CN114262677A - Lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel diseases - Google Patents
Lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel diseases Download PDFInfo
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Abstract
The invention discloses lactobacillus plantarum and application thereof in preventing and/or treating inflammatory bowel diseases. The invention separates and obtains plant Lactobacillus (Lactobacillus plantarum) Pm001, which is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of CCTCC NO: m2021926. In vivo and in vitro related experiments prove that the lactobacillus plantarum Pm001 can inhibit the growth of harmful intestinal bacteria and improve intestinal flora, has excellent resistance to inflammatory bowel diseases (including ulcerative colitis or Crohn's 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 diseases.
Background
Ulcerative Colitis (UC) and Crohn's Disease (CD) are two major types of Inflammatory Bowel Disease (IBD), both of which are global epidemic diseases. The current accepted pathogenesis hypothesis of IBD is 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 accumulation of microorganisms in the human gut, collectively known as the gut microflora. The gut microflora has physiological functions related to nutrition, the immune system and host defense. Recent advances in new generation sequencing technologies have identified alterations in the composition and function of the IBD gut microflora, 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. gut microbiota in the pathogenesis of infectious bone disease. clinical Journal of Gastroenterology 11,1-10 (2017); Franzosa, e.a.et. gut microbiome structure and metabolic activity in infectious bone disease. nat. Microbiol 4, 293-.
In China, the number and incidence rate of UC patients are obviously higher than those of CD, and the UC patients show a trend of increasing year by year. The medicines clinically used for treating UC at present are mainly divided into an aminosalicylic acid medicine, a glucocorticoid and a biological agent. Mainly acts on microorganisms and inflammatory cells in intestinal tracts to obtain the curative effects of resisting bacteria and diminishing inflammation. However, the three types of medicines have limited therapeutic effects on ulcerative colitis and have more side effects. In addition, although clinical symptoms can be improved to a great extent after treatment, they can still recur, and thus how to induce remission and maintain remission for a long period of time are critical to the treatment of UC (Guiling, Yanni, Chua Xiao Hua. maintenance of remission therapy is important in reducing the recurrence rate of patients with ulcerative colitis [ J ]. Chinese medicine, 2015, 27(21): 65-66).
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides lactobacillus plantarum and an application thereof in preventing and/or treating inflammatory bowel diseases.
In the first aspect of the invention, the lactobacillus plantarum Pm001 has a preservation number of CCTCC NO: m2021926.
In a second aspect of the invention, there is provided a composition comprising lactobacillus plantarum Pm001, a culture thereof, a suspension thereof or a lysate thereof, and one or more adjuvants.
In particular, the composition comprises lactobacillus plantarum Pm001 and one or more auxiliary materials.
Specifically, in the composition, the 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 gut flora homeostasis, preventing and treating intestinal disorders 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 live lactobacillus plantarum Pm001), 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, flavorants, scents, colorants, coatings, acidity regulators, preservatives, diluents, suspension stabilizers, and the like.
Specifically, the pharmaceutical composition may be in any suitable dosage form, particularly oral dosage forms, for example, liquid preparations (e.g., emulsions, suspensions, syrups, etc.), solid preparations (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 the lactobacillus plantarum Pm001 is a viable bacterium, and the effective viable count is more than or equal to 1 × 109CFU/mL, e.g., 1X 109To 10X 109CFU/mL，5×109CFU/mL。
In another embodiment of the invention, the composition is a food composition, which is a food taken by human beings in daily life, and can be a food composition directly added with lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (particularly, live lactobacillus plantarum Pm001), or a food composition produced by fermentation after being added with lactobacillus plantarum Pm 001.
Specifically, the food composition can be dairy products (such as fermented milk, yogurt, cream, cheese, etc.), bean products (such as soybean milk, fermented bean curd, fermented soybean, bean paste, etc.), fruit and vegetable products (such as vegetable beverage, pickled products (such as sauerkraut, sauerkraut)), meat products, seasonings, etc.
Specifically, the food composition may take any form, such as candy (e.g., tabletted candy, gel candy, gum candy, etc.), solid beverage (e.g., powder, granule, etc.), liquid beverage, etc.
In another embodiment of the invention, the composition is a nutraceutical composition, which can regulate the functions of the human body, but not for the purpose of treating diseases, comprising lactobacillus plantarum Pm001, a culture thereof, a suspension thereof or a lysate thereof (in particular, live lactobacillus plantarum Pm001), and one or more nutraceutical adjuvants.
Specifically, the nutraceutical excipients may be, for example, but not limited to, one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavorants, scents, colorants, coatings, acidity regulators, preservatives, diluents, suspension stabilizers, and the like.
