Anti-stress composition

Abstract

To provide an anti-stress composition.SOLUTION: A composition containing lactic acid bacteria of Lactococcus is given to a subject to exhibit an anti-stress action.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an anti-stress composition containing lactic acid bacteria of the genus Lactococcus. [Background technology]

[0002] Lactic acid bacteria is a general term for bacteria that produce lactic acid through metabolism, and are classified into lactobacilli and lactococci depending on their shape. The genus Lactococcus belongs to facultative anaerobic lactobacilli that are often used for fermenting raw milk and cheese (see Non-Patent Document 1). Health effects of ingesting lactobacilli of the genus Lactococcus have been reported, such as immunostimulatory effects (see Patent Documents 1 and 2, for example), fatigue recovery, or fatigue accumulation prevention effects (see Patent Document 3). On the other hand, there are many unknowns regarding the effectiveness of the genus Lactococcus against stress and depression, which are considered problems in modern society.

[0003] The interaction between the brain and the intestine is also called the brain-gut axis, and it is believed that information is transmitted between them via humoral factors (hormones, cytokines, etc.) and the pathways of the autonomic nervous system. Research to date has shown that the presence or absence of intestinal flora in the body changes the stress response, so it is believed that there is some kind of relationship between intestinal bacteria and stress (see Non-Patent Document 2). In addition, a composition for improving stress-induced decreased skin blood flow, which contains a fermented milk product as an active ingredient, has been disclosed (see Patent Document 4). However, it is not known at all that Lactococcus lactic acid bacteria themselves have an anti-stress effect. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2003-47462 A [Patent Document 2] JP 2003-88362 A [Patent Document 3] JP 2018-201487 A [Patent Document 4] Patent No. 3715640 [Non-patent literature]

[0005] [Non-Patent Document 1] The Science of Lactic Acid Bacteria and Bifidobacteria, edited by the Japanese Society for Lactic Acid Bacteria, Kyoto University Press, 2010 [Non-Patent Document 2] Clinical Nutrition Vol.128 No.6 (Special Issue), Hideho Ohata, Ishiyaku Publishing Co., Ltd., 2016 Summary of the Invention

[0006] The present inventors have found that administering a composition containing Lactococcus lactic acid bacteria to a subject exerts an anti-stress effect. The present invention is based on these findings.

[0007] Therefore, the present invention provides an anti-stress composition containing lactic acid bacteria of the genus Lactococcus.

[0008] According to the present invention, the following inventions are provided. (1) An anti-stress composition containing lactic acid bacteria of the genus Lactococcus. (2) The anti-stress composition according to (1), wherein the lactic acid bacteria of the genus Lactococcus are killed. (3) The anti-stress composition according to (1), wherein the lactic acid bacteria of the genus Lactococcus are heat-killed bacteria. (4) The anti-stress composition according to any one of (1) to (3), wherein the lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis NFRCL-11. (5) The anti-stress composition according to any one of (1) to (4), which is used for reducing stress. (6) The anti-stress composition according to any one of (1) to (5), which is used for promoting secretion of serotonin. (7) The anti-stress composition according to any one of (1) to (5), which is used for suppressing an increase in secretion of cortisol or corticosterone. (8) The anti-stress composition according to any one of (1) to (7), which is an anti-stress food composition. (9) An anti-stress agent containing lactic acid bacteria of the Lactococcus genus. (10) A method for reducing stress (excluding medical procedures on humans), comprising having a subject ingest lactic acid bacteria of the genus Lactococcus.

[0009] Use of the composition of the present invention is advantageous in that it exerts an anti-stress effect. [Brief description of the drawings]

[0010] [Figure 1] 1 shows the relative serotonin concentration (serotonin amount ratio) for 11 strains of lactic acid bacteria, with the serotonin concentration of the control taken as 1. In other words, a value greater than 1 indicates a higher serotonin concentration than when no lactic acid bacteria was added. [Diagram 2] Figure 2 shows the plasma corticosterone concentration (ng / ml) of mice after the forced swimming test. "*" indicates a significant difference (P<0.05) between the plasma corticosterone concentration of mice that were orally administered water and did not undergo the swimming test (shown as "Water (-FST)" in the figure) and the plasma corticosterone concentration of mice that were orally administered water (shown as "Water") or fluoxetine (shown as "FX" in the figure) and then performed the swimming test. Statistical analysis was performed using one-way ANOVA and Tukey test. In the figure, "N11" indicates the plasma corticosterone concentration of mice that were orally administered lactobacillus and then performed the swimming test. Description of the Invention

[0011] [Deposit of microorganisms] Lactococcus lactis subsp. lactis NFRCL-11 was deposited at the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (NITE) under the accession number NITE P-03141 on February 26, 2020 (date of deposit).

