JP5554175B2 - Metabolic syndrome prevention or improvement composition - Google Patents

Description

  The present invention relates to a composition for preventing or improving metabolic syndrome.

Metabolic syndrome generally refers to a state having two or more of hyperglycemia, hypertension, and lipid abnormality in addition to visceral obesity-type obesity. Patients who have symptoms such as hyperglycemia in preventing or improving metabolic syndrome, because it is said that excessive visceral fat is likely to cause lifestyle-related diseases such as diabetes, hypertension, and hyperlipidemia. High interest is gathered not only from healthy people.
While it is important to prevent and improve metabolic syndrome by appropriate exercise or the like, various components having effects that lead to prevention and improvement of metabolic syndrome have been developed.
For example, a preventive agent for metabolic syndrome containing astaxanthin and / or its ester as an active ingredient (Patent Document 1), a composition for improving metabolic syndrome using soybean germ protein as an active ingredient (Patent Document 2), and a specific treatment A metabolic syndrome improving agent containing a melon peel extract (Patent Document 3), a visceral fat accumulation inhibitor (Patent Document 4) characterized by containing L-arabinose and sucrose as active ingredients are known.

On the other hand, eels are rich in vitamins such as retinol and are well-known as nutrient-rich foods, but most of the most eaten Japanese eels in Japan currently depend on aquaculture. . Aquaculture is carried out by catching white eels, which are natural eel fry, but the yield is decreasing.
Because of this background, technologies such as nutritional supplements (for example, Patent Document 5) have been developed with a focus on the high nutritional value of eels. The physiological activity of eggs has not been clarified.
In addition, eel eggs are mainly used for aquaculture because of the difficulty of aquaculture of eels, and the idea of eating is not reached.

JP 2007-246504 A JP 2008-031121 A JP 2009-256234 A JP 2009-280501 A Japanese Patent No. 4314397

Accordingly, an object of the present invention is to provide a new metabolic syndrome preventing or improving composition as well as providing the use of eel eggs of the eel family Eelaceae.

The present invention is as follows.
[1] A composition for preventing or improving metabolic syndrome comprising an eel egg belonging to the eel family Eelaceae.
[2] The metabolic syndrome preventing or improving composition according to [1], wherein the eel egg is a mature egg, a dried product thereof, or an extract thereof.
[3] A composition for enhancing adipsin expression comprising an eel egg of the eel family Eelidae.

ADVANTAGE OF THE INVENTION According to this invention, while providing the use of the eel egg of the eel family Eelaceae, the novel metabolic syndrome prevention or improvement composition can be provided.

The metabolic syndrome prevention or improvement composition of the present invention is a metabolic syndrome prevention or improvement composition containing eel eggs.
The present invention is based on the finding that eel eggs have an adipsin gene expression enhancing action that is negatively correlated with obesity (adipogenesis). By including such eel eggs, a new metabolic A composition for preventing or improving syndrome can be provided.
However, as the eel egg of the present invention, an eel egg of the eel family Eelaceae is applied.

In the present invention, “prevention or improvement of metabolic syndrome” means suppression of occurrence of metabolic syndrome state or alleviation of metabolic syndrome state, but these should not be clearly recognizable. It is included in the term “metabolic syndrome prevention or improvement” in the invention.
In the present invention, “metabolic syndrome” means a state having any two or more of hyperglycemia, hypertension, and lipid abnormality in addition to visceral obesity-type obesity, but in the present invention, differentiation of lipid cells proceeds. Thus, any condition that can cause excessive accumulation of lipid in mast cells is included in the term “metabolic syndrome” in the present invention.

In this specification, the term “process” is not limited to an independent process, and is included in this term if the intended action of this process is achieved even when it cannot be clearly distinguished from other processes. .
Moreover, the numerical value range shown using "to" in this specification shows the range which includes the numerical value described before and behind "to" as a minimum value and a maximum value, respectively.
Further, in the present invention, when referring to the amount of each component in the composition, when there are a plurality of substances corresponding to each component in the composition, the plurality present in the composition unless otherwise specified. Means the total amount of substances.
The present invention will be described below.

