KR102257769B1 - Composition comprising betaglucan for improving intestinal microflora and use thereof - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating intestinal dyskinesia including mushroom-derived beta-glucan.
The mushroom-derived beta-glucan of the present invention increases the beneficial bacteria in the intestine, reduces harmful bacteria in the intestine, increases the water content and weight of the stool, and suppresses the intestinal motility and gastrointestinal damage, so intestinal motility disorders including it It can be effectively used as a composition for preventing, ameliorating or treating diseases.

Description

Composition comprising betaglucan for improving intestinal microflora and use thereof {Composition comprising betaglucan for improving intestinal microflora and use thereof}

The present invention relates to a composition for preventing, improving or treating intestinal dyskinesia including mushroom-derived beta-glucan.

Due to the recent westernization of diet, modern people's intake of dietary fiber has decreased, and intake of high-calorie foods such as meat and instant foods containing high fat and high protein has increased. It is deteriorating and the number of patients with constipation is increasing.

The large intestine breaks down food that has passed through the small intestine into a bacterial gun, absorbs nutrients such as moisture, short-chain fat, and vitamins, and makes the remaining debris into feces and excretes it. In other words, the control of the function of the large intestine refers to a series of actions that maintain and promote the body's bowel movement and bowel movements, and improve digestion and absorption. If the bowel function is deteriorated, the intestinal motility decreases, the mucous membrane of the intestine is damaged or inflamed, and the intestinal flora may become unbalanced.This may result in abdominal pain, difficulty in defecation, constipation, diarrhea, malnutrition, ulcerative colitis, diverticulitis or hemorrhoids, etc. Disease can occur.

Constipation refers to a symptom in which bowel movements do not occur normally, and the stool is hard and dry compared to when it is healthy, and the number of bowel movements and the amount of stool are reduced, resulting in symptoms that accompany discomfort or physiological disorders. Usually, if the number of bowel movements is less than twice a week, or if the amount of bowel movements is less than 30 g per day, it can be diagnosed as constipation. In case of constipation, not only the abdomen is in a swollen state, but also the toxins from the stool that could not be excreted are absorbed into the intestine and absorbed into the blood, thereby promoting skin aging, causing headaches, acne, and skin rashes. Hemorrhoids can cause hemorrhoids, such as prolapse. In addition, constipation is a cause of various serious secondary diseases such as bad breath, colon diseases such as colon cancer, arteriosclerosis, high blood pressure, stroke, and immune deficiency, so active prevention and treatment is required.

The intestinal flora in the human body is a total collection of intestinal microbes and is a biologically important member of the body. It is made up of hundreds of different microorganisms, and individual microorganisms may damage or benefit the host by producing or degrading substances. As a whole, the intestinal flora is involved in the supply of vitamins, prevention of infection, and intestinal function (interlocking movement and absorption), so the composition of the intestinal flora is known to be closely related to the occurrence of constipation and intestinal-related diseases.

Accordingly, a variety of medicines and functional foods for improving intestinal flora and constipation have been recently sold, but the effect is temporary and there is a problem of causing various side effects depending on the type. For example, when an stimulating drug containing anthraquinone derivatives such as senna and rhubarb is used as an herbal laxative, there are side effects such as abdominal pain and diarrhea, and problems with taking it during pregnancy or taking it continuously have been pointed out.

Beta-glucan is a generic term for polysaccharides in which glucose is polymerized around β-1,3 chemical bonds, and beta-glucans derived from microorganisms such as mushrooms and yeasts and dietary fibers of grains such as barley and oats are derived. There are vegetable beta glucans and the like. Beta-glucan enhances the overall immune function by enhancing the function of macrophages, or it has a protective effect on skin cells by helping to block UV rays or recover cells exposed to UV rays, and also has anticancer effects or cholesterol reduction effects. It is known to represent.

Korean Patent Laid-Open No. 10-2017-0061387 is for beta-glucan derived from microorganisms having an effect of improving gastrointestinal tract function, and using beta-glucan derived from Aureobasidium pullulans strains for enteritis or stomach damage It discloses that can be improved.

