CN112458020B - Probiotic composition for inhibiting helicobacter pylori and application thereof - Google Patents
Probiotic composition for inhibiting helicobacter pylori and application thereof Download PDFInfo
- Publication number
- CN112458020B CN112458020B CN202011412221.1A CN202011412221A CN112458020B CN 112458020 B CN112458020 B CN 112458020B CN 202011412221 A CN202011412221 A CN 202011412221A CN 112458020 B CN112458020 B CN 112458020B
- Authority
- CN
- China
- Prior art keywords
- helicobacter pylori
- lactobacillus
- probiotic composition
- parts
- cfu
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000590002 Helicobacter pylori Species 0.000 title claims abstract description 80
- 229940037467 helicobacter pylori Drugs 0.000 title claims abstract description 79
- 239000006041 probiotic Substances 0.000 title claims abstract description 75
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 75
- 239000000203 mixture Substances 0.000 title claims abstract description 74
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 67
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 28
- 241000218588 Lactobacillus rhamnosus Species 0.000 claims abstract description 39
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 34
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 34
- 241000186604 Lactobacillus reuteri Species 0.000 claims abstract description 34
- 241000186869 Lactobacillus salivarius Species 0.000 claims abstract description 34
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 34
- 229940001882 lactobacillus reuteri Drugs 0.000 claims abstract description 34
- 239000003814 drug Substances 0.000 claims abstract description 28
- 239000000843 powder Substances 0.000 claims description 46
- 230000001580 bacterial effect Effects 0.000 claims description 39
- 241000894006 Bacteria Species 0.000 claims description 27
- 238000004321 preservation Methods 0.000 claims description 21
- 238000002360 preparation method Methods 0.000 claims description 15
- 206010019375 Helicobacter infections Diseases 0.000 claims description 11
- 238000011282 treatment Methods 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims 1
- 230000002265 prevention Effects 0.000 claims 1
- 238000011321 prophylaxis Methods 0.000 claims 1
- 230000014509 gene expression Effects 0.000 abstract description 29
- 230000000694 effects Effects 0.000 abstract description 20
- 230000007923 virulence factor Effects 0.000 abstract description 20
- 239000000304 virulence factor Substances 0.000 abstract description 20
- 101100476480 Mus musculus S100a8 gene Proteins 0.000 abstract description 18
- 235000013305 food Nutrition 0.000 abstract description 11
- 229940079593 drug Drugs 0.000 abstract description 9
- 230000002195 synergetic effect Effects 0.000 abstract description 9
- 230000005764 inhibitory process Effects 0.000 abstract description 5
- 238000000855 fermentation Methods 0.000 description 17
- 230000004151 fermentation Effects 0.000 description 16
- 241000699670 Mus sp. Species 0.000 description 14
- 206010061218 Inflammation Diseases 0.000 description 13
- 230000004054 inflammatory process Effects 0.000 description 13
- 210000001156 gastric mucosa Anatomy 0.000 description 12
- 230000002829 reductive effect Effects 0.000 description 12
- 239000013589 supplement Substances 0.000 description 9
- 238000000034 method Methods 0.000 description 8
- 239000010802 sludge Substances 0.000 description 8
- 238000009777 vacuum freeze-drying Methods 0.000 description 8
- 229920002774 Maltodextrin Polymers 0.000 description 6
- 239000005913 Maltodextrin Substances 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 229940035034 maltodextrin Drugs 0.000 description 6
- 238000002560 therapeutic procedure Methods 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 238000003304 gavage Methods 0.000 description 4
- 238000009629 microbiological culture Methods 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000003828 downregulation Effects 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000000386 microscopy Methods 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000008520 organization Effects 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 208000002109 Argyria Diseases 0.000 description 2
- 238000009631 Broth culture Methods 0.000 description 2
- 239000004386 Erythritol Substances 0.000 description 2
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 2
- 241001473949 Helicobacter pylori NCTC 11637 = CCUG 17874 = ATCC 43504 Species 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 108010046334 Urease Proteins 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- -1 aqua Substances 0.000 description 2
- 229910052797 bismuth Inorganic materials 0.000 description 2
- JCXGWMGPZLAOME-UHFFFAOYSA-N bismuth atom Chemical compound [Bi] JCXGWMGPZLAOME-UHFFFAOYSA-N 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000006781 columbia blood agar Substances 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 2
- 229940009714 erythritol Drugs 0.000 description 2
- 235000019414 erythritol Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- FTSSQIKWUOOEGC-RULYVFMPSA-N fructooligosaccharide Chemical compound OC[C@H]1O[C@@](CO)(OC[C@@]2(OC[C@@]3(OC[C@@]4(OC[C@@]5(OC[C@@]6(OC[C@@]7(OC[C@@]8(OC[C@@]9(OC[C@@]%10(OC[C@@]%11(O[C@H]%12O[C@H](CO)[C@@H](O)[C@H](O)[C@H]%12O)O[C@H](CO)[C@@H](O)[C@@H]%11O)O[C@H](CO)[C@@H](O)[C@@H]%10O)O[C@H](CO)[C@@H](O)[C@@H]9O)O[C@H](CO)[C@@H](O)[C@@H]8O)O[C@H](CO)[C@@H](O)[C@@H]7O)O[C@H](CO)[C@@H](O)[C@@H]6O)O[C@H](CO)[C@@H](O)[C@@H]5O)O[C@H](CO)[C@@H](O)[C@@H]4O)O[C@H](CO)[C@@H](O)[C@@H]3O)O[C@H](CO)[C@@H](O)[C@@H]2O)[C@@H](O)[C@@H]1O FTSSQIKWUOOEGC-RULYVFMPSA-N 0.000 description 2
- 229940107187 fructooligosaccharide Drugs 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000002147 killing effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000000630 rising effect Effects 0.000 description 2
- 238000011895 specific detection Methods 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 241000589562 Brucella Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- 208000007882 Gastritis Diseases 0.000 description 1
- 102100028999 High mobility group protein HMGI-C Human genes 0.000 description 1
- 101000986379 Homo sapiens High mobility group protein HMGI-C Proteins 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 206010028116 Mucosal inflammation Diseases 0.000 description 1
- 241000283898 Ovis Species 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 208000007107 Stomach Ulcer Diseases 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000023652 chronic gastritis Diseases 0.000 description 1
- 229960002626 clarithromycin Drugs 0.000 description 1
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229960001305 cysteine hydrochloride Drugs 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 201000005917 gastric ulcer Diseases 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012134 rapid urease test Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009044 synergistic interaction Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
-
- 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
- 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
- A23V2400/173—Reuteri
-
- 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
- A23V2400/175—Rhamnosus
-
- 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
- A23V2400/181—Salivarius
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a probiotic composition for inhibiting helicobacter pylori and application thereof, belonging to the technical field of food and medicines. The probiotic composition comprises 4 probiotics: lactobacillus rhamnosus, lactobacillus plantarum, lactobacillus reuteri and lactobacillus salivarius. The probiotic composition has a synergistic inhibition effect on helicobacter pylori, can obviously inhibit the growth of the helicobacter pylori, reduce the gene expression level of virulence factors CagA, VacA and Ure, can enable the helicobacter pylori to infect the helicobacter pylori of a model mouse to turn negative, and can effectively reduce the number of the helicobacter pylori in a human body or enable the helicobacter pylori in the human body to turn negative. The four strains of the probiotic composition have mutual synergistic effect, and the effect of inhibiting the helicobacter pylori is obviously better than the single application effect of each strain.
