JP6752285B2 - Fat accumulation inhibitor, adipose progenitor cell differentiation inhibitor, visceral fat reducing agent, and food and drink for reducing visceral fat - Google Patents

Description

The present disclosure relates to a fat accumulation inhibitor, an adipose progenitor cell differentiation inhibitor, a visceral fat reducing agent, and a food or drink for reducing visceral fat.

Adipocytes are cells that develop from mesoderm stem cells and differentiate through adipose progenitor cells, etc., and are classified into white adipocytes that control fat synthesis, accumulation, and release, and brown adipocytes that control heat production. To.

Obesity is a condition in which fat is excessively accumulated in white adipocytes. Therefore, in order to prevent or improve obesity, it is effective to suppress the accumulation of fat in white adipocytes. In recent years, it has been clarified that the number of white adipocytes increases even after adulthood, and in order to prevent or improve obesity, it is also possible to suppress the differentiation of adipose progenitor cells into white adipocytes. It is valid.

Obesity is generally classified into subcutaneous fat obesity and visceral fat obesity. In particular, visceral fat obesity is known to increase the risk of diabetes, arteriosclerosis, hypertension, cerebral infarction, dyslipidemia and the like. Therefore, it is important to prevent or improve visceral fat obesity among obesity.

Conventionally, various techniques for preventing or improving obesity have been proposed.
For example, Patent Document 1 describes an adipocyte fat accumulation inhibitor containing an ethanol water extract of C. versicolor as an active ingredient.
Patent Document 2 describes an agent for suppressing differentiation of preadipocytes containing an extract of sacred lotus germ and yacon, and an anti-obesity agent containing this agent for suppressing differentiation as an active ingredient.
Patent Document 3 describes a body fat accumulation inhibitor, a visceral fat accumulation inhibitor, and the like containing egg white as an active ingredient.

Japanese Patent No. 4363825 Japanese Patent No. 4675781 Japanese Unexamined Patent Publication No. 2011-225496

The body fat accumulation inhibitor and the visceral fat accumulation inhibitor described in Patent Document 3 contain a food material as an active ingredient, and are considered to be excellent in safety in continuous administration or ingestion. However, the effect is not sufficient, and a search for a new active ingredient that is effective and excellent in safety has been desired.

Therefore, the present disclosure provides a novel fat accumulation inhibitor, an adipose progenitor cell differentiation inhibitor, a visceral fat reducing agent, and a food and drink for reducing visceral fat using an active ingredient that is effective and has excellent safety. Is an issue.

Specific means for solving the above problems include the following embodiments.
<1> A fat accumulation inhibitor containing a bird egg containing 4.2 mass ppm or more of iodine as an active ingredient.
<2> 4.2 An agent for suppressing the differentiation of adipose progenitor cells containing a bird egg containing iodine of 2 mass ppm or more as an active ingredient.
<3> A visceral fat reducing agent containing a bird egg containing 4.2 mass ppm or more of iodine as an active ingredient.
<4> A food or drink for reducing visceral fat containing a bird egg containing iodine of 4.2 mass ppm or more as an active ingredient.
<5> A visceral fat reducing agent that contains an egg yolk extract of a bird egg containing an iodide peptide as an active ingredient and is used so as to administer 300 μg or more of iodine per day.

According to the present disclosure, a novel fat accumulation inhibitor, an adipose progenitor cell differentiation inhibitor, a visceral fat reducing agent, and a food and drink for reducing visceral fat using an active ingredient that is effective and excellent in safety are provided. Can be done.

