CN115998777B

CN115998777B - Use of lactobacillus paracasei 207-27

Info

Publication number: CN115998777B
Application number: CN202211273484.8A
Authority: CN
Inventors: 迮晓雷; 李良; 董权锋; 汪玉芳; 奚彧; 张旭光; 贺瑞坤
Original assignee: BY Health Co Ltd
Current assignee: BY Health Co Ltd
Priority date: 2022-10-18
Filing date: 2022-10-18
Publication date: 2023-11-17
Anticipated expiration: 2042-10-18
Also published as: CN115998777A

Abstract

The present application relates to the use of lactobacillus paracasei (Lactobacillus paracasei) or a progeny thereof, or a composition comprising said lactobacillus paracasei or a progeny thereof, in the manufacture of a medicament or a food product, and to a method of increasing the concentration of lactobacillus paracasei in the gastrointestinal tract of a subject. Can promote small intestine movement, increase defecation amount and defecation weight of mammal under disease model, and also has effect in regulating intestinal flora (i.e. can restore intestinal flora under disease model to health state).

Description

Use of lactobacillus paracasei 207-27

Technical Field

The present application relates to the use of lactobacillus paracasei or its progeny or a composition comprising said lactobacillus paracasei or its progeny in the manufacture of a medicament or food, and to a method of increasing the concentration of lactobacillus paracasei in the gastrointestinal tract of a subject.

Background

Functional constipation is a clinically common functional disorder of the gastrointestinal tract. The prevalence rate of adult chronic constipation in China is 4% -6%, the prevalence rate of people over 60 years old can reach 22%, and the prevalence rate in western countries is about 27%. The pathogenesis of functional constipation mainly includes the following 4 types: insufficient intake of cellulose and water; the tension of intestinal smooth muscle is reduced, and the peristalsis is weakened; intestinal peristalsis is mechanically hindered; dyskinesia of the faecal muscle group. Research at home and abroad shows that the intestinal flora disturbance can participate in the occurrence and progress of chronic constipation except for the above 4 initial disease factors. In recent years, intestinal microecologics are used as auxiliary treatment for constipation, and have shown unique benefits in terms of efficacy and safety, so that the intestinal microecologics are increasingly valued.

The research on the action mechanism of probiotics on relaxing bowel and regulating intestinal flora is gradually going deep, but is still imperfect. Part of the theory includes: the probiotics can produce various organic acids through metabolism, so that the pH value in the intestinal cavity is reduced, and the intestinal peristalsis is regulated; part of probiotics can induce the rise of the plasma concentration of neurotransmitter dopamine and 5-HT, and can also accelerate the contraction of intestinal smooth muscle and strengthen the intestinal peristalsis; in addition, the healthy intestinal microbial environment is an important factor for promoting intestinal peristalsis, and the ingestion of probiotics can change the intestinal flora of a host, so that the healthy balanced intestinal flora is maintained, and the intestinal tract is beneficial to exerting normal absorption and excretion functions.

In addition, whether probiotics exert the above probiotic effects on the body depends on whether the strain can withstand the defense mechanisms of the gastrointestinal tract of the body, such as the low pH environment in gastric juice and bile acid in the small intestine, and the like, smoothly reach the intestinal tract in a viable state, adhere to intestinal epithelial cells, colonize the intestinal tract, generate beneficial metabolites, and interact with the host. Therefore, screening for strains that are able to enter the gut with high viability is also a primary concern for probiotic development.

According to the scientific consensus of probiotics: the current research shows that the genome of different probiotics has great difference, even the different strains of the same kind of probiotics have the difference. The document of IDF publication No.513/2021 Probiotics Strain level identification guide published by International Association of Dairy (International Dairy Federation, IDF) also states that the functionality and safety of probiotics are strain level specific. Since different strains of the same species of probiotics contain or express different functional genes, and can exert different probiotic effects, the probiotics need to be characterized and described at the microbial strain level. This difference in strain specificity is also reflected in the results of the related experimental studies of the present application: even though different strains are of the same source and similar species, the physiological characteristics and the probiotics efficacy of the strains are obviously different.

Therefore, a strain with the effect of relaxing bowel will have wide application potential in the production and application of probiotics.

Disclosure of Invention

The applicant screens out a probiotic strain, lactobacillus paracasei 207-27, at an early stage, and the microorganism preservation number is: GDMCC No.60960. In the subsequent research process, the strain is unexpectedly found to have outstanding efficacy in relaxing bowel. For example, it can promote small intestine movement, increase the defecation amount and defecation weight of mammals in disease models, and also has the function of positively regulating intestinal flora (i.e., can restore intestinal flora in disease states to a healthy state). Thus, the present application has been completed.