However, it is not directly disclosed that the use of mushroom-derived beta-glucan has an effect of improving the intestinal flora by increasing beneficial bacteria in the intestine and reducing harmful bacteria in the intestine.

Accordingly, the present inventors have made diligent efforts to provide a composition for improving the intestinal flora that is excellent in effect but not temporary. As a result, beta-glucan derived from mushrooms increases the beneficial bacteria in the intestine, reduces harmful bacteria in the intestine, and at the same time activates the function of the intestine, It has been confirmed that it can be effectively used as a composition for activating intestinal function and inhibiting intestinal damage as well as improving intestinal flora due to its excellent effect of inhibiting damage, and the present invention was completed.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing, improving or treating intestinal dyskinesia including mushroom-derived beta-glucan.

In order to achieve the above object, the present invention provides a composition for preventing, improving or treating intestinal dyskinesia, including mushroom-derived beta-glucan.

According to a preferred embodiment of the present invention, the mushroom may be Schizophyllum commune Fr. , and the beta-glucan may be 1,3-1,6-betaglucan.

According to a preferred embodiment of the present invention, the molecular weight of the beta-glucan may be 1.5 to 2 million daltons (dalton).

According to a preferred embodiment of the present invention, the composition is a fungus of the genus Anaerostipes, a fungus of the genus Coprobacillus, a fungus of the genus Allobaculum, a fungus of the genus Bifidobacterium , Roseburia (Roseburia) and Lactobacillus (Lactobacillus) may be to increase any one or more beneficial bacteria in the intestine selected from the group consisting of bacteria.

According to a preferred embodiment of the present invention, the composition is a fungus of the genus Akkermansia, a fungus of the genus Bilophila, a fungus of the genus Prevotella, a fungus of the genus Disulfovibrio, Lactococcus ) It may be to reduce any one or more intestinal harmful bacteria selected from the group consisting of the genus, Parabacteroides, Sutterella, and Dorea.

According to a preferred embodiment of the present invention, the bowel movement disorder disease may be any one or more selected from the group consisting of functional indigestion, irritable bowel syndrome, intestinal obstruction, constipation and diarrhea.

The mushroom-derived beta-glucan of the present invention increases the beneficial bacteria in the intestine and reduces harmful bacteria in the intestine, increases the water content and weight of the stool, and can suppress the intestinal motility and gastrointestinal damage, and thus prevents intestinal dyskinesia including this , Can be effectively used as a composition for improvement or treatment.

1 shows the results of analyzing the structure of the beta glucan of the present invention by ¹ H-NMR and C-NMR. It was confirmed that β-1,6 bonds and β-1,3 bonds exist in the beta glucan of the present invention.
Figure 2 shows the results of analyzing the structure of the beta-glucan of the present invention by H-NMR and C-NMR. It was confirmed that β-1,6 bonds and β-1,3 bonds exist in a ratio of 1:3 in the beta glucan of the present invention.
3 shows the results of analyzing the structure of the beta glucan of the present invention by FT-IR. It was confirmed that β-1,6 bonds and β-1,3 bonds exist in a ratio of 1:3 in the beta glucan of the present invention.
Figure 4 shows the results of measuring the molecular weight of the beta-glucan of the present invention by MALS. It was confirmed that the average molecular weight of the beta glucan of the present invention was 1.788 × 10 ^{6 Dalton.}
5 shows the results of measuring the molecular weight of beta-glucan of the present invention by GPC. It was confirmed that the average molecular weight of the beta-glucan of the present invention was 1.779 × 10 ^{6 Dalton.}
6 shows the results of analyzing the ecology of intestinal microorganisms. It was confirmed that the ecology of microorganisms significantly changed depending on the type of diet and the degree of beta-glucan intake.
7 shows the results of analyzing various microorganisms in the intestine according to the type of diet, the degree and duration of beta-glucan intake.
FIG. 8 shows the results of measuring changes in Lactobacillus genus known as beneficial bacteria in the intestine according to the type of diet, the degree and duration of beta-glucan intake. When the intake of beta-glucan was low (BG1 = 6mg/kg, BG2 = 15mg/kg, BG3 = 4mg/kg), it was confirmed that Lactobacillus bacteria did not increase, but rather decreased even if ingested continuously.
9 shows the change in body weight of the mouse according to the type of diet, the degree of intake of beta-glucan, and the duration.
Figure 10 shows the change in the amount of drinking water intake of mice according to the type of diet, the degree of beta-glucan intake, and the duration.
11 shows the change in dietary intake of mice according to the type of diet, the degree and duration of beta-glucan intake.
Figure 12 shows the change in the moisture content of the mouse part according to the type of diet, the intake of beta-glucan and the duration.
13 shows the change in weight of mouse feces according to the type of diet, the degree of intake of beta-glucan, and the duration.
14 shows the change in the intestinal migration rate of mice according to the type of diet, the degree of intake of beta-glucan, and the duration.
FIG. 15 shows changes in intestinal protein expression in mice according to the type of diet, the degree and duration of beta-glucan intake.
16 shows the effect of inhibiting gastrointestinal damage by beta glucan. It was confirmed that gastrointestinal ulcers or damage induced by indomethacin were suppressed by beta-glucan administration.