  The eel in the present invention is not particularly limited as long as it belongs to the eel family Anguillidae, and can include Japanese eel, giant eel, European eel, American eel, etc. From the viewpoint of the improvement effect, a eel or a Japanese eel is preferable, and a Japanese eel is particularly preferable.

  Eel eggs can be used in the present invention if they are present in the ovaries. Examples of eel eggs include immature eggs and mature eggs, and mature eggs are preferable from the viewpoint of reliable metabolic syndrome improvement or preventive effect. In the present invention, the eel egg may be an eel egg isolated from the ovary or may be used without isolating the ovary containing the eel egg. Preferably, the eel egg is isolated from the ovary. It is separated.

In the present invention, the immature egg means an egg having a mass of 10% or less with respect to the mass of the whole eel.
A mature egg means one collected from a mature ovary. The mature ovary may be a naturally-matured ovary or an artificially aged ovary. For ripening eel ovaries, the method applied to ripening fish ovaries can be applied as it is. For example, ripening may be performed by administering various gonadotropins or pituitary extracts of eels or other types of female adults to female eels. In addition, the eel can promote maturation by changing the water temperature.
Further, the immature egg and the mature egg may be an unfertilized egg or a fertilized egg, and may be an infertile egg or a fertilized egg. When using mature eggs, those collected from eel ovaries can be used as they are.

Further, from the viewpoint of reliably obtaining the metabolic syndrome prevention or improvement effect, the eel egg is preferably an immature egg or a mature egg immediately before maturation. These can be suitably used as a dry product or an extract (including an extract of a dry product). In addition, it is more preferable that it is a mature egg or its dried material, or these extracts.
The mature egg can be used in a state collected from the eel ovary, and may be used not only in the complete form of the egg but also as a crushed material, a pulverized material, a slice or the like. The dried product means a product obtained by drying mature eggs, and generally includes those having a water content of 25% by mass or less, preferably 15% by mass or less. The drying conditions for obtaining the dried product are not particularly limited as long as the conditions are applicable when producing a dried fish egg product. Applicable drying methods such as vacuum freeze drying, vacuum drying, spray drying and the like are applicable. What is necessary is just to apply on the conditions used normally. The dried eel egg is preferably a vacuum freeze-dried powder from the viewpoint of imparting stability of the components. When 100 g of mature egg is dried, it can be reduced to a mass of about 5 g to 20 g, depending on the degree of drying.

  A crushed product or a pulverized product means a product obtained by crushing or pulverizing an egg itself or a dried egg product. There is no restriction | limiting in particular about the size as a crushed material or a pulverized material. For crushing or crushing the egg or its dried product, a commonly used processing method such as a mill, a food processor, or a micro grinder may be applied as it is. When the powder is used as it is, it is used as it is, but in some cases, this powder may be formed into tablets, capsules, sugar-coated tablets, and the like.

  An extract means what was obtained by performing an extraction process with respect to a mature egg or its dried material. The extraction treatment includes all treatments that can be normally performed on mature eggs or dried products thereof, and in the case of mature eggs, it may be simply squeezed. In the case of a mature egg or a dried product thereof, an extraction process using a specific extraction solvent may be used. Examples of the extraction solvent used for obtaining the extract include water, ethanol, hydrous ethanol and the like in the case of food use. As extraction conditions, room temperature extraction, warm extraction, reflux, extraction after lyophilization, reduced pressure extraction, supercritical extraction (carbon dioxide) and the like can be used.

  The composition for preventing or improving metabolic syndrome of the present invention only needs to contain eel eggs. For this reason, it may be composed only of eel eggs. The metabolic syndrome preventing or improving composition of the present invention may be used for food and drink or for pharmaceutical use. For pharmaceutical use, it may contain eel eggs and a pharmaceutically acceptable carrier. In this case, examples of the pharmaceutically acceptable carrier include water and physiological saline.