Accordingly, in a first aspect, the present application provides the use of lactobacillus paracasei (Lactobacillus paracasei) or a progeny thereof, or a composition comprising same, in the manufacture of a medicament or foodstuff for maintaining and/or improving gastrointestinal health, or preventing and/or treating a disease or condition caused by a gastrointestinal abnormality, in a subject;

wherein the lactobacillus paracasei is deposited with the microorganism strain collection in Guangdong province with the deposit number of GDMCC No.60960.

In certain embodiments, the maintaining and/or improving gastrointestinal health comprises: positively regulate gastrointestinal microorganisms, and promote gastrointestinal motility.

In certain embodiments, improving gastrointestinal health refers to approximating a gastrointestinal status of a subject to a health status of the subject. In such embodiments, the subject is a subject with constipation.

In certain embodiments, maintaining gastrointestinal health refers to maintaining the gastrointestinal status of a subject in a relatively stable state. In such embodiments, the subject is a healthy or near healthy subject.

In certain embodiments, the upregulating gastrointestinal microorganisms comprises: stimulating the growth and/or activity of one or more microorganisms in the gastrointestinal tract that are beneficial to the health of the gastrointestinal tract, inhibiting the growth and/or activity of one or more pathogenic bacteria in the gastrointestinal tract, increasing the attachment of non-pathogenic bacteria to the mucosa on the surface of the gastrointestinal tract, increasing the ability of the intestinal tract to shield against pathogenic bacteria, and/or producing metabolites of microorganisms that are beneficial to the health of the gastrointestinal tract.

In certain embodiments, the promoting gastrointestinal motility comprises: the amount and/or weight of bowel movement by the subject is approximated to the health of the subject.

As used herein, the term "microorganism beneficial to gastrointestinal health" refers to any non-pathogenic microorganism that, when administered to a host in an effective amount, can have a beneficial effect on the health of the host.

In certain embodiments, the microorganism beneficial to gastrointestinal health is selected from bacteria, fungi.

In certain embodiments, the microorganism beneficial to gastrointestinal health is a probiotic.

In certain embodiments, the gastrointestinal disorder comprises: gastrointestinal infections, colonic dysfunction, gastrointestinal dysfunction caused by pathogenic bacteria (e.g. helicobacter pylori).

In certain embodiments, the disease or condition caused by an abnormality of the gastrointestinal tract comprises: abdominal pain, abdominal distention, constipation, diarrhea, anorexia, nausea, vomiting, and acid regurgitation.

As used herein, the term "medicament" encompasses medicaments for humans as well as medicaments for animals (i.e., veterinary applications). In certain embodiments, the medicament is for use in humans.

In certain embodiments, the pharmaceutical composition comprises a formulation of lactobacillus paracasei.

In certain embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

In certain embodiments, the pharmaceutical composition is formulated for oral administration.

In certain embodiments, the drug or food is a drug that targets gastrointestinal release, or is a drug that is released in the gastrointestinal tract in a controlled manner.

In certain embodiments, the medicament is in the form of a pill, powder, capsule, tablet (e.g., effervescent tablet), caplet, mouth-soluble granule, liquid, suppository, or enema.

In certain embodiments, the medicament is used alone or in combination with other antifungal agents, analgesics, anti-inflammatory agents, healing promoting agents, or moisturizers.

In the context of this document, the term "food" is used in a broad sense to include human foods and drinks, as well as animal foods and drinks (i.e., feeds). In certain embodiments, the food product is suitable and designed for human consumption.

It will be appreciated that the food product of the present application may be in the form of a liquid, solid, suspension or powder, depending on the application, mode of application or mode of administration.

In certain embodiments, the food product is selected from a solid beverage, a candy or a juice, or the food product is a dairy product (e.g., yogurt, flavored fermented milk, lactobacillus beverage, cheese).

In certain embodiments, the food product is a dietary supplement.

As used herein, the term "dietary supplement" refers to an edible product that is capable of providing a benefit (e.g., a nutritional effect, a prophylactic effect, a therapeutic effect, or other benefit) to a consumer. Dietary supplements in this context encompass health foods, biomedical foods, nutraceuticals, supplements, and the like.

In certain embodiments, the dietary supplement is formulated for oral administration.

In certain embodiments, the food product may further include (but is not limited to) one or any combination of the following: probiotics (e.g., probiotic bacteria), carbohydrates (e.g., dietary fibers), proteins (e.g., enzymes), lipids (e.g., fats), vitamins, minerals.

In certain embodiments, the food product may further comprise an immunomodulator.

In certain embodiments, the food product may also include a plant ingredient (e.g., flavonoids, polyphenol plant extracts, etc.), a milk substitute, or a metabolite or extract of bifidobacterium breve or progeny thereof.

In certain embodiments, the strains of the application may also be combined with various food acceptable adjuvants in sweeteners or flavors, color modulating substances, stabilizers, glidants, bulking agents, and the like.

