intestinal tracts;
wherein A: normal group; b: a model group; c: a positive control group; d: bifidobacterium longum 15708; e: bifidobacterium longum NX-4;
FIG. 2 is a graph showing statistics of inhibiting DSS-induced neutrophil aggregation to zebra fish intestinal tracts by bifidobacterium longum NX-4 according to the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Zebra fish are green fluorescent zebra fish and AB-series zebra fish with Tg (mpx: EGFP) centromere cells.
Dextran Sodium Sulfate (DSS) (MedChemExpress); mesalamine (MedChemExpress); bifidobacterium longum 15708 (ATCC 15708) was purchased from bio-technology limited of beijing Bai-o-borg.
EXAMPLE 1 preparation of Bifidobacterium longum NX-4 suspension (thallus)
Inoculating bifidobacterium longum NX-4 after activation culture in a BS liquid culture medium, culturing for 24 hours at 37 ℃, and centrifuging for 10 minutes at 4 ℃ and 6000r/min to obtain bacterial precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
EXAMPLE 2 preparation of Bifidobacterium longum 15708 suspension (thallus)
Inoculating bifidobacterium longum 15708 after activation culture in a BS liquid culture medium, culturing at 37 ℃ for 24 hours, and centrifuging at 4 ℃ for 10 minutes at 6000r/min to obtain bacterial precipitate; after the bacterial cell precipitate is washed twice by PBS, the bacterial cell is resuspended by PBS, and the cell concentration is regulated to be 1 multiplied by 10 6 CFU/mL gave a bacterial suspension (cell).
Example 3 Effect of Bifidobacterium longum NX-4 on DSS-induced neutrophil aggregation at the intestinal tract of zebra fish
Selecting healthy Tg (mpx: EGFP) zebra fish (3 dpf), placing in 6-well cell culture plate, 1 well per group, 15 wells per well (15 fish per well, counting neutrophil number in intestinal tract of each fish), adding 5mLPBS into normal group, model group, positive control group (mesalazine), and Bifidobacterium longum 15708 intervention group (1×10) 6 CFU/mL), bifidobacterium longum NX-4 intervention group (1X 10) 6 CFU/mL) were added with 5mL of 0.5% dss solution (w/w), incubated at 28 ℃ in a biochemical incubator for 72h, each group of zebra fish was washed 3 times with PBS, and then 5mL PBS was added to the normal and model groups; the positive control group was added with 5mL of 50. Mu.g/mL mesalamine solution; bifidobacterium longum 15708 intervention group addition 1X 10 6 CFU/mL bifidobacterium longum 15708, 5mL per well; bifidobacterium longum NX-4 intervention group addition 1X 10 6 CFU/mL bifidobacterium longum NX-4, 5mL per well, and placing in a biochemical incubator at 28 ℃ for further incubation for 48 hours, and placing in a fluorescence displayAnd observing the aggregation condition of neutrophils in the intestinal tract of the zebra fish under a micro-mirror, and photographing and recording.
SPSS19.0 software was used to statistically process the data, experimental data were all expressed as x+ -SEM data, analyzed by T-test, compared to normal group: #### p<0.001, compared to model group: * P:<0.001。
the experimental results of Bifidobacterium longum NX-4 inhibiting DSS-induced neutrophil aggregation to zebra fish intestinal tract are shown in FIGS. 1 and 2.
As can be seen from fig. 1 and 2, there is a small number of neutrophil aggregates in the intestinal tract of the normal group of zebra fish (13.79±1.46), while there is a large number of neutrophil aggregates in the intestinal tract of the model group of zebra fish. Meanwhile, the number of neutrophils in the intestinal tract of the zebra fish in the model group is 32.79+/-2.23, and compared with the normal group (13.79+/-1.46), the model group has obvious dissimilarity (P < 0.001), which indicates that the DSS-induced zebra fish intestinal inflammation model is successfully established.
As can be seen from fig. 1 and 2, only a small amount of neutrophils aggregated in the intestinal tract of zebra fish body spot fish of the positive control group (mesalamine). Meanwhile, the number of neutrophils in the intestinal tract of the zebra fish of the positive control group (mesalazine) is 16.21+/-1.25, and the difference is obvious compared with the model group (32.79+/-2.23) (P)<0.001). Therefore, the mesalamine has the effect of relieving the intestinal inflammation, and is consistent with clinical results, so that the test for evaluating the efficacy of relieving the intestinal inflammation is effective. Bifidobacterium longum 15708 intervention group (1×10 6 CFU/mL) of zebra fish with a large number of neutrophil aggregates in the intestine; bifidobacterium 15708 intervention group of the same duration (1×10 6 The number of neutrophils in the intestinal tract of the zebra fish (CFU/mL) was 27.43+ -2.35, and there was no significant difference (P) compared with the model group (32.79+ -2.23)>0.05). In addition, there was also only a small amount of neutrophil aggregation in the zebra fish intestinal tract of bifidobacterium longum NX-4, similar to the positive control group (mesalamine); co-time bifidobacterium NX-4 intervention group (1X 10) 6 The number of neutrophils in the intestinal tract of the zebra fish (CFU/mL) is 18.14+/-1.47, and the difference is obvious (P) compared with the model group (32.79+/-2.23)<0.001). Thus, the above results indicate that at the same concentration, bifidobacterium longum NX-4 induced accumulation of neutrophils to the intestinal tract of zebra fish against DSSThe inhibition effect of the compound is stronger than that of bifidobacterium longum 15708, and the compound shows good potential for relieving intestinal inflammation.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (2)
1. The application of bifidobacterium longum NX-4 in preparing a medicament for preventing or treating intestinal inflammation is characterized in that the preservation number of the bifidobacterium longum NX-4 is CGMCC No.20112.
2. Use of bifidobacterium longum NX-4 according to claim 1, in the preparation of a medicament for the prevention or treatment of intestinal inflammation, wherein said bifidobacterium longum NX-4 is a bacterial suspension.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2449890A1 (en) * | 2010-11-05 | 2012-05-09 | Nestec S.A. | Powdered cereal compositions comprising non-replicating probiotic microorganisms |
| CN108220206A (en) * | 2018-03-12 | 2018-06-29 | 江南大学 | A kind of bifidobacterium longum and its application |
| CN109152400A (en) * | 2016-03-07 | 2019-01-04 | 自然阳光生产公司 | Improve the method and composition of weight loss and health of heart in addition to diet and movement |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2449890A1 (en) * | 2010-11-05 | 2012-05-09 | Nestec S.A. | Powdered cereal compositions comprising non-replicating probiotic microorganisms |
| CN109152400A (en) * | 2016-03-07 | 2019-01-04 | 自然阳光生产公司 | Improve the method and composition of weight loss and health of heart in addition to diet and movement |
| CN108220206A (en) * | 2018-03-12 | 2018-06-29 | 江南大学 | A kind of bifidobacterium longum and its application |
Non-Patent Citations (2)
| Title |
|---|
| Faecalibacterium prausnitzii产物对大鼠脾细胞促炎因子IL-17分泌的抑制作用;张明明;杨晓彤;邱新运;洪娜;于成功;;胃肠病学(第07期);全文 * |
| 张明明 ; 杨晓彤 ; 邱新运 ; 洪娜 ; 于成功 ; .Faecalibacterium prausnitzii产物对大鼠脾细胞促炎因子IL-17分泌的抑制作用.胃肠病学.2012,(第07期),全文. * |
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