Specifically, the health product composition can be in any form, such as tablet, pill, capsule, candy (such as tablet candy, gel candy, gum 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 a small amount or trace amount during the production and use processes of feeds (such as livestock feed, livestock drinking water, pet ration, pet snacks, pet drinking water and the like), and comprises lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof (particularly, viable lactobacillus plantarum Pm001), and one or more additive adjuvants.
Specifically, the additive adjuvants may be, for example, but not limited to, one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, fragrances, colorants, coating agents, acidity regulators, preservatives, diluents, suspension stabilizers, and the like.
Specifically, the form of the feed additive composition may take any form, such as tablets, pills, capsules, powders, granules, liquids, and the like.
In another embodiment of the invention, the composition is a feed composition, which is a feed for human daily feeding animals (e.g. pets, livestock, laboratory animals), such as livestock feed, pet ration, pet treats, etc., which can be a feed composition directly added with lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof, or a lysate thereof (in particular, live lactobacillus plantarum Pm001), or a feed composition produced by fermentation after being added with 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 preparation of a medicament for the prevention and/or treatment of intestinal disorders.
In particular, the intestinal disease may be selected from duodenal, jejunal, ileum, caecum, colon or rectal diseases, in particular intestinal diseases affecting the ileum, rectum and 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).
Specifically, the inflammatory bowel disease may be a chronic inflammatory bowel disease or an acute inflammatory bowel disease, particularly an acute inflammatory bowel disease.
In another embodiment of the invention, the intestinal disease is a tumor of the intestine, such as, but not limited to, colon cancer, rectal cancer.
In particular, the symptoms of the intestinal disease are selected from: abdominal pain, diarrhea, hematochezia, vomiting, tenesmus, crypt abscesses, and one or more of weight loss, fever, anemia, hypoproteinemia, electrolyte abnormalities, polyps, bowel perforation, and the like.
In particular, the therapeutic effect may be to ameliorate one or more of the above symptoms of the bowel disease, reduce the rate of recurrence of the bowel disease, or even completely cure the bowel disease.
In particular, the preventive effect may be to reduce the incidence of intestinal diseases, or even to completely avoid such intestinal diseases.
In a fourth aspect of the present invention, there is provided a method for the prevention and/or treatment of intestinal disorders, comprising the step of administering to a subject in need thereof lactobacillus plantarum Pm001, a culture thereof, a suspension thereof or a lysate thereof (in particular live lactobacillus plantarum Pm001), or a pharmaceutical composition according to the second aspect of the present invention.
In particular, the intestinal disease may be selected from duodenal, jejunal, ileum, caecum, colon or rectal diseases, in particular intestinal diseases affecting the ileum, rectum and 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 disease is a tumor of the intestine, such as, but not limited to, colon cancer, rectal cancer.
In particular, the symptoms of the intestinal disease are selected from: abdominal pain, diarrhea, hematochezia, vomiting, tenesmus, crypt abscesses, and one or more of weight loss, fever, anemia, hypoproteinemia, electrolyte abnormalities, polyps, bowel perforation, and the like.
In particular, the therapeutic effect may be to ameliorate one or more of the above symptoms of the bowel disease, reduce the rate of recurrence of the bowel disease, or even completely cure the bowel disease.
In particular, the preventive effect may be to reduce the incidence of intestinal diseases, or even to completely avoid such intestinal diseases.
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 modulating gut flora homeostasis comprising the step of administering to a subject in need thereof a lactobacillus plantarum Pm001, a culture thereof, a suspension thereof or a lysate thereof (in particular a live lactobacillus plantarum Pm001), or a food, nutraceutical or feed additive, feed composition according to the second aspect of the invention.
In particular, the method of modulating gut 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., a pig, a cow, a sheep, a horse, a donkey, a fox, a racoon dog, a mink, a camel, etc.), a pet (e.g., a dog, a cat, a rabbit, a mouse (e.g., a guinea pig, a hamster, a gerbil, a dragon cat, a squirrel, etc.), an experimental animal (e.g., a monkey, a dog, a rabbit, a cat, a mouse (e.g., a mouse, a rat), etc.), and 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's disease), has no toxic or side effect, and has important application value. .
The Lactobacillus plantarum Pm001 disclosed by the invention has been preserved in the China center for type culture Collection (address: Wuhan, Wuhan university, China) at 23/7 in 2021, and the preservation number is as follows: CCTCC NO: m2021926.
Drawings
FIG. 1 shows the body weight change curves of mice in the saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treatment group and the Lactobacillus rhamnosus LGG-DSS control group during DSS-induced enteritis.
FIG. 2 shows fecal scoring curves during DSS-induced enteritis in mice from saline-DSS control group, Lactobacillus plantarum Pm001-DSS treatment group, and Lactobacillus rhamnosus LGG-DSS control group.