Description
Technical Field
The invention belongs to the technical field of food and medicines, and particularly relates to a probiotic composition for inhibiting helicobacter pylori and application thereof.
Background
Helicobacter pylori (Hp) can cause various digestive tract diseases, can be parasitic on gastric mucosa for a long time, is easy to cause chronic gastritis, gastric ulcer and even gastric cancer, and is classified as a type I carcinogenic factor by the World Health Organization (WHO).
The standard clarithromycin triple therapy for clearing Hp is proposed and widely applied in 1996, the currently recommended antibiotic quadruple therapy containing a bismuth agent is taken as a first choice for clinically eradicating Hp, but along with the continuous improvement of Hp to antibiotic resistance and the improvement of incidence rates of related side effects such as abdominal distension, diarrhea, constipation and the like caused by antibiotic and the bismuth agent, the radical treatment effect of the antibiotic therapy and the tolerance and drug compliance of patients are greatly reduced.
The probiotic therapy is a new therapy for treating Hp, and researches and clinical practices in recent years show that the application of probiotic preparations such as lactobacillus and bifidobacterium has certain curative effects on the aspects of improving the eradication rate of Hp, reducing the side effects of treatment and the like. The function of probiotics for treating Hp belongs to strain dependence, and the combination of different probiotic strains can have synergistic or antagonistic effect on the Hp treatment effect. The selection of probiotic bacterial strains for the treatment of Hp and the synergistic effect of different bacterial strain compositions for the treatment of Hp remain to be confirmed by a further considerable amount of research.
Disclosure of Invention
In view of the above, the present invention aims to provide a probiotic composition for inhibiting helicobacter pylori and its application, wherein the probiotic composition can exert a significant synergistic effect of inhibiting and killing helicobacter pylori.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a probiotic composition for inhibiting helicobacter pylori, which comprises the following bacterial powder in parts by weight: 10-30 parts of lactobacillus rhamnosus, 10-30 parts of lactobacillus plantarum, 10-30 parts of lactobacillus reuteri and 10-30 parts of lactobacillus salivarius.
Preferably, the probiotic composition comprises the following bacterial powder in parts by weight: 15-25 parts of lactobacillus rhamnosus, 15-25 parts of lactobacillus plantarum, 15-25 parts of lactobacillus reuteri and 15-25 parts of lactobacillus salivarius.
Preferably, the Lactobacillus rhamnosus comprises Lactobacillus rhamnosus LN56 with the preservation number of CGMCC No. 17370; the Lactobacillus plantarum comprises Lactobacillus plantarum (LN 66) with the preservation number of CGMCC No. 17369; the Lactobacillus reuteri comprises Lactobacillus reuteri LN83 with the preservation number of CGMCC No. 17541; the Lactobacillus salivarius comprises Lactobacillus salivarius LN12 with the preservation number of CGMCC No. 17542.
Preferably, the number of the live bacteria in the bacterial powder of the lactobacillus rhamnosus LN56, the bacterial powder of the lactobacillus plantarum LN66, the bacterial powder of the lactobacillus reuteri LN83 and the bacterial powder of the lactobacillus salivarius LN12 is 1.0 × 1010～3.0×1011CFU/g。
The invention also provides application of the probiotic composition in preparing a food or a medicament or a supplement for inhibiting the growth of helicobacter pylori.
The invention also provides application of the probiotic composition in preparation of foods, medicines or supplements for inhibiting the expression of helicobacter pylori virulence factor genes.
Preferably, the virulence factors include CagA, VacA and Ure.
The invention also provides application of the probiotic composition in preparation of a medicament for preventing and/or treating helicobacter pylori infection.
The invention also provides a medicament for preventing and/or treating helicobacter pylori infection, which takes the probiotic composition as an effective component.
The invention provides a probiotic composition for inhibiting helicobacter pylori, which has a remarkably higher inhibiting and killing effect on the helicobacter pylori than a single strain; compared with a single strain, the probiotic composition can effectively reduce the gene expression levels of helicobacter pylori virulence factors CagA, VacA and Ure, so that 4 bacteria in the probiotic composition generate a remarkable synergistic effect.
