JP2021059496A - CREG1 expression promoting composition - Google Patents

Abstract

To provide a novel CREG1 expression promoting composition.SOLUTION: A CREG1 expression promoting composition includes Dunaliella tertiolecta as an active ingredient. The CREG1 expression promoting composition also includes an extract from Dunaliella tertiolecta as an active ingredient.SELECTED DRAWING: Figure 1

Description

The present invention relates to a composition for promoting CREG1 expression.

As disclosed in Patent Document 1, the expression of CREG1 (cellular repressor of E1A-stimulated genes 1) increases with the differentiation of brown adipose tissue in the process of studying the mechanism of brown adipose tissue development in a cold environment. It is a gene found as one of the new genes. Further, Patent Document 1 discloses that CREG1 acts to induce the differentiation of brown adipocytes.

International Publication No. 2014-0277608

As a result of diligent research by the present inventors, the present invention has newly revealed that the components contained in the algae of the genus Dunariella of the order Chlorophycean vulgaris exert an action of promoting the expression of CREG1. It was made based on what was found in. An object of the present invention is to provide a novel composition for promoting CREG1 expression.

The composition for promoting CREG1 expression that solves the above problems contains Dunariella tertiolecta as an active ingredient.
The composition for promoting the expression of CREG1 that solves the above-mentioned problems contains an extract of Dunariella tertiolecta as an active ingredient, and the extract contains lower alcohol, ethyl acetate, hexane, dimethyl sulfoxide, acetonitrile, acetone, glycerin, and the like. And at least one organic solvent selected from propylene glycol, or an extract using the mixed solvent of the organic solvent and water as an extraction solvent.

According to the present invention, a novel composition for promoting CREG1 expression is provided.

The graph which shows the result of test 1. The graph which shows the result of test 2.

Hereinafter, the composition for promoting CREG1 expression of the present invention will be described.
The composition for promoting CREG1 expression contains Dunaliella.tertiolecta, which is an algae of the genus Dunaliella of the order Chlorophyceae and Volvocales, as an active ingredient. The active ingredient, Dunariella tatiolecta (hereinafter referred to as the above-mentioned algae), may be a naturally occurring algae or an artificially cultivated algae. It is industrially preferable to use artificially cultured algae from the viewpoints of stable supply and easy quality maintenance.

The compounding form of the algae in the CREG1 expression promoting composition is not particularly limited, and for example, the state as it is collected, the state of being crushed after collection, the state of being dried after collection, the state of being crushed after collection and It may be in any of the dried states. In addition, the algae may be blended in the form of an extract.

As the extraction solvent for extracting the extract from the algae, for example, an organic solvent and a mixed solvent of the organic solvent and water can be used. Examples of the organic solvent include lower alcohols, ethyl acetate, hexane, dimethyl sulfoxide, acetonitrile, acetone, glycerin, and propylene glycol. Examples of the lower alcohol include alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propanol, isopropanol and butanol.

Further, as the organic solvent, a single species may be used, or a mixed solvent in which a plurality of species are mixed may be used. When a mixed solvent of water and an organic solvent is used as the extraction solvent, the content of the organic solvent in the mixed solvent is preferably 50% by volume or more, more preferably 80% by volume or more. Further, additives such as organic salts, inorganic salts, buffers, emulsifiers and the like may be dissolved in the extraction solvent.

As the extraction method for extracting the extract from the algae, any known extraction method, for example, cold extraction, room temperature extraction, or heat extraction may be used. The extraction temperature can be appropriately set according to the type of solvent, extraction efficiency, deterioration of components, and the like.

The extraction operation is performed by immersing the above-mentioned algae, which is a raw material, in an extraction solvent for a predetermined time. At that time, the concentration of the algae in the extraction solvent can be appropriately set according to the type of the injection solvent, the extraction efficiency, the efficiency of the concentration treatment after the extraction, and the like. In such an extraction operation, in order to improve the extraction efficiency, further treatments such as reflux treatment, stirring treatment, pressurization treatment, and ultrasonic treatment may be performed, if necessary.

Further, the extraction operation may be performed only once for the same algae, or may be repeated a plurality of times. Then, the solid-liquid separation operation is performed after the extraction operation to separate the extract (extract) and the residue of the algae. As a method of solid-liquid separation operation, a known separation method such as filtration or centrifugation can be used. Moreover, the obtained extract (extract) may be concentrated or dried if necessary.

The CREG1 expression promoting composition containing the algae body increases the expression level of CREG1 in the living body by administering it to the living body. In particular, it increases the expression level of CREG1 in brown adipose tissue. Therefore, the CREG1 expression promoting composition can be applied as an experimental / research reagent for the purpose of increasing the expression level of CREG1 in a living body, for example, for studying the physiological process of CREG1. In addition, the CREG1 expression promoting composition can be applied as an additive for pharmaceuticals, quasi-drugs, and foods and drinks for the purpose of CREG1-based action such as induction of differentiation of brown adipocytes.

When the CREG1 expression promoting composition is applied to a drug or a quasi-drug, it is administered by ingestion (oral ingestion), intravascular administration, transdermal administration, mucosal administration, etc. It is possible to adopt a method of administration, a method of applying to the skin, an article, or the like. The dosage form is not particularly limited, and examples thereof include powders, powders, granules, tablets, capsules, pills, suppositories, liquids, injections, and coatings.

In addition, the composition for promoting CREG1 expression may contain other components other than the algae and the extract of the algae as long as the CREG1 expression promoting action is not impaired. Examples of other components include excipients, bases, emulsifiers, solvents, stabilizers, flavors, sweeteners and the like.

