JP7610808B2 - Expression enhancers and functional foods for promoting mental health - Google Patents

Description

The present invention relates to an expression promoter and a functional food for promoting mental health.

Dementia is a major social problem, and as Japan is facing an aging society, its incidence is expected to continue to increase, making addressing it an urgent task.

Depending on the severity of the symptoms, dementia can cause confusion in daily life and problems with those around you, and since understanding and assistance from family and acquaintances are important, it can also place a burden on these people.

Therefore, medical institutions have traditionally administered drug treatments to alleviate as many of the problems in daily life caused by dementia as possible (see, for example, Non-Patent Document 1).

Takeshi Tabira, "Treatment of Alzheimer's Disease: Current Status and Future", Journal of the Japan Geriatrics Society, Japan Geriatrics Society, 2012, Vol. 49, p. 402-418

Incidentally, preventing and improving dementia is extremely meaningful not only from the perspective of individual health issues, but also from the perspective of national medical finances.

In particular, if it is possible to prevent or improve dementia through daily eating habits, it will be possible to improve quality of life for the long term without relying on support from medical institutions, and it will be possible to reduce the personal and financial burden caused by the onset of dementia.

The present invention was made in consideration of these circumstances, and provides an expression promoter for mental health-related genes that can be expected to promote mental health, particularly to improve dementia, as well as functional foods for promoting mental health and topical skin care products.

In order to solve the above-mentioned conventional problems, the expression promoter of the present invention is (1) an expression promoter of mental health-related genes containing a dipeptide as a main component for inducing function , wherein the dipeptide contains at least Leu-Pro, and the mental health-related genes are a brain-derived neurotrophic factor gene and an arylalkylamine-N-acetyltransferase gene .

In addition, the functional food for promoting mental health according to the present invention is ( 2) a functional food for promoting mental health that contains a dipeptide as a main component for inducing function and promotes the expression of mental health-related genes , wherein the dipeptide contains at least Leu-Pro, and the mental health-related genes are a brain-derived neurotrophic factor gene and an arylalkylamine-N-acetyltransferase gene .

In addition, the topical skin product according to the present invention contains ( 3 ) the expression promoter described in (1) above.

In addition, in the present invention, ( 4 ) in a functional food for promoting mental health aimed at preventing or improving dementia by promoting the expression of brain-derived neurotrophic factor gene and improving sleep by promoting the expression of arylalkylamine-N-acetyltransferase gene, a dipeptide represented by Leu-Pro is used as a main component raw material for said prevention or improvement.

According to the expression promoter of the present invention, there is provided an expression promoter for mental health-related genes which contains a dipeptide as the main component for inducing function , wherein the dipeptide contains at least Leu-Pro and the mental health-related genes are the brain-derived neurotrophic factor gene and the arylalkylamine-N-acetyltransferase gene. Therefore, it is possible to provide an expression promoter for mental health -related genes which is expected to promote mental health, in particular improve dementia.

In addition, if the dipeptide contains at least Leu-Pro and the mental health-related genes are the brain-derived neurotrophic factor gene and the arylalkylamine-N-acetyltransferase gene, it is possible to provide an expression promoter that is expected to improve mental health by improving sleep in addition to improving dementia.

Furthermore, according to the mental health functional food of the present invention , a dipeptide is contained as a main component for inducing function, and the expression of mental health-related genes is promoted, and the dipeptide contains at least Leu-Pro, and the mental health-related genes are the brain-derived neurotrophic factor gene and the arylalkylamine-N-acetyltransferase gene , so that a mental health functional food that can be expected to promote mental health, particularly improve dementia, can be provided.Furthermore, a functional food that can be expected to promote mental health by improving sleep in addition to improving dementia can be provided.

In addition, the topical skin product according to the present invention contains the expression promoter described above, making it possible to provide a topical skin product that is expected to transdermally promote mental health, particularly improvement of dementia.

In addition, according to the present invention, in a functional food for promoting mental health aimed at preventing or improving dementia by promoting the expression of the brain-derived neurotrophic factor gene and improving sleep by promoting the expression of the arylalkylamine-N-acetyltransferase gene, a dipeptide represented by Leu-Pro is used as a main ingredient for said prevention or improvement, so that an expression promoter can be provided that is expected to improve mental health by improving sleep as well as improving dementia, and mental health can be improved through daily dietary habits.