Hereinafter, the present invention will be described in more detail.

As described above, conventional medicines used to improve intestinal flora or constipation have a limitation in that the effect is temporary and side effects such as abdominal pain occur. Accordingly, there is a need for a study on a composition that is excellent in effect on intestinal-related diseases such as constipation, which can occur due to changes in dietary habits of modern people, and that is consistently maintained.

In contrast, the mushroom-derived beta-glucan of the present invention can effectively improve the intestinal flora by increasing the beneficial bacteria in the intestine and reducing harmful bacteria in the intestine in the constipation-causing experimental group (Example 3), and when ingesting the beta-glucan, drinking water or dietary intake As a result, water content and weight of feces may be increased, thereby being effective in diseases of intestinal dyskinesia such as constipation (Examples 4 to 6). In addition, since the beta-glucan has an effect of suppressing gastrointestinal damage by affecting the expression level of intestinal inflammation-related proteins, it may be effective in intestinal inflammatory diseases such as enteritis (Example 8).

The "improvement of intestinal flora" of the present invention means that the intestinal flora is changed so that beneficial bacteria in the intestine are increased, harmful bacteria in the intestine are reduced, and as a result, the environment or function of the intestine can be restored normally.

The'beneficial bacteria in the intestine' of the present invention refer to the beneficial bacteria that live in the intestines and bacteria that have a beneficial effect on the intestinal environment when they are ingested and reached the intestines.

In the present invention, "intestinal harmful bacteria" refers to harmful bacteria living in the intestine and bacteria that have a harmful effect on the intestinal environment when ingested and reached the intestine.

Therefore, the present invention can provide a composition for improving the intestinal flora comprising a mushroom-derived beta glucan (betaglucan).

The mushroom may mean all mushrooms containing beta glucan, preferably, it is preferably a chima mushroom (Schizophyllum commune Fr.).

The beta-glucan may mean all beta-glucans known to be contained in mushrooms, but is preferably 1,3-1,6-betaglucan. The molecular weight of the beta-glucan is preferably 1.5 to 2 million daltons, more preferably 160 to 1.9 million daltons, and most preferably 170 to 1.8 million daltons. desirable.

The composition of the present invention has the effect of increasing the beneficial bacteria in the intestine, and the beneficial bacteria in the intestine may contain all bacteria that can help restore normal intestinal environment or function, but preferably Anaerostipes ), Coprobacillus, Allobaculum, Bifidobacterium, Roseburia, and Lactobacillus It is preferable to increase any one or more beneficial bacteria in the intestine selected from the group, and more preferably, bacteria of the genus Anaerostipes, bacteria of the genus Coprobacillus, bacteria of the genus Allobaculum, and Bifidobak It is preferable to increase any one or more beneficial bacteria in the intestine selected from the group consisting of bacteria of the genus Bifidobacterium and bacteria of the genus Lactobacillus.