Next, the effect of the above embodiment will be described.
(1) The composition for promoting CREG1 expression contains Dunariella tertiolecta as an active ingredient. In addition, the composition for promoting CREG1 expression contains an extract of Dunariella tertiolecta as an active ingredient.

By ingesting the CREG1 expression promoting composition having the above constitution, the expression of CREG1 in the living body can be promoted.
The present embodiment can be modified and embodied as follows.

-The CREG1 expression promoting composition of the above embodiment may contain other components as long as the desired action is not impaired.
-The intake amount and the intake period of the CREG1 expression promoting composition of the above embodiment are not particularly limited, and are appropriately determined in consideration of the state of physical function of the ingestor, age, sex, and other conditions.

-The composition for promoting CREG1 expression of the above embodiment can be applied not only to humans but also to feeds, drugs and the like for domestic animals such as livestock.
Next, the technical idea that can be grasped from the above-described embodiment and modified example will be described.

(A) A composition for inducing brown adipose cell differentiation containing an extract of Dunariella tatiolecta as an active ingredient.
(B) Contains an extract of Dunariella tertiolecta as an active ingredient, the extract being at least one organic selected from lower alcohols, ethyl acetate, hexane, dimethyl sulfoxide, acetonitrile, acetone, glycerin, and propylene glycol. A composition for inducing brown fat cell differentiation, which is an extract using a solvent or a mixed solvent of the organic solvent and water as an extraction solvent.

(C) It has an extraction step of extracting an extract from Dunariella tertiolecta using an extraction solvent, and the extraction solvent is lower alcohol, ethyl acetate, hexane, dimethyl sulfoxide, acetonitrile, acetone, glycerin, and propylene glycol. A method for producing a composition for promoting CREG1 expression, which is at least one organic solvent selected from the above, or a mixed solvent of the organic solvent and water.

The above embodiment will be described in more detail with reference to test examples below. The present invention is not limited thereto.
<Test 1>
(Preparation of extract)
100 ml of ethanol was added to 6 g of a dried product of Dunariella tatiolecta, and the mixture was reflux-extracted in a hot water bath at 90 ° C. for 1 hour. After centrifuging the extract at 3200 rpm for 5 minutes, the supernatant was expressed as No. 2 Naturally filtered with filter paper. The filtrate was concentrated with an evaporator to a volume of 50 ml, and this was used as a test extract.

After differentiating C3H10T1 / 2 cells into _{brown adipose cells in the presence of 5% CO 2} by a conventional method, the test extract was prepared at 0,0.03,0.1,0.3,1.0,3.0. The cells were cultured overnight in a serum-free culture medium containing% (v / v). The next day, the culture supernatant was removed, the cells were washed with cold PBS, and then the cell protein was collected and centrifuged in a cell lysate to prepare a cell extract. The CREG1 level contained in this cell extract was measured by Western blotting.

The measured CREG1 level is corrected using the expression level of tubulin measured in the same manner as a control protein, and the relative CREG1 level is set to "1" when the addition amount of the test extract is 0%. I asked for a level. The result is shown in FIG. In addition, "DT (%)" of FIG. 1 means the concentration of the test extract in the culture medium used.

From the results shown in FIG. 1, it can be seen that the intracellular CREG1 level increases in a dose-dependent manner in the test extract. The CREG1 level under the condition that the test extract was added so as to be 0.3% was about 30 times the CREG1 level of the control (0%) to which the DT extract was not added.

<Test 2>
Two-month-old male mice (C57BL / 6J) bred in a neutral temperature environment of 30 ° C. were divided into a control group (5 mice), a first test group (6 mice), and a second test group (5 mice). It was bred for 3 months. During the breeding period, the mice in the control group were fed a high-fat diet (B15040: manufactured by Claire Japan) as a feed. Mice in the first test group were fed a high-fat diet (B15040: manufactured by Claire Japan, Ltd.) with a diet containing 1% by mass of dry powder of Dunariella tertiolecta. Mice in the second test group were fed a high-fat diet (B15040: manufactured by Claire Japan, Ltd.) with a diet supplemented with a dry powder of Dunariella tertiolecta in an amount of 2.5% by mass.

After 3 months of breeding, brown adipose tissue was collected from each group of mice, and cDNA was synthesized using the total RNA fraction prepared from the brown adipose tissue. Using this cDNA, the mRNA expression level of CREG1 and the mRNA expression level of 36B4 as an internal standard were measured by a quantitative PCR method. The measured mRNA expression level of CREG1 was corrected with the mRNA expression level of 36B4, and the relative mRNA expression level of CREG1 with the mRNA expression level of CREG1 in the control group being "1" was determined. The result is shown in FIG.

From the results shown in FIG. 2, it can be seen that the mRNA expression level of CREG1 increases in a dose-dependent manner of Dunariella tertiolecta in the feed. The mRNA expression level of CREG1 in the second test group fed with the diet supplemented with Dunariella tertiolecta so as to be 2.5% by mass was the mRNA expression level of CREG1 in the control group without the addition of Dunariella tertiolecta. It was about 1.7 times the amount.

Claims (2)

A composition for promoting CREG1 expression containing Dunariella tartiolecta as an active ingredient. Contains the extract of Dunariella tertiolecta as an active ingredient,
The extract is an extraction using at least one organic solvent selected from lower alcohol, ethyl acetate, hexane, dimethyl sulfoxide, acetonitrile, acetone, glycerin, and propylene glycol, or a mixed solvent of the organic solvent and water as an extraction solvent. A composition for promoting the expression of CREG1 which is a substance.

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