In certain embodiments, the food product further comprises a prebiotic.

In certain embodiments, the prebiotic is selected from the group consisting of fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, isomaltooligosaccharides, soy oligosaccharides, inulin, spirulina, arthrospira, coriolus versicolor polysaccharides, nitrogen-containing polysaccharides from the group consisting of the plant, casein hydrolysates, alpha-lactalbumin, lactoferrin, or any combination thereof.

In certain embodiments, the food product is in the form of a pill, powder, capsule, tablet (e.g., effervescent tablet), caplet, mouth-soluble granule, liquid.

In certain embodiments, the subject is a mammal. In certain embodiments, the mammal is selected from the group consisting of a mouse, pig, rabbit, monkey, sheep, human.

In certain embodiments, the lactobacillus paracasei in the food product is at 10 ⁶ To 10 ¹² CFU/dose amount is present.

In certain embodiments, the lactobacillus paracasei in the food product is at 10 ⁸ To 10 ¹² CFU/dose (e.g., 10 ⁸ CFU/dose, 10 ⁹ CFU/dose, 10 ¹⁰ CFU/dose, 10 ¹¹ CFU/dose, 10 ¹² CFU/dose) is present.

It will be appreciated that it will be within the ability of one skilled in the art to administer an amount of lactobacillus paracasei effective for a subject, depending on the specific situation of the subject.

In certain embodiments, the composition comprises the lactobacillus paracasei or progeny thereof, and a microorganism selected from the group consisting of: bacteria, fungi, or any combination thereof.

In certain embodiments, the microorganism is a probiotic.

In certain embodiments, the microorganism is a yeast.

In certain embodiments, the yeast is selected from the group consisting of Saccharomyces cerevisiae (Saccharomyces cerevisiae), saccharomyces boulardii (Saccharomyces boulardii), kluyveromyces marxianus (Kluyveromyces marxianus), or any combination thereof.

In certain embodiments, the bacteria are selected from the group consisting of Lactobacillus (Lactobacillus spp.), bifidobacterium (Bifidobacterium spp.), bacillus (Bacillus spp.), propionibacterium spp.), streptococcus (Streptococcus spp.), lactococcus (Lactococcus spp.), pediococcus (Pediococcus spp.), enterococcus (Enterococcus spp.), staphylococcus (Staphylococcus spp.), or any combination thereof.

In certain embodiments, the bacterium of the genus lactobacillus is selected from the group consisting of: lactobacillus paracasei, lactobacillus acidophilus (Lactobacillus acidophilus), lactobacillus brevis (Lactobacillus brevis), lactobacillus jensenii (Lactobacillus jensenii), lactobacillus inerticus (Lactobacillus iners), lactobacillus casei (Lactobacillus casei), lactobacillus crispatus (Lactobacillus crispatus), lactobacillus curvatus (Lactobacillus curvatus), lactobacillus delbrueckii (Lactobacillus delbrueckii), lactobacillus fermentum (Lactobacillus fermentum), lactobacillus gasseri (Lactobacillus gasseri), lactobacillus helveticus (Lactobacillus helveticus), lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus plantarum (Lactobacillus plantarum), lactobacillus reuteri (Lactobacillus reuteri), lactobacillus rhamnosus (Lactobacillus rhamnosus), lactobacillus sake (Lactobacillus sakei), lactobacillus salivarius (Lactobacillus salivarius), or any combination thereof.

In certain embodiments, the bacterium of the genus bifidobacterium is selected from the group consisting of: bifidobacterium animalis (Bifidobacterium animalis), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium breve (Bifidobacterium breve), bifidobacterium infantis (Bifidobacterium infantis), bifidobacterium longum (Bifidobacterium longum), bifidobacterium adolescentis (Bifidobacterium adolescentis), or any combination thereof.

In certain embodiments, the bacteria of the genus bacillus are selected from the group consisting of: bacillus subtilis (Bacillus subtilis), bacillus coagulans (Bacillus coagulans), or any combination thereof.

In certain embodiments, the bacteria of the genus propionibacterium are selected from the group consisting of: propionibacterium scheelite (Propionibacterium shermanii), propionibacterium freudenreichii (Propionibacterium freudenreichii), propionibacterium propionicum (Propionibacterium acidipropionici), or any combination thereof.

In certain embodiments, the bacteria of the streptococcus genus are selected from: streptococcus thermophilus (Streptococcus thermophilus), streptococcus salivarius (Streptococcus salivarius), or any combination thereof.

In certain embodiments, the bacterium of the genus lactococcus is lactococcus lactis (Lactococcus lactis).

In certain embodiments, the bacterium belonging to the genus Enterococcus (Enterococcus spp.) is selected from the group consisting of: enterococcus faecalis (Enterococcus faecalis), enterococcus faecium (Enterococcus faecium), enterococcus mundtii (Enterococcus mundtii), or any combination thereof.