The embodiment of the invention proves that after a certain amount of probiotic composition is orally taken, the growth of helicobacter pylori can be effectively inhibited in vivo, the gene expression level of helicobacter pylori virulence factors CagA, VacA and Ure is reduced, the cavitation damage to gastric mucosa cells and the like is reduced, the gene expression of urease Ure is inhibited, the ammonia production capacity of the helicobacter pylori is reduced, the pH of the surrounding environment is reduced, the growth of the helicobacter pylori is not facilitated, and the helicobacter pylori is easy to be killed by antibiotics. The effect of eliminating the helicobacter pylori is obviously better than that of the single application of each strain when the helicobacter pylori infects a model mouse by feeding the probiotic composition, and the application of the probiotic composition can eradicate the helicobacter pylori on the gastric mucosa of the mouse and eliminate inflammation. The results of taking the probiotic composition by human bodies also show that the composition can obviously reduce the number of helicobacter pylori in the human bodies, the effective rate reaches 90 percent, and the negative conversion rate of the helicobacter pylori reaches 70 percent.
Biological preservation information
Lactobacillus rhamnosus LN56, deposited in China general microbiological culture Collection center, with the deposit Address of Beijing, North Cheng Yang district, West Lu No.1 Hospital No. 3, and the deposit organization is abbreviated as: CGMCC with preservation date of 2019, 03 and 20 months and biological preservation number of CGMCC No. 17370;
lactobacillus plantarum LN66, deposited in China general microbiological culture Collection center, having the deposit address of Beijing province, No.1 Xilu-Beichen province, the rising district, No. 3, and the short name of the deposit institution: CGMCC with preservation date of 2019, 03 and 20 months and biological preservation number of CGMCC No. 17369;
lactobacillus reuteri LN83, deposited in China general microbiological culture Collection center, with the deposit address Beijing, North Chen-Yang district, West Lu No.1 Hospital No. 3, the deposit organization is abbreviated as: CGMCC with the preservation date of 2019, 04 months and 10 days and the biological preservation number of CGMCC No. 17541;
lactobacillus salivarius LN12, deposited in China general microbiological culture Collection center, with the deposit address of Beijing province, the rising district, Xilu No.1 Hospital No. 3, and the deposit institution abbreviated as: CGMCC with preservation date of 2019, 04 months and 10 days and biological preservation number of CGMCC No. 17542.
Detailed Description
The invention provides a probiotic composition for inhibiting helicobacter pylori, which comprises the following bacterial powder in parts by weight: 10-30 parts of lactobacillus rhamnosus, 10-30 parts of lactobacillus plantarum, 10-30 parts of lactobacillus reuteri and 10-30 parts of lactobacillus salivarius.
In the probiotic composition, the lactobacillus rhamnosus is preferably 15-25 parts by weight, and more preferably 20 parts by weight. The Lactobacillus rhamnosus provided by the invention preferably comprises Lactobacillus rhamnosus LN56 with the preservation number of CGMCC No. 17370. The number of the live bacteria in the bacterial powder of the lactobacillus rhamnosus LN56 is preferably 1.0 x 1010～3.0×1011CFU/g, more preferably 5.0X 1010～2.0×1011CFU/g, most preferably 8.0X 1010CFU/g. The preparation method of the lactobacillus rhamnosus LN56 powder is not particularly limited, and preferably comprises the following steps: 1) inoculating the lactobacillus rhamnosus LN56 into an MRS culture solution or an improved MRS culture solution, and fermenting at 36-38 ℃ for 12-24 h to obtain lactobacillus rhamnosus LN56 fermentation liquor; the improved MRS culture solution takes the MRS culture solution as a basic culture medium and contains cysteine hydrochloride with the mass concentration of 0.05%;
2) centrifuging the lactobacillus rhamnosus LN56 fermentation liquor, and collecting precipitates to obtain lactobacillus rhamnosus LN56 bacterial sludge;
3) and (3) carrying out vacuum freeze drying on the lactobacillus rhamnosus LN56 bacterial sludge to obtain bacterial powder of lactobacillus rhamnosus LN 56.
The inoculation amount of the inoculation in the step 1) of the method is preferably 1 per thousand, the fermentation temperature is preferably 37 ℃, and the fermentation time is preferably 20 hours. The fermentation according to the invention is preferably an anaerobic fermentation. The rotating speed of the centrifugation in the step 2) of the invention is preferably 8000-12000 r/min, and more preferably 10000 r/min; the time for centrifugation is preferably 60-120 min, and more preferably 90 min. The vacuum freeze-drying in step 3) of the present invention preferably comprises: mixing the bacterial sludge, water and freeze-drying carrier, and then carrying out vacuum freeze-drying; the mass ratio of the bacterial sludge, the water and the freeze-drying carrier is preferably 1: (1-6): (1-5), more preferably 1: 3: 2. the freeze-dried carrier of the present invention preferably comprises skimmed milk powder or maltodextrin. The vacuum freeze drying instrument is not particularly limited, a Toffia LYO-20 vacuum freeze drying machine is preferably adopted, and the temperature of a heat transfer clapboard is preferably set to be-10 ℃, and more preferably-5 ℃ when the vacuum freeze drying is carried out; the degree of vacuum in the drying oven is preferably set to 10 to 16Pa, and more preferably to 13 Pa. The time of the vacuum freeze drying is preferably 20-48 h, and more preferably 28 h. The lactobacillus rhamnosus LN56 can inhibit the growth of helicobacter pylori and reduce the gene expression level of helicobacter pylori virulence factors CagA, VacA and Ure.
The probiotic composition comprises lactobacillus plantarum powder, wherein the lactobacillus plantarum powder is preferably 15-25 parts by weight, and more preferably 20 parts by weight. The preparation method of the lactobacillus plantarum powder is preferably the same as that described above, and is not described herein again. The Lactobacillus plantarum provided by the invention preferably comprises Lactobacillus plantarum LN66 with the preservation number of CGMCC No. 17369. The number of the live bacteria in the bacterial powder of the lactobacillus plantarum LN66 is preferably 1.0 x 1010～3.0×1011CFU/g, more preferably 5.0X 1010～2.0×1011CFU/g, most preferably 8.0X 1010CFU/g. In the invention, the lactobacillus plantarum LN66 can inhibit the growth of helicobacter pylori and reduce the gene expression level of helicobacter pylori virulence factors CagA, VacA and Ure.
