JP2010083881A - Intestinal tract protectant - Google Patents

Abstract

[Problem] To provide an intestinal protective agent capable of suppressing intestinal mucosal injury through a decrease in intestinal barrier function and the initiation of an inflammatory reaction in the intestinal tract when the intestinal defense mechanism fails. To do.
[MEANS FOR SOLVING PROBLEMS] An intestinal tract protecting agent containing, as an active ingredient, cells of a strain belonging to Lactobacillus brevis or a processed product thereof. Moreover, the intestinal protective agent which contains the culture supernatant of the strain which belongs to Lactobacillus brevis (Lactobacillus brevis), or its processed material as an active ingredient.
[Selection figure] None

Description

  The present invention relates to an intestinal protective agent.

  In the normal intestinal tract of mammals, various defense mechanisms work to protect the intestinal mucosa from external stimuli. When such a defense mechanism fails for some reason, the intestinal tract (intestinal mucosa) is damaged, and in some cases, it is the onset of various intestinal diseases including inflammatory bowel disease (Crohn's disease, ulcerative colitis). Will be involved.

  Regarding the intestinal protective agent, for example, an intestinal protective agent containing one or more substances selected from whey protein, casein, serum albumin and degradation products thereof as an active ingredient is known (see Patent Document 1). In addition, an inflammatory bowel disease preventive / therapeutic agent containing a Lactobacillus genus bacterium (particularly, Lactobacillus casei, Lactobacillus gasseri) or a polysaccharide fraction derived therefrom as an active ingredient is known (see Patent Document 2).

JP-A-9-241177 JP 2003-73286 A

  An intestinal protective agent that suppresses intestinal mucosal injury by protecting the intestinal tract (intestinal mucosa) is considered to be effective in preventing or ameliorating various intestinal diseases accompanied by intestinal mucosal injury. Although some enteroprotective agents are known, there are still not enough options to meet the diverse demands of consumers.

  Then, this invention makes it a subject to provide a novel intestinal protective agent.

  The present invention provides an intestinal protective agent containing a bacterial cell of a strain belonging to Lactobacillus brevis or a processed product thereof as an active ingredient. Moreover, this invention provides the intestinal protective agent which contains the culture supernatant of the strain which belongs to Lactobacillus brevis (Lactobacillus brevis), or its processed material as an active ingredient.

  The intestinal protective agent of the present invention suppresses, for example, a decrease in the intestinal (intestinal mucosa) barrier function and the initiation of an inflammatory reaction (production of inflammatory substances) in the intestinal (intestinal mucosa) when the defense mechanism in the intestinal tract breaks down. This makes it possible to suppress intestinal mucosal injury. In addition, by inhibiting intestinal mucosal injury, it is possible to prevent or ameliorate (treat or reduce) various intestinal diseases (eg, inflammatory bowel disease (Crohn's disease, ulcerative colitis)) associated with intestinal mucosal injury. And

  Since the intestinal tract protecting agent of the present invention exhibits the effects as described above, it can be used, for example, to suppress a decrease in intestinal (intestinal mucosa) barrier function. It can also be used to suppress the induction of an inflammatory reaction (production of inflammatory substances) in the intestinal tract (intestinal mucosa). Moreover, it can be used as a preventive or ameliorating agent for various intestinal diseases (for example, inflammatory bowel disease (Crohn's disease, ulcerative colitis)) accompanied by intestinal mucosal injury.

  Lactobacillus brevis is a type of lactic acid bacterium that has been used for fermented foods for a long time, and has been established to be safe for living organisms. Therefore, the intestinal tract protecting agent of the present invention is highly safe to living bodies and can be ingested continuously for a long period of time, and is used as a pharmaceutical ingredient, food / drink ingredient, food / drink additive, feed ingredient, feed additive, etc. be able to.

  There are four subspecies of Lactobacillus brevis: brevis, grevesensis, otakiensis, and coagulans. As the strain in the intestinal protective agent of the present invention, a strain belonging to subspecies brevis is preferable, and among the strains belonging to subspecies brevis, for example, Lactobacillus brevis (Lactobacillus brevis) SBC8803 strain (accession number: FERM BP-). 10632) is particularly suitable.