The present invention relates to an expression promoter for mental health-related genes, and in particular provides an expression promoter for mental health-related genes that is expected to promote mental health, in particular to improve dementia.

Here, mental health refers to mental health and mental health, and in this specification is a concept that includes issues related to dementia and sleep.

The expression promoter according to this embodiment is characterized in that it contains a dipeptide represented by Gly-Pro and/or Leu-Pro as the main component for inducing function.

Gly-Pro is a dipeptide (glycylproline) formed by the peptide bond between glycine and proline, and is a compound represented by the following structural formula (i).

Leu-Pro is a dipeptide (leucylproline) formed by the peptide bond between leucine and proline, and is a compound represented by the following structural formula (ii).

As will be explained later with reference to experimental results, these dipeptides have the effect of promoting the expression of mental health-related genes, making it possible to provide an agent for promoting the expression of mental health-related genes that is expected to promote mental health, particularly improvement of dementia.

This mental health-related gene may also be a brain-derived neurotrophic factor gene.

Promoting the expression of the brain-derived neurotrophic factor (BDNF) gene is expected to affect the brain function control mechanism through intestinal activation, enhancing the growth and maintenance of the nervous system, and further alleviating and suppressing dementia through the brain function control mechanism.

It is also expected to contribute to improving mental health by improving memory, increasing energy (preventing or improving depression), and suppressing appetite (preventing overeating and obesity).

The dipeptide may also include at least Leu-Pro, and the mental health-related genes may be a brain-derived neurotrophic factor gene and an arylalkylamine-N-acetyltransferase (AANAT) gene.

By configuring in this way, in addition to the effect derived from the increased expression of the brain-derived neurotrophic factor gene described above, it is possible to expect a sleep-improving effect derived from the increased expression of the arylalkylamine-N-acetyltransferase gene.

In other words, melatonin is known as a substance that has the effect of switching between wakefulness and sleep and inducing natural sleep, and arylalkylamine-N-acetyltransferase is the rate-limiting enzyme in melatonin biosynthesis. Promoting the expression of the arylalkylamine-N-acetyltransferase gene can increase the amount of melatonin, so using it at the right time can improve sleep, including making it easier to fall asleep.

This specification also provides functional foods for promoting mental health that contain dipeptides represented by Gly-Pro and/or Leu-Pro as the main ingredient for inducing function.

In other words, the functional food for promoting mental health according to this embodiment contains dipeptides represented by Gly-Pro and/or Leu-Pro, and due to the above-mentioned effects derived from these dipeptides, it is expected to improve dementia and sleep.

The phrase "contains as a major ingredient for inducing a function" means "contains as an active ingredient" in the context of pharmaceuticals. In other words, when considering health functional foods, it means that the above dipeptide is contained as an ingredient for exerting a function or efficacy, although the expressions may differ: functional ingredients in functional foods, nutritional ingredients in nutrient functional foods, and ingredients involved in foods for specified health uses. Additionally, functional foods for promoting mental health also include beverages.

This specification also provides topical skin products containing the above-mentioned expression promoter. Here, topical skin products are toiletries and the like that are applied to the skin or hair by coating or spraying, and conceptually include quasi-drugs and cosmetics.

For example, cosmetics include not only basic cosmetics (skin care cosmetics) and finishing cosmetics (makeup cosmetics), but also hand soap, shampoo, body soap, etc. Furthermore, examples of quasi-drugs include hair dyes, permanent wave agents, bath agents, mouth fresheners, hair growth agents, hair removers, etc.

Such topical skin products allow the dipeptide to be taken in transdermally, and can provide topical skin products that are expected to promote mental health, particularly dementia, transdermally. In other words, it is possible to provide products that allow the dipeptide to be absorbed transdermally, such as quasi-drugs and cosmetics.

This specification also provides a method for using dipeptides as a main ingredient in functional foods for promoting mental health.

Specifically, in the first aspect, we provide a method for using a dipeptide represented by Gly-Pro and/or Leu-Pro as a main ingredient for the improvement of mental health functional foods aimed at preventing or improving dementia by promoting the expression of brain-derived neurotrophic factor genes.

This method of use makes it possible to provide a functional food for promoting mental health that is expected to improve dementia, and to promote mental health through daily dietary habits.

In addition, as a second aspect, a method is also provided for using a dipeptide represented by Leu-Pro as a main ingredient for preventing or improving dementia by promoting the expression of the brain-derived neurotrophic factor gene and for improving sleep by promoting the expression of the arylalkylamine-N-acetyltransferase gene in a functional food for promoting mental health.

