JPH1149697A - Stress-suppressant and stress suppressant food - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a stress-suppressant and a stress-suppressant food that is excellent in safety and can effectively suppress the stress by using a specific substance obtained from glutene as an active ingredient. SOLUTION: This stress suppressant agent and stress suppressant food are obtained by using a glutene hydrolyzate (suitably containing peptides with a molecular weight of 100-3,000). Glutene is hydrolyzed with an enzyme (particularly an endo type protease is suitable) under mild conditions. The daily dose of the glutene hydrolyzate to an adult human is 5-100 grams.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a stress suppressor using a gluten hydrolyzate and a food for suppressing stress.

[0002]

2. Description of the Related Art Recently, in a complicated modern society, there is a concern that the application of stress may adversely affect the human body.
When the living body receives an excessive stress stimulus, the adaptation to the stress cannot be maintained, and a severe stress state, that is, a state in which the mind and body are broken down or a resistance to another stress stimulus is weakened is exhibited. It is in such cases that stress is closely linked to the onset and progression of the disease, and therefore several stress suppressants have been proposed for the prevention and treatment of the disease.

[0003]

However, the conventional stress suppressants are not always expected to have a sufficient effect, and also have a problem in safety, and in particular, side effects cannot be overlooked. . In fact, little was known about foods for suppressing stress.

[0004] Accordingly, an object of the present invention is to provide a stress inhibitor and a food for stress suppression that are excellent in safety and can effectively suppress stress.

[0005]

Means for Solving the Problems In view of the above-mentioned situation, the present inventors have conducted intensive studies and have surprisingly found that gluten hydrolyzate is
The present inventors have found that the present invention is effective for prevention and treatment of various diseases caused by stress, and completed the present invention.

That is, the present invention provides a stress suppressant containing a gluten hydrolyzate as an active ingredient and a food for suppressing stress containing a gluten hydrolyzate.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION Wheat is generally classified into spring wheat, winter wheat, hard wheat, soft wheat, and red wheat and white wheat depending on the seed color. There is no particular limitation on the wheat used as the raw material of the product, and any type of wheat can be used. Gluten can be produced from wheat flour according to a conventional method.

As a method for decomposing gluten, various methods such as a method using an enzyme, a method using an acid, and a method using an alkali are known. In the present invention, any of these methods can be used. The decomposition method is desirable because the desired decomposition product can be obtained efficiently and reliably under mild conditions. When degraded by enzymes, endo-type proteases are suitable.Examples of the endo-type proteases include pepsin of animal origin, papain and bromelain of plant origin, aspartic protease of microbial origin, metalloprotease and the like. Can be.
Among them, so-called acidic proteases having an optimal pH on the acidic side, such as pepsin and aspartic protease of microbial origin, are preferred. The protease may be used in a free state or may be used in a fixed state. The amount of protease used is 25-500 units per gram of gluten.
s, especially 50 to 150 units. The decomposition reaction is usually
Gluten is dispersed in water or the like to form an aqueous dispersion, the pH of the solution is adjusted to about 1.5 to 5.0, and the temperature is adjusted to about
It is good to perform for 40 hours. By appropriately selecting the reaction conditions, a decomposition product having a desired molecular weight distribution can be obtained. The obtained decomposed product may be fractionated and purified by means such as filtration with an ultrafiltration membrane or gel filtration. Of these, the molecular weight is 100 to 30,000, preferably 50
The peptide preferably contains 70% by weight or more, particularly 80% by weight or more, of the peptide in the range of 0 to 10,000.

Here, the molecular weight and molecular weight distribution of the gluten hydrolyzate in the present invention were measured by the following methods. (Method for Measuring Molecular Weight and Molecular Weight Distribution) (1) Preparation of Calibration Curve Size exclusion high performance liquid chromatography (S) using the following three kinds of substances having known molecular weights as molecular weight markers:
E-HPLC) was performed under the following measurement conditions to prepare a calibration curve. Molecular weight marker ribonuclease A (RNase A) (molecular weight 13,7
00; Pharmacia Fine Chemicals Co., Ltd.) Aprotinin (molecular weight 6,500; Sigma Chemical Co.) Bradykinin (molecular weight 1,060; Peptide Research Laboratories) Measurement conditions used Column: Asahipak320 (Asahi Kasei Corporation) Eluent: 40% acetonitrile solution containing 0.1% trifluoroacetic acid Eluent flow rate: 1.0 ml / min Detection method: 2 using L4000UV (manufactured by Hitachi, Ltd.)
Measure UV absorbance at 20 nm. Preparation of calibration curve As a result of the above measurement, RNase A (molecular weight 13,700)
At the eluent volume of 8.14 ml, aprotinin (molecular weight 6,500) at the eluent volume of 8.88 ml,
Since bradykinin (molecular weight 1,060) had an ultraviolet absorption of 220 nm at 10.50 ml of the elution volume solution, the molecular weight [log (MW)] was plotted on the semi-log paper, and the eluent volume (ml) was plotted. When the values were plotted on the horizontal axis, linearity was recognized. Therefore, linear regression was performed and a straight line was drawn. This calibration straight line was used as a calibration curve.