[0012] The anti-stress composition of the present invention contains lactococcus lactic acid bacteria, and may contain them as active ingredients. The lactococcus lactic acid bacteria contained in the composition of the present invention may be either live or killed, but is preferably killed, more preferably heat-killed, and even more preferably heat-killed powder. Heat-killed bacteria can be obtained by, for example, heating live bacteria at 95°C or higher for about 10 minutes.

[0013] The lactic acid bacteria of the genus Lactococcus contained in the composition of the present invention are not particularly limited, but examples include Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, and Lactococcus lactis subsp. hordniae, and of these, Lactococcus lactis subsp. lactis is preferred.

[0014] When the composition of the present invention contains Lactococcus lactis subsp. lactis, the Lactococcus lactis subsp. lactis is not particularly limited as long as it is Lactococcus lactis subsp. lactis, but Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NFRC3, Lactococcus lactis subsp. lactis NFRC4, Lactococcus lactis subsp. lactis NFRC7, Lactococcus lactis subsp. lactis NFRC9, Lactococcus lactis subsp. lactis NFRC10, or Lactococcus lactis subsp. lactis NFRC11 is preferred, and Lactococcus lactis subsp. lactis NBRC12007, Lactococcus lactis subsp. lactis NFRC10, or Lactococcus lactis subsp. lactis NFRC11 is more preferred, and Lactococcus lactis subsp. lactis NFRC11 is particularly preferred.

[0015] In the present invention, "stress" means functional changes occurring in the mind and body when various external stimuli act as a burden, and "anti-stress" means not causing such functional changes. "Anti-stress" means returning or bringing the functional changes to the original state after various external stimuli act as a burden and cause functional changes in the mind and body, making the functional changes in the mind and body not occur or difficult to occur even when various external stimuli act as a burden, or making the functional changes in the mind and body not occur or difficult to occur even when various external stimuli act as a burden on the mind and body in the first place. In this specification, "stress reduction" means making the functional changes in the mind and body not occur or difficult to occur even when various external stimuli act as a burden. According to a preferred embodiment of the present invention, the composition is an anti-stress composition used for stress reduction, containing lactic acid bacteria of the genus Lactococcus. According to a more preferred embodiment of the present invention, the composition is an anti-stress composition used for acute stress reduction. Here, acute stress refers to functional changes occurring in the mind and body due to short-term external stimuli, for example, those caused by external stimuli being continuously or intermittently loaded within one day, preferably within one hour, and more preferably within 10 minutes. Examples of acute stress include relatively short-term stress such as a written test, a crowded train in the morning, and stress from long hours of overtime work. Meanwhile, here, chronic stress refers to functional changes in the mind and body caused by long-term external stimuli, for example, those caused by external stimuli being continuously or intermittently loaded for a period longer than one day. Examples of chronic stress include long-term stress such as relatively long-term interpersonal conflicts at school or work. In addition, according to another more preferred embodiment of the present invention, the composition is an anti-stress composition used to reduce stress caused by mental and / or physical external stimuli.Here, factors that cause stress include mental (interpersonal conflict, overcrowded environment, etc.), physical (long hours of overtime work, excessive exercise, etc.), physical (ultraviolet rays, noise, thermal environment, etc.), chemical (chemical substances such as formaldehyde), and biological (viruses, bacteria, parasites) external stimuli, and according to a preferred embodiment of the present invention, the composition of the present invention can further reduce stress caused by mental and / or physical external stimuli among these. Note that the forced swimming test in the examples of the present application below is considered to be a mixed mental and physical stimulus.

[0016] According to a more preferred embodiment of the present invention, the anti-stress composition contains lactococcus lactic acid bacteria and is used to promote the secretion of serotonin. Here, serotonin is a kind of physiologically active amine, synthesized from tryptophan in the body, and is abundant in the brain, gastrointestinal tract, blood, etc., and acts on the neurotransmission of the brain and also has a mental stabilizing effect. In other words, serotonin can prevent or make it difficult for changes in the physical and mental functions to occur even when various external stimuli act as a burden.

[0017] According to a further preferred embodiment of the present invention, there is provided an anti-stress composition containing Lactococcus lactic acid bacteria for use in suppressing an increase in secretion of cortisol or corticosterone, where cortisol and corticosterone are types of glucocorticoids, which are adrenal cortical hormones, and are closely related to mental stability, with cortisol being a hormone found in primates such as humans, and corticosterone being a hormone found in rodents such as rats and mice.

[0018] The content of Lactococcus lactic acid bacteria in the composition of the present invention is not particularly limited. For example, the content of Lactococcus lactic acid bacteria in the composition of the present invention is 1×10 9 1×10 or more, preferably 1×10 9 ~1×10 12 1×10 9 ~1×10 11 1×10 9~1×10 10 1×10 9 ~5×10 9 These contents may be the contents when heat-treated killed cell powder is included.

