CN114262677B - Lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel disease - Google Patents
Lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel disease Download PDFInfo
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Abstract
The invention discloses lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel disease. The lactobacillus plantarum (Lactobacillus plantarum) Pm001 is isolated and preserved in China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: m2021926. The in vivo and in vitro related experiments prove that the lactobacillus plantarum Pm001 can inhibit the growth of intestinal harmful bacteria, improve intestinal flora, has excellent resistance to inflammatory bowel diseases (including ulcerative colitis or Crohn disease), has no toxic or side effect, and has important application value.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus plantarum Pm001 and application thereof in preventing and/or treating inflammatory bowel disease.
Background
Ulcerative colitis (ulcerative colitis, UC) and Crohn's Disease (CD) are two major types of inflammatory bowel disease (Inflammatory bowel disease, IBD), both of which are global epidemic diseases. The currently accepted pathogenesis of IBD is hypothesized that complex interactions between genetic, environmental factors and the host immune system lead to abnormal immune responses and chronic intestinal inflammation. There is a complex and abundant region of microbial accumulation in the human gut, collectively referred to as the gut microbiota. The intestinal microflora has physiological functions related to nutrition, the immune system and host defenses. Recent advances in new generation sequencing technology have established changes in the composition and function of the intestinal microflora of IBD, which is known as dysbiosis. Clinical and experimental data suggest that dysbiosis may play a critical role in the pathogenesis of IBD (Nishida, A.et al, gut microbiota in the pathogenesis of inflammatory bowel disease.clinical Journal of Gastroenterology, 1-10 (2017); franzosa, E.A.et al, gut microbiome structure and metabolic activity in inflammatory bowel disease.Nat Microbiol 4,293-305 (2019)).
In China, the number and incidence rate of UC patients are obviously higher than those of CD, and the trend of the UC patients is rising year by year. The current clinical drugs for treating UC are mainly classified into an amino salicylic acid drug, a glucocorticoid, a biological agent and the like. Mainly through acting on the microorganism and inflammatory cells in the intestinal tract to obtain the curative effects of resisting bacteria and diminishing inflammation. However, the three medicines have more limited therapeutic effects and more side effects on ulcerative colitis. In addition, although significant improvement in clinical symptoms can be achieved after treatment, it is still reproducible, so how to induce relief and maintain relief for a long period of time is critical for the treatment of UC (Lu Guiling, ni, cai Xiaohua. The importance of maintenance relief treatment in reducing the recurrence rate of ulcerative colitis patients [ J ]. Chinese health medicine, 2015, 27 (21): 65-66).
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel diseases.
In a first aspect of the present invention, there is provided lactobacillus plantarum Pm001 having a accession number cctccc NO: m2021926.
In a second aspect of the present invention, there is provided a composition comprising lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof, and one or more excipients.
In particular, the composition comprises lactobacillus plantarum Pm001 and one or more auxiliary materials.
Specifically, in the composition, lactobacillus plantarum Pm001 is a living bacterium or a non-living bacterium, particularly a living bacterium.
In particular, the composition may also contain other probiotics and/or prebiotics, for example other microorganisms and/or prebiotics that are beneficial in regulating intestinal flora homeostasis, preventing and treating the occurrence of intestinal diseases such as inflammatory bowel disease.
In one embodiment of the invention, the composition is a pharmaceutical composition comprising lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (in particular lactobacillus plantarum Pm001 viable bacteria), and one or more pharmaceutically acceptable excipients.
Specifically, the pharmaceutically acceptable excipients may be, for example, but not limited to, one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, coloring agents, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers, and the like.
In particular, the pharmaceutical composition may be in any suitable dosage form, in particular an oral dosage form, for example, liquid formulations (e.g., emulsions, suspensions, syrups, etc.), solid formulations (e.g., powders, tablets, pills, granules, capsules, lozenges, etc.), and the like.
In some embodiments of the invention, the pharmaceutical composition is a liquid preparation, wherein Lactobacillus plantarum Pm001 is a viable organism with an effective viable organism count of greater than or equal to 1×10 9 CFU/mL, e.g., 1X 10 9 Up to 10X 10 9 CFU/mL，5×10 9 CFU/mL。
In another embodiment of the present invention, the composition is a food composition, which is a food for daily intake by human beings, and may be a food composition directly added with lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (particularly lactobacillus plantarum Pm001 viable bacteria), or may be a food composition produced by fermentation after adding lactobacillus plantarum Pm 001.
Specifically, the food composition may be dairy products (such as fermented milk, yogurt, cream, cheese, etc.), bean products (such as soy milk, fermented bean curd, fermented soybean, bean paste, etc.), fruit and vegetable products (such as vegetable drinks, pickled products (such as kimchi, pickle)), meat products, seasonings, etc.
