JP5592640B2 - Antistress agent containing lactic acid bacteria fermented royal jelly, method for producing the same, hypothalamus-pituitary-adrenocortical activity inhibitor, and sympathetic-adrenal medullary activity inhibitor - Google Patents

Description

The present invention uses a royal jelly as a raw material, an anti-stress agent containing a lactic acid bacteria fermented royal jelly obtained by fermenting using lactic acid bacteria as an active ingredient , a production method thereof , an activity inhibitor of the hypothalamus-pituitary-adrenocortical system, And a sympathetic-adrenal medullary activity inhibitor .

  Generally, royal jelly is a jelly-like substance made by mixing secretions secreted from the hypopharyngeal gland and the greater vagina by female honeybees 3 to 15 days after emergence, and contains specific proteins, fatty acids, minerals, etc. It is known that Royal jelly is known to have various physiological effects such as blood pressure lowering action, antitumor action, wound healing promotion, antibacterial action and the like. Therefore, royal jelly has been used not only for health foods with high nutritional value but also for applications such as pharmaceuticals and cosmetics.

  Conventionally, various processes have been performed in order to improve the function of royal jelly. For example, methods disclosed in Patent Documents 1 and 2 are known. Patent Document 1 discloses the production of a royal jelly extract obtained by adding, for example, water-containing ethanol to fresh royal jelly or dried royal jelly to prepare a mixed solution, dissolving the soluble components in the royal jelly in water-containing ethanol, and then filtering the solution. A method is disclosed. Royal jelly extract contains fatty acids peculiar to royal jelly such as decenoic acid, lipids composed of esters thereof, special water-soluble proteins contained only in royal jelly, amino acids, carbohydrates, minerals and the like.

  Patent Document 2 discloses a cosmetic containing a fermented product obtained by fermenting royal jelly with microorganisms. Such a fermented product improves the skin beautifying effect by a fibroblast activation effect and an intracellular tyrosinase activity suppression effect.

JP 2000-60455 A JP 2006-219434 A

  By the way, modern humans are irritating to various living organisms (stress), for example, physical and chemical stress such as noise and air pollution, physiological stress such as overwork and lack of sleep, and psychological such as anxiety and tension.・ Exposed to social stress. When a living body continuously receives stress or receives a large amount of stress at once, the body and mind may be distorted, and the mind and body cannot be tolerated and stress disease may develop.

  It is known that when a living body receives stress, the activities of the sympathetic nerve-adrenal medullary system and the hypothalamus-pituitary-adrenal cortex system increase. The increase in the sympathetic-adrenal medullary system causes symptoms such as increased blood pressure, sweating, salivation, and arousal. Increased hypothalamic-pituitary-adrenocortical system, for example, promotes the secretion of adrenocorticotropic hormone (ACTH) from the anterior pituitary gland, and levels of glucocorticoids such as cortisol, corticosterone, and cortisone in the blood Rises. Glucocorticoids exert, for example, a gluconeogenesis-promoting action and an anti-inflammatory action, but if they are secreted excessively, they may worsen diabetes or worsen infectious diseases.

  The present invention is based on the discovery that ingesting lactic acid-fermented royal jelly alleviates the stress reaction that occurs in vivo and exhibits an excellent anti-stress effect.

The object of the present invention is to provide an anti-stress agent that can be used for various uses such as foods and beverages and pharmaceuticals , a method for producing the same , a hypothalamus-pituitary-adrenocortical activity inhibitor, and a sympathetic nerve- It is to provide an adrenal medullary activity inhibitor .

To achieve the above object, the anti-stress agent of claim 1, royal jelly and lactic acid bacteria fermentation royal jelly obtained by fermenting with lactic acid bacteria, the lactic acid bacteria Gala Kutobachirusu sakei, La Kutobachirusu Acid Filth, Bae At least one selected from Diococcus acidilacticity, Streptococcus thermophilus, and Bifidobacterium longum, and a hypothalamic-pituitary-adrenocortical activity inhibitor or sympathetic nerve in a psychological stress state- It is applied as an activity inhibitor of adrenal medullary system.