It is a figure which shows the amount of fat accumulation (relative value) in adipocyte when the yolk extract (IY) of an egg containing high iodine or the yolk extract (OY) of a normal egg is added to a medium. It is a figure which shows the amount of fat accumulation (relative value) in adipocyte when sodium iodide is added to the culture medium. It is a figure which shows the glycerol-3-phosphate dehydrogenase (GPDH) activity of adipocytes when the yolk extract (IY) of the egg containing high iodine or the yolk extract (OY) of a normal egg is added to the medium. is there. Expression level (relative value) of PPAR (Peroxisome Proliferator Activated Receptor) -γ2 gene in adipocytes when egg yolk extract (IY) of high iodine content egg or egg yolk extract (OY) of normal egg is added to the medium. It is a figure which shows. It is a figure which shows the expression level (relative value) of the adipocyte adipogenin gene in the case of adding the yolk extract (IY) of an iodine-rich egg or the yolk extract (OY) of a normal egg into a medium. It is a figure which shows the expression level (relative value) of the leptin gene of adipocyte when the egg yolk extract (IY) of the iodine-rich egg or the yolk extract (OY) of a normal egg is added to the culture medium. It is a figure which shows the expression level (relative value) of the adiponectin gene of adipocyte when the yolk extract (IY) of the egg containing high iodine or the yolk extract (OY) of a normal egg is added to the culture medium.

Hereinafter, embodiments of the present invention will be described in detail. However, the present invention is not limited to the following embodiments.
The numerical range indicated by using "~" in the present specification indicates a range including the numerical values before and after "~" as the minimum value and the maximum value, respectively.

<Adipose accumulation inhibitor, adipose progenitor cell differentiation inhibitor, visceral fat reducing agent, and food and drink for reducing visceral fat>
The fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, the visceral fat reducing agent, and the food and drink for reducing visceral fat of the first embodiment are all bird eggs containing 4.2% by mass or more of iodine (hereinafter,). , Also referred to as "iodine-rich egg") is contained as an active ingredient.

Examples of the bird egg include eggs of birds such as chickens, quails, silkie chickens, and ducks, and chicken eggs are preferable.
Eggs with a high iodine content can be obtained by feeding birds with a feed having an increased iodine content by blending an iodine source and increasing the iodine content in the eggs.
As an iodine source, iodine compounds such as calcium iodate, potassium iodate, potassium iodide, sodium iodide, timol iodide, copper iodide, hyposalicylic acid, calcium periodate, calcium iodobihemate; kelp, Examples thereof include seaweeds having a high iodine content such as kelp or processed products thereof. From the viewpoint of bird health, the transfer rate of iodine to eggs, and the like, it is preferable to use at least one iodine source selected from the group consisting of calcium iodate, potassium iodate, and potassium iodide. Further, it is more preferable to use seaweeds in combination with at least one selected from the group consisting of calcium iodate, potassium iodate, and potassium iodide.

The amount of iodine given to birds can be adjusted as appropriate depending on the type of bird. In the case of spawning chickens, for example, an iodine intake of 5 mg to 250 mg per chicken per day is preferable, and an iodine intake of 5 mg to 15 mg per chicken per day is more preferable. Assuming that the laying chicken ingests about 100 g of feed per chicken per day, the iodine content in the feed is preferably 50 mass ppm to 2500 mass ppm, preferably 50 mass ppm to 150 mass ppm. Is more preferable.

When a bird is fed a diet containing a high iodine content, the desired iodine-rich egg is produced after about one week. For example, when a feed containing about 50% by mass of iodine is given to laying hens, eggs containing about 300 μg of iodine are laid.

The iodine content of the iodine-rich egg is not particularly limited as long as it is 4.2 mass ppm or more with respect to the total amount of the edible portion, and is preferably 4.2 mass ppm to 20.0 mass ppm, for example. By setting the iodine content of the iodine-rich eggs to 20.0 mass ppm or less, the decrease in the egg-laying rate tends to be suppressed. Since the edible portion of shelled chicken eggs mainly distributed in the market is about 45 g to 66 g per egg, the iodine content of 4.2 mass ppm to 20.0 mass ppm is about 200 μg per egg. It corresponds to an iodine content of ~ 1400 μg.