[0019] The composition of the present invention may be used with Lactococcus lactic acid bacteria alone or in combination with other ingredients, such as amino acids such as γ-aminobutyric acid (GABA) and theanine, and antioxidants such as ubiquinol (reduced coenzyme Q10).

[0020] According to another preferred embodiment of the present invention, an anti-stress food composition containing Lactococcus genus lactic acid bacteria is provided. The composition containing Lactococcus genus lactic acid bacteria can be contained in food and provided as a food composition. The food containing the composition of the present invention is not particularly limited, but examples thereof include supplements, beverages, fermented foods, rice, bread, noodles, and confectionery.

[0021] According to another preferred embodiment of the present invention, there is provided an anti-stress agent comprising a lactic acid bacterium belonging to the genus Lactococcus. The anti-stress agent of the present invention may contain a lactic acid bacterium belonging to the genus Lactococcus as an active ingredient.

[0022] According to another preferred embodiment of the present invention, there is provided an anti-stress pharmaceutical composition (drug) containing Lactococcus genus lactic acid bacteria. A pharmaceutical (pharmaceutical composition) is prepared as an oral or parenteral formulation in accordance with a conventional method by using additives acceptable for formulation. When the pharmaceutical is an oral formulation, it can take the form of a solid formulation such as a tablet, powder, fine granules, granules, capsules, pills, or sustained release agent, or a liquid formulation such as a solution, suspension, or emulsion. From the viewpoint of ease of ingestion (administration) to patients, oral formulations are preferred for pharmaceuticals. Examples of additives acceptable for formulation include excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, flavorings, buffers, antioxidants, and pH adjusters.

[0023] According to another aspect of the present invention, there is provided a method for reducing stress, comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of stress relief). The lactic acid bacterium of the genus Lactococcus may be ingested once or multiple times, but even a single ingestion is advantageous in that the effect of the present invention can be achieved. According to a preferred aspect of the present invention, there is provided a method for reducing stress (excluding medical procedures for humans) comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of stress relief). In this specification, "medical procedures for humans" refers to the act of having a human ingest (administer) a medicine, etc., as prescribed by a doctor or the like. Subjects also include animals other than humans (livestock animals such as horses and cows, pet animals such as dogs and cats, ornamental animals kept in zoos, etc.).

[0024] According to another preferred embodiment of the present invention, there is provided a method for promoting serotonin secretion, comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of anti-stress). According to another more preferred embodiment of the present invention, there is provided a method for promoting serotonin secretion (excluding medical procedures for humans), comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of anti-stress).

[0025] According to another preferred embodiment of the present invention, there is provided a method for suppressing increased secretion of cortisol or corticosterone, comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of stress relief). According to another more preferred embodiment of the present invention, there is provided a method for suppressing increased secretion of cortisol or corticosterone (excluding medical procedures for humans), comprising ingesting a lactic acid bacterium of the genus Lactococcus to a subject (preferably a subject in need of stress relief).

[0026] According to another aspect of the present invention, there is provided use of a lactic acid bacterium of the genus Lactococcus for the manufacture of a food composition or pharmaceutical composition for anti-stress.

[0027] According to another aspect of the present invention, there is provided a use of a lactic acid bacterium of the genus Lactococcus for anti-stress purposes.

[0028] According to another aspect of the present invention, there is provided a lactic acid bacterium of the genus Lactococcus for anti-stress purposes.

[0029] The Lactococcus lactic acid bacteria in the anti-stress agent and anti-stress pharmaceutical composition of the present invention may be the same as those in the composition of the present invention. EXAMPLES

[0030] The present invention will be described in more detail below with reference to examples, but the technical scope of the present invention is not limited to these examples.

[0031] Test Example 1: Evaluation of serotonin (5-HT) secretion effect of lactic acid bacteria As cultured cells, RIN-14B cells (intestinal secretory cell-like cultured cells) (ATCC A American T type C ulture C The lactic acid bacteria used were obtained from the Niigata Prefectural Agricultural Research Institute Food Research Center (NFRC strains) and 5 strains from the National Institute of Technology and Evaluation (NBRC strains), for a total of 11 strains. Killed bacterial powders that had been heat-treated at 95°C or higher for 10 minutes were used in the following tests. The 11 strains of lactic acid bacteria used in the tests are listed in Table 1 and Figure 1 below.

[0032] Add 2 x 10 RIN-14B cells to each well of a cell culture plate. 5The cells were seeded at 100 μg / mL per well, and after seeding, the killed lactic acid bacteria powder was added at 100 μg / mL. The control was a sample to which the killed lactic acid bacteria powder was not added. After the addition of the killed lactic acid bacteria powder, the sample was cultured in an incubator at 37°C and 5% CO2 for 1 hour. After the culture, the concentration of serotonin secreted into the cell supernatant was measured by ELISA using an ELISA kit (Enzo) and compared with the control. The results are shown in Table 1 and Figure 1 below.