The present metabolic syndrome preventing or improving composition can contain additives ordinarily used in oral compositions and the like as long as the effects of eel eggs are not impaired.
Examples of such additive components include excipients, diluents, additives, disintegrants, binders, coating agents, wetting agents, lubricants, lubricants, flavors, sweeteners, solubilizers, and the like. Specific examples include magnesium carbonate, titanium dioxide, lactose, mannitol and other sugars, talc, gelatin, starch, cellulose and cellulose derivatives, animal oils and vegetable oils, polyethylene glycol, water, monohydric or polyhydric alcohols. be able to.

  Moreover, it does not restrict | limit especially as a form of the metabolic syndrome prevention or improvement composition of this invention, According to the form of the eel egg used, it selects suitably. Examples of selectable forms include capsules, tablets, pills, solutions, syrups, troches, powders, granules, infusions and injections. Among these, from the viewpoint of easy intake, capsules, tablets, granules or powders are preferable.

  The metabolic syndrome preventing or ameliorating composition of the present invention is administered at a predetermined dose to animals including humans within a dose range that is regarded as safe. The specific dose is appropriately selected in consideration of the type of dosage form, administration method, patient age and weight, patient symptoms, and the like. For example, as an oral dosage form, generally, the amount of eel eggs per day for an adult can be 0.5 mg / kg body weight to 1000 mg / kg body weight.

  As described above, the composition for preventing or improving metabolic syndrome of the present invention is used for alleviating or stopping the progress rate of metabolic syndrome, particularly for improving lipid abnormalities. Examples of indicators of the progress rate of metabolic syndrome include serum lipid, blood pressure, blood glucose level, visceral fat accumulation, and the like.

The present metabolic syndrome prevention or improvement composition has an adipsin expression enhancing action or a metallothionein expression enhancing action. For this reason, prevention or improvement of the present metabolic syndrome may be evaluated based on enhancement of gene expression of adipsin, metallothionein, or both.
Adipsin is a protein known to suppress the differentiation of adipocytes, improve insulin resistance and promote the metabolism (combustion) of saccharides. Enhancement of adipsin gene expression suppresses the differentiation of mast cells having an action of accumulating fat, and is thus considered to contribute to the suppression of obesity.
Metallothionein is a kind of metal binding protein having a molecular weight of about 6000 to 7000, and has a heavy metal binding site in the molecule and has a heavy metal detoxification action. In addition, in experiments using metallothionein-deficient mice, it has been reported that fat uptake into fat cells is significantly increased. The enhanced expression of metallothionein is thought to contribute to the suppression of obesity gene expression.

  The composition for preventing or improving metabolic syndrome is not limited to human use, and may be used for other mammals that can cause metabolic syndrome. Examples of such mammals include dogs, cats and other pets.

  Since this metabolic syndrome prevention or improvement composition has the effect of prevention or improvement of metabolic syndrome, it may be used as a food or drink as described above. That is, the food / beverage products of this invention are food / beverage products containing an eel egg. Examples of such foods and beverages include soft drinks, fruit drinks, vegetable drinks, effervescent drinks, milk drinks, lactic acid bacteria drinks, and alcoholic drinks. Examples of foods include liquid, solid and powdered taste foods, seasonings and spices, or cooked and processed foods, health foods, functional foods, foods for specified health use, nutritional supplements, etc. Can do.

The composition for enhancing expression of adipsin according to the present invention includes an eel egg.
As described above, compositions comprising eel eggs induce enhanced expression of adipsin. Therefore, according to the present invention, various symptoms caused by suppression of adipsin expression can be prevented or improved by administering an adipsin expression enhancing composition to a subject. Examples of such improvement of symptoms include lipid abnormalities, glucose intolerance, and metabolic syndrome including hypertension.

  In addition, the composition containing the eel egg which concerns on this invention shows metallothionein and CYP450 expression enhancement as mentioned above. Therefore, in addition to the composition for preventing or improving metabolic syndrome, various symptoms caused by suppression of expression of metallothionein and CYP450 can be prevented or improved. Examples of the various symptoms include gene damage and antioxidant effect, cell destruction by radiation and amyotrophic lateral sclerosis (ALS), and the composition containing eel eggs according to the present invention is a gene. It can be used for repair of damage, repair of cell destruction by antioxidant action or radiation, prevention of amyotrophic lateral sclerosis (ALS).