In a second aspect, the application provides a method of increasing the concentration of lactobacillus paracasei in the gastrointestinal tract of a subject, the method comprising administering to the subject a therapeutically effective amount of lactobacillus paracasei or a progeny thereof, or a composition comprising the same;

In certain embodiments, the composition is as defined in the first aspect.

In a third aspect, the application provides a method of maintaining and/or improving gastrointestinal health, or preventing and/or treating a disease or condition caused by a gastrointestinal disorder in a subject, comprising administering to the subject an effective amount of lactobacillus paracasei (Lactobacillus paracasei) or a progeny thereof, or a composition comprising the same;

In certain embodiments, the composition is as defined in the first aspect.

The application aims to provide lactobacillus paracasei 207-27 (Lactobacillus paracasei 207-27) from intestinal tracts of healthy infants in Sichuan China, and the microorganism preservation number is as follows: GDMCC No.60960. The classification is named: lactobacillus paracasei. Preservation unit: the collection of microorganism strains in Guangdong province. Preservation time: 2020, 1 month and 15 days. Preservation address: building 5 of Guangzhou City, guangdong, first, china, no. 100, university, 59, and Guangdong province.

Another object of the application is to provide the use of Lactobacillus paracasei 207-27 as a probiotic for relaxing bowel.

The lactobacillus paracasei 207-27 has good tolerance to gastric acid and bile salts, and meets the basic requirements of probiotics.

The lactobacillus paracasei 207-27 disclosed by the application can improve the ink propulsion rate of a small intestine movement experiment of a constipation model mouse, increase the defecation quantity and weight of the constipation model mouse and regulate intestinal flora.

Lactobacillus paracasei 207-27 can be used in a microecological formulation for the above functions, including solids and liquids. Including powders, tablets, film formulations, granules, lozenges, pills, suspensions, capsules, liquids, or other dosage forms suitable for oral administration.

A method of preparing a lactobacillus paracasei 207-27 product formulation, the method comprising: the lactobacillus paracasei 207-27 is directly mixed with the remaining filling. Other fillers include, but are not limited to: maltodextrin, resistant dextrin, lactose, starch, dietary fiber, microcrystalline cellulose, sugar alcohol (such as mannitol), glidant (such as magnesium stearate), sweetener, essence, pigment, etc. For liquid dosage forms, such as liquid suspensions, including oils, as well as necessary emulsifying and suspending aids, and the like.

The lactobacillus paracasei 207-27 can be used as a main component alone or in combination with other probiotics and related functional raw materials.

The Lactobacillus paracasei 207-27 can also be used as a starter for fermenting dairy products and plant fermented products. In particular, the lactobacillus paracasei 207-27 may be used as a starter for dairy products, for example, fermented milk, flavored fermented milk, milk-containing beverages, or plant-based fermented products.

In addition, lactobacillus paracasei 207-27 can be added as a probiotic to various foods.

Definition of terms

In the present application, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Further, the procedures of molecular genetics, nucleic acid chemistry, molecular biology, biochemistry, cell culture, microbiology, cell biology, genomics and recombinant DNA, etc., as used herein, are all conventional procedures widely used in the corresponding field. Meanwhile, in order to better understand the present application, definitions and explanations of related terms are provided below.

As used herein, the term "prebiotic" refers to a substance capable of promoting the growth or amplification of at least one probiotic strain. In certain embodiments, the prebiotic is capable of promoting the growth or expansion of at least one probiotic strain in a subject (e.g., in the human gastrointestinal tract). In certain preferred embodiments, the prebiotic is capable of promoting the growth or amplification of lactobacillus paracasei in the gastrointestinal tract of a human.

As used herein, the term "microorganism beneficial to gastrointestinal health" refers to any non-pathogenic microorganism that, when administered to a host in an effective amount, can have a beneficial effect on the health of the host. In certain embodiments, the microorganism beneficial to gastrointestinal health is selected from bacteria, fungi. In certain embodiments, the microorganism beneficial to gastrointestinal health is a probiotic.

As used herein, the term "progeny" refers to daughter cells produced by a microorganism grown (e.g., in culture in a medium). It will be appreciated that genetic material may change (e.g., mutation, substitution or deletion of 1 or several bases) during the cultivation of a microorganism, while the presence or absence of the same function may be significantly differentiated for different strains (see, e.g., IPAinternational probiotics association). Thus, in certain embodiments, the offspring have the same function (e.g., are capable of enhancing immunity, ameliorating allergic reactions, etc.) as the strain from which they were derived. In certain embodiments, the offspring have the same physiological and biochemical characteristics (e.g., the same colony morphology, the same 16s DNA) and the same function (e.g., promoting small bowel movement, increasing the number and weight of bowel movement in a mammal in a disease model, and also having a positive regulation of intestinal flora) as the strain from which they were derived. In certain embodiments, the lactobacillus paracasei progeny is a gram positive bacterium, the colony morphology is rod-like or bar-like, the edges and corners are clear, and there is no spore. In certain embodiments, the offspring of lactobacillus paracasei have the ability to promote small bowel movement, increase the number and weight of bowel movements in mammals in disease models, and positively regulate intestinal flora.