The required amount of iodine in spawning chickens is 0.2 mg per kg of feed (according to the Japanese feed standard poultry (2011 version)), and the actual iodine content in commercial feed is 0.3 mg to 2.0 mg per kg of feed. It has become. The iodine content of normal eggs produced from spawning chickens bred using this commercially available feed is about 9 μg per egg (according to the Standard Tables of Food Composition in Japan 2015 (7th edition)), and at most one egg. It is about 30 μg per unit.

Eggs high in iodine are also available as commercial products. For example, as a commercially available product of chicken eggs containing 4.2 mass ppm or more of iodine, "iodine egg light" ("iodine egg" is a registered trademark) of Nippon Agricultural Products Co., Ltd. can be mentioned.

The fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, the visceral fat reducing agent, and the food and drink for reducing visceral fat of the first embodiment may contain whole eggs of iodine-rich eggs as an active ingredient. In addition, since most of the iodine in the iodine-rich egg is contained in the yolk as an iodide peptide bound to the peptide, the fat accumulation inhibitor, the adipose precursor cell differentiation inhibitor, and the visceral fat reduction of the first embodiment. The agent and the food and drink for reducing visceral fat may contain the egg yolk of an egg containing a high iodine content as an active ingredient, or may contain an egg yolk extract as an active ingredient. The egg yolk extract is not particularly limited as long as it contains the iodide peptide in the egg yolk.
In addition, whole eggs, egg yolks, and egg yolk extracts of iodine-rich eggs may be subjected to treatments such as drying, concentration, powdering, and granulation, if necessary.

The dosage form of the fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, and the visceral fat reducing agent of the first embodiment is not particularly limited, and examples thereof include powders, granules, tablets, capsules, syrups, and emulsions. .. The fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, and the visceral fat reducing agent of the first embodiment further contain various excipients, binders, disintegrants, solvents, etc., depending on the dosage form. You may.

The dose and duration of the fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, and the visceral fat reducing agent of the first embodiment, and the intake amount and the intake period of the food and drink for reducing visceral fat of the first embodiment are There is no particular limitation as long as the desired effect is achieved. As an example, it is preferable to continue the dose or intake at which the daily iodine amount is 300 μg or more for one month or more, and more preferably for three months or more. The upper limit of dose or intake can be set based on the tolerable upper limit of iodine. As an example, the dose or intake may be such that the daily amount of iodine is 3000 μg or less.

Prevention or improvement of obesity by administering the fat accumulation inhibitor, the adipose progenitor cell differentiation inhibitor, or the visceral fat reducing agent of the first embodiment, or by ingesting the visceral fat reducing food or drink of the first embodiment. It is possible to do. That is, according to the first embodiment, the following method is also provided.
(1) A method for suppressing fat accumulation, which comprises administering to a subject an effective amount of a bird egg containing 4.2 mass ppm or more of iodine.
(2) A method for suppressing differentiation of adipose progenitor cells, which comprises administering to a subject an effective amount of a bird egg containing 4.2 mass ppm or more of iodine.
(3) A method for reducing visceral fat, which comprises administering to a subject an effective amount of a bird egg containing 4.2 mass ppm or more of iodine.
(4) A method for reducing visceral fat, which comprises ingesting an effective amount of bird eggs containing 4.2 mass ppm or more of iodine.
(5) A method for preventing or ameliorating obesity, which comprises administering to a subject an effective amount of a bird egg containing 4.2 mass ppm or more of iodine.
(6) A method for preventing or ameliorating obesity, which comprises ingesting an effective amount of bird eggs containing 4.2 mass ppm or more of iodine.

<Visceral fat reducing agent>
The visceral fat reducing agent of the second embodiment is used so that an egg yolk extract of a bird egg containing an iodide peptide is used as an active ingredient and 300 μg or more of iodine is administered per day.

Examples of the bird egg include eggs of birds such as chickens, quails, silkie chickens, and ducks, and chicken eggs are preferable. The bird egg may be a normal egg or an iodine-rich egg described above, and the iodine-rich egg described above is preferable in that it contains a high iodine content.