  Hereinafter, the present invention will be described in detail with reference to examples. However, the present invention is not limited to them. Unless otherwise specified, “%” or “part” is based on mass.

[Example 1]
Analysis of components of eel eggs For eel ovaries, we used artificial ripened ovaries (eel mature eggs, EME), which were obtained by administering pituitary glands of female salmon immediately after egg laying to captured natural Japanese eels for 14 weeks. did.
The EME was freeze-dried and immediately pulverized with a mill at room temperature, and the 60-mesh sieve was placed in a sealed container containing an oxygen scavenger and a desiccant and stored in a freezer until use.
This dry powder was used in the following experiments.
About the obtained dry powder of EME, each of the general component, retinol, α-tocopherol, amino acid composition, fatty acid composition, and polyamine composition was measured as follows.

(1) General components General components were quantified according to the following method.
That is, the moisture content was calculated from the mass before and after drying using a freeze-drying method. The protein was calculated by multiplying the nitrogen amount determined by the modified Kjeldahl method by “nitrogen-protein conversion factor” 6.25.
Lipids were determined by the chloroform-methanol improved extraction method. Ash content was determined by the direct ashing method (550 ° C.).
Carbohydrates were calculated using the anthrone-sulfuric acid method according to the Japanese food standard composition table. In order to ensure consistency in composition during animal experiments, the carbohydrate value obtained by the subtraction method was used.

(2) Measurement of the amount of vitamin A / E contained in the sample Entrusted to the Japan Food Analysis Center, the freeze-dried product was measured by the HPLC method.
(3) Measurement of amino acid composition contained in sample An analysis using the ninhydrin method was performed.
(4) Measurement of fatty acid composition by gas chromatography Lipids extracted by the chloroform-methanol modified extraction method were converted to fatty acid methyl esters by the KOH-methanol method, and subjected to gas chromatography analysis under the following conditions to obtain the fatty acid composition. .
GC14-B Gas Chromatography (Shimadzu Corporation)
Make-up gas: Nitrogen.
Carrier gas: helium (flow rate: 180 kPa)
Column: SPB-PUFA, 30 m × 0.25 mm ID, 0.2 μm
(Sigma Aldrich Japan Co., Ltd.)
Column temperature: rising temperature from 50 ° C to 210 ° C.
Temperature increase rate: 4 ° C./min. Final retention time: 40 minutes.
Detector: FID (hydrogen flame ionization detector)
Split ratio: 1:60 Chart speed is 10mm / min
Integrator: CR-8A Chromatopack (Shimadzu Corporation)

(5) Preparation of fatty acid composition measurement sample Using a chloroform-methanol improved extraction method from eel mature eggs (EME), eel meat (M: eel meat) or eel liver (EL: eel liver), Lipid was extracted, 0.1 mM internal standard product (tricosanoic acid) was added, and after volatilizing and drying sufficiently with an evaporator, 0.2 ml of 2 ml of hexane and 2M potassium hydroxide-methanol was added and vortexed at 15000 rpm. After centrifugation for 5 minutes, a supernatant was obtained. The fatty acid composition was measured using the supernatant.
For identification of fatty acid composition, 37 kinds of Superco FAME mix (Sigma Aldrich Japan Co., Ltd.) were used as standard products.

(6) Preparation of polyamine measurement sample About 100 mg of EME, M and EL were weighed and placed in a 15 ml test tube, 0.9 ml of 0.1 M hydrochloric acid and an internal standard product. 20 μl was added, homogenized, and centrifuged at 12,000 rpm for 10 minutes. The supernatant was taken, an equal amount of 20% TCA was added, and the mixture was centrifuged at 12000 rpm for 10 minutes. The supernatant was measured for polyamine concentration using HPLC.