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier that is pharmacologically and/or physiologically compatible with the subject and active ingredient, is well known in the art (see, e.g., remington's Pharmaceutical sciences. Mediated by Gennaro AR,19th ed.Pennsylvania:Mack Publishing Company,1995), and includes, but is not limited to: pH adjusters, surfactants, adjuvants, ionic strength enhancers. For example, pH modifiers include, but are not limited to, phosphate buffers; surfactants include, but are not limited to, cationic, anionic or nonionic surfactants, such as Tween-80; ionic strength enhancers include, but are not limited to, sodium chloride.

As used herein, the term "food" includes human foods and drinks, as well as animal foods and drinks (i.e., feed). In certain embodiments, the food product is suitable and designed for human feeding.

As used herein, the term "drug" encompasses drugs in human and veterinary medicine for use by both humans and animals, as well as drugs for incorporation into animal feed (e.g., livestock feed and/or pet food). Furthermore, the term "drug" as used herein means any substance that provides a therapeutic, prophylactic and/or beneficial effect.

As used herein, the term "CFU (color-Forming Units)" refers to the total population of microorganisms such as bacteria, fungi, yeast, etc., in a product, typically used as a viable count calculation.

As used herein, the term "CFU/dose" means the amount of bacteria present in a composition/food/pharmaceutical composition provided to a subject daily or each time. For example, in certain embodiments, the Lactobacillus paracasei in the food product is at 10 ⁶ To 10 ¹² The CFU/dose amount is present (e.g. 10 ⁸ To 10 ¹² CFU/dose). In such embodiments, if lactobacillus paracasei is administered in a food product (e.g., in a solid beverage, yogurt), the food product provided to the subject daily or each time (e.g., solid beverage, yogurt) may contain about 10 ⁶ To 10 ¹² Lactobacillus paracasei of CFU. Of course, alternatively, the amount of such bacteria may be divided into multiple administrations, provided that the total amount of lactobacillus paracasei received by the subject at any particular time (e.g., every 24 hours) is from about 10 ⁶ To about 10 ¹² Bacteria of CFU, namely Lactobacillus paracasei satisfying the above food, were added at 10 ⁶ To 10 ¹² The CFU/dose amount is present (e.g. 10 ⁸ To 10 ¹² CFU/dose).

As used herein, the term "pathogen" refers to a microorganism that is capable of invading a host to cause an infection, including viruses, bacteria, fungi, mycoplasma, chlamydia, and the like. In general, they produce pathogenic agents that cause host infection. In certain embodiments, the pathogenic bacteria are microorganisms capable of causing infection in a mammal (e.g., mouse, human). In certain embodiments, the pathogenic bacteria are bacteria capable of causing gastrointestinal tract infections in a mammal (e.g., mouse, human).

As used herein, the term "upregulating gastrointestinal microorganisms" refers to bringing the gastrointestinal microorganisms of a subject towards the health state of the subject.

Advantageous effects of the application

The applicant has previously screened a probiotic Lactobacillus paracasei 207-27 having accession number GDMCC No.60960. In the subsequent research process, the strain is unexpectedly found to have outstanding efficacy in relaxing bowel. For example, the intestinal motility can be promoted, the defecation amount and defecation weight of the mammal in the disease model can be increased, and the function of the intestinal flora can be positively regulated (i.e., the intestinal flora in the disease model can be restored to a healthy state). Meanwhile, in the previous study, the strain has been found to have good tolerance to gastric acid and bile salts. Thus, the strain has a high potential in maintaining the health of healthy subjects and/or restoring the health of gastrointestinal tract in subjects suffering from gastrointestinal disorders (e.g., constipation).

Embodiments of the present application will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only for illustrating the present application and are not intended to limit the scope of the present application. Various objects and advantageous aspects of the present application will become apparent to those skilled in the art from the following detailed description of the preferred embodiments.

Description of preservation of biological Material

Lactobacillus paracasei 207-27 (Lactobacillus paracasei 207-27) has been deposited with the guangdong province microorganism strain collection (GDMCC, guangdong Microbial Culture Collection Center) located in building 5 of institute 100, first middle road, guangzhou city, having deposit number GDMCC No.60960 and a deposit time of 2020, month 1 and 15.

Detailed Description

The application will now be described with reference to the following examples, which are intended to illustrate the application, but not to limit it.