  ADVANTAGE OF THE INVENTION According to this invention, the novel intestinal protective agent which has high safety | security to a biological body and can be used also as a component of food-drinks is provided. Moreover, the pharmaceutical, food / beverage products, food / beverage additive, feed, etc. containing such an intestinal protective agent are provided.

2 is a Western blot of the protein extracted in Example 1. 4 is a graph showing intestinal mannitol flux stimulated with monochloramine in Example 2. FIG. It is a Western blot of the protein extracted in Example 3. It is a Western blot of the protein extracted in Example 4. 2 is an SDS-Page photograph of the culture supernatant and precipitated fraction obtained in Reference Example 1. It is a box-and-whisker plot which shows distribution of the inflammation score of the mouse | mouth which administered dextran sulfate sodium in Example 5. FIG.

  Hereinafter, embodiments of the present invention will be described.

  In one aspect, the intestinal tract protecting agent of the present invention contains a bacterial cell belonging to Lactobacillus brevis or a processed product thereof as an active ingredient. Moreover, in another one aspect | mode, the culture supernatant of the strain which belongs to Lactobacillus brevis (Lactobacillus brevis), or its processed material is contained as an active ingredient.

  Lactobacillus brevis has four subspecies [Lactobacillus brevis, Gravesensis, Otakiensis] based on differences in the base sequence of 16S ribosomal DNA, the rate of acid production from the consumed sugar, and the like. And coagulans].

  As a strain belonging to Lactobacillus brevis, a strain belonging to subspecies brevis is preferable, and among strains belonging to subspecies brevis, for example, Lactobacillus brevis SBC8803 strain is preferable. The Lactobacillus brevis SBC8803 strain was founded on June 28, 2006 at the National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center (1st, 1st East, 1st Street, Tsukuba City, Ibaraki Prefecture, Japan, 6th postal code 305-8586). The deposited strain has the deposit number FERM BP-10632. The SBC8803 strain can grow even in the presence of alcohol, and is suitable for use as a component of alcoholic beverages.

  The strain in the intestinal protective agent of the present invention may be any strain belonging to Lactobacillus brevis, and may be, for example, one that can be separated from the natural world or one that can be obtained from a cell bank such as ATCC.

  As a microbial cell of the said strain, only the microbial cell of 1 type of a strain may contain, or the microbial cells of 2 or more types of strains may be contained together. In addition, the microbial cells may be either live cells or dead cells. Bacteria can be produced in large quantities by culturing viable cells. The medium may be either a liquid medium or a solid medium, but preferably contains a nitrogen source and a carbon source. Nitrogen sources include meat extract, peptone, gluten, casein, yeast extract, amino acids, etc., and carbon sources include glucose, xylose, fructose, inositol, maltose, water candy, soup, starch, bacus, bran, Molasses, glycerin and the like can be used. Further, ammonium sulfate, potassium phosphate, magnesium chloride, sodium chloride, iron, manganese, molybdenum and the like can be added as inorganic substances, and vitamins and the like can be further added. Suitable media include MRS media, LBS media, Rogosa media, WYP media, GYP media and the like. The culture temperature is usually about 25 to about 40 ° C, preferably about 30 to about 38 ° C (particularly about 37 ° C). The culture time is usually about 6 to about 62 hours. The pH of the medium is usually about 3 to about 6, preferably about 4 to about 6. Cultivation may be performed in an incubator, or aeration and shaking may be performed during the cultivation.

  As the treated product of the microbial cells, for example, a treated product obtained by heating the microbial cells at 100 ° C. or more for several minutes or more (for example, the microbial cells are subjected to autoclave treatment at a temperature of 110 to 125 ° C. for 10 minutes or more. Treated product), a treated product obtained by freeze-drying, spray-drying, etc. on the bacterial cells, or a treated product obtained by physically destroying the bacterial cells with an ultrasonic wave, a French press or the like. Such a treated product of microbial cells is suitable as an active ingredient of an intestinal tract protecting agent in terms of easy handling as compared with untreated microbial cells (particularly live microbial cells).