The probiotic composition preferably comprises 15-25 parts by weight of lactobacillus reuteri, and preferably 20 parts by weight of lactobacillus reuteri. The preparation method of the lactobacillus reuteri powder is preferably the same as that described above, and is not described herein again. The Lactobacillus reuteri preferably comprises Lactobacillus reuteri LN83 with the preservation number of CGMCC No. 17541. The number of the live bacteria in the bacterial powder of the lactobacillus reuteri LN83 is 1.0 multiplied by 1010～3.0×1011CFU/g, preferably 1.0X 1010～1.5×1011CFU/g, more preferably 2.0X 1010～1.0×1011CFU/g, most preferably 5.0X 1010CFU/g. The Lactobacillus reuteri LN83 can inhibit the growth of helicobacter pylori, and reduce the gene expression level of helicobacter pylori virulence factors CagA, VacA and Ure.
The probiotic composition preferably comprises 15-25 parts by weight of lactobacillus salivarius, and more preferably 20 parts by weight of lactobacillus salivarius. Preparation method of lactobacillus salivarius powderThe method is preferably the same as described above and will not be described further herein. The Lactobacillus salivarius preferably comprises Lactobacillus salivarius LN12 with the preservation number of CGMCC No. 17542. The number of viable bacteria in the powder of the lactobacillus salivarius LN12 is 1.0 multiplied by 1010～3.0×1011CFU/g, preferably 1.0X 1010～1.5×1011CFU/g, more preferably 2.0X 1010～1.0×1011CFU/g, most preferably 5.0X 1010CFU/g. The lactobacillus salivarius LN12 can inhibit the growth of helicobacter pylori and reduce the gene expression level of helicobacter pylori virulence factors CagA and VacA.
According to the invention, the lactobacillus rhamnosus, the lactobacillus plantarum, the lactobacillus reuteri and the lactobacillus salivarius are preferably mixed according to the mass ratio of 1:1:1:1, so that the synergistic interaction effect can be generated: for example, the diameters of the inhibition zones of the 4 single strains LN56, LN66, LN83 and LN12 are 18.33mm, 18.43mm, 17.33mm and 17.02mm, respectively, while the diameters of the inhibition zones of the probiotic composition are 24.48mm, which has very significant difference; 4 single strains LN56, LN66 and LN83 have slight down-regulation effect on the gene expression level of virulence factors CagA, VacA and Ure, LN12 has no down-regulation effect, but the probiotic composition can respectively down-regulate the gene expression level of CagA, VacA and Ure by 20 times, 10 times and 8 times; the above four strains alone can only reduce the amount of helicobacter pylori and the degree of inflammation without eradicating and eliminating it, but the probiotic composition can eradicate helicobacter pylori on the gastric mucosa of mice and eliminate inflammation, and thus the probiotic composition can be used for preventing and/or treating helicobacter pylori infection.
The invention also provides application of the probiotic composition in preparing a food or a medicament or a supplement for inhibiting the growth of helicobacter pylori. The formulation of the food or drug or supplement of the present invention is not particularly limited, but is preferably a powder, tablet, granule, aqua, pill, capsule or gel, and more preferably a powder. In the present invention, when the formulation of the food or drug or supplement is a powder, it is preferable to further include an adjuvant, which is preferably maltodextrin, erythritol, and fructooligosaccharide. The preparation method of the food or the medicament or the supplement is not particularly limited, and the preparation method is conventional in the field. The content of the probiotic composition in the food or medicament or supplement is preferably 20-100% by weight, and more preferably 90% by weight.
The invention also provides application of the probiotic composition in preparation of foods, medicines or supplements for inhibiting the expression of helicobacter pylori virulence factor genes. The virulence factors of the invention preferably comprise CagA, VacA and Ure, and compared with blank control, the gene expression levels of the CagA, the VacA and the Ure are respectively reduced by 20 times, 10 times and 8 times after the probiotic composition is eaten. The formulation and preparation method of the food or drug or supplement of the present invention are preferably the same as those described above, and are not described herein again.
The invention also provides application of the probiotic composition in preparation of a medicament for preventing and/or treating helicobacter pylori infection. After a certain amount of the medicine is orally taken, the growth of helicobacter pylori can be effectively inhibited in vivo, and the gene expression level of virulence factors CagA, VacA and Ure of the helicobacter pylori is reduced, so that the cavitation damage to gastric mucosa cells and the like are reduced, the gene expression of urease Ure is inhibited, the ammonia production capacity of the helicobacter pylori is reduced, the pH value of the surrounding environment is reduced, the growth of the helicobacter pylori is not facilitated, and the helicobacter pylori is easy to be killed by antibiotics; the effect of eliminating helicobacter pylori by treating helicobacter pylori infection model mice by using the medicament is obviously better than the effect of singly applying LN56, LN66, LN83 and LN12 strains, the application of 4 probiotics can eradicate the helicobacter pylori on the gastric mucosa of the mice and eliminate inflammation, and the independent application of the four probiotics can only reduce the amount and the degree of inflammation of the helicobacter pylori but can not eradicate and eliminate the helicobacter pylori. The results of human body taking the probiotic composition also show that the medicament can obviously reduce the number of helicobacter pylori in human body, the effective rate reaches 90%, and the negative conversion rate of helicobacter pylori reaches 70%.
The invention also provides a medicament for preventing and/or treating helicobacter pylori infection, which takes the probiotic composition as an effective component.