This method of use can provide an expression promoter that is expected to improve mental health by improving sleep as well as improving dementia, making it possible to promote mental health through daily dietary habits.

The following will further explain the expression promoter and mental health promotion functional food of this embodiment, including the results of tests to confirm the action of the dipeptide added to these preparations and foods.

1. Anti-dementia evaluation test using brain-derived neurotrophic factor gene expression as an indicator
In this study, we used two types of dipeptides (Gly-Pro and Leu-Pro) to examine changes in brain-derived neurotrophic factor gene expression.

First, human colon cancer-derived cells (Caco-2) were cultured. Caco-2 cells were precultured in a φ10 cm dish using Dulbecco's Modified Eagle Medium (DMEM) (high glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) until they became confluent.

Thereafter, the cells were washed with phosphate-buffered saline (PBS) and resuspended in medium, then seeded onto a 24-well plate at a density of 1.0×10 ⁵ cells/well and cultured overnight in a CO ₂ incubator (37° C., 5% CO ₂ ).

Next, the medium of the cells cultured overnight was replaced with serum-free DMEM medium (containing 1% penicillin-streptomycin) containing one of the dipeptides, and the cells were cultured for 24 hours in a _CO2 incubator. The dipeptides were added to the medium at a final concentration of 400 μg/mL.

Next, we compared the gene expression levels using real-time PCR. Specifically, we harvested Caco-2 cells after culturing them with the addition of the dipeptide, and extracted total RNA using the PureLink RNA Mini kit (Invitrogen).

Next, cDNA was synthesized from the extracted total RNA using ReverTra Ace qPCR RT Master Mix with gDNA Remover (TOYOBO).

Next, real-time PCR was performed using the synthesized cDNA as a template with an AriaMX real-time PCR device (Agilent Technologies).

For the real-time PCR reaction, THUNDERBIRD SYBR qPCR Mix (TOYOBO) was used. 5'-GTCAAGTTGGGAGCCTGAAATAGTG-3' (SEQ ID NO: 1) and 5'-AGGATGCTGGTCCAAGTGGTG-3' (SEQ ID NO: 2) were used as primers for the brain-derived neurotrophic factor gene, and 5'-GCACCGTCAAGGCTGAGAAC-3' (SEQ ID NO: 3) and 5'-ATGGTGGTGAAGACGCCAGT-3' (SEQ ID NO: 4) were used as primers for the internal standard GAPDH. The real-time PCR reaction conditions were 40 cycles of a two-step PCR reaction consisting of an initial denaturation at 95°C for 3 minutes, followed by denaturation at 95°C for 3 seconds and annealing/extension at 60°C for 30 seconds. The results are shown in Table 1.

Table 1 shows the change in expression of the brain-derived neurotrophic factor gene in Caco-2 cells after addition of the dipeptide (n=3, mean ±SD., *p<0.05, **p<0.01). As can be seen from Table 1, it was confirmed that the expression of the brain-derived neurotrophic factor gene was significantly increased by approximately 1.5 times when Gly-Pro was added and approximately 1.2 times when Leu-Pro was added, compared to the control (water).

These results suggest that Gly-Pro and Leu-Pro may alleviate or suppress dementia by enhancing the production of brain-derived neurotrophic factor, which then promotes the activation of nerve cells through a brain function control mechanism mediated by intestinal activation.

In other words, it has been shown that the expression promoter and mental health-promoting functional food according to this embodiment are expected to alleviate and suppress dementia by containing the dipeptide represented by Gly-Pro and/or Leu-Pro.

2. Sleep evaluation test using the expression of melatonin biosynthesis-related enzyme genes as an index
In this study, we used Leu-Pro as the dipeptide and investigated the changes in expression of the arylalkylamine-N-acetyltransferase gene.

First, SH-SY5Y cells derived from human neuroblastoma were cultured. SH-SY5Y cells were precultured in a φ10 cm dish using Dulbecco's Modified Eagle Medium (DMEM) (high glucose) (containing 1% penicillin-streptomycin and 10% fetal bovine serum (FBS)) until they became confluent.

Thereafter, the cells were washed with phosphate-buffered saline (PBS) and resuspended in medium, then seeded onto a 24-well plate at a density of 1.0×10 ⁵ cells/well and cultured overnight in a CO ₂ incubator (37° C., 5% CO ₂ ).