  The culture supernatant is a supernatant of a culture obtained by culturing viable cells of a strain belonging to Lactobacillus brevis in a liquid medium. As a liquid culture medium, what contains a nitrogen source and a carbon source is preferable. Nitrogen sources include meat extract, peptone, gluten, casein, yeast extract, amino acids, etc., and carbon sources include glucose, xylose, fructose, inositol, maltose, water candy, soup, starch, bacus, bran, Molasses, glycerin and the like can be used. Further, ammonium sulfate, potassium phosphate, magnesium chloride, sodium chloride, iron, manganese, molybdenum and the like can be added as inorganic substances, and vitamins and the like can be further added. Suitable liquid media include MRS media, LB media, Rogosa media, WYP media, GYP media and the like. The culture temperature is usually about 27 to about 40 ° C, preferably about 30 to about 38 ° C (particularly about 37 ° C). The culture time is usually about 6 to about 62 hours, preferably about 12 to about 48 hours. The pH of the medium is usually about 3 to about 6, preferably about 4 to about 6. Cultivation may be performed in an incubator, or aeration and shaking may be performed during the cultivation. The culture supernatant can be separated from the culture by, for example, filtering the culture with a filter.

  As is apparent from the above, suitable culture methods for obtaining the culture supernatant include, for example, about 30 to about 38 ° C. (especially using MRS medium, LB medium, etc.) having a pH of about 4 to about 6 in an incubator. And a method of culturing at about 37 ° C. for about 12 to about 48 hours (particularly about 12 hours, about 24 hours, about 36 hours or about 48 hours).

  As the processed product of the culture supernatant, for example, a processed product obtained by solid-liquid separation of the culture supernatant by centrifugation, a processed product obtained by removing water from the culture supernatant by freeze drying, an evaporator, etc. Processed product obtained by concentrating the culture supernatant under reduced pressure, processed product obtained by concentrating the culture supernatant using an ultrafiltration membrane or the like, or obtained by solid-liquid separation of the culture supernatant using a filter or the like A processed material is mentioned. Also, for example, after centrifuging the culture supernatant (eg, 3000 rpm, 10 minutes), the supernatant is filtered with a syringe filter (eg, 0.45 μm), and the filtrate is separated with ammonium sulfate (eg, 65% saturated ammonium sulfate). And a precipitate fraction obtained by dialyzing the precipitate with distilled water.

  The intestinal protective agent of the present invention may be in any form such as solid (for example, powder obtained by freeze-drying), liquid (water-soluble or fat-soluble solution or suspension), paste, etc., and powders, granules Any dosage form such as an agent, a tablet, a syrup, a troche, or a capsule may be used. In addition, the intestinal protective agent of the present invention may be composed of a bacterial cell belonging to Lactobacillus brevis or a processed product thereof, or a culture supernatant of a strain belonging to Lactobacillus brevis or a processed product thereof. .

  The above-mentioned various preparations consist of a bacterial strain belonging to Lactobacillus brevis or a processed product thereof or a culture supernatant or a processed product thereof, and pharmaceutically acceptable additives (excipients, binders, lubricants, disintegrations). Agents, emulsifiers, surfactants, bases, solubilizers, suspending agents, and the like).

  For example, the excipient includes lactose, sucrose, starch, dextrin and the like. Examples of the binder include polyvinyl alcohol, gum arabic, tragacanth, gelatin, hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone and the like. Examples of the lubricant include magnesium stearate, calcium stearate, talc and the like. Examples of the disintegrant include crystalline cellulose, agar, gelatin, calcium carbonate, sodium bicarbonate, dextrin and the like. Examples of the emulsifier or surfactant include Tween 60, Tween 80, Span 80, and glyceryl monostearate. Examples of the base include cetostearyl alcohol, lanolin, polyethylene glycol, rice bran oil, fish oil (DHA, EPA, etc.), olive oil and the like. Examples of the solubilizer include polyethylene glycol, propylene glycol, sodium carbonate, sodium citrate, Tween 80 and the like. Examples of the suspending agent include Tween 60, Tween 80, Span 80, glyceryl monostearate, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, hydroxymethyl cellulose, sodium alginate and the like.