The egg yolk extract is not particularly limited as long as it contains the iodide peptide in the egg yolk. The egg yolk extract may be subjected to treatments such as drying, concentration, powdering, and granulation, if necessary.

The dosage form of the visceral fat reducing agent of the second embodiment is not particularly limited, and examples thereof include powders, granules, tablets, capsules, syrups, and emulsions. The visceral fat reducing agent of the second embodiment may further contain various excipients, binders, disintegrants, solvents and the like, depending on the dosage form.

The dose of the visceral fat reducing agent of the second embodiment is not particularly limited as long as the daily iodine amount is 300 μg or more. The upper limit of the dose can be set based on the tolerable upper limit of iodine. As an example, the dose can be such that the amount of iodine per day is 3000 μg or less.
The administration period of the visceral fat reducing agent of the second embodiment is not particularly limited as long as the desired effect is achieved. As an example, continuous administration for 1 month or longer is preferable, and continuous administration for 3 months or longer is more preferable.

Obesity can be prevented or ameliorated by administering the visceral fat reducing agent of the second embodiment. That is, according to the second embodiment, the following method is also provided.
(1) A method for reducing visceral fat, which comprises administering an egg yolk extract of a bird egg containing an iodide peptide in an amount of 300 μg or more per day.
(2) A method for preventing or ameliorating obesity, which comprises administering an egg yolk extract of a bird egg containing an iodide peptide in an amount of 300 μg or more per day.

Hereinafter, the present invention will be specifically described with reference to Examples, but the present invention is not limited to the Examples.

[Preparation Example 1]
Eggs containing about 1300 μg of iodine per 100 g of edible portion (Nosan Corporation, “Iodine Egg Light”) are used as iodine-rich eggs, and only egg yolks are freeze-dried (-10 ° C, 180 minutes) in a sterile condition. ) To obtain egg yolk. 5 mL lysis buffer (20 mM Tris-HCl (pH 7.6), 5 mM DTT (Dithiothreitol), 0.1 w / v% SDS (Sodium Dodecyl Sulfate), 1 v / v% protease inhibitor" per 1 g of the obtained egg powder. After adding the cocktail) and suspending it, centrifugation was performed at 17,000 G for 15 minutes. PBS (Phosphate Buffered Saline) was added to the obtained supernatant to adjust the protein concentration to 10 μg / μL, and the protein was used as an egg yolk extract (IY) of iodine-rich eggs. The protein concentration was measured by the Bradford method.
In addition, an egg yolk extract (OY) of a normal egg was prepared in the same manner as above except that a commercially available chicken egg was used instead of the iodine-rich egg.

[Experimental Example 1]
In Experimental Example 1, mouse adipose progenitor cells 3T3-L1 were differentiated into adipocytes, and the amount of fat accumulated in the adipocytes was measured. 3T3-L1 cells show a fibroblast-like morphology under normal culture conditions, but when they are brought into a confluent state and then given insulin stimulation, they form fat granules inside the cells and differentiate into adipocytes. ..

First, 3T3-L1 cells were seeded in 6-well plates and maintained by DMEM (Dulbecco's Modified Eagle Medium) containing 10 v / v% FBS (Fetal Bovine Serum) until 2 days after confluence. The medium was changed every 48 hours. The medium was then replaced with DMEM containing 10 v / v% FBS, 0.5 mM isobutylmethylxanthine, 1 μM dexamethasone, and 1.7 μM insulin, and cultured for 48 hours to induce adipocyte differentiation. The medium was then replaced with DMEM containing 10 v / v% FBS. Further, egg yolk extract (IY) of high iodine-containing eggs or egg yolk extract (OY) of normal eggs was added to a final concentration of 5 μg / mL, 10 μg / mL, or 50 μg / mL, and cultured for 10 days. .. The medium was changed every 48 hours.