Specifically, the food composition may take any form, such as a candy (e.g., a tabletted candy, a gel candy, a gum base candy, etc.), a solid beverage (e.g., a powder, granules, etc.), a liquid beverage, etc.
In another embodiment of the invention, the composition is a nutraceutical composition which is capable of modulating the functioning of the human body, but not for the purpose of treating a disease, comprising lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (in particular lactobacillus plantarum Pm001 viable bacteria), and one or more nutraceutical adjuvants.
Specifically, the health-care product auxiliary materials can be one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, colorants, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers and the like, for example, but not limited to.
Specifically, the health product composition can be in any form, such as tablet, pill, capsule, candy (such as tabletting candy, gel candy, gum base candy, etc.), solid beverage (such as powder, granule, etc.), liquid beverage, etc.
In another embodiment of the invention, the composition is a feed additive composition, which is a substance added in small or trace amounts during the production and processing, use of feed (such as livestock feed, livestock drinking water, pet ration, pet treat, pet drinking water, etc.), comprising lactobacillus plantarum Pm001, its culture, its bacterial suspension or its lysate (in particular lactobacillus plantarum Pm001 live bacteria), and one or more additive adjuvants.
Specifically, the additive adjuvant may be, for example, but not limited to, one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, coloring agents, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers, and the like.
Specifically, the feed additive composition may take any form, such as tablets, pills, capsules, powders, granules, liquids, and the like.
In another embodiment of the present invention, the composition is a feed composition which is a feed for daily human fed animals (e.g., pets, livestock, experimental animals), for example, livestock feed, pet ration, pet treat, etc., and may be a feed composition directly added with lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (particularly lactobacillus plantarum Pm001 live bacteria), or may be a feed composition produced by fermentation after adding lactobacillus plantarum Pm 001.
In a third aspect of the invention, there is provided the use of lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof for the manufacture of a medicament for the prevention and/or treatment of an intestinal disease.
In particular, the intestinal disease may be selected from duodenal, jejunal, ileal, cecal, colonic or rectal diseases, in particular intestinal diseases involving the ileum, rectum, colon.
In one embodiment of the invention, the intestinal disease is an inflammatory bowel disease, such as, but not limited to, ulcerative Colitis (UC), crohn's Disease (CD).
In particular, the inflammatory bowel disease may be a chronic inflammatory bowel disease or an acute inflammatory bowel disease, in particular an acute inflammatory bowel disease.
In another embodiment of the invention, the intestinal disorder is an intestinal tumor, such as, but not limited to, colon cancer, rectal cancer.
Specifically, the symptoms of the intestinal disease are selected from: abdominal pain, diarrhea, hematochezia, vomiting, tenesmus, crypt abscess, and one or more of weight loss, fever, anemia, hypoalbuminemia, electrolyte abnormality, polyps, intestinal perforation, etc.
In particular, the therapeutic effect may be to ameliorate one or more of the above symptoms of the intestinal disease, reduce the recurrence rate of the intestinal disease, or even completely cure the intestinal disease.
In particular, the prophylactic effect may be to reduce the incidence of intestinal disease, even completely avoid it.
In a fourth aspect of the present invention, there is provided a method for preventing and/or treating an intestinal disease comprising the step of administering lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (in particular lactobacillus plantarum Pm001 live bacteria), or a pharmaceutical composition according to the second aspect of the present invention, to a subject in need thereof.
In particular, the intestinal disease may be selected from duodenal, jejunal, ileal, cecal, colonic or rectal diseases, in particular intestinal diseases involving the ileum, rectum, colon.
In one embodiment of the invention, the intestinal disease is an inflammatory bowel disease, such as, but not limited to, ulcerative Colitis (UC), crohn's Disease (CD).
In another embodiment of the invention, the intestinal disorder is an intestinal tumor, such as, but not limited to, colon cancer, rectal cancer.
Specifically, the symptoms of the intestinal disease are selected from: abdominal pain, diarrhea, hematochezia, vomiting, tenesmus, crypt abscess, and one or more of weight loss, fever, anemia, hypoalbuminemia, electrolyte abnormality, polyps, intestinal perforation, etc.
In particular, the therapeutic effect may be to ameliorate one or more of the above symptoms of the intestinal disease, reduce the recurrence rate of the intestinal disease, or even completely cure the intestinal disease.
In particular, the prophylactic effect may be to reduce the incidence of intestinal disease, even completely avoid it.
In particular, the subject may be a mammal, in particular a human.
In a fifth aspect of the invention, there is provided a method of regulating intestinal flora homeostasis comprising the step of administering to a subject in need thereof lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (in particular lactobacillus plantarum Pm001 viable bacteria), or a food, nutraceutical or feed additive composition, a feed composition according to the second aspect of the invention.
In particular, the method of modulating intestinal flora homeostasis is for non-disease prevention or treatment purposes.
