JP2001231495A - Food having activity for inhibiting accumulation of eosinophil - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a food obtained by using a food as a raw material, having no side effect and further having activities for inhibiting the accumulation of eosinophils. SOLUTION: The accumulation of the eosinophils can be prevented by using a material obtained by subjecting the cell walls of lactic bacteria to pulverization. The pulverized material of the cell walls of the lactic material is not only used singly but also used by mixing the material with an existing food because the pulverized material of the cell walls of the lactic bacteria has no side effect and can be formed into a powder.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a food having an eosinophil accumulation-suppressing action, comprising a substance having a cell wall disrupted by lactic acid bacteria as a main component.

[0002]

2. Description of the Related Art Eosinophils are leukocytes which are a small percentage of the total amount of leukocytes contained in the blood of normal humans in about 2 to 5%, but are greatly involved in inflammation of allergic diseases. It has been known. When classifying leukocytes that collect at sites of inflammation caused by allergic antigens, the percentage of eosinophils is high. Eosinophils are activated by cytokines and certain types of T cells, move through the gaps in blood vessels and move to mucosal sites such as the trachea, causing new inflammation.
In type 1 allergy, after an allergic reaction occurs within minutes after exposure to the antigen, when the inflammatory reaction occurs again within several hours to over ten hours, eosinophils are abundantly present in the inflamed area. Therefore, various studies have reported that eosinophils cause chronic allergic inflammation.

At present, an eosinophil accumulation-suppressing action has been reported as one of the mechanisms of action of histamine-added human γ-globulin which has long been used as a remedy for allergy. It is also known that steroids frequently used for allergic diseases have a function of reducing eosinophils.

[0004]

It is thought that even if an allergen enters the body, allergic reactions can be reduced by suppressing the accumulation of eosinophils that cause inflammation at the reaction site. However, depending on the drug, there are concerns about side effects. Therefore, they try to prevent the accumulation of eosinophils in the daily diet.

[0005]

A wide variety of effects have been reported for lactic acid bacteria. In particular, it has been reported that lactic acid bacteria belonging to the genus Enterococcus have an immunopotentiating effect. It has been reported in an ACA experimental system that the anti-allergic action is enhanced by decomposing the cell wall of the cells with an enzyme (Japanese Patent Application Laid-Open No. H11-124336).
Therefore, a lactic acid bacterium whose cell wall was crushed beforehand was orally administered to an allergic model animal in which a ragweed antigen induced a type 1 allergic inflammatory reaction (peritonitis) in the mouse abdominal cavity and caused accumulation of eosinophils. However, the present invention was completed because the accumulation of eosinophils was significantly suppressed.

[0006] The total number of leukocytes in the abdominal cavity and the classification of various leukocytes were measured in the animals to which the cell wall-crushed product of the lactic acid bacterium used in the present invention was administered. It turns out that it is decreasing.
Therefore, it can be said that the product of the present invention does not have an immunosuppressive effect of reducing the amount of eosinophils due to total leukopenia, but suppresses only the accumulation of eosinophils in some way.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION As a method for crushing cell walls of lactic acid bacteria used in the present invention, any of enzyme treatment, physical crushing such as ultrasonic waves, or a combination thereof can be used. As the enzyme to be used, actinase, zymolyase, chitarase, lysozyme, mutanolysin, achromopeptidase and the like can be used, regardless of the kind, as long as they are commonly used to lyse bacteria. It is also possible to use a mixture. The purpose is to crush the lactic acid bacteria and completely remove the contents of the cytoplasm.

The lactic acid bacterium used in the present invention is a strain isolated from the stool of food or from the stool of a healthy person, so there is no risk of side effects.

In order to formulate the processed product of the cells, carriers such as starch, lactose and soy protein, excipients, binders, disintegrants, lubricants, stabilizers, and additives such as flavoring agents are used. And into tablets or granules by a well-known method.

[0010] The amount used varies depending on symptoms, age, etc.
As an active ingredient, 0.002 to 0.1 g / kg body weight per day can be usually administered to an adult once or several times a day.

[0011]

[Embodiment 1] Enterococcus faecalis NF-1011 (No. 12564, manufactured by B.I.) is inoculated into Rogosa liquid medium having the following composition (the number of bacteria: 106 cells / ml),
After culturing for 16 hours, a culture solution of about 10 9 viable cells / ml was obtained. The obtained culture was centrifuged at 12,000 rpm for 20 minutes to collect cells, and the cells were washed twice with distilled water to obtain cells.
The cells were suspended in distilled water, and mutanolysin was added to a final concentration of 30.
μg / ml, and treated at 37 ° C for 4 hours.
After heat treatment in minutes, a freeze-drying method was performed to obtain a dried bacterial cell sample.