The invention according to claim 2, royal jelly and lactic acid bacteria fermentation royal jelly obtained by fermenting with lactic acid bacteria, the lactic acid bacteria Gala Kutobachirusu sakei, La Kutobachirusu Acid Filth, Pediococcus acidilactici, Streptococcus A hypothalamic-pituitary-adrenocortical activity inhibitor in a psychological stress state, which is at least one selected from Thermophilus and Bifidobacterium longum.

The invention according to claim 3, royal jelly and lactic acid bacteria fermentation royal jelly obtained by fermenting with lactic acid bacteria, the lactic acid bacteria Gala Kutobachirusu sakei, La Kutobachirusu Acid Filth, Pediococcus acidilactici, Streptococcus A sympathetic-adrenal medullary activity inhibitor in a psychological stress state, which is at least one selected from Thermophilus and Bifidobacterium longum.

Invention of Claim 4 in the manufacturing method of the anti-stress agent applied as activity inhibitor of hypothalamus-pituitary-adrenal cortex system or activity inhibitor of sympathetic nerve-adrenal medullary system in psychological stress state, the royal jelly, without the addition of glutamic acid, Lactobacillus sakei, La Kutobachirusu acid Filth, Pediococcus acidilactici, Streptococcus thermophilus, and with at least one of lactic acid bacteria selected from the group consisting of Bifidobacterium longum It is characterized by being fermented.

ADVANTAGE OF THE INVENTION According to this invention, the anti-stress agent which can be utilized for various uses, such as food / beverage products and a pharmaceutical , its manufacturing method , the hypothalamus-pituitary-adrenal cortex activity inhibitor, and the sympathetic-adrenal medullary system It is possible to provide an activity inhibitor .

The data which shows content of GABA in 100g of solid content of the lactic acid fermentation royal jelly in Experiment 1. FIG. The vertical axis represents GABA content (mg). Data showing the corticosterone concentration in serum in Test Example 2. The vertical axis represents the serum corticosterone concentration (ng / mL). Data showing the increase rate of amylase activity in saliva in Test Example 3. The vertical axis indicates the value (%) when the value before stress loading is 100%.

Hereinafter, embodiments embodying the antistress agent of the present invention will be described.
The anti-stress agent of this embodiment contains royal jelly as a raw material, and contains lactic acid bacteria fermented royal jelly obtained by fermenting the royal jelly with lactic acid bacteria as an active ingredient.

  As the royal jelly as a raw material, both raw royal jelly and royal jelly powder (FD-RJ) obtained by drying and pulverizing raw royal jelly by freeze drying or the like can be used. The production area of the raw royal jelly and the dried royal jelly used in the present embodiment is not particularly limited, and may be any of Asian countries such as China, Taiwan and Japan, European countries, North American countries, and South American countries such as Brazil.

  The lactic acid bacteria used for fermentation are not particularly limited, and known lactic acid bacteria used in general foods such as dairy products, fish / livestock products, brewed products, and pickles can be used. Examples include Lactobacillus, Pediococcus, Streptococcus, Bifidobacterium, and Leuconostoc. Among these, Lactobacillus genus, Pediococcus genus, Streptococcus genus, and Bifidobacterium genus are preferable from the viewpoint of excellent antistress action. Examples of the genus Lactobacillus include Lactobacillus plantarum, Lactobacillus sakei, Lactobacillus delbruecki, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus brevis, and Lactobacillus bulgaricus. Examples of the genus Pediococcus include Pediococcus acidilacticity, Pediococcus cerevisiae, Pediococcus sogee, and Pediococcus halophyllus. Examples of the genus Streptococcus include Streptococcus thermophilus, Streptococcus lactis, Streptococcus cremoris, and Streptococcus faecalis. Examples of the genus Bifidobacterium include Bifidobacterium longum, Bifidobacterium infantis, and Bifidobacterium breve.