The composition of the present invention has the effect of reducing harmful bacteria in the intestine, and the harmful bacteria in the intestine may contain all bacteria that are not helpful in recovering the intestinal environment or function normally, but preferably, Akkermansia genus Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabacteroides, Sutterella It is preferable to reduce any one or more harmful bacteria in the intestine selected from the group consisting of bacteria and Dorea sp., more preferably Akkermansia sp., Prevotella sp., Sutter It is preferable to reduce any one or more harmful bacteria in the intestine selected from the group consisting of bacteria of the genus Sutterella and bacteria of the genus Dorea.

Therefore, the composition for improving intestinal flora of the present invention may be used as a composition for increasing beneficial bacteria in the intestine or a composition for inhibiting harmful bacteria in the intestine.

The present invention may also provide a pharmaceutical composition for preventing or treating intestinal inflammatory diseases or intestinal dyskinesia including mushroom-derived betaglucan.

Since the mushroom-derived beta-glucan is the same as that used in the composition for improving the intestinal flora, the description is replaced with the above description.

The beta glucan of the present invention is anerostifes (Anaerostipes) genus bacteria, Coprobacillus (Coprobacillus) genus bacteria, Allobaculum genus bacteria, Bifidobacterium (Bifidobacterium) genus bacteria, Roseburia (Roseburia ) To increase any one or more beneficial bacteria in the intestine selected from the group consisting of genus bacteria and Lactobacillus genus bacteria,

Akkermansia, Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabacteroides , It is preferable to reduce any one or more harmful bacteria in the intestine selected from the group consisting of bacteria of the genus Sutterella and bacteria of the genus Dorea.

The intestinal inflammatory disease of the present invention may mean any disease caused by the occurrence of inflammation in the large intestine or small intestine, but is preferably selected from the group consisting of intestinal Behcet's disease, Crohn's disease, ulcerative enteritis, acute enteritis, and chronic enteritis. It is preferable that it is any one or more.

The bowel movement disorder disease of the present invention may mean any disease that may occur due to a decrease in the motor function of the large intestine or small intestine, but is preferably selected from the group consisting of functional indigestion, irritable bowel syndrome, intestinal obstruction, constipation and diarrhea. It is preferable that it is any one or more.

The pharmaceutical composition of the present invention may be in various oral or parenteral dosage forms. When formulating the composition, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostatic agents, chelating agents (e.g., EDTA or glutathione), fillers, bulking agents, binders, adjuvants (e.g., Aluminum hydroxide), suspending agents, thickening agents, wetting agents, disintegrants or surfactants, diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in one or more compounds, such as starch (corn starch, wheat starch, rice starch, potato Starch), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl -It is prepared by mixing cellulose or gelatin. For example, tablets or dragees can be obtained by blending the active ingredient with a solid excipient, pulverizing it, adding a suitable auxiliary, and processing it into a granule mixture.

In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions or syrups, and various excipients, such as wetting agents, sweeteners, fragrances or preservatives, are included in addition to water and liquid paraffin, which are commonly used simple diluents. I can. In addition, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, and an anti-coagulant, a lubricant, a wetting agent, a fragrance, an emulsifying agent and a preservative, etc. may be additionally included. .

Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations or suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, and the like may be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, it is used externally to the skin; Intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dura mater or intracerebrovascular injection; Transdermal administration; Alternatively, it may be formulated in the form of a nasal inhalant according to a method known in the art.

In the case of the injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. Examples of suitable carriers for injections include, but are not limited to, water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof, and/or solvents or dispersion media containing vegetable oils. I can. More preferably, suitable carriers include isotonic solutions such as Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanol amine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. Etc. can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be additionally included. In addition, the injection may further contain an isotonic agent such as sugar or sodium chloride in most cases.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. A pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type of disease, severity, drug activity, drug sensitivity, administration time of the patient. , Route of administration and rate of excretion, duration of treatment, factors including concurrent drugs and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. That is, the total effective amount of the composition of the present invention may be administered to a patient in a single dose, and may be administered by a fractionated treatment protocol that is administered for a long period in multiple doses. . It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

The dosage of the pharmaceutical composition of the present invention varies according to the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. As a daily dosage, when parenterally administered, it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per 1 kg of body weight per day based on beta-glucan, and when oral administration of the present invention Based on beta-glucan, per 1 kg of body weight per day, preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per day, may be administered in divided doses from 1 to several times. However, since it may increase or decrease depending on the route of administration, the severity of obesity, sex, weight, age, etc., the dosage amount does not limit the scope of the present invention in any way.