Next, the medium of the cells cultured overnight was replaced with serum-free DMEM medium (containing 1% penicillin-streptomycin) containing the dipeptide, and the cells were cultured for 24 hours in a _CO2 incubator. The dipeptide was added to the medium at a final concentration of 400 μg/mL.

Next, we compared gene expression levels using real-time PCR. Specifically, SH-SY5Y cells were harvested after culturing with the addition of dipeptides, and total RNA was extracted using the PureLink RNA Mini kit (Invitrogen).

Next, cDNA was synthesized from the extracted total RNA using ReverTra Ace qPCR RT Master Mix with gDNA Remover (TOYOBO).

Next, real-time PCR was performed using the synthesized cDNA as a template with an AriaMX real-time PCR device (Agilent Technologies).

For the real-time PCR reaction, THUNDERBIRD SYBR qPCR Mix (TOYOBO) was used. 5'-TCCTGTGGAGATACCTTCACCA-3' (SEQ ID NO: 5) and 5'-CACAGTTCAGAAGGCAAGAGGT-3' (SEQ ID NO: 6) were used as primers for the arylalkylamine-N-acetyltransferase gene, and 5'-GCACCGTCAAGGCTGAGAAC-3' (SEQ ID NO: 3) and 5'-ATGGTGGTGAAGACGCCAGT-3' (SEQ ID NO: 4) were used as primers for the internal standard GAPDH. The real-time PCR reaction conditions were 40 cycles of a two-step PCR reaction consisting of initial denaturation at 95°C for 3 minutes, denaturation at 95°C for 3 seconds, and annealing/extension at 60°C for 30 seconds. The results are shown in Table 2.

Table 2 shows the change in expression of the arylalkylamine-N-acetyltransferase gene in SH-SY5Y cells after the addition of dipeptides (n=3, mean ±SD., *p<0.05, **p<0.01). As can be seen from Table 2, it was confirmed that when Leu-Pro was added, the expression of the arylalkylamine-N-acetyltransferase gene significantly increased by about 1.2-fold compared to the control (water).

These results suggest that Leu-Pro is expected to increase melatonin levels and improve sleep by promoting the expression of the arylalkylamine-N-acetyltransferase gene, the rate-limiting enzyme in melatonin biosynthesis.

In other words, it was shown that the expression promoter and mental health functional food according to this embodiment contain a dipeptide represented by Leu-Pro, and thus can be expected to improve sleep.

As described above, the expression promoter according to this embodiment is an expression promoter for mental health-related genes, and contains a dipeptide represented by Gly-Pro and/or Leu-Pro as a main component for inducing function, so that it is possible to provide an expression promoter for mental health-related genes that can be expected to promote mental health, particularly improve dementia.

In addition, the functional food for promoting mental health according to the present invention contains a dipeptide represented by Gly-Pro and/or Leu-Pro as the main component for inducing function, and therefore it is possible to provide a functional food for promoting mental health that is expected to promote mental health, particularly improve dementia.

Finally, the above-mentioned explanations of each embodiment are merely examples of the present invention, and the present invention is not limited to the above-mentioned embodiments. Therefore, even if the embodiment is different from the above-mentioned embodiments, various modifications can be made depending on the design, etc., as long as they do not deviate from the technical concept of the present invention.

Claims (4)

An agent for promoting the expression of a mental health-related gene , comprising a dipeptide as a main component for inducing a function ,
The expression promoter, characterized in that the dipeptide contains at least Leu-Pro, and the mental health-related genes are a brain-derived neurotrophic factor gene and an arylalkylamine-N-acetyltransferase gene . A functional food for promoting mental health, which contains a dipeptide as a main component for inducing a function and promotes the expression of mental health-related genes ,
A functional food for promoting mental health, characterized in that the dipeptide contains at least Leu-Pro, and the mental health-related genes are a brain-derived neurotrophic factor gene and an arylalkylamine-N-acetyltransferase gene . A skin care product containing the expression promoter according to claim 1 . The use of a dipeptide represented by Leu-Pro as a main ingredient for preventing or improving dementia by promoting the expression of the brain-derived neurotrophic factor gene, and for improving sleep by promoting the expression of the arylalkylamine-N-acetyltransferase gene in a functional food for promoting mental health, with the objectives of preventing or improving dementia by promoting the expression of the brain-derived neurotrophic factor gene, and for improving sleep by promoting the expression of the arylalkylamine-N-acetyltransferase gene.