1 shows the composition of Rogosa liquid medium. Trypticase 10 g Yeast extract 5 g Tryptose 3 g Monopotassium phosphate 3 g Dipotassium phosphate 3 g Triammonium citrate 2 g Tween 80 (surfactant) 1 g Glucose 20 g Cysteine hydrochloride 0.2 g Salt solution (as per 1) 5 ml Distilled water 1000 ml (adjusted to pH 7.0, heat sterilized at 121 ° C. for 15 minutes) (1) Salt solution: 11.5 g MgSO4.7H2O 0.68 g FeSO4.7H2O 2.4 g MnSO4.2H2O 2.4 g distilled water 100 ml

Embodiment 2 FIG. Preparation of Ultrasonic Treated Cell Preparation Enterococcus faecali (Enterococcus faecali)
s) ATCC 19433T, Enterococcus faecium (Enteroc
occus faecium) ATCC19434T, Lactobacillus casei (L
(actobacillus casei) ATCC393, Lactobacillus brevis JCM1059T, and Bifidobacterium longum JCM1217T were cultured in an MRS medium (Difco) for 24 hours. After completion of the culture, the cells were collected by centrifugation (12,000 × g, 20 minutes) and washed twice with distilled water to obtain the respective cells. These cells were suspended or dissolved in physiological saline (0.85% saline). This bacterial solution is treated with an ultrasonic crusher (Kubota 201M type)
The cells were treated at a frequency of 9 KHz at a low temperature (5 to 7 ° C.) for 50 minutes to crush the cell wall, and dried to obtain each dried cell.

Embodiment 3 FIG. Eosinophil accumulation inhibition test using mouse peritonitis model 4-week-old female BALB / C mice (Japan SLC) were divided into 7 groups after one week of preliminary breeding, and one group (control group) was diluted 10 times with ragweed antigen (Torii). The drug was subcutaneously administered on days 0, 7, and 14 of the experiment and three times during the 21-day breeding period to enhance sensitization.
On day 21, ragweed antigen was administered intraperitoneally to promote induction,
Thereafter, the total number of leukocytes and the number of eosinophils obtained by intraperitoneal washing with PBS were measured. Lactic acid bacteria cell wall crushed product is PB
The cells were suspended in S and administered daily at a dose of 60 mg / mouse using a gastric tube during the breeding period. Induction and sensitization were performed in the same manner as the control group.

FIG. 1 shows the total leukocyte count in the abdominal cavity of the mouse.
It can be seen that there is no great difference in the total leukocyte amount appearing in the abdominal cavity in any group.

FIG. 2 shows the number of eosinophils in the mouse abdominal cavity. It can be seen that the number of eosinophils decreased only by oral administration of the cells every day, suppressing allergic reactions.

No significant weight loss was observed in any of the groups during the experimental period, and no abnormalities considered to be side effects were found.

[0018]

From the above results, it can be seen that a food containing a lactic acid bacterium cell wall crushed product as a main component has an effect of preventing allergic symptoms by preventing accumulation of eosinophils and has no side effect. It can be taken for a long time with peace of mind. Moreover, since the lactic acid bacteria cell wall crushed product is physically stable and in a powdery form, it can be added to various foods.

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[Procedure amendment]

[Submission date] March 10, 2000 (200.3.1.1)
0)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Brief description of drawings

[Correction method] Change

[Correction contents]

[Brief description of the drawings]

FIG. 1 shows the total leukocyte count The number of leukocytes leached into the peritoneal cavity is shown in terms of the number per 1 ml. In the control group, only saline was orally administered daily, and ~
Administered orally 60 mg / animal of each lactic acid bacterium preparation.
No clear difference was observed in each group as compared to the control group. The vertical axis indicates the number of white blood cells leached into the abdominal cavity, cells / m.
1 and the abscissa indicates the control group and each lactic acid bacteria group.

FIG. 2 shows the number of eosinophils leached into the peritoneal cavity in terms of the number per 1 ml. Each lactic acid bacteria administration group shows a lower value compared to the control group,
In particular, significant differences were observed between and lactic acid bacteria. The vertical axis is the number of eosinophils leached into the abdominal cavity cells / ml.
, And the horizontal axis shows the control group and each lactic acid bacteria group.

[Description of Signs] *; p <0.05, **; p <0.01 vs. control group; Enterococcus faecalis NF-1011; Enterococcus faecalis ATCC1943
3T; Enterococcus faecium ATCC1943
Lactobacillus casei ATCC393; Lactobacillus brevis JCM1059T; Bifidobacterium longum JCM1217
T

[Procedure amendment 2]

[Document name to be amended] Drawing

[Correction target item name] All figures

[Correction method] Change

[Correction contents]

FIG.

FIG. 2

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Claims (2)

[Claims] Claims: 1. A food having an eosinophil accumulation-suppressing action, comprising a substance obtained by destroying the cell wall of a lactic acid bacterium as a main raw material. 2. The food having an eosinophil accumulation-inhibiting action according to claim 1, wherein the cell wall destruction method of the lactic acid bacterium is one or both of an enzyme treatment and a physical destruction treatment such as ultrasonic waves.