Specifically, the subject may be a mammal, for example, a human, a domestic animal (e.g., pig, cow, sheep, horse, donkey, fox, raccoon dog, marten, camel, etc.), a pet (e.g., dog, cat, rabbit, mouse (e.g., guinea pig, hamster, gerbil, dragon cat, squirrel, etc.), an experimental animal (e.g., monkey, dog, rabbit, cat, mouse (e.g., mouse, rat), etc.), or the like, particularly a human.
The lactobacillus plantarum Pm001 is obtained through separation, and related experiments in vivo and in vitro prove that the lactobacillus plantarum Pm001 can inhibit the growth of harmful intestinal bacteria, improve intestinal flora, has excellent resistance to inflammatory bowel diseases (including ulcerative colitis or Crohn disease), has no toxic or side effect, and has important application value. .
Lactobacillus plantarum (Lactobacillus plantarum) Pm001 described in the present invention was deposited at the China center for type culture Collection (address: university of Wuhan, china) at 2021, 7 months, 23 days, accession number: cctccc NO: m2021926.
Drawings
FIG. 1 shows the body weight change curves of mice in the saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treated group and the Lactobacillus rhamnosus LGG-DSS control group during DSS induced enteritis.
FIG. 2 shows stool score curves during DSS induced enteritis in saline-DSS control, lactobacillus plantarum Pm001-DSS treated and Lactobacillus rhamnosus LGG-DSS control mice.
FIG. 3 shows the length of the colon of day 11 mice in the saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treated group and the Lactobacillus rhamnosus LGG-DSS control group.
FIG. 4 shows the distal colon H & E staining of mice in saline-DSS control, lactobacillus plantarum Pm001-DSS treated and Lactobacillus rhamnosus LGG-DSS control, showing intestinal epithelial destruction.
Detailed Description
Unless defined otherwise, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention relates.
As used herein, the term "culture" refers to a population or growth of microorganisms within a certain space over a certain period of time, and particularly to a liquid or solid medium in which a population of microorganisms (lactobacillus plantarum Pm 001) has grown after artificial inoculation and culture.
As used herein, the term "lysate" refers to the product obtained after cell lysis of a microorganism (lactobacillus plantarum Pm 001).
As used herein, the term "bacterial suspension" refers to a suspension containing cells of a microorganism (lactobacillus plantarum Pm 001).
As used herein, the term "treating" includes eradicating, removing, reversing, alleviating, altering or controlling the disease after its onset.
As used herein, the term "preventing" refers to the ability to avoid, minimize, or make difficult the onset or progression of a disease or condition by treatment prior to the onset of the disease.
As used herein, the terms "patient" or "subject" and the like are used interchangeably herein to refer to any animal or cell thereof, whether in vitro or in situ, treated according to the methods described herein. Specifically, the aforementioned animals include mammals, for example, humans, domestic animals (i.e., animals that are domesticated by human feeding and whose reproduction can be controlled artificially, for functions such as eating, working, fur, pets, experiments, etc., such as economic animals, pets, experimental animals, etc.), and particularly humans.
Various publications, patents, and published patent specifications cited herein are incorporated by reference in their entirety.
The technical solutions of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments 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.
Example 1: lactobacillus plantarum Pm001 can alleviate mice DSS induced enteritis phenotype
(1) Preparation of WT mice
30 wild type BL/C57 mice, 8 weeks old, were purchased from Shanghai Ling company, housed in SPF barrier system, and after one week adaptation, the mice were randomly divided into 3 groups, which were respectively a saline-DSS control group (negative control group), a Lactobacillus plantarum Pm001-DSS treated group, and a Lactobacillus rhamnosus LGG-DSS control group (positive control group).
(2) Lactobacillus plantarum Pm001 intestinal tract field planting
Inoculating Lactobacillus plantarum Pm001 and Lactobacillus rhamnosus LGG into liquid MRS culture medium, respectively, anaerobic culturing at 37deg.C for 20-22 hr, centrifuging at 8000rpm for 5min to collect thallus, washing twice with physiological saline, re-suspending the bacteria in physiological saline to a final concentration of about 5×10 9 CFU/mL. WT mice were gavaged at a dose of 200. Mu.L bacteria per mouse, and the treatment was repeated every other day for 2 weeks, while the saline-DSS control group was only gavaged with saline. After the 2-week gastric lavage treatment is finished, the mice can be subjected to an enteritis induction experiment, and the same bacterial load can be irrigated in every other day during the enteritis induction period.
(3) Establishing a mouse DSS induced enteritis model
The normal saline-DSS control group, the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group are changed into 3 percent (mass percent) Dextran Sodium Sulfate (DSS) water solution, and after 9 days of treatment, the normal water is changed into normal water until the experiment is finished. Namely, mice in the physiological saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treatment group and the Lactobacillus rhamnosus LGG-DSS control group drink 3% DSS aqueous solution on days 1-9 of DSS-induced enteritis, and ordinary drinking water on days 10-12.