  The fermentation treatment can be performed by inoculating lactic acid bacteria into a solution obtained by diluting a royal jelly raw material with a suitable solvent such as water at a predetermined magnification and culturing at a predetermined temperature. The dilution ratio of the royal jelly raw material is not particularly limited, but in the case of powdered (freeze-dried) royal jelly (FD-RJ), the dilution is preferably 5 to 20 times, more preferably 8 to 15 times. When the dilution ratio is less than 5 times, the increase in the viscosity of the reaction solution due to royal jelly cannot be sufficiently suppressed, so that it is difficult to rapidly proceed the fermentation treatment. On the other hand, when the dilution rate exceeds 20 times, it may take a lot of time to concentrate the obtained fermentation-treated royal jelly. In the case of raw royal jelly, the dilution is preferably 1 to 10 times, more preferably 2 to 5 times. When the dilution factor is less than 1, the increase in the viscosity of the reaction solution due to royal jelly cannot be sufficiently suppressed, so that it is difficult to rapidly proceed the fermentation treatment. On the other hand, when the dilution rate exceeds 10 times, it may take a lot of time to concentrate the obtained fermentation-treated royal jelly.

  The culture conditions for lactic acid bacteria can be appropriately set depending on the type of lactic acid bacteria. The culture temperature is not particularly limited as long as it is a temperature at which lactic acid bacteria can grow, but is preferably 20 to 45 ° C, more preferably 25 to 40 ° C. It is. The culture time can be appropriately set according to the type of lactic acid bacteria and the culture temperature, but preferably 1 to 5 days, more preferably about 2 to 4 days. If the culture period is short, the lactic acid bacteria cannot grow sufficiently, and if the culture period is long, the growth of the lactic acid bacteria enters a decline period and the fermentation treatment rate decreases. For the culture, either shake culture or static culture can be used.

  Lactic acid bacteria fermented royal jelly after fermentation treatment may be applied to food and drink, pharmaceuticals, etc. as it is, or may be applied by evaporating the solvent and concentrated, or applied after drying and powdering Good. Moreover, the lactic acid bacteria fermented royal jelly after the fermentation treatment may be applied to foods and drinks, pharmaceuticals, etc. with the lactic acid bacteria in a live bacteria state, and may be applied to various uses after sterilization treatment, for example, boiling.

  The lactic acid bacteria fermented royal jelly obtained as described above has a high anti-stress effect. Therefore, it can be applied as an anti-stress agent for the purpose of obtaining those effects. As a specific blending form, it can be applied as a food / beverage product, a pharmaceutical product, a quasi-drug, and the like for the purpose of obtaining those effects.

  When applying the anti-stress agent of this embodiment to food-drinks, it can be used by adding to various food materials or beverage materials. The form of the food or drink is not particularly limited, and may be any of liquid, powder, gel, and solid, and the dosage form is any of tablets, capsules, granules, and drinks. May be. Among these, a capsule is preferable because hygroscopicity is suppressed. As said food-drinks, you may mix | blend gelatinizer containing foodstuffs, saccharides, a fragrance | flavor, a sweetener, fats and oils, a base material, an excipient | filler, a food additive, a subsidiary material, a bulking agent etc. suitably as another component.

  When using the anti-stress agent of this embodiment as a pharmaceutical or a quasi-drug, it is possible to employ administration methods such as enteral administration in addition to administration by taking (oral intake). The anti-stress agent of this embodiment is desirably administered by oral intake. Although it does not specifically limit as a dosage form, For example, a powder, a powder, a granule, a tablet, a capsule, a pill, a liquid agent etc. are mentioned. Moreover, you may mix | blend an excipient | filler, a base, an emulsifier, a solvent, a stabilizer etc. as an additive.