The dosage form of the drug is not particularly limited, and is preferably powder, tablet, granule, aqua, pill, capsule or gel, and more preferably powder. In the present invention, when the dosage form of the medicament is powder, the medicament preferably further comprises an auxiliary material, and the auxiliary material is preferably maltodextrin, erythritol and fructo-oligosaccharide. The preparation method of the medicine is not particularly limited, and the conventional preparation method in the field is adopted. The content of the probiotic composition in the medicament is preferably 20-100% by mass, and more preferably 90% by mass. When the medicine is used for preventing and/or treating helicobacter pylori infection, the dosage is preferably 1-20 g/day, and more preferably 8 g/day based on the dosage of the probiotic composition.
The present invention provides a helicobacter pylori inhibiting probiotic composition and its use, which will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Adopting lactobacillus rhamnosus LN56, lactobacillus plantarum LN66, lactobacillus reuteri LN83 and lactobacillus salivarius LN 12; performing facultative anaerobic activated culture in MSR culture medium at 37 deg.C for 24 hr, centrifuging at 10000rpm for 10min to remove thallus to obtain cell-free fermentation supernatant, and respectively detecting the cell-free fermentation supernatant (CFS) of single strain and the inhibitory effect of the composition of CFS of the 4 strains on helicobacter pylori. Helicobacter pylori (Helicobacter pylori ATCC 43504) was purchased from American type culture Collection. The helicobacter pylori solid culture adopts 10% sterile defibrinated goat blood Columbia blood agar culture medium to culture for 3-4 d under the microaerophilic condition at 37 ℃, and the liquid culture adopts 10% fetal calf serum Brookfield broth culture medium to culture for 2-3 d under the microaerophilic condition at 37 ℃ and at 80 rpm.
The specific detection method comprises the following steps: the Oxford cup diffusion experiment method is characterized in that the bacterium density of the helicobacter pylori bacterium liquid in the middle logarithmic phase is adjusted to 2 x 10 by using a broth culture solution6CFU/mL, collecting 200 μ L of bacteria solution to 10% sterile defibered sanguis Caprae Seu Ovis and Columbia blood agar culture medium plate, uniformly coating with sterile glass rake, carefully placing Oxford cup on the solid plate with tweezers, and respectively collecting the above prepared Lactobacillus rhamnosus L200. mu.L of each of the CFS of N56, the CFS of Lactobacillus plantarum LN66, the CFS of Lactobacillus reuteri LN83, the CFS of Lactobacillus salivarius LN12, and the CFS of the 4 strains were placed in an Oxford cup, and MRS liquid medium was used as a negative control. Firstly diffusing the solid flat plate in a refrigerator at 4 ℃ for 3-4 h, then transferring the flat plate to an incubator at 37 ℃ for microaerobic culture, observing the bacteriostatic result after 48h, measuring the diameter of a bacteriostatic ring by using a vernier caliper, and repeating for 3 times.
The bacteriostatic effect of the CFS of the single probiotic and the equal proportion composition of the 4 CFSs on the helicobacter pylori is shown in Table 1, the CFS of the single probiotic has different degrees of inhibition on the helicobacter pylori, and the inhibitory effect of the equal proportion composition of the CFS of the 4 probiotics on the helicobacter pylori is obviously greater than that of the CFS of the single probiotic, so that the synergistic effect is obvious.
TABLE 1 inhibition of helicobacter pylori by probiotic CFS
Note: indicates that the ratio of MRS is significantly different (P <0.05), and indicates that the ratio of 4 bacteria is significantly different from the ratio of single bacteria (P <0.05)
Example 2
The effect on the gene expression levels of the virulence factors CagA, VacA and Ure of H.pylori was examined using the cell-free fermentation supernatants (CFS) of Lactobacillus rhamnosus LN56, Lactobacillus plantarum LN66, Lactobacillus reuteri LN83, Lactobacillus salivarius LN12 obtained in example 1 and a combination of CFS of these 4 bacteria.
The specific detection method comprises the following steps: culturing Helicobacter pylori (Helicobacter pylori ATCC 43504) in 10% culture medium of fetal calf serum Brucella broth for 48-72 h, and diluting the broth to bacterial density of 1 × 107CFU/mL. 5mL LN56 CFS, 5mL LN66 CFS, 5mL LN83 CFS, 5mL LN12 CFS, and 5mL of a proportional combination of 4 or more bacterial CFS (each 1.25mL) were added to each flask, followed by 5mL of 10% fetal bovine serum Brookfield broth and 5mL of each flask10mL 1×107And (3) carrying out microaerobic culture on the CFU/mL Hp bacterial liquid at 37 ℃ for 48-72 h at 80 rpm. Control group replaced CFS with 5mL MRS.
After completion of the culture, the contents of each flask were centrifuged to remove the supernatant, and the cells were carefully washed with sterile PBS buffer, centrifuged, followed by addition of 10mL of sterile PBS buffer, repeated pipetting, and transferred to a centrifuge tube without rnase contamination. Total RNA extraction was performed using the TransZol Up Plus RNA Kit. After obtaining total RNA, the RNA was purified by PrimeScriptTMThe RT reagent Kit with gDNA Eraser reverse transcribes to synthesize cDNA. Adding TB
Premix Ex TaqTMII (Tli RNaseH plus) dye in qTOWER3G fluorescent quantitative PCR instrument for fluorescent quantitative detection. The primers used in the experiments are listed in table 2. The parameters of the fluorescent quantitative PCR instrument are set as follows: pre-denaturation at 95 ℃ for 2 min; and (4) circulating for 40 times: 95 ℃ for 15 s; 15s at 55 ℃; 68 ℃ for 20 s. Finally, Hp 16S rRNA is taken as an internal reference gene, and 2-ΔΔCtThe experimental data are processed by the method, the relative expression quantity of the genes is determined, the results are shown in the table 3, the inhibiting effect of the 4 CFS equal proportion compositions on the gene expression level of the main virulence factor secreted by Hp is obviously stronger than the inhibiting effect of the CFS of a single bacterium, the results show that the inhibiting effect of the CFS compositions of the 4 bacteria on the gene expression of the virulence factor has a synergistic effect, and the difference has statistical significance (P is the difference of the gene expression level of the virulence factor)<0.05). Compared with a control group, the CFS composition of the 4 bacteria respectively down-regulates the gene expression levels of CagA, VacA and Ure by about 20 times, 10 times and 8 times, while the CFS of a single bacterium down-regulates the gene expression level of CagA by 2.31 times to 4.76 times, the gene expression level of VacA by 2.20 times to 2.82 times and the gene expression level of Ure by 1 time to 3.31 times, wherein the LN12 CFS has no down-regulation effect on the gene expression level of Ure. It can be seen that the CFS of the individual bacteria LN56, LN66, LN83 and LN12 down-regulated the gene expression levels of 3 virulence factors CagA, VacA and Ure of Hp, but were all significantly less than the CFS composition of the 4 bacteria in general. The CFS equal proportion composition of 4 bacteria can inhibit Hp virulence factors CagA, VacA andthe gene expression level of Ure indicates that the 4 bacteria CFS composition can better attenuate Hp toxicity.