(7) Measurement of polyamine amount by cation exchange HPLC The OPA post-column HPLC method using a citrate buffer as a mobile phase was used.
The flow rate was citrate buffer (0.28M citrate hydrate, 2M NaCl adjusted to pH 5.5 with NaOH and 12.5% MeOH added), 0.5 ml / min, OPA reaction solution (ο-phthalate) Aldehyde (OPA) 6 mM, 0.4 M Boric acid adjusted to pH 10.4 with KOH and 0.2% mercaptoethanol and 0.1% Brij-35 added) 0.25 ml / min, excitation wavelength 345 nm, emission wavelength The measurement was performed at 450 nm and a column temperature of 65 ° C.
LC-6A (Shimadzu Corporation) was used as the mobile phase pump, and NP-FX-3 (Nippon Seimitsu Co., Ltd.) and LC-9A (Shimadzu Corporation) were used as the OPA reaction liquid pump. As the fluorescence detector, RF-530 (Shimadzu Corporation) was used. The column resin used was MCl GEL CK10S (Mitsubishi Chemical Corporation).

  For some items, we also measured the body and liver of eels collected for comparison. Eel liver was freeze-dried and then ground in a mill and stored in a freezer until use.

The results are shown in Tables 1 to 4.
Further, “DM” in Tables 1 and 3 means dry matter. The numerical value in Table 4 means the amount of fatty acid (g) in 100 g total fatty acid.

[Example 2]
Evaluation of functionality of eel eggs Seven-week-old C57BL / 6J male mice were divided into three groups, and the control (C) group was given a purified feed based on AIN-93G. Furthermore, the sample which replaced 2.5 mass% or 5.0 mass% of protein amount with the eel egg dry powder, respectively was given (EME2.5 group and 5.0 group).
In addition, in each group, in order to adjust the difference of the nutrient component by EME addition, it was set as the feed which prepared the feed which adjusted the amount of protein and lipid to the same level as the control (C) group (refer Table 5). In addition, food was freely consumed with water. In addition, the numerical value of Table 5 represents g.

Blood was collected after 3 weeks of feed administration. Further, total RNA was extracted from the liver according to a conventional method and used for analysis of mRNA expression level.
For the DNA microarray slide, Whole Mouse Genome Oligo Microarray Kit (Agilent Technologies) was used. For labeling, Low RNA Input linear amplification & labeling Kit Puls, one-color, RNA Spike-In Kit (Agilent Technologies) was used. For the hybridization, Gene Expression Hybridization Kit (Agilent Technologies) was used. For the incubation during the experiment, Gene Amp PCR system 9700 from Applied Biosystem was used. Gene Expression Wash (Agilent Technologies) was used for washing. The experimental operation was performed according to the product manual. DNA Microarray Scanner (Agilent Technologies) was used for imaging, and Feature Extraction (Agilent Technologies) was used for digitization. The analysis was performed according to the attached manual of Agilent Technologies. The results are shown in Table 6. The numerical values in Table 6 indicate the relative intensity with respect to the fluorescence intensity of each gene in the control group.

As a result of the analysis, the number of genes whose expression was more than doubled as compared with the control group was 143 in the EME2.5 group and 151 in the EME5.0 group. In particular, as shown in Table 6, the increased levels of expression of adipsin and metallothionein 1 and 2 were significant compared to the control group.
Since adipsin is thought to contribute to the suppression of adipocyte differentiation and metallothionein is thought to contribute to the suppression of obesity gene expression, the composition containing the eel egg of this example is administered to the subject. By this, the prevention or improvement effect of metabolic syndrome, especially the lipid abnormality improvement effect can be expected. Moreover, since the expression of adipsin and metallothionein is enhanced, the improvement of other symptoms based on the suppression of the expression of these genes can be expected in addition to the prevention or improvement of metabolic syndrome.

  Therefore, according to the present invention, metabolic syndrome can be effectively prevented or improved.

Claims (3)

A composition for preventing or improving metabolic syndrome comprising an eel egg of the eel family Eelaceae.   The composition for preventing or improving metabolic syndrome according to claim 1, wherein the eel egg is a mature egg, a dried product thereof, or an extract thereof. An adipsin expression-enhancing composition comprising an eel egg of the eel family Eelaceae.