FIG. 3 shows the length of colon of 11 days of mice in the normal saline-DSS control group, the Lactobacillus plantarum Pm001-DSS treatment group and the Lactobacillus rhamnosus LGG-DSS control group.
FIG. 4 shows H & E staining of distal colon of mice in saline-DSS control group, Lactobacillus plantarum Pm001-DSS treatment group and Lactobacillus rhamnosus LGG-DSS control group to show 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 pertains.
As used herein, the term "culture" refers to a cell population or growth of a microorganism in a space over time, and in particular to a liquid or solid medium in which a population of microorganisms (lactobacillus plantarum Pm001) has grown after artificial inoculation and culture.
The term "lysate" as used in the present invention refers to the product obtained after the cell lysis of the microorganism (Lactobacillus plantarum Pm 001).
The term "bacterial suspension" as used in the present invention 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 "prevention" refers to the ability to prevent, 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 and refer to any animal or cell thereof, whether in vitro or in situ, that is treated according to the methods described herein. Specifically, the aforementioned animals include mammals, such as humans, domestic animals (i.e., animals domesticated by human breeding and whose reproduction can be controlled artificially, for functions such as eating, working, fur, pets, experiments, such as economic animals, pets, laboratory animals, etc.), and particularly humans.
The disclosures of the various publications, patents, and published patent specifications cited herein are hereby incorporated by reference in their entirety.
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: lactobacillus plantarum Pm001 can relieve mouse DSS-induced enteritis phenotype
(1) Preparation of WT mice
30 8-week-old wild-type BL/C57 mice were purchased from Shanghai Ling, Inc., and bred in an SPF barrier system, and after one week of acclimation, the mice were randomly divided into 3 groups, namely, a normal saline-DSS control group (negative control group), a Lactobacillus plantarum Pm001-DSS treatment group, and a Lactobacillus rhamnosus LGG-DSS control group (positive control group).
(2) Intestinal colonization with lactobacillus plantarum Pm001
Inoculating Lactobacillus plantarum Pm001 and Lactobacillus rhamnosus LGG in liquid MRS culture medium, performing anaerobic and static culture at 37 deg.C for 20-22h, centrifuging at 8000rpm for 5min to collect thallus, washing with normal saline twice, resuspending the thallus in normal saline, and standingAt a concentration of about 5X 109CFU/mL. WT mice were treated by gavage at a dose of 200. mu.L of bacteria per mouse, and treatment was repeated every other day for 2 weeks, while the saline-DSS control group was only treated by gavage with saline. After the 2-week gavage treatment is finished, the mice can be subjected to an induced enteritis experiment, and the same bacterial amount of the gavage is still performed every other day during the induced enteritis period.
(3) Model for establishing mouse DSS-induced enteritis
The drinking water of mice in the normal saline-DSS control group, the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group is changed into 3 percent (mass percent) Dextran Sodium Sulfate (DSS) aqueous solution, and the drinking water is changed into normal drinking water after 9 days of treatment until the experiment is finished. Namely, mice in a normal saline-DSS control group, a lactobacillus plantarum Pm001-DSS treatment group and a lactobacillus rhamnosus LGG-DSS control group drink 3% DSS aqueous solution on days 1-9 of DSS induced enteritis, and mice drink ordinary drinking water on days 10-12.
Mice body weight change, stool dryness, and hematochezia were recorded on days 1-12 of induced enteritis. And (3) grading the feces: 0, solid stool; 1, solid stool, easy deformation; 2, unformed excrement; 3, liquid stool. A comparison of body weight changes in mice is shown in figure 1 and a comparison of stool scores is shown in figure 2.
According to the figure, mice in the normal saline-DSS control group, the lactobacillus plantarum Pm001-DSS treatment group and the lactobacillus rhamnosus LGG-DSS control group all have weight loss after drinking DSS, which indicates that the DSS enteritis induction is 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 difference 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 feces 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 difference between the scores of the mouse feces of the lactobacillus plantarum Pm001-DSS treatment group and the mouse feces of the lactobacillus rhamnosus LGG-DSS control group is not obvious, so that the enteritis symptoms of the mice 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, the mice were sacrificed by cervical dislocation, and the colon part of the mice was removed and measured for length, and the results are shown in fig. 3. A1 cm section of the distal colon tissue is taken for formalin fixation, and then HE staining and tissue morphology analysis are carried out, and the obtained result is shown in FIG. 4.
From fig. 3, it can be seen that: in the DSS induced enteritis model, the intestinal lengths of mice in a lactobacillus plantarum Pm001-DSS treatment group and a lactobacillus rhamnosus LGG-DSS control group are respectively (7.16 +/-0.38 cm) and (6.52 +/-0.53 cm), and the intestinal lengths are obviously longer than those of the mice in a normal saline-DSS group (5.2 +/-0.35 cm); from fig. 4, it can be seen that: the intestinal epithelial cells of mice in the lactobacillus plantarum Pm001-DSS group and the lactobacillus rhamnosus LGG-DSS control group can still see a relatively 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 figure morphologically reflects that the incidence of enteritis of mice in the lactobacillus plantarum Pm001-DSS group is remarkably reduced.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and the like that are within the spirit and principle of the present invention are included in the present invention.