Mice were recorded for weight changes, stool dryness, and hematochezia on days 1-12 of enteritis induction. Fecal scoring: 0, solid state stool; 1, solid stool, which is easy to deform; 2, not forming feces; 3, liquid excrement. Comparison of body weight changes in mice is shown in figure 1 and comparison of stool scores is shown in figure 2.
From this figure, it can be seen that weight loss occurred in mice of the saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treated group and the Lactobacillus rhamnosus LGG-DSS control group after drinking DSS, indicating that DSS enteritis induction was successful. Further comparing, the weight loss amplitude of the mice in the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group is obviously smaller than that of the mice in the normal saline-DSS control group, and the weight loss amplitude of the mice in the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group is not obvious; meanwhile, the scores of the mouse faeces of the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group are obviously lower than those of the normal saline-DSS group, and the differences of the scores of the mouse faeces of the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group are not obvious, so that the symptoms of the mouse enteritis of the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group are obviously improved compared with those of the normal saline-DSS group.
On day 12 of DSS treatment, mice were sacrificed by cervical dislocation, colon portions of the mice were removed, and their lengths were measured, and the results are shown in fig. 3. The distal colon tissue was formalin-fixed at about 1cm of the intestinal segment and subsequently HE stained and histomorphometric analyzed, with the results shown in fig. 4.
From fig. 3, it can be known that: in the DSS induced enteritis model, the intestinal length of the lactobacillus plantarum Pm001-DSS treated group and the lactobacillus rhamnosus LGG-DSS control group mice is respectively (7.16+/-0.38 cm) and (6.52+/-0.53 cm), and is obviously longer than that of normal saline-DSS group mice (5.2+/-0.35 cm); from fig. 4, it can be known that: the intestinal epithelial cells of the mice in the Lactobacillus plantarum Pm001-DSS group and the Lactobacillus rhamnosus LGG-DSS control group can still see a more complete crypt structure, while the intestinal epithelial cells of the mice in the normal saline-DSS group are completely destroyed, and have no complete crypt structure. The graph reflects that the enteritis incidence of the lactobacillus plantarum Pm001-DSS mice is obviously reduced in morphology.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is to be construed as including any modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
The foregoing embodiments and methods described in this invention may vary based on the capabilities, experience, and preferences of those skilled in the art.
The listing of the steps of a method in a certain order in the present invention does not constitute any limitation on the order of the steps of the method.
Claims (18)
1. Lactobacillus plantarum Pm001, which is classified and namedLactobacillus plantarumThe preservation number is CCTCC NO: m2021926.
2. A composition comprising lactobacillus plantarum Pm001 or a bacterial suspension thereof according to claim 1, and one or more excipients.
3. The composition of claim 2, wherein the composition is a pharmaceutical composition.
4. A composition according to claim 3, wherein the pharmaceutical composition comprises lactobacillus plantarum Pm001 viable bacteria and one or more pharmaceutically acceptable excipients.
5. The composition of claim 4, wherein the pharmaceutically acceptable adjuvant is selected from the group consisting of: one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, coloring agents, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers.
6. The composition of claim 3, wherein the pharmaceutical composition is in an oral dosage form.
7. The composition of claim 6, wherein the oral dosage form is an emulsion, suspension, syrup, powder, tablet, pill, granule, capsule, or lozenge.
8. The composition of claim 2, wherein the composition is a food composition.
9. The composition of claim 8, wherein the food composition is a food composition directly added with lactobacillus plantarum Pm001 or a food composition produced by fermentation after adding lactobacillus plantarum Pm 001.
10. The composition of claim 9, wherein the food composition is a dairy product, a soy product, a fruit and vegetable product, a meat product, or a flavoring.
11. The composition of claim 2, wherein the composition is a nutraceutical composition.
12. The composition of claim 11, wherein the nutraceutical composition comprises live lactobacillus plantarum Pm001 and one or more nutraceutical excipients.
13. The composition of claim 12, wherein the nutraceutical adjunct is selected from the group consisting of: one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, coloring agents, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers.
14. The composition of claim 12, wherein the nutraceutical composition is a tablet, a pill, a capsule, a candy, a solid beverage, a liquid beverage.
15. The composition of claim 2, wherein the composition is a feed additive composition or a feed composition.
16. Use of lactobacillus plantarum Pm001 or a bacterial suspension thereof according to claim 1 for the manufacture of a medicament for the prevention and/or treatment of intestinal diseases.
17. The use according to claim 16, wherein the intestinal disorder is inflammatory bowel disease or an intestinal tumor.
18. The use of claim 17, wherein the inflammatory bowel disease is ulcerative colitis or crohn's disease.
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