(2) Measurement of molecular weight and molecular weight distribution of gluten hydrolyzate Gluten hydrolyzate was subjected to size exclusion high performance liquid chromatography in the same manner as for each of the above molecular weight markers. Was measured for ultraviolet absorbance. Then, the vertical axis is the ultraviolet absorbance, and the horizontal axis is the ultraviolet absorption curve as the eluent volume (ie, molecular weight) on the same scale as the calibration curve created above,
The molecular weight distribution of the gluten hydrolyzate was measured.

The gluten hydrolyzate thus obtained has an excellent stress-suppressing action, and is useful as a stress-suppressing agent for preventing or treating stress, or as a food for stress-suppression by being added to food. . In addition, such a gluten hydrolyzate is obtained by simply decomposing gluten, which has been generally used as food, by the above-described method or the like, and is excellent in safety.

The stress suppressor of the present invention may contain excipients, stabilizers, preservatives, binders, disintegrants, and the like, which are mixed with ordinary drugs. The dosage form is not particularly limited, and may be an appropriate dosage form such as a liquid, capsule, granule, pill, powder, tablet and the like. The method of administration is not particularly limited, and an appropriate administration method such as oral administration and enteral administration can be selected.

The food for suppressing stress of the present invention can be obtained by appropriately mixing a gluten hydrolyzate with other foods. The form is not particularly limited, and may be any form such as liquid, paste, and solid.

The administration of gluten hydrolyzate to humans depends on the age,
The dosage and administration should be determined in consideration of body weight, symptoms, etc. In many cases, the effective dose is 5 to 10 per adult per day.
It is preferably 0 g, particularly preferably 10 to 50 g. It is appropriate that these are divided and administered before or after a meal or with a meal.

[0015]

Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.

Example 1 Preparation of Gluten Decomposed Product 534 g of powdered gluten was dispersed in 3500 ml of 0.02N citric acid aqueous solution with stirring to prepare an aqueous gluten dispersion having a pH of 4.0. 0.8 g (75 units) of acidic protease (Amano M: manufactured by Amano Pharmaceutical Co., Ltd.)
/ Gluten 1 g) and reacted at 45 ° C. for 5 hours. After the reaction, the pH of the solution was adjusted to 5.0,
The acid protease was inactivated by heating for minutes. After cooling,
Centrifugation was performed at 5,000 G for 20 minutes to remove undegraded precipitate. To 3200 ml of the supernatant, 12 g of activated carbon (Takecol: manufactured by Takeda Pharmaceutical Co., Ltd.) and 60 g of diatomaceous earth (Radiolite: manufactured by Showa Chemical Co., Ltd.) were added, and the mixture was added at room temperature.
Stirred for hours. After filtration through filter paper, the pore size was further increased to 0.
Membrane filtration was performed using a 45 μm membrane filter. The obtained clear liquid was spray-dried to obtain 300 g of a powder of a gluten decomposition product. It has a molecular weight of 500-1.
Peptides (weight average molecular weight 50
00) was 81.5% by weight.

A 25% egg white protein diet shown in Table 1 was used as a control diet, and two thirds of the egg white protein of the control diet was replaced with the above-prepared gluten hydrolyzate. Invention).

[0018]

[Table 1]

Test Example 1 Twelve Wistar male rats (Kudo, Kumamoto) weighing about 200 g were divided into two groups of 6 rats each at 25 ± 2 ° C. and a humidity of 5
They were bred under the conditions of 0 to 70% and lighting time of 8 to 20 hours / day. First, the above-mentioned rats were bred with commercially available feed (Oriental Yeast Co., Ltd.) for one week after being brought into the same facility, and adapted to the environment. Next, the rats of the first group were given a 25% egg white protein diet freely at 17 to 18 o'clock every evening and bred for 28 days (control), and the rats of the second group were given the above-mentioned food for stress suppression. They were fed and bred in the same manner as in the control section (section of the present invention). Next, the inhibitory effect of gastric ulcer caused by stress load was measured from the ratio of ulcer area by the following method.

Method for measuring gastric ulcer inhibitory effect: Taka
gi et al. (Takagi, K. et al: Jpn. J. Pharmacol, 1).
8; 9-18 (1968)) to reduce the water immersion restraint stress by a stress cage (stainless steel wire cage).
Loaded for hours. That is, the rats were restrained in a standing position so that they could not move in the stress cage, and were immersed in water at a water temperature of 21 to 23 ° C. to the neck height of the rats. The laboratory was dark and quiet during stress loading, and the room temperature was 25 ± 2 ° C. Fasting was performed during stress loading. After the end of the stress load, the rat was quickly released from the restraint, the occiput was hit hard, and the rats were stunned by concussion. Thereafter, the rat was decapitated with a rat guillotine, and the stomach was separated from the trunk into physiological saline. Then Kanazawa's method (Hiroshi Kanazawa, Journal of the Japanese Society of Gastroenterology, 77, 1
345-1354 (1980)), the ratio of the gastric ulcer area was calculated from the stomach extracted. That is, first, the esophagus is ligated using forceps, and 10% formalin 10
ml was injected, and the same part was ligated using forceps. After leaving it to stand for about 30 minutes and fixing, it was incised from the duodenum to the esophagus along the greater curvature side, attached to a styrofoam plate with a pin, and further fixed in 10% formalin. A photograph of the fixed stomach was taken, 1 mm ² of bleeding spots remaining on the gastric mucosa was counted and counted, and the total area was defined as the ulcer area. Further, the mucous membrane of the whole stomach was quantified by the same method, and the ratio of the ulcer to the entire mucous membrane (the ratio of the ulcer area) was calculated.

The results are shown as an average value of 6 rats ± standard error. In the first group (control group), 26.5 ± 3.8.
% In the second group (section of the present invention), 10.8 ± 2.4%
In the plot of the present invention, the ratio of the ulcer area was significantly lower than that in the control plot (p <0.05).

Test Example 2 In the rats of the first group (control group) and the second group (group of the present invention) in Test Example 1, suppression of the grooming time, which is considered to be increased by stress, is performed by the following method. The effect was measured.

Method for measuring the effect of suppressing grooming: Before stress, rats of group 1 (control group) and group 2 (invention group) perform intermittent grooming for 10 minutes. The time was measured. Next, except that the load time was set to 3 hours,
A stress load was applied in the same manner as in Test Example 1. Thereafter, the rats were quickly released from the restraint, and given a one-hour rest in the same room, and the grooming time of each group of rats was measured in the same manner as described above. The results are shown in Table 2 as an average value of 6 rats ± standard error.

[0024]

[Table 2]

As shown in Table 2, due to the stress load,
In the first group (control group), the grooming time increased. On the other hand, in the second group (section of the present invention), no increase in grooming time was observed. There is no statistically significant difference between the two groups (p> 0.
05), Table 2 shows that the addition of the gluten hydrolyzate can suppress an increase in grooming time caused by stress load.

Reference Example 1 Table 3 shows the body weight of each group of rats immediately before starting administration of a 25% egg white protein diet or the like and immediately after administration for 28 days.

[0027]

[Table 3]

From Table 3, no significant difference was observed in the growth of rats between the two groups (p> 0.05), indicating that there was a difference between the 25% egg white protein diet and a part thereof replaced with gluten hydrolyzate. In addition, it is considered that there was no effect on rat growth.

[0029]

EFFECTS OF THE INVENTION The use of the stress suppressant and the food for suppressing stress according to the present invention makes it possible to provide excellent safety and effectively suppress stress.

Claims (4)

[Claims] 1. A stress inhibitor comprising a gluten hydrolyzate as an active ingredient. 2. The gluten hydrolyzate has a molecular weight of 100 to 30.
The stress inhibitor according to claim 1, which contains a peptide having a molecular weight of 000. 3. A food for suppressing stress containing a gluten hydrolyzate. 4. The gluten hydrolyzate has a molecular weight of 100 to 30.
The food for stress suppression according to claim 3, which contains a peptide of 000.