The foregoing embodiments and methods described in this disclosure may vary based on the abilities, experience, and preferences of those skilled in the art.
The mere order in which the steps of a method are listed in the present invention does not constitute any limitation on the order of the steps of the method.
Claims (10)
1. The lactobacillus plantarum Pm001 has a preservation number of CCTCC NO: m2021926.
2. A composition comprising lactobacillus plantarum Pm001, culture thereof, suspension thereof or lysate thereof, according to claim 1, together with one or more adjuvants.
3. The composition of claim 2, wherein the composition is a pharmaceutical composition;
preferably, the pharmaceutical composition comprises viable lactobacillus plantarum Pm001 and one or more pharmaceutically acceptable auxiliary materials;
more preferably, the pharmaceutically acceptable excipients are selected from: one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, fragrances, colorants, coatings, acidity regulators, preservatives, diluents, and suspension stabilizers;
more preferably, the pharmaceutical composition is in an oral dosage form, such as an emulsion, suspension, syrup, powder, tablet, pill, granule, capsule or lozenge.
4. The composition of claim 2, wherein the composition is a food composition;
preferably, the food composition is a food composition directly added with lactobacillus plantarum Pm001, or a food composition produced by fermentation after being added with lactobacillus plantarum Pm 001;
more preferably, the food composition is a dairy product, a soy product, a fruit and vegetable product, a meat product, or a condiment.
5. The composition of claim 2, wherein the composition is a nutraceutical composition;
preferably, the health-care product composition comprises viable lactobacillus plantarum Pm001 and one or more health-care product auxiliary materials;
more preferably, the health product auxiliary material is selected from: one or more of fillers, binders, wetting agents, disintegrants, lubricants, flavoring agents, fragrances, colorants, coatings, acidity regulators, preservatives, diluents, and suspension stabilizers;
more preferably, the health care product composition is a tablet, a pill, a capsule, a candy, a solid beverage or a liquid beverage.
6. The composition of claim 2, wherein the composition is a feed additive composition or a feed composition.
7. Use of lactobacillus plantarum Pm001, a culture thereof, a bacterial suspension thereof or a lysate thereof according to claim 1 for the preparation of a medicament for the prevention and/or treatment of intestinal diseases.
8. The use of claim 7, wherein the intestinal disorder is inflammatory bowel disease or intestinal tumor.
9. The use of claim 8, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.
10. A method of modulating gut 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 of claim 1, or a composition of any one of claims 4-6.
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Cited By (2)
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| CN114990014A (en) * | 2022-05-31 | 2022-09-02 | 成都医学院 | Lactobacillus plantarum for preventing and treating inflammatory enteritis and application thereof |
| CN115737689A (en) * | 2022-11-01 | 2023-03-07 | 中国疾病预防控制中心传染病预防控制所 | Application of lactobacillus in preparing medicine for preventing and treating inflammatory bowel disease |
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| CN111235070A (en) * | 2020-03-18 | 2020-06-05 | 河北农业大学 | Breast milk infant source lactobacillus plantarum BF _15 and application thereof |
| CN111529553A (en) * | 2020-05-28 | 2020-08-14 | 东北农业大学 | Application of plant lactobacillus capable of degrading tryptophan and tryptophan mixture |
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| CN107619811A (en) * | 2017-11-08 | 2018-01-23 | 光明乳业股份有限公司 | Lactobacillus plantarum CCFM200 bacterial strains and application |
| CN111235070A (en) * | 2020-03-18 | 2020-06-05 | 河北农业大学 | Breast milk infant source lactobacillus plantarum BF _15 and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114990014A (en) * | 2022-05-31 | 2022-09-02 | 成都医学院 | Lactobacillus plantarum for preventing and treating inflammatory enteritis and application thereof |
| CN114990014B (en) * | 2022-05-31 | 2023-05-23 | 成都医学院 | Lactobacillus plantarum for preventing and treating inflammatory enteritis and application thereof |
| CN115737689A (en) * | 2022-11-01 | 2023-03-07 | 中国疾病预防控制中心传染病预防控制所 | Application of lactobacillus in preparing medicine for preventing and treating inflammatory bowel disease |
| CN115737689B (en) * | 2022-11-01 | 2023-05-05 | 中国疾病预防控制中心传染病预防控制所 | Application of lactobacillus in preparation of medicines for preventing and treating inflammatory bowel disease |
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