After culturing for 10 days, the differentiated adipocytes were washed 3 times with PBS and fixed with a 10 v / v% paraformaldehyde solution for 10 minutes or longer. After fixation, adipocytes were washed twice with PBS and stained with 0.5 w / v% Oil Red O solution (Sigma Aldrich) under 37 ° C. conditions. Then, the amount of fat accumulated in adipocytes was evaluated by measuring the absorbance at a wavelength of 570 nm.

The evaluation result of the amount of fat accumulated in adipocytes is shown in FIG. FIG. 1 shows the amount of fat accumulated in adipocytes (relative value) as an average value ± standard error (n = 6 in each case). As shown in FIG. 1, when the egg yolk extract (IY) of an iodine-rich egg was added to the medium so as to have a final concentration of 50 μg / mL, the egg yolk extract (OY) of a normal egg was added to the medium. The amount of fat accumulated in adipocytes was significantly reduced as compared with the case of addition (p <0.05).

[Experimental Example 2]
In Experimental Example 2, the amount of fat accumulated in adipocytes was measured in the same manner as in Example 1 except that sodium iodide was used instead of the egg yolk extract. Sodium iodide was added to the medium so that the final concentration of iodide ion was ^10-9 M, ^10-8 M, ^10-7 M, or ^10-6 M.

The evaluation result of the amount of fat accumulated in adipocytes is shown in FIG. As shown in FIG. 2, addition of sodium iodide to the medium did not show a significant change in the amount of fat accumulated in adipocytes. From the results of Experimental Examples 1 and 2, the fat accumulation inhibitory effect of the egg yolk extract (IY) of the iodine-rich egg is due to the iodide peptide contained in the iodine-rich egg, not the ionic iodine. It is suggested that there is.

[Experimental Example 3]
In Experimental Example 3, 3T3-L1 cells were differentiated into adipocytes and GPDH activity was measured. GPDH is an enzyme involved in adipose synthesis and is also an index of differentiation of adipose progenitor cells into adipocytes.

First, as in Example 1, 3T3-L1 was used in a medium supplemented with an egg yolk extract (IY) of an iodine-rich egg or an egg yolk extract (OY) of a normal egg so as to have a final concentration of 50 μg / mL. The cells were cultured for 10 days to obtain adipocytes. After culturing for 10 days, adipocytes (Day 10) were washed with PBS, the cells were disrupted, and GPDH activity was measured for the cell disruption solution using the GPDH activity measurement kit (Cell Garage Co., Ltd.). The obtained measurement results were corrected by the amount of protein in the cell disruption solution. For comparison, GPDH activity was measured in the same manner for cells (Day 0) before culturing for 10 days.

The measurement result of GPDH activity is shown in FIG. As shown in FIG. 3, when the yolk extract (IY) of an iodine-rich egg was added to the medium, the GPDH activity was compared with the case where the yolk extract (OY) of a normal egg was added to the medium. Was significantly reduced (p <0.05).

[Experimental Example 4]
In Experimental Example 4, 3T3-L1 cells were differentiated into adipocytes, and the expression level of a gene that is an index of differentiation from adipose progenitor cells to adipocytes was measured.

First, as in Example 1, 3T3-L1 was used in a medium supplemented with an egg yolk extract (IY) of an iodine-rich egg or an egg yolk extract (OY) of a normal egg so as to have a final concentration of 50 μg / mL. The cells were cultured for 10 days to obtain adipocytes. Total RNA was extracted from adipocytes (Day 10) after culturing for 10 days, and the expression levels of PPAR-γ2, adipogenin, leptin, and adiponectin genes were measured by real-time reverse transcription PCR (Polymerase Chain Reaction) according to a conventional method. did. The obtained measurement results were corrected by the expression level of the β-actin gene, which is an internal standard. For comparison, the gene expression level was measured in the same manner for the cells (Day 0) before culturing for 10 days.