The effects exhibited by this embodiment will be described below.
(1) In this embodiment, lactic acid bacteria fermented royal jelly obtained by fermenting royal jelly with lactic acid bacteria can alleviate various stress reactions that occur in vivo. Therefore, the lactic acid bacteria fermented royal jelly of this embodiment can be preferably applied to the fields of foods and drinks and pharmaceuticals as an anti-stress agent for the purpose of exerting an anti-stress action.

  (2) The anti-stress agent of this embodiment exhibits the effect | action which relieve | moderates the stress reaction which occurs in the living body. Specifically, the activity of the sympathetic nerve-adrenal medulla system and the hypothalamus-pituitary-adrenal cortex system that occur when excessive stress is applied can be suppressed from increasing. Therefore, it is expected to relieve symptoms caused by an increase in the sympathetic nerve-adrenal medullary system, such as an increase in blood pressure, sweating, saliva secretion, arousal and the like. It is also expected to suppress symptoms caused by increased activity of the hypothalamus-pituitary-adrenal cortex, for example, an increase in blood glucocorticoid concentration.

  (3) In this embodiment, lactic acid bacteria fermentation royal jelly obtained by fermenting royal jelly with lactic acid bacteria can increase the content of γ-aminobutyric acid (GABA). Therefore, it is expected to exert a stress reduction effect by ingestion.

  (4) In the present embodiment, preferably, at least one selected from Lactobacillus, Pediococcus, Streptococcus, and Bifidobacterium is used as the lactic acid bacterium. Therefore, the anti-stress action can be further improved.

  (5) In this embodiment, preferably, Lactobacillus plantarum, Lactobacillus sakei, Lactobacillus delbruecki, Lactobacillus acidophilus, Pediococcus acidilacty, Streptococcus thermophilus, and lactic acid bacteria, At least one selected from Bifidobacterium longum is used. Therefore, the anti-stress action can be further improved.

  (6) The antistress agent of this embodiment uses royal jelly as a naturally derived raw material. Therefore, it can be ingested safely and without causing side effects as compared with known synthetic pharmaceuticals such as tranquilizers.

  (7) The antistress agent of this embodiment can be preferably applied as a blood glucocorticoid elevation inhibitor. Therefore, especially when glucocorticoid is secreted excessively, it can be preferably applied to patients who may worsen diabetes or worsen infectious diseases.

  (8) The antistress agent of this embodiment uses royal jelly as a raw material. Therefore, intake of nutrients necessary for the growth of proteins, vitamins, minerals, amino acids, fatty acids and the like can also be expected. In addition, in addition to the physiological actions derived from royal jelly, such as improvement of immune function and nutritional tonic, the effects of improving neuralgia, hepatitis, climacteric disorder and osteoporosis can be expected by ingestion.

In addition, you may change the said embodiment as follows.
-The lactic acid-fermented royal jelly in the above embodiment can be applied not only to foods and drinks and medicines taken by humans, but also to livestock feeds as supplements, dietary supplements, medicines and the like. For example, it is known that secretion of glucocorticoids is promoted when stress is applied to animals other than humans. Since the increase in the glucocorticoid concentration stimulates the degradation of skeletal muscle, it is considered that lactic acid fermentation royal jelly is applied as a meat deterioration inhibitor for the purpose of exerting an inhibitory action on glucocorticoid secretion.

Examples are given below to describe the above embodiments more specifically, but the present invention is not limited to these examples. Hereinafter, Example 1 shall be read as Reference Example 1.
Example 1
First, 90 g of purified water was added to 10 g of lyophilized royal jelly (FD-RJ) (diluted 10 times), and stirred with a stirrer to prepare a 10% FD-RJ solution. Next, this solution was adjusted to pH 7.0 with a sodium hydroxide solution for food addition, and then sterilized by autoclaving. Then, after adjusting the pH to 7.0 again, 50 mg of Lactobacillus plantarum (LP115, manufactured by Kyowa High Foods Co., Ltd.) as a lactic acid bacterium was added as a dry weight of the product, and static culture was performed at 30 ° C. for 48 hours. . The cultured sample was pulverized by freeze-drying to obtain the lactic acid fermentation RJ of Example 1.