  The intestinal tract protecting agent of the present invention can be used as a pharmaceutical ingredient, a food / beverage product component, a food / beverage product additive, a feed component, a feed additive, and the like.

  For example, the intestinal tract protecting agent of the present invention is applied to food and drink such as water, soft drinks, fruit juice drinks, milk drinks, alcoholic drinks, breads, noodles, rice, tofu, dairy products, soy sauce, miso, and confectionery. It can be used as an additive. These foods and drinks may further contain other additives usually used in the art, and examples of such additives include bitters, flavors, apple fibers, soybean fibers, meat extracts, blacks. Vinegar extract, gelatin, corn starch, honey, animal and vegetable oils; monosaccharides such as glucose and fructose; disaccharides such as sucrose; polysaccharides such as dextrose and starch; sugar alcohols such as erythritol, xylitol, sorbitol, and mannitol; vitamin C and the like Vitamins. The intestinal tract protecting agent of the present invention can also be used as a component for food for specified health use, food for special use, dietary supplement, health food, functional food, food for the sick, and the like. The food and drink containing the intestinal tract protecting agent of the present invention may be a fermented product obtained by fermenting milk, skim milk, soy milk or the like with a strain belonging to Lactobacillus brevis.

  The intestinal protective agent of the present invention may be administered to humans or non-human mammals. The dosage and administration method can be appropriately determined according to the condition, age, etc. of the individual to be administered. Suitable administration methods include, for example, oral administration.

  Hereinafter, based on an Example, this invention is demonstrated more concretely. However, the present invention is not limited to the following examples.

[Example 1]
(Preparation of test sample)
The cells of Lactobacillus brevis SBC8803 strain were sterilized by autoclaving (105 ° C., 30 minutes) and then freeze-dried. The lyophilized cells were suspended in RPMI medium and the cell suspension [0.1% (w / V), 1% (w / v)]. In addition, an RPMI medium containing no bacterial cells was prepared as a control.

(Irritation to the intestinal tract)
The small intestine was taken out from male, C57BL / 6J mice (Charles River Japan) 6 to 10 weeks old, and the intestine was washed with PBS, and then the intestine was divided into three equal parts. For each of the obtained three intestinal tracts, one end was tied with a surgical thread, and a cell suspension [0.1% (w / v), 1% (w / v)] or RPMI medium (control) The other end was tied with a surgical thread. Three intestinal tracts were soaked in RPMI medium, incubated for 2 hours at 37 ° C. in a CO ₂ incubator, and proteins were extracted from the intestinal epithelium using a Mammalian Cell Extraction Kit (BioVision).

(Western blotting)
The extracted protein was subjected to Western blotting as follows. First, 30 μg of the extracted protein was separated by SDS-PAGE and transferred to a PVDF membrane using a semi-driving device. The PVDF membrane was then blocked by shaking for 1 hour in 5% skim milk containing 0.1% Tween20. Then, the PVDF membrane was reacted with a primary antibody [anti-Hsp25 antibody, anti-Hsp70 antibody or anti-Hsc70 antibody]. Further, the PVDF membrane was sufficiently washed with PBS containing 0.1% Tween 20, and then reacted with a secondary antibody [HRP-conjugated anti-rabbit antibody or HRP-conjugated anti-mouse antibody]. Finally, the PVDF membrane was thoroughly washed with PBD containing 0.1% Tween 20, and then the antigen protein was detected by the chemiluminescence method.

  Hsc70 is one of the proteins that are constantly expressed in intestinal epithelial cells. On the other hand, Hsp25 and Hsp70 inhibit the production of inflammatory substances (interleukin-1β, interleukin-6, tumor necrosis factor (TNF) -α, etc.) and the induction of the inflammatory reaction associated therewith, thereby damaging cells. It is a heat shock protein that suppresses

(result)
The results are shown in FIG. FIG. 1 is a Western blot of extracted proteins. Table 1 is a table | surface which shows the intensity | strength of the band on the western blot of FIG. In FIG. 1 and Table 1, the concentration [% (w / v)] indicates the cell concentration in the cell suspension.