The composition of the present invention may be used alone or in combination with surgery, radiation therapy, hormone therapy, chemotherapy, and methods using biological response modifiers.

The present invention may also provide a health functional food composition for preventing or improving intestinal inflammatory diseases or intestinal dyskinesia including mushroom-derived betaglucan.

Since the mushroom-derived beta-glucan is the same as that used in the composition for improving the intestinal flora, the description is replaced with the above description.

The beta glucan of the present invention is anerostifes (Anaerostipes) genus bacteria, Coprobacillus (Coprobacillus) genus bacteria, Allobaculum genus bacteria, Bifidobacterium (Bifidobacterium) genus bacteria, Roseburia (Roseburia ) To increase any one or more beneficial bacteria in the intestine selected from the group consisting of genus bacteria and Lactobacillus genus bacteria,

Akkermansia, Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabacteroides , It is preferable to reduce any one or more harmful bacteria in the intestine selected from the group consisting of bacteria of the genus Sutterella and bacteria of the genus Dorea.

Since the intestinal inflammatory disease to intestinal motility disorder disease of the present invention is the same as the target disease of the pharmaceutical composition, the description is replaced with the above description.

The health functional food composition according to the present invention can be prepared in various forms according to conventional methods known in the art. General foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (e.g., canned fruit, canned food, jam, marmalade, etc.), fish, meat and processed foods thereof (e.g. ham, sausage) Corn beef), bread and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), fruit juice, various drinks, cookies, sweets, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine , Vegetable protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.), etc., can be prepared by adding the beta glucan of the present invention. In addition, as a nutritional supplement, it is not limited thereto, but may be prepared by adding the beta glucan of the present invention to capsules, tablets, pills, and the like. In addition, the health functional food is not limited thereto, but for example, the beta-glucan of the present invention itself is prepared in the form of tea, juice, and drink, and liquefied, granulated, encapsulated and powdered so that it can be consumed (health beverage). Can be consumed. In addition, in order to use the beta-glucan of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate. In addition, it can be prepared in the form of a composition by mixing with the beta-glucan of the present invention and a known active ingredient known to have an effect of preventing or improving intestinal inflammatory diseases or intestinal dyskinesia.

When using the beta-glucan of the present invention as a health drink, the health drink composition may contain various flavoring agents or natural carbohydrates as an additional ingredient, like a normal drink. The natural carbohydrates described above include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; It may be a sugar alcohol such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumatin and stevia extract; Synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition, the beta-glucan of the present invention may be contained as an active ingredient of a food composition for preventing and improving intestinal inflammatory diseases or intestinal dyskinesia, the amount of which is effective for achieving prevention and improvement of intestinal inflammatory diseases or intestinal dyskinesia. The amount is not particularly limited, but is preferably 0.01 to 100% by weight based on the total weight of the composition. The food composition of the present invention may be prepared by mixing beta-glucan with other active ingredients known to be effective in intestinal inflammatory diseases or intestinal dyskinesia.

In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, It may contain preservatives, glycerin, alcohols or carbonation agents, and the like. In addition, the health functional food composition of the present invention may contain flesh for the manufacture of natural fruit juice, fruit juice beverage, or vegetable beverage. These ingredients may be used independently or in combination. Although the ratio of these additives is not very important, it is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the health functional food composition of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are for illustrative purposes only, and it is obvious to those of ordinary skill in the art that the scope of the present invention is not construed as being limited by these examples.

Beta glucan analysis

<1-1> Beta glucan structure analysis

The structure of the beta glucan of the present invention ^{was analyzed by 1} H-NMR (nuclear magnetic resonance), C-NMR, and FT-IR (fourier transform infrared).

As a result, ^{β-1,3 bonds (A1C1, B1C1, B2C1) and β-1,6 bonds by comparing Proton (1} H) NMR (top of x-axis) and Carbon (C) NMR (left of y-axis) The presence of (C1C1) was confirmed (Fig. 1). In addition, by comparing the size of β-1,6 carbon bonds and β-1,3 carbon bond peaks and HSQC NMR spectrum in ^{1 H-NMR and C-NMR, β-1,6 bonds and β-1} It was confirmed that the ratio of ,3 binding was 1:3 (FIG. 2). Using FT-IR, the valley value of the identifiable wavelength of beta-glucan was measured, and β-1,3-glucan having β-1,6 structures were identified, and β-1,6 bonds and β- It was confirmed that the ratio of 1,3 binding was 1:3 (FIG. 3).

<1-2> Beta-glucan average molecular weight

When the average molecular weight of beta-glucan was measured using multi-angle light scattering (MALS) and gel permeation chromatography (GPC), it was confirmed that it was a molecular weight of 1.77 to 1.79 million daltons (FIGS. 4 and 5).

Experimental animals and experimental group settings

5-week-old ICR mice (Daehan Biolink) were subjected to a laboratory acclimation period for 1 week, followed by an experiment for 12 weeks, and the number of individuals for each condition was 10, grouped by a random method so that the average weight and standard deviation were uniform for each condition. Were classified.

In order to confirm the effect of improving intestinal function or inhibiting intestinal damage by beta-glucan, a constipation inducing experimental group was prepared. Specifically, the general treatment group (Normal diet, ND) fed with general feed, the low fiber diet (LFD) fed with a low dietary fiber feed, and the high fat diet fed with high fat feed , HFD). To each experimental group, the beta-glucan identified in [Example 1] was administered daily for 6 to 12 weeks at a dose of 6 mg/kg (BG1, low) or 15 mg/kg (BG2, high) once a day. . Beta glucan was not administered as a control (CTL). Each experimental group was allowed to freely consume diet and drinking water, and the experiment was conducted under conditions of 12 hours (light/dark cycle), temperature 23±2℃, and 50±5 relative humidity.

Intestinal flora analysis

<3-1> Intestinal microbial ecology analysis

Non-metric multidimensional scaling (NMDS) analysis results for each diet. As a result of NMDS analysis (Non-metric multidimensional scaling analysis) of the experimental groups prepared in [Example 2], the beta-glucan feeder, as shown in [Fig. 6], beta It was confirmed that most of the intestinal microbial ecology was significantly changed upon ingestion of glucan.

In particular, compared to the low fiber treatment group, the microbial ecology changes due to the intake of beta glucan (BG) were significantly observed in the high fat treatment group (High Fat) or the general treatment group (Normal). The intestinal microbial ecology changed more significantly in the kg BG intake group (BG2, High conc. Beta-glucan) than in the 6 mg/kg BG intake group (BG1, Low conc. Beta-glucan).

<3-2> Changes in intestinal microflora

LefSe analysis was performed through Linear Discriminant Analysis (LDA) analysis of LEfSe software in order to confirm changes in the intestinal microflora of the experimental groups prepared in [Example 2]. LEfSe is for univariate analysis, and it is an analysis tool that identifies microbial biomarkers through Kruskal-Willis analysis and then verifies initial analysis results through Wilcoxon rank-sum post-mortem analysis. In addition, in addition to the experimental group prepared in [Example 2], in order to specifically confirm the change of the bacteria of the genus Larctobacillus , which is known as a beneficial bacteria in the intestine, an experimental group in which 4 mg/kg of beta glucan was administered to male ICR mice ( BG3) was prepared. Each dose of beta glucan was orally administered to each experimental group (BG1, BG2, and BG3) once a day for 7 or 11 days, and then DNA was extracted from mouse fecal samples and qPCR was performed. C1 and C2 were not administered with beta glucan as controls.

As a result, as shown in [Fig. 7], the intestinal flora varied according to the type of diet, the dose of beta-glucan, and the intake period, and was particularly evident in the high fat diet. The bacteria marked in blue are the reduced intestinal harmful bacteria, and the bacteria marked in red are the increased beneficial bacteria in the intestines.

Specifically, bacteria of the genus Anaerostipes, which are beneficial bacteria in the intestines, bacteria of the genus Coprobacillus, bacteria of the genus Allobaculum, bacteria of the genus Bifidobacterium, Losebu Including the genus Roseburia and the genus Lactobacillus, Clostridiales, Collinsella, Helicobacteraceae, Mucispirillum, Copro Coprococcus, Turicibacter genus and Luminococcaceae genus were increased; Pathogenic bacteria of the genus Akkermansia, Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabak Including the genus Parabacteroides, the genus Sutterella, and the genus Dorea, the genus Oscillospira, the genus Enterococcus, and the genus Ruminococcus The bacteria decreased.

In addition, as shown in [Fig. 8], in the control group (C1, C2) without beta glucan intake, there was almost no change in the bacteria of the genus Lactobacillus. On the other hand, it was confirmed that the Lactobacillus bacteria increased about 2.8 times and 3.4 times, respectively, in the group that consumed 6mg/kg or 15mg/kg of beta glucan for 11 days. When beta-glucan was ingested at 4mg/kg, there was no increase in the Lactobacillus genus, but rather Lactobacillus bacteria decreased.

Physiological analysis

<4-1> weight change

The change in body weight of the mice in the experimental group prepared in [Example 2] was measured once a week for a total of 12 weeks.

As a result, as shown in [Fig. 9], in the normal diet group (Normal Diet), the body weight increased according to the beta glucan intake, and in the high fat treatment group (High fat), the body weight decreased according to the beta glucan intake. there was.

<4-2> Drinking water intake

The amount of drinking water intake of the mice in the experimental group prepared in [Example 2] was measured once a week for a total of 12 weeks.

As a result, as shown in [Fig. 10], it was confirmed that the drinking water intake increased as the intake of beta-glucan increased.

<4-3> Comparison of dietary intake

The daily dietary intake of the mice in the experimental group prepared in [Example 2] was measured once a week for a total of 12 weeks.

As a result, as shown in [Fig. 11], dietary intake decreased by intake of beta-glucan in the normal diet group, and low fiber feed or high fat diet in the two constipation inducing experimental groups (low fiber and high fat). It was confirmed that the dietary intake decreased due to the increase due to beta-glucan intake.

Intestinal fecal index analysis

<5-1> Fecal moisture content comparison

The feces of the experimental mice prepared in [Example 2] were obtained, their weight was measured, dried in a dryer at 60° C. for 24 hours, and then the weight was measured, and feces using the difference in weight before and after drying The moisture content contained in was measured.

As a result, it was confirmed that the water content was higher in the beta-glucan intake group (BG1, BG2) than in the beta-glucan non-intake group in the constipation inducing experimental group (low fiber and high fat) as shown in [Fig. 12]. This could be interpreted as an increase in stool water content by intake of beta-glucan in the constipation inducing experimental group.

<5-2> Fecal weight comparison

The feces of the mice in the experimental group prepared in [Example 2] were obtained, and the weight thereof was measured once a week for a total of 12 weeks.

As a result, as shown in [Fig. 13], changes between groups began to be clearly seen from 6 weeks after ingestion of beta-glucan (BG). In the normal diet group, the weight of feces increased according to BG intake, and in the constipation inducing experimental group (low fiber and high fat), the weight of feces decreased according to BG intake.

In general, it is known that when the water content in the feces decreases, it leads to a decrease in the weight of the feces and causes constipation. However, in the case of the present invention, when analyzed together with the results of Example <5-1>, in the general treatment group, the water content of the stool decreased according to the BG intake, but the weight of the stool increased, and in the low dietary fiber treatment group, according to the BG intake. The moisture content of the stool increased, but the weight of the stool decreased. In the high fat treatment group, both the moisture content and weight of feces increased. That is, in the case of constipation caused by high fat intake, the water content and weight of the stool increase with BG intake, so constipation can be improved.

Intestinal motor performance analysis

The purpose of this study was to determine the effect of beta-glucan on intestinal motility using intestinal mobility.

Specifically, the experimental groups prepared in [Example 2] were orally administered 0.2 ml of 3% activated carbon diet (charcoal meal, C3345, Sigma-Aldrich) dissolved in 0.5% CMC-Na solution. It was killed after 30 minutes and the gastrointestinal tract was excised, and the rate of movement of activated carbon from the pylorus to the entrance of the appendix was measured. The minister movement rate was calculated by the formula of [Equation 1] below.

[Equation 1]

Intestinal movement rate (%) = Intestinal movement distance of activated carbon diet / Distance from pylorus to cecum × 100

As a result, as shown in [Fig. 14], in the general treatment group (Normal Diet) and the low fiber treatment group (Low fiber) intake of beta glucan (BG) decreased intestinal movement compared to the non-intake group, but It was significantly increased with increasing dose. In the high fat treatment group (HFD), the intestinal migration rate was increased in the BG intake group compared to the control group. In general, it is known that when the water content of the stool decreases, the mobility of the intestine also decreases. As in the result of [Example 5], it was determined that the water content of the stool increased when ingesting BG in the HFD group, so that the intestinal mobility was also fast.

Intestinal tissue changes

The intestinal tissues of each of the experimental groups of [Example 2] were separated and protein changes were confirmed using Western Blot analysis.

As a result, as shown in [Fig. 15], beta-glucan changes the intestinal cell proliferation rate regardless of diet, but the intestinal migration efficiency of beta-glucan seems to vary depending on the diet. Overall, the expression of COX2 was reduced, and it was judged that the effect of improving inflammation was high.

Effects on acute enteritis

Specifically, PBS, indomethacin (Indomethacin), beta glucan (BG) was administered once daily for 5 days to 6-week-old C57BL mice, and then 100 mg/kg of indomethacin dissolved in ethanol was added to experimental mice that were fasted for 24 hours. It is administered orally using respect. After 4 hours, after fasting and water saving, 24 hours later, the stomach was opened to separate the stomach, incised and unfolded, rinsed in PBS, and unfolded to take a picture. For gastric tissue staining, H/E staining was performed by fixing in 4% formaldehyde and observed under a microscope.

As a result, it was confirmed that, as shown in [Fig. 16], the group administered orally with beta-glucan and indomethacin decreased the depth or range of ulcers than in the group treated with indomethacin alone, resulting in decreased gastric wall damage. This is a result showing that beta-glucan is effective in inhibiting intestinal damage.

Claims (8)

As a pharmaceutical composition for the prevention or treatment of intestinal dyskinesia comprising 1,3-1,6-betaglucan derived from Schizophyllum commune Fr.,
The molecular weight of the beta glucan is 1.5 to 2 million daltons,
The beta-glucan is a fungus of the genus Anaerostipes, a fungus of the genus Coprobacillus, a fungus of the genus Allobaculum, a fungus of the genus Bifidobacterium, a fungus of the genus Roseburia And to increase any one or more beneficial bacteria in the intestine selected from the group consisting of bacteria of the genus Lactobacillus,
Akkermansia, Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabacteroides , Sutterella (Sutterella) and Dorea (Dorea) a pharmaceutical composition characterized in that reducing any one or more harmful intestinal bacteria selected from the group consisting of bacteria.
delete delete delete The pharmaceutical composition according to claim 1, wherein the intestinal motility disorder is at least one selected from the group consisting of functional indigestion, irritable bowel syndrome, intestinal obstruction, constipation and diarrhea.
As a health functional food composition for preventing or improving intestinal dyskinesia disease comprising 1,3-1,6-betaglucan derived from Schizophyllum commune Fr.,
The molecular weight of the beta glucan is 1.5 to 2 million daltons,
The beta-glucan is a fungus of the genus Anaerostipes, a fungus of the genus Coprobacillus, a fungus of the genus Allobaculum, a fungus of the genus Bifidobacterium, a fungus of the genus Roseburia And to increase any one or more beneficial bacteria in the intestine selected from the group consisting of bacteria of the genus Lactobacillus,
Akkermansia, Bilophila, Prevotella, Disulfovibrio, Lactococcus, Parabacteroides , Sutterella (Sutterella) and Dorea (Dorea) a health functional food composition, characterized in that reducing any one or more harmful intestinal bacteria selected from the group consisting of bacteria.
delete The health functional food composition according to claim 6, wherein the intestinal motility disorder is at least one selected from the group consisting of functional indigestion, irritable bowel syndrome, intestinal obstruction, constipation and diarrhea.

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