TABLE 2 real-time fluorescent quantitative PCR primers
TABLE 3 suppression of Hp virulence factor gene expression levels by probiotic CFS
Note: the values in the table are relative mRNA expression, indicating significant differences between the 4 CFS compositions and the single CFS (P <0.05)
Example 3
Respectively inoculating lactobacillus rhamnosus LN56, lactobacillus plantarum LN66, lactobacillus reuteri LN83 and lactobacillus salivarius LN12 into an MRS culture solution, and fermenting at 37 ℃ for 16-24 h to respectively obtain lactobacillus rhamnosus LN56 fermentation liquor, lactobacillus plantarum LN66 fermentation liquor, lactobacillus reuteri LN83 fermentation liquor and lactobacillus salivarius LN12 fermentation liquor;
respectively centrifuging the fermentation liquor of lactobacillus rhamnosus LN56, the fermentation liquor of lactobacillus plantarum LN66, the fermentation liquor of lactobacillus reuteri LN83 and the fermentation liquor of lactobacillus salivarius LN12 at 10000r/min for 90min, collecting precipitates, and respectively obtaining lactobacillus rhamnosus LN56 bacterial sludge, lactobacillus plantarum LN66 bacterial sludge, lactobacillus reuteri LN83 bacterial sludge and lactobacillus salivarius LN12 bacterial sludge;
respectively mixing the obtained lactobacillus rhamnosus LN56 bacterial mud, lactobacillus plantarum LN66 bacterial mud, lactobacillus reuteri LN83 bacterial mud or lactobacillus salivarius LN12 bacterial mud with 3 times of water and 2 times of freeze-drying carrier by weight of the bacterial mud, and carrying out vacuum freeze-drying for 28h at-5 ℃ to respectively obtain bacterial powder of lactobacillus rhamnosus LN56, bacterial powder of lactobacillus plantarum LN66, bacterial powder of lactobacillus reuteri LN83 or bacterial powder of lactobacillus salivarius LN 12.
Example 4
Taking 10g of the powder of Lactobacillus rhamnosus LN56 prepared in example 3 (viable count is 1.0 × 10)11CFU/g), 30g of powder of Lactobacillus plantarum LN66 (viable count 1.0X 10) prepared in example 311CFU/g), 10g of powder of Lactobacillus reuteri LN83 prepared in example 3 (viable count 3.0X 10)10CFU/g), 30g of powder of Lactobacillus salivarius LN12 (viable count 3.0X 10) prepared in example 310CFU/g) and 60g maltodextrin are stirred for 10min at the rotating speed of 1000r/min to obtain the probiotic composition for inhibiting helicobacter pylori.
Example 5
30g of the powder of Lactobacillus rhamnosus LN56 prepared in example 3 (viable count 3.0X 10)10CFU/g), 10g of powder of Lactobacillus plantarum LN66 (viable count 3.0X 10) prepared in example 310CFU/g), 30g of powder of Lactobacillus reuteri LN83 prepared in example 3 (viable count 1.0X 10)11CFU/g), 10g of powder of Lactobacillus salivarius LN12 prepared in example 32 (viable count 1.0X 10)11CFU/g) and 60g maltodextrin are stirred for 10min at the rotating speed of 1000r/min to obtain the probiotic composition for inhibiting helicobacter pylori.
Example 6
20g of the powder of Lactobacillus rhamnosus LN56 prepared in example 3 (viable count 8.0X 10)10CFU/g), 20g of powder of Lactobacillus plantarum LN66 (viable count 8.0X 10) prepared in example 310CFU/g), 20g of powder of Lactobacillus reuteri LN83 prepared in example 3 (viable count 5.0X 10)10CFU/g), 20g of powder of Lactobacillus salivarius LN12 (viable count 5.0X 10) prepared in example 310CFU/g) and 60g maltodextrin are stirred for 10min at the rotating speed of 1000r/min to obtain the probiotic composition for inhibiting helicobacter pylori.
Example 7
Selecting 188 healthy male SPF-grade BABL/C mice at the age of 8-10 weeks, and administering helicobacter pylori (Hp) (content 10) to the other mice except 20 mice in the negative control group9cfu/ml) fresh suspension 1ml was intragastrically administered 1 time every other day for 8 times. After the last gastric lavage for 1 week, 8 mice are killed, the gastric mucosa is taken for rapid urease test, the results are positive, HE staining microscopy of gastric mucosa tissues shows that epithelial cells of the gastric mucosa are exfoliated and infiltrated by inflammatory cells, special silver staining microscopy of gastric mucosa tissues shows Hp, and both prove that the Hp infection model is established.