(Example 2)
Lactic acid fermentation RJ was obtained in the same manner as in Example 1 except that Pediococcus acidilactici (PA122, manufactured by Sanei Saccharification Co., Ltd.) was used as the lactic acid bacterium.

(Example 3)
Lactobacillus Streptococcus thermophilus, Lactobacillus delbrucky SSP bulgaricus, Lactobacillus delbrucky SSP lactis, Bifidobacterium, and Lactobacillus acidophilus (ABY-2C, Kyowa) A lactic acid fermentation RJ was obtained in the same manner as in Example 1 except that Hyphys) was used.

<Test Example 1: Measurement of γ-aminobutyric acid (GABA) in lactic acid fermentation RJ>
About lactic acid fermentation RJ of each Example, content of GABA was measured using HPLC. As a control, a sample obtained by treating in the same manner as in Example 1 without adding lactic acid bacteria was used. The HPLC conditions were as follows: mobile phase: 100 mM phosphate buffer: acetonitrile = 88.6: 11.4, column L-column ODS CERI, measurement wavelength Ex. 365 nm, Em. It was 490 nm. The results are shown in FIG.

  As shown in FIG. 1, it was confirmed that the GABA content in RJ was increased by the lactic acid fermentation treatment of each example. Since GABA is expected to exert a stress reduction effect, the intake of lactic acid fermentation RJ is expected to improve the stress reduction effect.

<Test Example 2: Measurement of serum corticosterone concentration>
(Animal used)
ddy strain (SLC) male mice (Chubu Science Materials) 4 weeks old, body weight 16-19 g were used. Animals were housed by placing 4 animals in plastic gauges under conditions of room temperature 23 ± 1 ° C., humidity 55 ± 5%, and illumination time 14 hours / day (6:00 to 20:00). As the solid feed, CE-2 manufactured by CLEA Japan was used, and tap water was freely consumed as drinking water. After arrival, pre-breeding was conducted for one week, and animals with no abnormal health condition were used for the test. The animals were divided into the following groups A to C with 6 animals per group.

As group A, no stress load, no intake of lactic acid fermentation RJ, ingesting water instead As group B, there is a stress load, no intake of lactic acid fermentation RJ, ingesting water instead As group C, there is a stress load, lactic acid fermentation RJ was ingested.

The lactic acid fermentation RJ to be administered was that of Example 1.
(Stress load)
In the case of A and B groups, water was applied in the case of Group C, and in the case of Group C, a stress load (restraint) for 30 minutes was performed immediately after administration of fermentation RJ (implemented between 14:00 and 16:00). Restraint was performed by placing the mouse in a 50 mL centrifuge tube, filling the excess space with absorbent cotton and closing the lid (restraint enough to move the head. Maintaining a state where the body orientation and the like cannot be changed).

(Measurement of stress markers)
Corticosterone which is a kind of glucocorticoid as a stress hormone secreted by stress was measured as a stress marker. Blood was collected immediately after 30 minutes of restraint. The obtained blood was put into a small test tube containing heparin and centrifuged (3000 rpm × 10 minutes: KUBOTA RS3010M) to obtain plasma. Using the obtained plasma, the serum corticosterone concentration was measured. The measurement was performed with Corticodterone ELISA Kit (EC 3001-1 manufactured by Assay Max). In addition, ether anesthesia may cause stress and affect the results. In addition, blood collection from the tail vein takes time to obtain the required amount, and it is expected to cause stress on animals. The blood was decapitated. The results are shown in FIG.

  As shown in FIG. 2, the increase in corticosterone concentration was confirmed in the B group given the stress load by about 8-9 times compared to the A group not given the stress load. On the other hand, it was confirmed that group C administered with lactic acid fermentation RJ significantly suppressed the increase in corticosterone concentration compared to group B. As described above, lactic acid fermentation RJ is expected to be able to alleviate the stress reaction that occurs in vivo.