The experiments and methods described in the examples were performed substantially in accordance with conventional methods well known in the art and described in various references unless specifically indicated. For example, for the conventional techniques of immunology, biochemistry, chemistry, molecular biology, microbiology, cell biology, genomics and recombinant DNA used in the present application, reference may be made to Sambrook (Sambrook), friech (Fritsch) and manitis (Maniatis), molecular cloning: laboratory Manual (MOLECULAR CLONING: A LABORATORY MANUAL), edit 2 (1989); the handbook of contemporary molecular biology (CURRENT PROTOCOLS IN MOLECULAR BIOLOGY) (edited by f.m. ausubel (f.m. ausubel) et al, (1987)); series (academic publishing company) of methods in enzymology (METHODS IN ENZYMOLOGY): PCR2: practical methods (PCR 2: A PRACTICAL APPROACH) (M.J. MaxPherson (M.J. MacPherson), B.D. Thoms (B.D. Hames) and G.R. Taylor (G.R. Taylor) editions (1995)), and animal cell CULTURE (ANIMAL CELL CULTURE) (R.I. French Lei Xieni (R.I. Freshney) editions (1987)).

In addition, the specific conditions are not specified in the examples, and the process is carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention. Those skilled in the art will appreciate that the examples describe the application by way of example and are not intended to limit the scope of the application as claimed. All publications and other references mentioned herein are incorporated by reference in their entirety.

Example 1 animal experiments with Lactobacillus paracasei 207-27 for relaxing bowel (Small intestine exercise experiment)

The applicant has screened a probiotic strain Lactobacillus paracasei 207-27 in the early stage and filed a patent. During subsequent studies, it was unexpectedly found that Lactobacillus paracasei 207-27 has a prominent effect in relaxing bowel. Therefore, in this example, another 5 strains (all Lactobacillus paracasei) in the same strain pool were selected as a comparison, and experiments were performed together with Lactobacillus paracasei 207-27 of the present application (all of these strains showed better acid and bile salt resistance and better cell adhesion in the earlier probiotic property study).

All Lactobacillus paracasei referred to in the present application were derived from a Shang Chen-fold healthy own strain pool, which was isolated from normal term neonatal stool samples born at the Hospital of child-birth, university of Sichuan Hua Xifu. Among these, the inclusion criteria for newborns are: residing in five urban areas of Chengdu city; the birth weight is 2500-4000g after 37-42 weeks, and the infant has no congenital abnormality or birth defect. The exclusion criteria were: the mother used antibiotics within one month before birth; parents have infectious diseases such as AIDS, tuberculosis, hepatitis B and the like; the neonate cannot collect the feces due to severe diseases such as neonatal pneumonia.

Collecting fresh feces within 1-4 months after birth of the infant is convenient for the aseptic feces collection tube. Immediately after sampling, the strain was stored temporarily at 4℃and immediately after sampling, the sample was sent to the laboratory at low temperature for dilution culture of fecal samples, if no immediate operation was possible, the strain was stored at 4℃anaerobically, cultured on the same day, then isolated and purified by plate method to give single strain, and specific species of the isolated strain were identified by API 50CH and 16S rDNA sequencing of Mei Liai, numbered (207-33, 207-35, 207-27, 208-24, 208-29, 206-11) and deposited in Shang Chen-fold healthy own strain library, respectively.

1.1 Experimental methods

1.1.1 laboratory animals

Beijing Wanghua Fukang Biotechnology Co., ltd. [ license number: 24 g-28 g SPF grade Kunming healthy clean grade male mice bred by SCXK (Beijing) 2019-0008 are grouped according to 12 groups, and small intestine movement experiments are carried out.

1.1.2 doses

The experiment was performed in 6 intervention groups, constipation model control group and blank control group. Intervention group test substance: 10.00g of fungus powder is weighed, sterile water is added to 25.0mL (the concentration of viable bacteria is 2 multiplied by 10) ⁹ CFU/mL); equivalent amounts of sterile water were administered to the blank and model controls (0 g /). The administration was once daily and each index was measured after 14d of continuous gastric lavage. Mice were perfused with a volume of 0.5 mL/day. Both the intervention group and the control group were given maintenance feed. The preparation method of the bacterial powder (probiotic bacterial powder) comprises the following steps: resuscitates the strain with MRS culture medium, cultures with liquid MRS to the end of logarithmic phase, centrifugally collects thalli, freeze-dries in vacuum, and adjusts the concentration of viable bacteria by adding auxiliary materials such as maltodextrin, thus obtaining the bacterial powder.