The Hp-infected mice were randomly divided into 8 groups (20 mice per group) by a random table method, and then 20 negative control groups, 20 negative control groups and Hp-infected positive control groups were perfused with PBS buffer, and the remaining 7 groups were separately perfused with the probiotic compositions prepared in examples 4, 5 and 6 (4.0X 10)9CFU/day), and lactobacillus rhamnosus LN56(4.0 × 10) prepared in example 3 (see below)9CFU/day), Lactobacillus plantarum LN66(4.0 × 10)9CFU/day), Lactobacillus reuteri LN83(4.0 × 10)9CFU/day) and Lactobacillus salivarius LN12 (4.0X 10)9CFU/day). The mice are completely sacrificed 1 week after the last gavage after the continuous gavage treatment for 14d for 1 time every day, the Hp distribution condition is evaluated by taking histopathological special silver staining of the gastric mucosa, and the degree score of the gastric mucosal inflammation is observed by HE (high intensity ultrasound) staining microscopy. Specific results are shown in table 4, the 4 probiotic compositions can significantly inhibit helicobacter pylori infection of model mice, and particularly, example 6 can ensure that 95% of mice have gastric mucosa with helicobacter pylori turning negative and inflammation disappeared; the 4 strains are used alone, but the quantity of the helicobacter pylori is reduced, the inflammation degree is reduced, and the helicobacter pylori cannot turn negative and disappear. Therefore, the combined application of the 4 probiotics has the effect of inhibiting the helicobacter pylori obviously better than the single application of LN56, LN66, LN83 and LN12 strains, and the 4 probiotics have synergistic effect and can inhibit the helicobacter pylori more effectively than any one strain alone.
TABLE 4 Effect of probiotics on H.pylori-infected model mice
Note: the data in the table are the number of mice, -, +, ++, +++, respectively, which represent H.pylori/negative/no inflammation of the degree of inflammation, mild positive/mild inflammation, moderate positive/moderate inflammation and severe positive/severe inflammation. Mice in the growth process died after gavage, so there were fewer than 20 mice in different groups. After grouping, 10 mice were sacrificed before the first gavage, and Hp distribution and inflammation score were determined.
Example 8
Respectively applying the probiotic compositions prepared by the invention (the content of live bacteria of 4 strains is respectively 2.0 multiplied by 10)10CFU/day, total viable bacteria content 8.0 × 1010CFU/day) and low dose of the 4-strain composition (viable bacteria content of 4 strains is 2.0 × 109CFU/day, total viable bacteria content 8.0 × 109CFU/day) were administered to 20 helicobacter pylori positive patients. By using13C-Urea breath test for changes in H.pylori: the patient holds breath for 15s, half of the gas is discharged, and the other part of the gas is blown into the gas collecting bag, which is called as zero breath; 13G capsule 1 is taken with mineral water, exhaled gas is collected after sitting still for 30min, the gas is a sample gas, the sample gas of 0min and 30min is detected by an infrared spectrometer, the detection result is recorded, the specific result is shown in table 5, after helicobacter pylori infected patients take the probiotic composition for 14 days, the infection degree begins to have a descending trend, after taking for 28 days, 90% of the patients have obvious curative effect, wherein, 70% of the patients have helicobacter pylori turning to negative. Meanwhile, by taking the probiotic composition with the dosage lower than the regulated dosage of the invention as a control, the result shows that after the probiotic composition is taken by a patient for 28 days, the effect of inhibiting the helicobacter pylori is obviously lower than that of the probiotic composition, and no patient can convert the helicobacter pylori to negative. The result shows that the content of the viable bacteria in the probiotic composition is not less than 2.0 multiplied by 10 for each bacterium10CFU/day, total viable count not less than 8.0 × 1010CFU/day.
And (3) judging the DOB value: measurement in sample gas for 30min13C-CO2The difference between the δ permillage values of the δ permillage breath samples at zero subtracted by δ permillage: delta thousandths (30min) -delta thousandths (0min), 4.0% -4.0% of DOB is negative (-), 4% -10% of DOB is slight positive (+), 10% -20% of DOB is medium positive (+), and DOB is medium positive (+), (0min)>20% are strongly positive (+++).
TABLE 5 therapeutic Effect of different viable bacteria content for the treatment of helicobacter pylori infection
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. The probiotic composition for inhibiting helicobacter pylori is characterized by comprising the following bacterial powder in parts by weight: 10-30 parts of lactobacillus rhamnosus, 10-30 parts of lactobacillus plantarum, 10-30 parts of lactobacillus reuteri and 10-30 parts of lactobacillus salivarius;
the Lactobacillus rhamnosus is Lactobacillus rhamnosus LN56 with the preservation number of CGMCC No. 17370; the Lactobacillus plantarum is Lactobacillus plantarum (LN 66) with the preservation number of CGMCC No. 17369; the Lactobacillus reuteri is Lactobacillus reuteri LN83 with the preservation number of CGMCC No. 17541; the Lactobacillus salivarius is Lactobacillus salivarius LN12 with the preservation number of CGMCC No. 17542;
the number of viable bacteria in the bacterial powder of the lactobacillus rhamnosus LN56, the bacterial powder of the lactobacillus plantarum LN66, the bacterial powder of the lactobacillus reuteri LN83 and the bacterial powder of the lactobacillus salivarius LN12 is 1.0 multiplied by 1010～3.0×1011CFU/g。
2. The probiotic composition according to claim 1, characterized in that the probiotic composition consists of the following bacterial powder in parts by weight: lactobacillus rhamnosus LN 5615-25 parts, lactobacillus plantarum LN 6615-25 parts, lactobacillus reuteri LN 8315-25 parts and lactobacillus salivarius LN 1215-25 parts.
3. Use of a probiotic composition according to claim 1 or 2 in the manufacture of a medicament for inhibiting the growth of helicobacter pylori.
4. Use of a probiotic composition according to claim 1 or 2 for the preparation of a medicament for the prevention and/or treatment of helicobacter pylori infection.