[0033] [Table 1]

[0034] From the results in Table 1 and Figure 1, Lactococcus lactis subsp. Lactis NBRC12007, Lactococcus lactis subsp. Lactis NFRC3, Lactococcus lactis subsp. Lactis NFRC4, Lactococcus lactis subsp. Lactis NFRC7, Lactococcus lactis subsp. Lactis NFRC9, Lactococcus lactis subsp. Lactis NFRC10, and Lactococcus lactis subsp. Lactis NFRC11 have a serotonin ratio higher than 1.2 and thus have a high amount of serotonin secretion, and therefore have a high anti-stress effect, while Lactococcus lactis subsp. Lactis NBRC12007, Lactococcus lactis subsp. Lactis NFRC10, and Lactococcus lactis subsp. Lactis NFRC11 has a serotonin ratio higher than 2.0, resulting in a high level of serotonin secretion, and therefore a stronger anti-stress effect, while Lactococcus lactis subsp. Lactis NFRC11 has a serotonin ratio higher than 3.0, resulting in a high level of serotonin secretion, and therefore a particularly strong anti-stress effect.

[0035] Test example 2: Forced swimming test Eight-week-old BALB / c mice (obtained from CLEA Japan) were divided into four groups (n=6-7 per group), and the test was carried out according to the test procedure in Table 2 below. [Table 2]

[0036] The test groups are as follows: Group 1: Oral administration of water (forced swim test (FST) not performed) (-FST) Group 2: Oral administration of water (forced swimming test) (+FST) Group 3: Oral administration of fluoxetine (20 mg / kg-BW) (positive control; commercially available as an antidepressant) (forced swimming test) (+FST) Group 4: Oral administration of lactic acid bacteria (Lactococcus lactis subsp. lactis NFRCL-11) (forced swimming test) (+FST) * The lactic acid bacteria of Group 4 were heat-sterilized and administered at a dose of 1 x 10 per mouse. 9 The tablets were prepared so that each dose was one piece and administered orally.

[0037] The forced swimming test was performed by placing the mice in a water tank (cylinder size: φ20 × 500 mm, water temperature: 23-25 ​​°C, water volume: 15 cm). Immediately after the forced swimming test, blood was collected and the plasma corticosterone concentration (ng / ml) was measured using an ELISA kit (Enzo). The results are shown in Figure 2.

[0038] According to the results of the forced swimming test, the plasma corticosterone concentration was significantly increased in Group 2 (oral administration of water (forced swimming test performed)) and Group 3 (oral administration of fluoxetine (forced swimming test performed)) compared to Group 1 (oral administration of water (forced swimming test not performed)). On the other hand, the plasma corticosterone concentration was lower in Group 4 (oral administration of lactobacillus (forced swimming test performed)) compared to Groups 2 and 3, and no significant increase was observed compared to Group 1.

[0039] Therefore, it was found that by using a composition containing Lactococcus lactic acid bacteria (in particular, Lactococcus lactis subsp. lactis NFRCL-11) as an active ingredient, the plasma corticosterone concentration was reduced and the stress caused by swimming was alleviated, and that the composition can be used as an anti-stress composition.

Claims

1. An anti-stress composition comprising a lactic acid bacterium of the genus Lactococcus, The anti-stress composition, wherein the lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis NFRCL-11, which is deposited at the Patent Microorganisms Depositary Center under the accession number NITE P-03141.

2. The anti-stress composition according to claim 1 , wherein the lactic acid bacteria of the genus Lactococcus are killed.

3. The anti-stress composition according to claim 1, wherein the lactic acid bacteria of the genus Lactococcus are heat-killed bacteria.

4. The anti-stress composition according to any one of claims 1 to 3, which is used for reducing stress.

5. The anti-stress composition according to any one of claims 1 to 4, which is used for promoting secretion of serotonin.

6. The anti-stress composition according to any one of claims 1 to 4, which is used for suppressing an increase in secretion of cortisol or corticosterone.

7. The anti-stress composition according to any one of claims 1 to 6, which is an anti-stress food composition.

8. An anti-stress agent comprising a lactic acid bacterium of the genus Lactococcus, The anti-stress agent, wherein the lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis NFRCL-11, which is deposited at the Patent Microorganisms Depositary Center under the accession number NITE P-03141.

9. A method for reducing stress, comprising administering a lactic acid bacterium of the genus Lactococcus to a subject (excluding humans), The method for reducing stress, wherein the lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis NFRCL-11, which is deposited at the Patent Microorganisms Depositary Center under the accession number NITE P-03141.

10. Lactococcus lactis subsp. lactis NFRCL-11 strain, deposited at the Patent Microorganisms Depositary Center under the accession number NITE P-03141.

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