The measurement results (relative values) of the expression levels of the PPAR-γ2, adipogenin, leptin, and adiponectin genes are shown in FIGS. 4A to 4D. "ND" in FIGS. 4A-4C means no detection. As can be seen from FIGS. 4A to 4D, when the egg yolk extract (IY) of the iodine-rich egg was added to the medium, it was compared with the case where the egg yolk extract (OY) of the normal egg was added to the medium. As a result, the expression levels of the PPAR-γ2, adiponectin, leptin, and adiponectin genes were significantly reduced (PPAR-γ2, adiponectin, and leptin: p <0.05, adiponectin: p <0.1).

[Experimental Example 5]
In Experimental Example 5, the subjects were divided into two groups to continuously ingest iodine-rich eggs or normal eggs, and the effect on body fat was evaluated by a double-blind method. As the iodine-rich egg, a chicken egg containing about 1300 μg of iodine per 100 g of the edible portion (Nosan Corporation, “Iodine Egg Light”) was used. In addition, a commercially available chicken egg was used as a normal egg.

The subjects were 56 men and women aged 40 to 65 years old and having a BMI (Body Mass Index) of 25 kg / m ² or more and less than 30 kg / m ² , and the test group (male: 14 and female: 14) and the control group ( It was divided into two groups (male: 14 people, female: 14 people). For the test group, one iodine-rich egg per day was continuously ingested as a boiled egg for 12 weeks. As for the control group, one normal egg per day was continuously ingested as a boiled egg for 12 weeks. Then, the body fat area was measured using the abdominal CT (Computed Tomography) image, and various tests such as blood test and urinalysis were performed and compared before and after the test.

Table 1 shows the changes in the total fat area, visceral fat area, and subcutaneous fat area before and after the test. Table 1 shows total cholesterol (T-cho), triglyceride (TG), LDL (Low Density Lipoprotein) cholesterol (LDL-cho), HDL (High Density Lipoprotein) cholesterol (HDL-cho), and free blood. The changes in the amounts of triiodothyronine (FT3) and free thyroxin (FT4) before and after the test are also shown. Table 1 shows the mean ± standard error for each group.

As shown in Table 1, in the test group, the total fat area and the visceral fat area significantly decreased before and after the test (total fat area: p = 0.037, visceral fat area: p = 0.001), but subcutaneous fat. No significant change was shown in area. On the other hand, in the control group, none of the total fat area, the visceral fat area, and the subcutaneous fat area showed a significant change.
In the comparison between the groups, the change amount and the rate of change in the visceral fat area were significantly larger in the test group (change amount: p = 0.049, change rate: p = 0.031), and a difference between the groups was observed.

The amounts of total cholesterol, triglyceride, LDL cholesterol, and HDL cholesterol in blood related to lipid metabolism did not show significant changes in either the test group or the control group.
In addition, the amounts of free triiodothyronine (FT3) and free thyroxine (FT4) in blood related to thyroid function did not show any significant changes in either the test group or the control group, and remained within the normal range.
From these results, the visceral fat reduction effect of the iodine-rich egg is not due to the improvement of lipid metabolism function or the improvement of thyroid function, but to the iodide peptide contained in the iodine-rich egg. Is suggested.

All documents, patent applications, and technical standards described herein are to the same extent as if the individual documents, patent applications, and technical standards were specifically and individually stated to be incorporated by reference. Incorporated herein by reference.

Claims (5)

4.2 A fat accumulation inhibitor containing a bird egg containing iodine of 2 mass ppm or more as an active ingredient. 4.2 An agent for suppressing the differentiation of adipose progenitor cells containing a bird egg containing iodine of 2 mass ppm or more as an active ingredient. 4.2 A visceral fat reducing agent containing a bird egg containing iodine of 2 mass ppm or more as an active ingredient. 4.2 Food and drink for reducing visceral fat containing bird eggs containing iodine of 2 mass ppm or more as an active ingredient. A visceral fat reducing agent that contains an egg yolk extract of a bird egg containing an iodide peptide as an active ingredient and is used so as to administer 300 μg or more of iodine per day.