1.1.3 small intestine movement experiments

Following continuous administration of test subjects for 14d, each group of mice was fasted without water withdrawal for 16h. Loperamide hydrochloride (4 mg/kg BW) was given intragastrically to the constipation model control and to the six intervention groups, and distilled water was given intragastrically to the blank control. After 0.5h of loperamide hydrochloride, each intervention group was given ink (5% charcoal, 10% gum arabic) with the corresponding test sample, and the blank and model control groups were given ink, which was infused at 0.5 mL/dose. Immediately after 25 minutes, the animals were sacrificed by cervical vertebra removal, the celiac separation mesentery was opened, the upper end from the pylorus, the lower end to the ileocecum were cut off, the intestinal canal was placed on a tray, the small intestine was gently pulled into a straight line, the length of the intestinal canal was measured as the "total length of the small intestine", and the front edge from the pylorus to the ink was the "push length of the ink".

Ink advance = ink advance length (cm)/small intestine total length (cm) x100%.

And converting the obtained ink propelling rate into data. The obtained data are metering data, and on the premise that the constipation model of the small intestine is established, the ink propelling rate of the mice in the tested sample group is obviously higher than that of the mice in the model control group, so that the positive result of the index can be judged.

1.1.3 experimental results

The body weights of the groups before and after the intervention are not different, which indicates that the strains of the intervention groups have no adverse effect on the normal development of mice. The model control group showed a significant decrease in ink push rate (P < 0.001) compared to the blank control group, indicating successful model establishment. Animal experiments prove that the lactobacillus paracasei 207-27 can obviously improve the intestinal ink propulsion rate of each group of mice (table 1), and compared with a model control group, the propulsion rate of the ink of the mice in the group 207-27 is improved, and the improvement has obvious difference, so that the lactobacillus paracasei 207-27 can be judged to have the effect of relaxing bowel. Meanwhile, the intestinal ink thrust rates of mice of other strains in the same strain library can not reach statistically significant difference, which indicates that the strain specificity exists in the capability of lactobacillus paracasei 207-27 for relaxing bowel.

TABLE 1 animal experiment results of the influence of Lactobacillus paracasei 207-27 on the ink Propulsion rate in the intestinal tract

#: compared with the blank control group, has obvious difference

X: compared with the model control group, has obvious difference

EXAMPLE 2 animal experiments with Lactobacillus paracasei 207-27 for relaxing bowel (defecation amount and weight)

2.1 Experimental methods

2.1.1 mice stool particle number and weight experiments

Animals were grouped and fed in the same manner as 1.1.1 and 1.1.2, and after 14d of continuous administration of the test subjects, each group of mice was fasted without water inhibition for 16h. The blank control group was given distilled water by gavage, the constipation model control group and the intervention group were given by gavage with loperamide hydrochloride (5 mg/kgBW). After administration of the lo butylamine hydrochloride for 0.5h, mice in the blank control group and the model control group were filled with ink, and the dose group was administered with ink containing the test sample, and the ink was filled with ink at a dose of 0.5 mL/dose. Animals were kept in single cages and fed with normal drinking water. The number and weight of black bowel movements discharged within 6h from the ink filling were recorded for each animal. The obtained data are metering data, and on the premise that the constipation model of the small intestine is established, the number of the black bowel movement particles discharged by the mice in the tested sample group in 6 hours is obviously higher than that of the mice in the model control group, and the positive result of the index can be judged. The weight of the black stool discharged in 6 hours is obviously higher than that of a model control group, and the positive result of the index can be judged.

2.2 experimental results

The mice constipation model was caused with loperamide hydrochloride. The model control group showed a significant decrease in stool size and weight (P < 0.001)) compared to the placebo group, indicating successful model establishment. Animal experiments prove that the lactobacillus paracasei 207-27 can obviously improve the defecation quantity (table 2) and the defecation weight (table 3) of mice in the intervention group, and compared with a model control group, the defecation granule quantity and the defecation weight of the mice in the 207-27 group are obviously increased, and obvious differences exist, so that the bifidobacteria 207-27 can be judged to have the effect of relaxing bowel. Meanwhile, the number of the other strains in the same strain library for defecation and the weight of the other strains for defecation can not reach statistically significant difference, which indicates that the strain specificity exists in the capacity of lactobacillus paracasei 207-27 for loosening bowel to relieve constipation.

TABLE 2 Effect of Lactobacillus paracasei 207-27 on the number of bowel movements in mice

TABLE 3 influence of Lactobacillus paracasei 207-27 on the weight of the bowel movement of mice

#: compared with the blank control group, has obvious difference

X: compared with the model control group, has obvious difference

EXAMPLE 3 animal experiments with Lactobacillus paracasei 207-27 to modulate intestinal flora

3.1 Experimental methods

40 four week old male Balb/c mice were randomly divided into 4 groups of 10. The blank control group and the OVA model control group were perfused with 0.2ml of physiological saline per day, and the intervention group (207-27+OVA group) was perfused with 0.2ml of 207-27 bacterial suspension per day. The absolute dose of the bacterial suspension is 10 ⁹ CFU/0.2ml, stomach was irrigated continuously for 42 days. Wherein the OVA model control group and the intervention group were subjected to intraperitoneal OVA injection molding on days 7, 21, 28 and 35. Preparation of OVA: 40 μg OVA+0.2ml Al (OH) ₃ An adjuvant.