  As is clear from FIG. 1 and Table 1, the expression level of Hsc70 was not significantly different between the cell suspension and the control, whereas the expression levels of Hsp25 and Hsp70 were There was significantly more in the intestine where the body suspension was injected.

[Example 2]
(Preparation of test sample)
Test samples were prepared in the same manner as in Example 1. In addition, an RPMI medium containing no bacterial cells was prepared as a control.

(Irritation to the intestinal tract)
The small intestine was taken out from male, C57BL / 6J mice (Charles River Japan) 6 to 10 weeks old, and the intestine was washed with PBS, and then the intestine was divided into three equal parts. For each of the obtained three intestinal tracts, one end was tied with a surgical thread, and a cell suspension [0.1% (w / v), 1% (w / v)] or RPMI medium (control) The other end was tied with a surgical thread. Three intestinal tracts were immersed in RPMI medium and incubated at 37 ° C. for 2 hours in a CO ₂ incubator.

(Measurement of mannitol flux)
Next, the contents were discharged from each intestinal tract, and the intestinal tract was further divided into two halves, and then one tritium labeled mannitol aqueous solution (1 μCi [ ³ H] mannitol / mL) added with monochloramine (NH ₂ Cl) was added to one intestinal tract. Into the other intestine, a tritium-labeled mannitol aqueous solution (1 μCi [ ³ H] mannitol / mL) without addition of monochloramine was injected, and the intestine was immersed in an RPMI medium and left still. The medium was sampled after 5 minutes, 20 minutes, and 35 minutes, and the amount of tritium was measured with a scintillation counter. “Amount of tritium after 20 minutes−amount of tritium after 5 minutes” and “amount of tritium after 35 minutes−amount of tritium after 5 minutes” were calculated, and the mannitol fluxes were 15 minutes and 30 minutes, respectively.

  Monochloramine is a substance known to oxidatively damage the gastrointestinal mucosa. The mannitol flux of the intestinal tract can be used as an index of intestinal mucosal injury (decrease in intestinal (intestinal mucosa) barrier function).

(result)
The results are shown in FIG. FIG. 2 is a graph showing intestinal mannitol flux (15 minutes, 30 minutes) stimulated with monochloramine. In FIG. 2, the concentration [% (w / v)] indicates the cell concentration in the cell suspension.

  As is clear from FIG. 2, the mannitol flux in the intestinal tract stimulated with monochloramine was significantly smaller in the intestinal tract into which the cell suspension was injected. The mannitol flux in the intestine that was not stimulated with monochloramine was small enough to be ignored in any intestine.

Example 3
(Preparation of culture supernatant)
10% (w / v) Lactobacillus brevis SBC8803 cells in MRS medium were cultured at 37 ° C. for 12 hours, 36 hours and 60 hours in an incubator to obtain a culture supernatant. Moreover, the MRS culture medium which does not contain a culture supernatant was prepared as control.

(Stimulation of intestinal epithelial cells)
In the obtained culture supernatant, Caco-2 cells (intestinal epithelial cells derived from human colon cancer) were incubated at 37 ° C. for 24 hours, and proteins were extracted from Caco-2 cells using a Mammarian Cell Extraction Kit (BioVision).

(Western blotting)
The extracted protein was subjected to Western blotting in the same manner as in Example 1 except that anti-Hsp27 antibody or anti-Hsc70 antibody was used as the primary antibody.

  Hsc70 is one of the proteins that are constantly expressed in intestinal epithelial cells. On the other hand, Hsp27 suppresses the production of inflammatory substances (interleukin-1β, interleukin-6, tumor necrosis factor (TNF) -α, etc.) and the induction of the inflammatory reaction associated therewith, thereby suppressing cell damage. It is a heat shock protein.

(result)
The results are shown in FIG. FIG. 3 is a Western blot of the extracted protein. Table 2 is a table | surface which shows the intensity | strength of the band on the western blot of FIG. In FIG. 3 and Table 2, time indicates the culture time of the cells.

  As is clear from FIG. 3 and Table 2, the expression level of Hsc70 did not show a large difference between the culture supernatant and the control, whereas the expression level of Hsp27 was found in the culture supernatant. Significantly more in the incubated Caco-2 cells.