<Test Example 3: Salivary amylase activity measurement>
(subject)
The test was conducted on 24 healthy normal persons who were not allergic to chronic disease or RJ. The target age was 20 to 65 years. Each group was divided into 8 persons and each group received the following samples.

As group D, placebo, ingestion 2000 mg (maltodextrin, trade name Pinex: made by Matsutani Chemical Co., Ltd.),
As group E, FD-RJ powder, intake 2000 mg,
As F group, it was set as the fermentation RJ powder of Example 1, and intake amount 2000mg.

(Stress load)
One hour before stress loading, each sample was ingested and gargled. As the stress load, Uchida-Kraepelin test was used (obtained from Japan Institute of Psychiatric Technology). The Kraepelin test is a simple one-digit addition. This was performed for 15 minutes and stressed the subjects.

(Measurement of amylase activity)
When stressed, sympathetic-adrenal medullary activity increases. Thereby, the salivary line secreting amylase is stimulated and the secretion of amylase is increased. As a result, amylase (hereinafter referred to as sAMY) activity in saliva increases due to stress load. Therefore, it is said that sAMY activity can be used as an index of the degree of strength of the exchanged nerve excitation state with respect to stress load. SAMY activity was measured 15 minutes before stress loading and after 15 minutes stress loading. The measurement was performed using a salivary amylase monitor (manufactured by Nipro Corporation). The increase rate of sAMY activity was determined from comparison with values before and after stress loading. The increase rate indicates a value (%) when the value before stress loading is 100%. The results are shown in FIG.

  As shown in FIG. 3, it is confirmed that the increase in sAMY activity is suppressed when lactic acid fermentation RJ is ingested. Lactic acid fermentation RJ is expected to alleviate various symptoms caused by an increase in the sympathetic-adrenal medullary system.

  Next, the technical idea that can be grasped from the above embodiment and other examples will be described below. (A) A meat deterioration inhibitor comprising lactic acid-fermented royal jelly as an active ingredient.

Claims (4)

Contains lactic acid bacteria fermented royal jelly obtained by fermenting royal jelly with lactic acid bacteria,
The lactic acid bacteria Gala Kutobachirusu sakei, is at least one selected La Kutobachirusu Acid Filth, Pediococcus acidilactici, Streptococcus thermophilus, and from Bifidobacterium longum,
1. An anti-stress agent characterized by being applied as an activity inhibitor of hypothalamus-pituitary-adrenal cortex system or sympathetic-adrenal medulla activity inhibitor in a psychological stress state. Contains lactic acid bacteria fermented royal jelly obtained by fermenting royal jelly with lactic acid bacteria,
The lactic acid bacteria Gala Kutobachirusu sakei, La Kutobachirusu Acid Filth, Pediococcus acidilactici, Streptococcus thermophilus, and is at least one selected from Bifidobacterium longum, hypothalamus in psychological stress state -Pituitary-adrenocortical activity inhibitor. Contains lactic acid bacteria fermented royal jelly obtained by fermenting royal jelly with lactic acid bacteria,
The lactic acid bacteria Gala Kutobachirusu sakei, La Kutobachirusu Acid Filth, is at least one selected from Pediococcus acidilactici, Streptococcus thermophilus, and Bifidobacterium longum, sympathetic in psychological stress state -Adrenal medullary activity inhibitor. In a method for producing an anti-stress agent applied as an activity inhibitor of hypothalamus-pituitary-adrenocortical system or sympathetic-adrenal medullary activity inhibitor in a psychological stress state, royal jelly is added without adding glutamic acid , Lactobacillus sakei, La Kutobachirusu acid Filth, Pediococcus acidilactici, Streptococcus thermophilus, and at least one anti-stress, characterized in that fermenting with lactic acid bacteria selected from Bifidobacterium longum Manufacturing method.

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