3.2 sample collection and index determination

After the experiment is finished, a mouse fecal sample is collected, fecal DNA is extracted by using a fecal bacterium total DNA extraction kit, and the fecal flora diversity and composition are analyzed by adopting second-generation sequencing.

3.3 experimental results

3.3.1 analysis of the composition of the fecal flora of mice

As shown in table 4, the OVA model group mice (mouse allergic disease model) had fecal bacteria Chao1 index and ACE index higher than the blank control group (P < 0.05); the fecal bacteria Chao1 index and ACE index of the mice in the intervention group were significantly lower than those in the model group (P < 0.05). The Chao1 index and the ACE index are important indexes for measuring the richness of communities. The intestinal flora of mice after OVA injection is significantly changed, the change is derived from allergy-induced disturbance of the intestinal flora, the experimental result shows that the model is successfully constructed, and two indexes of the intervention group are significantly lower than those of the model group and are closer to the blank control group, so that the lactobacillus paracasei 207-27 has a certain regulation and recovery effect on the change of the intestinal flora, namely, the lactobacillus paracasei 207-27 has the function of positively regulating the intestinal flora (namely, can recover the intestinal flora in a disease model to a healthy state).

Table 4. Analysis of the fecal flora α -diversity of mice of each group (n=6)

Note that: "a" means P <0.05 compared to the OVA model group

Conclusion: after the lactobacillus paracasei 207-27 live bacteria preparation is infused into experimental rats of a disease model, the intestinal movement (P < 0.05) of the mice can be promoted, the defecation quantity (P < 0.05) and the weight (P < 0.05) of the mice can be increased, and the intestinal flora composition of the mice can be positively regulated. It is explained that Lactobacillus paracasei 207-27 has the effect of relaxing bowel. Meanwhile, the lactobacillus paracasei 207-27 can maintain the intestinal health of healthy mice and the richness of intestinal flora.

EXAMPLE 4 Lactobacillus paracasei 207-27 human experiments to smooth the bowel to relieve constipation

4.1 Experimental methods

After the constipation degree was determined by the scale for 46 subjects with constipation, 1 bag of probiotic powder sample was administered daily (the preparation method was as described in example 1, the number of viable bacteria in each bag was 6×10 ¹⁰ Lactobacillus paracasei 207-27 of CFU). Daily follow-up, take condition was recorded and re-tested by the same scale on days 7, 14. The crowd effectiveness of the sample was judged by comparing the symptoms of constipation before probiotic administration with the constipation level at days 7 and 14. And simultaneously recording various adverse reactions in the experimental process.

4.2 experimental results

TABLE 5 statistics of constipation status of subjects

Recording of the number of bowel movements	Before eating (human)	7 days (person)	14 days (person)
				Constipation (less than 3 times/1 times/week)	21	7	4
Normal (once daily)	21	34	38
				Diarrhea (more than 3 times per day)	4	5	4

As shown in table 5, the experimental results show that 67% of constipation subjects have significantly relieved constipation after taking the probiotic powder for 7 days; 81% of constipation subjects have significantly improved constipation 14 days after taking the probiotic powder. Meanwhile, during the experiment period of 14 days, adverse reactions such as diarrhea, flatulence and the like are not observed in the subjects who normally defecation.

Conclusion: lactobacillus paracasei 207-27 can effectively relieve constipation of constipation people, and improve intestinal health of tested people.

Although specific embodiments of the application have been described in detail, those skilled in the art will appreciate that: many modifications and variations of details may be made to adapt to a particular situation and the application is intended to be within the scope of the application. The full scope of the application is given by the appended claims together with any equivalents thereof.

Claims

1. Use of lactobacillus paracasei (Lactobacillus paracasei) in preparing a medicament for preventing and/or treating constipation or a health food with bowel relaxing effect;

2. The use of claim 1, wherein the drug is a drug that targets gastrointestinal release or is a drug that is controlled to release in the gastrointestinal tract.

3. The use of claim 1, wherein the medicament further comprises a pharmaceutically acceptable carrier.

4. The use of claim 1, wherein the medicament is in the form of a pill, powder, capsule, tablet, mouth-soluble granule, liquid or suppository.

5. The use of claim 1, wherein the medicament is in the form of an effervescent tablet or enema.

6. The use according to any one of claims 1 to 5, wherein lactobacillus paracasei is present in the health food at 10 ⁶ To 10 ¹² CFU/dose amount is present.

7. The use according to claim 6, wherein lactobacillus paracasei is used as a food for health care in an amount of 10 ⁸ To 10 ¹² CFU/dose amount is present.