Example 4
(Preparation of culture supernatant)
A culture supernatant of Lactobacillus brevis SBC8803 strain was obtained in the same manner as in Example 3 except that the culture time was 24 hours. Moreover, the MRS culture medium which does not contain a culture supernatant was prepared as control.

(Ammonium sulfate fraction)
The obtained culture supernatant was centrifuged (3000 rpm, 10 minutes), and the supernatant was filtered with a syringe filter (0.45 μm). The filtrate was fractionated with 65% saturated ammonium sulfate, and the precipitate was dialyzed with distilled water to obtain a precipitate fraction.

(Stimulation of intestinal epithelial cells)
In the obtained precipitate fraction, Caco-2 cells (human colon cancer-derived intestinal epithelial cells) were incubated at 37 ° C. for 24 hours, and proteins were extracted from Caco-2 cells using a Mammarian Cell Extraction Kit (BioVision).

(Western blotting)
The extracted protein was subjected to Western blotting in the same manner as in Example 3.

(result)
The results are shown in FIG. FIG. 4 is a western blot of the extracted protein. In FIG. 4, “× 100”, “× 10”, and “× 1” indicate that the dilution rate of the precipitated fraction is 100 times, 10 times, and 1 time, respectively.

  As is clear from FIG. 4, the Hsc70 expression level did not show a large difference between the precipitate fraction and the control, whereas the Hsp27 expression level was incubated in the precipitate fraction. It was significantly higher in Caco-2 cells.

  According to Examples 1 to 4, the intestinal protective agent of the present invention causes a decrease in the intestinal (intestinal mucosa) barrier function and the induction of an inflammatory reaction in the intestinal tract (intestinal mucosa) when the intestinal defense mechanism fails (inflammatory substance It was confirmed that intestinal mucosa injury could be suppressed through this.

[Reference Example 1]
In the same manner as in Example 4, preparation of the culture supernatant and ammonium sulfate fractionation were performed. The obtained culture supernatant and precipitate fraction were subjected to SDS-Page, and the protein was detected by silver staining.

  The results are shown in FIG. FIG. 5 is an SDS-Page photograph of the culture supernatant and the precipitated fraction. In FIG. 5, “× 1”, “× 5”, “× 25”, “× 125”, and “× 625” are dilution ratios of 1 ×, 5 ×, and 25 × of the culture supernatant or the precipitate fraction, respectively. , 125 times, and 625 times.

  Reference Example 1 showed that the culture supernatant of Lactobacillus brevis SBC8803 strain contained 25 kDa and 50 kDa proteins.

Example 5
(Preparation of test sample)
The cells of Lactobacillus brevis SBC8803 strain were sterilized by autoclaving (105 ° C., 30 minutes), freeze-dried, the freeze-dried cells were suspended in distilled water, and 0.1% (w / v) cell suspension was obtained. A turbid liquid was obtained.

(Ulcerative colitis induction test)
Test sample administration:
After dividing 8 C57BL / 6 mice (male, body weight 18-25 g) (Nippon Charles River) into a fungus body administration group (3 animals) and a fungus body non-administration group (5 animals), daily for 10 days, 1 mL of the cell suspension was orally administered to the mice in the fungus group, and 1 mL of distilled water was orally administered to the mice in the non-cell group. During this period, all groups of mice had free access to normal food. The body weight of each mouse was measured on the 10th day, and the body weight (mean ± standard deviation) was as follows.

Body weight (g):
Fungus body administration group: 21.7 ± 2.2
Non-administration group: 22.2 ± 1.1

Administration of DSS:
Next, each group of mice was allowed to freely drink 2% (w / v) dextran sodium sulfate (DSS) aqueous solution for 14 days. During this period, 1 mL of the bacterial cell suspension was orally administered every other day to the mice in the bacterial cell administration group. In addition, during the above period, all groups of mice were allowed to freely take normal food. DSS is known to induce intestinal inflammation similar to ulcerative colitis histologically and immunologically.

  On the 14th day, the weight of each mouse was measured, and after sacrifice (cervical dislocation), the intestinal tract was removed and the intestinal length (length from the anus to the cecum) was measured. Body weight (mean ± standard deviation) and intestinal length (mean ± standard deviation) were as follows.

Body weight (g):
Bacterial administration group: 17.3 ± 4.8
Non-administration group: 18.8 ± 1.9
Intestinal length (cm):
Fungus body administration group: 6.2 ± 1.9
Non-administration group: 6.2 ± 0.5

Histological evaluation of inflammation:
A tissue of the distal large intestine (near the rectum) was further collected from the intestine, and formalin-fixed and paraffin-embedded. Then, it was sliced into 4 μm thicknesses, and hematoxylin and eosin staining was performed on tissue sections (2 sections / animal) to obtain tissue specimens (2 specimens / animal). Each specimen was observed with an optical microscope, and the degree of inflammation was evaluated histologically according to the following criteria (see Berg DJ et al., J Clin Invest, 1998).

Evaluation criteria:
Score 0: Inflammatory cell infiltration is not observed.
Score 1: 1 to several mononuclear cells infiltrate the mucosal epithelium. There is no decrease in mucin production from goblet cells.
Score 2: Mild inflammatory cell infiltration is observed in the mucosal epithelium. Inflammatory cell infiltration is mononuclear cell dominant, but neutrophil infiltration is also seen.
Score 3: A decrease in mucin production is observed. Often inflammatory cell infiltration into the submucosa.
Score 4: Inflammatory cell infiltration extends to the muscle layer and mucin production is almost lost. A crypt abscess or ulcer is observed.

(result)
The evaluation results (inflammation scores) were as shown below and in FIG. FIG. 6 is a box-and-whisker diagram showing the distribution of inflammation scores of mice in each group. In FIG. 6, the tips of the upper and lower whiskers indicate the positions of the maximum value and the minimum value of the inflammation score, respectively. Moreover, the upper end and lower end of a box show the position of a 25% point and a 75% point, respectively, and the center point of a box shows the position of a median value. In both the fungus body administration group and the fungus body non-administration group, the lower whiskers overlap the lower end of the box.

Evaluation results:
Fungus administration group: 1, 1, 2, 2, 2, 3
Non-administration group: 2, 2, 2, 3, 3, 3, 3, 3, 4, 4

  As is clear from the above results, inflammation was remarkably suppressed in the bacterial cell administration group. The inflammation score of the fungus body administration group was significantly lower than that of the fungus body non-administration group (Mann Whitney test, p <0.05). In addition, there was no significant difference between groups regarding body weight and intestinal length (Mann Whitney test).

  According to Example 5, the intestinal protective agent of the present invention causes a decrease in the intestinal (intestinal mucosa) barrier function and the induction of an inflammatory reaction in the intestinal (intestinal mucosa) when the intestinal defense mechanism fails (production of inflammatory substances). It was confirmed that it was possible to suppress intestinal mucosa injury through this.

  The intestinal protective agent of the present invention can be used for prevention and improvement of various intestinal diseases.

Claims (9)

  An intestinal protective agent containing, as an active ingredient, a bacterial cell of a strain belonging to Lactobacillus brevis or a processed product thereof.   An intestinal protective agent containing a culture supernatant of a strain belonging to Lactobacillus brevis or a processed product thereof as an active ingredient.   The intestinal tract protecting agent according to claim 1 or 2, wherein the strain belongs to Lactobacillus brevis subspecies brevis (Lactobacillus brevis subspecies brevis).   The intestinal protective agent according to claim 3, wherein the strain is Lactobacillus brevis SBC8803 (accession number: FERM BP-10632) strain.   The intestinal tract protecting agent according to any one of claims 1 to 4, which is used for suppressing a decrease in intestinal barrier function in a mammal.   The intestinal protective agent according to any one of claims 1 to 5, which is used for suppressing the induction of an inflammatory reaction in a mammalian intestinal tract.   The pharmaceutical containing the intestinal tract protective agent as described in any one of Claims 1-6.   Food / beverage products containing the intestinal protective agent as described in any one of Claims 1-6.   The food-drinks additive containing the intestinal tract protective agent as described in any one of Claims 1-6.