5. A medicament for the prophylaxis and/or treatment of helicobacter pylori infection, which comprises the probiotic composition according to claim 1 or 2 as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011412221.1A CN112458020B (en) | 2020-12-04 | 2020-12-04 | Probiotic composition for inhibiting helicobacter pylori and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011412221.1A CN112458020B (en) | 2020-12-04 | 2020-12-04 | Probiotic composition for inhibiting helicobacter pylori and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112458020A CN112458020A (en) | 2021-03-09 |
| CN112458020B true CN112458020B (en) | 2022-02-18 |
Family
ID=74805619
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011412221.1A Active CN112458020B (en) | 2020-12-04 | 2020-12-04 | Probiotic composition for inhibiting helicobacter pylori and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112458020B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113384662A (en) * | 2021-07-13 | 2021-09-14 | 镇江市天益生物科技有限公司 | Helicobacter pylori-resistant probiotic composition and preparation method and application thereof |
| CN115153026B (en) * | 2022-04-13 | 2024-01-26 | 微康益生菌(苏州)股份有限公司 | Application of lactobacillus rhamnosus strain LRa05 in preparation of helicobacter pylori and/or streptococcus mutans preparation inhibition |
| CN115029265B (en) * | 2022-06-20 | 2023-06-20 | 自然资源部第一海洋研究所 | Lactobacillus plantarum M503, preparation compounded with lactobacillus rhamnosus and application of lactobacillus plantarum M503 in helicobacter pylori resistant medicines |
| CN115094001B (en) * | 2022-06-24 | 2023-09-26 | 上海利康精准医疗技术有限公司 | Leuconostoc mesenteroides and application thereof in preparation of helicobacter pylori inhibiting drugs |
| CN116731913B (en) * | 2023-05-30 | 2024-03-15 | 江西仁仁健康微生态科技有限公司 | Application of lactobacillus plantarum in preparation of helicobacter pylori resistant products |
| CN117562930B (en) * | 2024-01-15 | 2024-04-16 | 山东中科嘉亿生物工程有限公司 | Compound bacterial agent for preventing helicobacter pylori infection, and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105995972A (en) * | 2016-05-13 | 2016-10-12 | 晶叶（青岛）生物科技有限公司 | Symbiotic microflora enzyme, application thereof and food prepared from symbiotic microflora enzyme |
| CN111053237A (en) * | 2019-12-30 | 2020-04-24 | 广州市中食科康保健食品有限公司 | Health food composition for reducing gastric helicobacter pylori and preparation method thereof |
| CN111956671A (en) * | 2020-07-06 | 2020-11-20 | 薛松晓 | Composite probiotics for treating helicobacter pylori infection |
-
2020
- 2020-12-04 CN CN202011412221.1A patent/CN112458020B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105995972A (en) * | 2016-05-13 | 2016-10-12 | 晶叶（青岛）生物科技有限公司 | Symbiotic microflora enzyme, application thereof and food prepared from symbiotic microflora enzyme |
| CN111053237A (en) * | 2019-12-30 | 2020-04-24 | 广州市中食科康保健食品有限公司 | Health food composition for reducing gastric helicobacter pylori and preparation method thereof |
| CN111956671A (en) * | 2020-07-06 | 2020-11-20 | 薛松晓 | Composite probiotics for treating helicobacter pylori infection |
Non-Patent Citations (1)
| Title |
|---|
| Efficacy and safety of probiotic-supplemented triple therapy for eradication of Helicobacter pylori in children: a systematic review and network meta-analysis;Feng Jue-Rong等;《Eur J Clin Pharmacol》;20170705;第73卷;第1199-1208页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112458020A (en) | 2021-03-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112458020B (en) | Probiotic composition for inhibiting helicobacter pylori and application thereof | |
| TWI652343B (en) | Lactobacillus crimper (LACTOBACILLUS Crispatus) and application thereof | |
| CN107299065B (en) | Lactobacillus plantarum and application thereof in preparation of vagina bacteriostatic drugs | |
| CN113832077B (en) | Lactobacillus rhamnosus and application thereof | |
| WO2017020784A1 (en) | Bacteroides fragilis and application thereof | |
| CN107267415B (en) | Lactobacillus reuteri and application thereof in preparing vagina bacteriostatic drug | |
| CN114350578B (en) | Lactobacillus plantarum LP1Z for producing lysozyme and efficiently antagonizing multidrug-resistant helicobacter pylori and application thereof | |
| CN113293118B (en) | Lactobacillus rhamnosus LR3001 and application thereof | |
| CN116083325B (en) | Lactobacillus rhamnosus for improving helicobacter pylori related gastrointestinal diseases and application thereof | |
| CN110023486B (en) | Lactobacillus acidophilus and culture method and application thereof | |
| CN116731891B (en) | Bifidobacterium breve B2798 and application thereof in preparation of probiotic preparation | |
| EP3808357A1 (en) | Composition and uses thereof | |
| CN114774315B (en) | Application of lactobacillus rhamnosus strain LRa05 in preparation of immunity enhancing product and/or eczema relieving product | |
| CN114806958B (en) | Stomach-derived bacillus coagulans BCF-01 and application thereof | |
| CN113637603B (en) | Lactobacillus entericus and application thereof | |
| CN113575758A (en) | Composite probiotics and preparation method thereof | |
| CN111471623A (en) | Composition of three lactobacilli and application thereof | |
| CN114836349B (en) | Lactobacillus acidophilus LA16 for antagonizing helicobacter pylori and application thereof | |
| CN110938563B (en) | Lactobacillus BJ-REBORN001 and application thereof in preparation of helicobacter pylori inhibiting fermentation broth | |
| CN117402794B (en) | Lactobacillus gasseri and application thereof | |
| CN116103197B (en) | Streptococcus thermophilus with helicobacter pylori inhibiting effect and application thereof | |
| CN117343880B (en) | Saliva host-associated lactobacillus and application thereof | |
| CN116478874B (en) | Lactobacillus paracasei for improving chronic low-grade inflammation and application thereof | |
| CN110538201A (en) | Lactobacillus composition and application thereof | |
| CN116004475B (en) | Lactobacillus plantarum for preventing and assisting in treating stomach deficiency-cold and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |