JP7171264B2 - Intestinal barrier improving composition - Google Patents

Description

The present invention relates to an intestinal barrier improving composition and the like.

Vertebrates including humans sustain life by ingesting and absorbing water and food. The absorption takes place through the gastrointestinal tract, particularly the intestine, through which foreign foreign substances, including food taken by mouth, pass. At the same time, various bacteria invade the intestinal tract, settle in the intestinal tract in a nutrient-rich environment, proliferate, and form a bacterial flora called intestinal flora. In order for the host to maintain a good relationship with intestinal bacteria, an immune system specific to the intestinal tract has been developed. For example, intestinal epithelial cells separate intestinal bacteria from intestinal epithelial tissue by constructing a mucosal barrier, and control the intestinal immune system by transmitting signals to immunocompetent cells.

Absorptive epithelial cells present on the inner surface of the intestinal tract are specialized cells for absorbing nutrients and water, and occupy the majority of the intestinal epithelial cells. The surface of the apical surface of absorptive epithelial cells is covered with an aggregate of sugar chains called glycocalyx, which prevents intestinal bacteria from entering the intestinal epithelial cells. Similarly, goblet cells present on the inner surface of the intestinal tract maintain the homeostasis of the mucus that covers the intestinal mucosa by producing and secreting large amounts of glycoproteins (mucins), which are the main components of mucus, and are responsible for intestinal epithelial cells. Prevents invasion of intestinal bacteria.

Thus, maintaining the intestinal barrier function (particularly the intestinal mucosal barrier) is important for maintaining the intestinal environment and maintaining health. Therefore, there is a need for methods to improve the intestinal barrier.

JP 2014-14372 A WO 12/099216 JP 2014-187948 A JP 2014-187949 A JP 2017-066097 A

An object of the present invention is to provide a method for improving the intestinal barrier.

The present inventors have discovered the possibility that proteoglycan and lactic acid fermented figs can improve the intestinal barrier, and have made further improvements to complete the present invention.

The invention includes, for example, the subject matter described in the following sections.
Section 1.
A composition for improving the intestinal barrier, containing proteoglycan and/or lactic acid fermented fig.
Section 2.
Item 2. The composition for improving the intestinal barrier according to Item 1, which is for intestinal regulation.
Item 3.
Item 3. The composition for improving the intestinal barrier according to Item 1 or 2, for increasing the amount of intestinal mucin.
Section 4.
Item 3. The composition for improving the intestinal barrier according to Item 1 or 2, for improving intestinal flora.
Item 5.
Item 5. The intestinal barrier-improving composition according to any one of Items 1 to 4, wherein the fermented product of figs with lactic acid bacteria is a fermented product of Lactobacillus pentosus of figs.
Item 6.
Item 6. The intestinal barrier improving composition according to any one of Items 1 to 5, which is an oral composition.
Item 7.
Item 7. The composition for improving the intestinal barrier according to any one of Items 1 to 6, containing proteoglycan and a fermented product of figs with lactic acid bacteria.

The composition for improving the intestinal barrier function according to the present invention not only has the effect of improving the intestinal barrier function when it is unsatisfactory and returning it to a normal state, but also improves the intestinal barrier function from the normal state to a more excellent state. It also has an effect. That is, the composition for improving the intestinal barrier according to the present invention can be used not only as a composition for repairing the intestinal barrier but also as a composition for enhancing the intestinal barrier. Furthermore, the intestinal barrier-improving composition according to the present invention can also have the effect of preventing deterioration of the intestinal barrier function in advance. For example, when the intestinal barrier function is lowered by an intestinal inflammation-inducing substance or the like, the composition for improving the intestinal barrier of the present invention is ingested before being exposed to the inducing substance, thereby preventing the intestinal barrier function from being exposed to the inducing substance. The effect of suppressing or preventing the deterioration of the barrier function that occurs in some cases can be obtained. That is, the composition for improving intestinal barrier according to the present invention can also be used as a composition for preventing intestinal barrier deterioration. Further, for example, the intestinal barrier-improving composition can be preferably used as a composition for intestinal regulation.

An overview of the small intestinal epithelial model produced is shown. Fig. 1 shows the results of evaluating the intestinal barrier by measuring the membrane potential difference by transepithelial electrical resistance (TER) measurement using the small intestinal epithelial model shown in Fig. 1 . The horizontal axis indicates time (h), and the vertical axis indicates the membrane potential difference change rate (%). 1 shows the results of a fecal mucin amount test for humans who ingested proteoglycan or a fermented product of lactic acid bacteria of figs. 1 shows the aggregated results of questionnaires to humans who ingested proteoglycan or fermented figs with lactic acid bacteria. 1 shows the results of analysis of bacterial species in the intestinal flora of humans who ingested proteoglycan or fermented figs with lactic acid bacteria. Fig. 2 shows the results of examining the inflammation-preventing effects of proteoglycan and fermented fig lactic acid bacteria on colitis-induced model hairless mice. Fig. 2 shows the analysis results of phenol content in feces of humans who ingested proteoglycan.

Each embodiment of the present invention will be described in further detail below.

The intestinal barrier-improving composition included in the present invention contains proteoglycan and/or Lactobacillus pentosus fermented fig. The intestine whose barrier function is improved by the composition is preferably the small intestine and/or the large intestine, more preferably the small intestine.

Proteoglycans are complex polysaccharides in which glycosaminoglycans such as chondroitin sulfate and dermatan sulfate are covalently bound to a protein core, and are abundant in animal tissues, particularly cartilage tissues. It is also known that proteoglycan exists in vivo in a structure in which the core protein is further bound to hyaluronic acid, and its molecular weight is as large as tens of thousands to tens of millions. Among them, those having a molecular weight of about 200,000 to 20,000,000 are preferable, those having a molecular weight of about 1,000,000 to 18,000,000 are more preferable, those having a molecular weight of about 1,500,000 to 16,000,000 are more preferable, and those having a molecular weight of about 2,500,000 to 15,000,000 are more preferable. Preferred, and even more preferred is between 5 million and 15 million. Not only the separated and purified proteoglycan but also the crudely purified proteoglycan can be used. Quantitation of proteoglycan is carried out by molecular weight fractionation using gel chromatography (molecular weight marker: dextran with known molecular weight), and the fraction (or dried product thereof) with a molecular weight of 10,000 or more and the presence of acidic sugars and proteins is confirmed. This can be done by measuring The presence and amount of acidic sugar can be confirmed by measuring the amount of uron by the carbazole-sulfuric acid method. Also, the presence and amount of protein can be measured by absorbance at around 280 nm. Such preferred proteoglycans can be prepared, for example, by the methods described in WO 12/099216 and JP-A-2017-066097, and the molecular weight can be measured.

The amount of proteoglycan ingested by an adult per day from the intestinal barrier-improving composition according to the present invention is not particularly limited as long as the effects of the present invention can be obtained. For example, in terms of dry mass, it is preferably 10 to 10000 mg/day, more preferably 50 to 5000 mg/day, even more preferably 100 to 1000 mg/day, and even more preferably 100 to 500 mg/day.

As the lactic acid bacteria fermented fig, one obtained by fermenting crushed figs with lactic acid bacteria is preferable.

Ficus (Fig) is a deciduous tall tree of the genus Ficus of the Moraceae family native to the Middle and Near East. there is In the present invention, it is preferable to use cryptids, in which flowers grow inside the fruits, which are produced by Fig-type strains of figs. Although the variety is not limited, 'Horai persimmon' and 'Masui Dauphine' can be preferably used.

Crushed figs are obtained by thinning figs (especially cryptocarps), removing the flower stalks if necessary, and crushing them (whole figs) as they were harvested (crushed whole figs). is preferred. The crushing method is not particularly limited, and a conventional method can be used. The degree of crushing is not particularly limited as long as the crushed product has fluidity to the extent that it does not interfere with the fermentation treatment, and can be set as appropriate. The temperature during the crushing treatment may be room temperature, but it is preferable to perform the treatment at a temperature of 15°C or less, more preferably 10°C or less. Further, crushed figs obtained by crushing frozen fruits as they are is also one of the preferred embodiments.

Lactobacillus pentosus, Lactobacillus plantarum , Lactobacillus acidophilus and the like are preferably used as lactic acid bacteria, and Lactobacillus pentosus is particularly preferable.

Fermentation treatment includes, for example, a method of inoculating 10 ⁶ or more, preferably 10 ⁷ to 10 ⁹ figs per 1 g of crushed whole figs. A preferred embodiment includes a method of inoculating a starter containing 10 ⁹ to 10 ¹¹ CFU/ml of lactic acid bacteria by adding 0.5 to 10% by mass to the crushed whole fig fruit. During the fermentation process, the pH of the fermented liquid gradually decreases. In the present invention, the fermentation is preferably carried out until the pH becomes 5.0 or less, more preferably 4.6 or less, and even more preferably 4.3 or less. Fermentation can be either aerobic fermentation or anaerobic fermentation, but anaerobic fermentation is preferred. In the case of aerobic fermentation, standing fermentation is preferable to stirring fermentation. Lactic acid fermentation can be carried out at a temperature of 15°C to 45°C, preferably 20°C to 40°C, more preferably 25°C to 37°C. When the temperature of the whole fig crushed product to be fermented is high due to heat treatment for the purpose of sterilization, etc., the residual heat can be used to start fermentation by adding a lactic acid bacteria starter within the above temperature. The time for lactic acid fermentation is, for example, 5 hours to 72 hours, preferably 10 hours to 36 hours. In addition, if fermentation is performed for a long time, residual microorganisms other than lactic acid bacteria may proliferate and deteriorate the quality. It is preferable to adjust the species and number of lactic acid bacteria.

The amount of fermented fig fermented with lactic acid bacteria per day for an adult to be taken by the composition for improving the intestinal barrier according to the present invention is not particularly limited as long as the effects of the present invention can be obtained. For example, in terms of dry mass, it is preferably 10 to 10000 mg/day, more preferably 50 to 5000 mg/day, even more preferably 100 to 1000 mg/day, and even more preferably 200 to 800 mg/day.

In addition, in the composition for improving the intestinal barrier according to the present invention, the content ratio of proteoglycan to the lactic acid fermented fig (proteoglycan: lactic acid fermented fig) is preferably about 1:0.1 to 10, for example. About 1:0.5 to 5 is more preferable, about 1:1 to 4 is more preferable, and about 1:2 to 3 is even more preferable.

The contents of proteoglycan and the lactic acid fermented product of figs in the intestinal barrier-improving composition according to the present invention can be appropriately set within a range that does not impair the effects of the present invention. For proteoglycan, for example, about 1 to 90% by mass, 10 to 80% by mass, or about 20 to 50% by mass. Further, for example, about 0.1 to 99% by mass, about 10 to 80% by mass, or about 20 to 50% by mass, etc. can be mentioned for figs fermented with lactic acid bacteria.

The composition for improving the intestinal barrier function according to the present invention not only has the effect of improving the intestinal barrier function when it is unsatisfactory and returning it to a normal state, but also improves the intestinal barrier function from the normal state to a more excellent state. It also has an effect. That is, the composition for improving the intestinal barrier according to the present invention can be used not only as a composition for repairing the intestinal barrier but also as a composition for enhancing the intestinal barrier. Furthermore, the intestinal barrier-improving composition according to the present invention can also have the effect of preventing deterioration of the intestinal barrier function in advance. For example, when the intestinal barrier function is lowered by an intestinal inflammation-inducing substance or the like, the composition for improving the intestinal barrier of the present invention is ingested before being exposed to the inducing substance, thereby preventing the intestinal barrier function from being exposed to the inducing substance. The effect of suppressing or preventing the deterioration of the barrier function that occurs in some cases can be obtained. That is, the composition for improving intestinal barrier according to the present invention can also be used as a composition for preventing intestinal barrier deterioration. Further, for example, the intestinal barrier-improving composition can be preferably used as a composition for intestinal regulation.

In addition, the composition for improving the intestinal barrier can exhibit the effect of improving the intestinal barrier by containing the proteoglycan or the lactic acid fermented product of the fig. Here, the proteoglycan has a relatively long period of time from when it is orally ingested to when the effect of improving the intestinal barrier (particularly, the effect of regulating the intestinal function) is exhibited. In other words, the effect develops slowly. Therefore, the intestinal barrier-improving composition containing the proteoglycan can also be called a sustained-release intestinal barrier-improving composition. On the other hand, the fermented product of figs with lactic acid bacteria takes a relatively short time to exhibit an intestinal barrier improving effect (especially an intestinal regulation effect) after oral intake. That is, the effect is immediately manifested. Therefore, the intestinal barrier-improving composition containing the fermented product of lactic acid bacteria of fig can be said to be a rapid-acting intestinal barrier-improving composition. For this reason, a composition for improving the intestinal barrier containing the proteoglycan and the fermented product of lactic acid bacteria of fig can exhibit both rapid-acting and sustained-releasing effects, and is particularly preferable.

When ingested, the intestinal barrier-improving composition can also exert an effect of improving the intestinal flora. More preferably, the balance of bacterial species in the intestinal flora can be improved. Subjects with imbalanced bacterial species in their intestinal flora, as well as subjects that are not so imbalanced, can be encouraged to have a more favorable bacterial species balance.

In particular, ingestion can have the effect of increasing bacteria that contribute to improving the intestinal barrier. More specifically, the effect of increasing Bifidobacterium and/or the effect of increasing Bacteroides can be preferably exhibited. Bifidobacterium is a representative of so-called good bacteria and is excellent in intestinal regulation. Bacteroides are believed to have immunomodulatory effects on the intestinal immune system.

In addition, the regulating effect of the intestinal barrier-improving composition can be maintained even if the intestinal barrier-improving composition is not ingested for a while after continuous ingestion. In particular, it can have an effect of suppressing or reducing the increase of bacterial groups (so-called bad bacteria) that may contribute to deterioration of the intestinal environment. For example, it can exhibit an effect of suppressing or decreasing bacterial groups such as Clostridium subcluster XIVa and Clostridium cluster IX. Clostridium subcluster XIVa is a bacterium that produces deoxycholic acid, and deoxycholic acid is believed to be associated with cellular senescence.

Furthermore, the intestinal barrier-improving composition can have the effect of increasing the amount of intestinal mucin by ingestion. This may enhance the intestinal mucosal barrier.

The intestinal barrier-improving composition according to the present invention is not particularly limited, but is preferably an oral composition, particularly preferably a food or drink composition. Most preferably, it is a food or drink composition for the purpose of regulating intestinal flora and intestinal environment. Also, the form of the composition is not particularly limited, and may be solid, liquid, gel, or the like. Also, for example, tablets, capsules, drinks, jelly, granules, and the like. Each such composition can be prepared by a conventional method.

The composition for improving the intestinal barrier can also contain components other than the proteoglycan and the fermented product of figs with lactic acid bacteria within a range that does not impair the effects of the present invention. Examples of such other ingredients include pH adjusters, thickeners, surfactants, flavoring agents, sweetening agents, wetting agents and the like, but are not particularly limited. Such a component can be used individually by 1 type or in combination of 2 or more types.

When the composition of the present invention is liquid or gel and can be measured using a pH composite electrode, the pH of the composition is, for example, preferably 3.0 to 8.0, more preferably 3 .0-7. It is more preferably 3.0 to 5.5. The measurement with the pH composite electrode is performed with the composition as it is without dilution or the like. The measurement temperature is 20° C. and the measurement time is 2 minutes. When adjusting the pH of the composition, commonly used pH adjusters such as citric acid, phosphoric acid, malic acid, gluconic acid, maleic acid, aspartic acid, succinic acid, glucuronic acid, fumaric acid, glutamic acid , adipic acid, salts thereof, sodium bicarbonate, hydrochloric acid, sodium hydroxide, potassium hydroxide, and the like can be used.

Examples of thickening agents include guar gum, agar, carrageenan, xanthan gum, and gum arabic. A thickener can be used individually by 1 type or in combination of 2 or more types.

As surfactants, nonionic surfactants, anionic surfactants or amphoteric surfactants can be blended. Specifically, nonionic surfactants include sugar fatty acid esters such as sucrose fatty acid esters, maltose fatty acid esters and lactose fatty acid esters, fatty acid alkanolamides, sorbitan fatty acid esters, fatty acid monoglycerides, polyoxyethylene addition coefficients of 4 to 15. , a polyoxyethylene alkyl ether system having an alkyl group having 10 to 18 carbon atoms or a polyoxyethylene alkylphenyl ether having a polyoxyethylene addition coefficient of 10 to 18 and an alkyl group having 9 carbon atoms, diethyl sebacate, poly Oxyethylene hydrogenated castor oil, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene glycerin fatty acid ester, polyoxyethylene sorbitol fatty acid ester, polyethylene glycol fatty acid ester, polyethylene lanolin, polyethylene sterol, polyethylene lanolin alcohol, alkyl glucoside, polyoxyethylene polyoxy and propylene block copolymers. Examples of anionic surfactants include sulfuric acid ester salts such as sodium lauryl sulfate and sodium polyoxyethylene lauryl ether sulfate, sulfosuccinates such as sodium lauryl sulfosuccinate and sodium polyoxyethylene lauryl ether sulfosuccinate, sodium cocoyl sarcosinate, and lauroyl methylalanine. Acyl amino acid salts such as sodium, sodium cocoyl methyl taurate, and the like. Examples of amphoteric surfactants include betaine acetate surfactants such as betaine lauryldimethylaminoacetate and betaine coconut oil fatty acid amidopropyldimethylaminoacetate, and imidazoline surfactants such as sodium N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine. Active agents, amino acid-type active agents such as N-lauryldiaminoethylglycine, and the like. These surfactants can be blended singly or in combination of two or more. However, when used as a food composition, blending a large amount of surfactant leads to increased irritation, which is not preferable and should be noted.

Flavoring agents include menthol, carboxylic acid, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronol, α-terpineol, methyl acetate, citronenyl acetate, methyl eugenol, cineol, linalool, ethyl Linalool, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, perilla oil, wintergreen oil, clove oil, eucalyptus oil, pimento oil, etc. More than one species can be used in combination. The blending amount of the flavoring agent is, for example, 0.01 to 5% by mass, preferably 0.03 to 1% by mass, based on the total amount of the composition.

Sweeteners include, for example, saccharin, saccharin sodium, acesulfame potassium, stevia extract, stevioside, neohesperidyl dihydrochalcone, glycyrrhizin, perillartine, soumatin, aspartylphenylalanine methyl ester, methoxycinnamic aldehyde, palatinose, palatinite, xylitol, lactitol. etc. These sweeteners can be used alone or in combination of two or more.

Wetting agents include, for example, glycerin, ethylene glycol, propylene glycol, 1,3-butylene glycol, sorbitol, low molecular weight polyethylene glycol, monosaccharides, disaccharides, oligosaccharides, sugars such as reduced starch syrup, and the like, either singly or in combination. Any combination of the above can be used.

In this specification, the term "comprising" includes "consisting essentially of" and "consisting of."

The present invention will be described in more detail below, but the present invention is not limited to the following examples.

Intestinal barrier evaluation test using an artificial small intestinal epithelial model A small intestinal epithelial model was prepared using Caco cells, the model was stimulated with inflammatory substances (TNF-α and IFN-γ), and the degree of barrier disruption due to stimulation was evaluated. It was examined to what extent the proteoglycan and the fermented product of lactobacilli of fig can be improved. Fig. 1 shows an outline of the small intestinal epithelial model that was prepared. Details of the materials used are shown below.
<Materials used>
Cells: CACO-2 (DSP ECACC cells) P=45
Culture medium:
・D-MEM high glucose (Sigma-Aldrich)
・FBS (Biowest)
・Antibiotics Anti-Anti (gibco)
* Reagents for passage and plating prepared so that FBS is 10% and antibiotics are 1%:
- 0.25% Trypsin-EDTA (gibco) diluted to 0.1% * Neutralization was performed in medium. Neutralization was carried out within 5 minutes of the addition of trypsin.
Plating:
・Transwell polycarbonate membrane permeable support
24 well insert set (COSTAR)
Membrane diameter: 6.5 mm, culture area: 0.33 cm ² , pore size: 0.4 μm
Reagent used (additive material):
・TNF-α, INF-γ (gibco): 10 ng/mL each
・Proteoglycan (preparation method described below)
・Lactic acid fermented fig (preparation method is described below)
・D-MEM medium (10% FBS)

[Preparation of proteoglycan]
Proteoglycan was prepared according to the method for extracting proteoglycan from frozen salmon nasal cartilage in the example of WO 12/099216. Specifically, the frozen salmon nasal cartilage was coarsely pulverized to a size of about 2 mm by a crusher, washed with water and warm water, added with 2.5 times the amount of distilled water, and heated at 95° C. for 3.5 hours. After that, unnecessary substances (residues) were removed by filtration and centrifugation at 16000 rpm, and the resulting supernatant was concentrated by vacuum concentration to obtain a concentrate. A freeze-dried product obtained by freeze-drying the concentrate was used for the following small intestinal epithelial model or human test. In addition, the concentrate was diluted with water and used for the colitis model mice described below.

In addition, when the proteoglycan molecular weight contained in the obtained concentrate (confirmed by the uronic acid amount chromatogram measured by the carbazole-sulfuric acid method) was measured by the method described in the examples of JP-A-2017-066097, the concentration was The liquid contained proteoglycan with a molecular weight of 1,800,000 or more. The molecular weight distribution was about 1,800,000 to 20,000,000.

[Preparation of lactic acid fermented product of figs]
Masui Dauphine fig flower stalks were removed and washed with water. After removing moisture, the whole fig was pulverized using a food processor to obtain a pulverized whole fig. 10 g of the lactic acid bacteria solution was added to 200 g of the whole fig crushed product, lightly mixed so as to be uniform, and then left to stand at 30° C. for 24 hours for fermentation under anaerobic conditions. Lactobacillus pentosus was used as the lactic acid bacterium. In the fermentation process, the end point of the fermentation process was confirmed by measuring the pH and acidity of the fermented product. The resulting fermented product was freeze-dried and used as a lactic acid bacteria fermented product of figs.

<Preparation and evaluation test of small intestinal epithelial model>
Caco cells were seeded and cultured on a Transwell polycarbonate membrane permeable support to prepare a small intestinal epithelial model (Fig. 1). After seeding, the cells were cultured for 13 days (the medium was changed every 2 to 3 days), IFN-γ was added on the 14th day, and on the 15th day proteoglycan or fermented fig lactic acid bacteria or quercetin was added together with TNF-α. did. Quercetin has the effect of suppressing inflammatory substances, and therefore, when quercetin is added, the action of stimulating substances is suppressed and the effect of improving the intestinal barrier is obtained (positive control).

TNF-α and IFN-γ are applied to the Basal (basement membrane side) of the small intestinal epithelial model for the purpose of stimulating them. were added respectively (see FIG. 1). More specifically, IFN-γ was added to a final concentration of 10 ng/mL 24 hours before evaluation. One hour before the start of evaluation (-1h), proteoglycan, lactic acid fermented fig or quercetin was added. Quercetin was added to a final concentration of 200 μM. In addition, TNF-α was added to a final concentration of 10 ng/mL just before the start of evaluation (0h). The evaluation was performed by measuring the membrane potential difference over time using a transepithelial electrical resistance (TER) measuring device: Milli-cell ERS (Millipore). If the barrier function is lowered, the membrane potential difference is reduced. Moreover, the same test was performed without applying any stimulus.

These results are shown in FIG. In FIG. 2, "cont" indicates a negative control, in which neither stimulation nor evaluation substance was added. "Stimulus" refers to the addition of stimulus only. In FIG. 2, "PG" indicates addition of proteoglycan, and "Fig" indicates addition of lactic acid fermented fig.

In the absence of stimulation (i.e., the intestinal barrier is normal) (right side of FIG. 2), the intestinal barrier was enhanced in the group added with proteoglycan and fermented fig lactic acid bacteria compared to no addition (cont). rice field. In addition, when there is stimulation (that is, when the intestinal barrier is poor) (left side of FIG. 2), the proteoglycan addition group is equivalent to the case where no stimulation is given (cont), and the lactic acid fermented fig addition group It improved the intestinal barrier more than when no stimulation was given.

Human study 1 (examination of fecal mucin content and intestinal regulation effect)
Three types of hard capsules containing the following ingredients were prepared.
Capsule 1 (PG): 200 mg of proteoglycan (dry matter) and 472.5 mg of dextrin per 6 capsules Capsule 2 (fig): 472.5 mg of lactic acid fermented fig (dry matter) and 472.5 mg of dextrin per 6 capsules Capsule 3 (placebo): 472.5 mg dextrin per 3 capsules

Then, 33 women in their 30s to 50s were divided into 3 groups (capsule 1 (PG) intake group, capsule 2 (fig) intake group, and capsule 3 (placebo) intake group), and a placebo-controlled double-blind parallel group. In an interim study, each capsule was orally ingested for 6 weeks. The Capsule 1 (PG) intake group and the Capsule 2 (fig) intake group were allowed to ingest 6 tablets per day, and the Capsule 3 (placebo) intake group was allowed to ingest 3 tablets per day.

Feces were collected at the start of intake and 4 weeks after the start of intake, and the amount of mucin contained was examined. At the end of the examination, the subjects were asked to answer a questionnaire about changes in their intestinal functions compared to when they started ingestion. The question items of the questionnaire are as shown at the bottom of FIG. The higher the number, the higher the number of times or amount, or the better the condition), and the average value was calculated.

Fig. 3 shows the results of the fecal mucin content test, and Fig. 4 shows the results of the questionnaire survey. 3 and 4, * indicates p<0.05 and ** indicates p<0.01, respectively.

From FIG. 3, it was found that the amount of mucin in the intestine increased in both the groups that took proteoglycan and the fermented product of lactic acid bacteria of figs. Moreover, from FIG. 4, it was found that both proteoglycan and the fermented product of figs with lactic acid bacteria tend to exhibit an intestinal regulation effect. In particular, it was found that proteoglycan exerts an intestinal regulation effect relatively late after ingestion, while fig fermented with lactic acid bacteria exerts an intestinal regulation effect immediately after ingestion.

Furthermore, in the test, immediately after the end of the intake and 6 weeks after the end of the intake (hereinafter sometimes referred to as "after washout"), feces were collected from each subject, and using the stool specimen, Bacterial species of the intestinal flora were analyzed. Bacterial species of the intestinal flora were analyzed using the T-RFLP (Terminal Restriction Fragment Length Polymorphism Analysis) method. The analysis was outsourced to Techno Suruga Lab Co., Ltd.

FIG. 5 shows the average abundance ratio of each bacterial species for each subject in the Capsule 1 (PG) intake group, the Capsule 2 (fig) intake group, and the Capsule 3 (placebo) intake group.

In addition, in FIG. 5, * indicates that there was a significant difference compared with the placebo group. More specifically, Bifidobacterium: Fig>Placebo (p=0.075), Bacteroides: PG>Placebo (p=0.024), Fig>Placebo (p=0.015) immediately after ingestion. Also, after washout, Fig<placebo (p=0.001) for Clostridium subcluster XIVa and PG<placebo (p=0.062) for Clostridium cluster IX.

Investigation using colitis model mice The anti-inflammatory effects of proteoglycan and fermented fig lactic acid bacteria were examined in colitis-induced hairless mice.

Animals used: 40 female HR-1 mice (Japan SLC, 8 weeks old) (n=10)
Pathogen: dextran sulfate (DSS) (molecular weight 36000-50000; MP Biomedicals)
Test group: 4 groups (grouped by administered substance)
Distilled water group (negative control group),
1.5% DSS group (positive control group),
1.5% DSS + fig lactic acid fermented product group (test substance A administration group),
1.5% DSS + proteoglycan group (test substance B administration group)
Measurement item: colitis score (DAI score: Disease Activity Index Score)

Specifically, the mice were divided into 4 groups and made to ingest each of the above-described administration substances using a probe for 14 consecutive days. In addition, 1 g of lactic acid fermented figs and 5 mg of proteoglycans were ingested per day. Thereafter, they were allowed to take DSS for 7 consecutive days, followed by 7 days without the administered substance and DSS, and then again with DSS for 7 consecutive days.

The last day of the period in which each substance was administered (day 0), the last day of the period in which DSS was continuously ingested (day 7), and the last day of the period in which neither the substance nor DSS was ingested (day 7). The DAI scores measured on day 14), and also on days 1, 2, 3, and 4 (days 15, 16, 17, and 18) of the period in which DSS intake was resumed thereafter are shown in FIG. shown in The DAI score is an index showing the severity of colitis, and the higher the number, the more severe the injury.

In the group ingested first with proteoglycan or lactic acid fermented fig, colitis was suppressed during the period after resuming DSS intake. From this, it was found that proteoglycan or the group of fermented figs with lactic acid bacteria has the effect of preventing colitis caused by exposure to a colitis-inducing substance (here, DSS).

Human study 2 (examination of phenol amount in feces)
Two types of hard capsules containing the following ingredients were prepared.
Capsule 1 (PG): 200 mg of the proteoglycan (dry matter) and 472.5 mg/3 capsules of dextrin Capsule 3 (placebo): 472.5 mg of dextrin/3 capsules

Then, 31 women in their 30s to 50s were divided into 2 groups (capsule 1 (PG) intake group and capsule 3 (placebo) intake group), and in a placebo-controlled double-blind parallel group study, each for 6 weeks. The capsule was orally ingested. The Capsule 1 (PG) intake group was allowed to ingest 3 tablets per day, and the Capsule 3 (placebo) intake group was allowed to ingest 3 tablets per day. One subject who took a laxative during the test period was excluded from the analysis. For this reason, 15 subjects in the Capsule 1 intake group and 15 subjects in the Capsule 3 intake group were subjected to the following result analysis.

Feces were collected at the start of intake, 2 weeks after the start of intake, and 6 weeks after the start of intake, and the amount of phenol contained was measured. The measurement of the amount of phenol in feces was outsourced to Techno Suruga Labo Co., Ltd.

FIG. 7 shows the results of inter-group comparison of the amount of phenol in feces. In FIG. 7, * indicates p<0.05 and ** indicates p<0.01, respectively. It was found that proteoglycan has the effect of significantly reducing the amount of phenol in feces, and thus improves the intestinal environment.

Claims (6)

proteoglycans, and
A composition for improving an intestinal barrier containing a fermented product of lactic acid bacteria of figs . The composition for improving the intestinal barrier according to claim 1, which is for intestinal regulation. The composition for improving the intestinal barrier according to claim 1 or 2, for increasing the amount of intestinal mucin. The composition for improving the intestinal barrier according to claim 1 or 2, for improving intestinal flora. The intestinal barrier-improving composition according to any one of claims 1 to 4, wherein the fermented product of figs with lactic acid bacteria is a fermented product of Lactobacillus pentosus of figs. The composition for improving the intestinal barrier according to any one of claims 1 to 5, which is an oral composition.

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