WO2018159522A1 - Food composition and medical composition for preventing or alleviating metabolic syndrome - Google Patents

Abstract

Provided is a food composition for preventing or alleviating metabolic syndrome, the food composition containing, as active ingredients, a 67 kDa laminine receptor (67LR) agonist and a sulfur-containing compound found from an allium plant extract.

Description

Food composition and pharmaceutical composition for preventing or improving metabolic syndrome

The present invention relates to a food composition and a pharmaceutical composition for preventing or improving metabolic syndrome. This application claims priority on March 1, 2017 based on Japanese Patent Application No. 2017-038595 filed in Japan, the contents of which are incorporated herein by reference.

The metabolic syndrome is defined as a state corresponding to the following (1) and further corresponding to two or more of the following (2) to (4).
(1) The waist circumference is 85 cm or more for men and 90 cm or more for women (corresponding to a visceral fat area of 100 cm ² or more).
(2) Blood triglyceride is 150 mg / dL or higher and / or blood HDL cholesterol is 40 mg / dL or lower (3) Systolic blood pressure is 130 mmHg or higher and / or diastolic blood pressure is 85 mmHg or higher (4) Fasting Blood glucose level is 110mg / dL or more

In the state of metabolic syndrome, it is known that arteriosclerosis that causes myocardial infarction and cerebral infarction proceeds rapidly. For this reason, consumers are highly interested in metabolic syndrome, and various supplements and the like for anti-metabolic syndrome are commercially available.

Incidentally, the 67 kDa laminin receptor (hereinafter sometimes referred to as “67LR”) is a cell membrane protein that binds to laminin, which is a main component of the basement membrane (see, for example, Non-Patent Document 1). The inventors of the present invention bind to 67LR on the cell membrane, and epigallocatechin-gallate (hereinafter, sometimes referred to as “EGCG”), which is one of the main catechins contained in green tea, has anticancer activity. Has been clarified to exhibit (see, for example, Non-Patent Document 2).

An object of the present invention is to provide a composition effective for prevention, improvement or treatment of metabolic syndrome.

The present invention includes the following aspects.
[1] A food composition for preventing or improving metabolic syndrome, comprising as an active ingredient a 67LR agonist and a sulfur-containing compound found in an Allium plant extract.
[2] The food composition for preventing or improving metabolic syndrome according to [1], wherein the sulfur-containing compound found in the Allium plant extract contains alliin, allicin, diallyl disulfide, diallyl trisulfide, or diallyl tetrasulfide. object.
[3] The food composition for preventing or improving metabolic syndrome according to [1] or [2], wherein the 67LR agonist comprises EGCG, oolong tea polymerized polyphenol, procyanidin C1, or a derivative thereof.
[4] A method for preventing or improving metabolic syndrome, comprising a step of causing a human or animal to ingest the food composition according to any one of [1] to [3] (excluding medical practice for humans).
[5] A pharmaceutical composition for preventing or treating metabolic syndrome, comprising as an active ingredient a 67LR agonist and a sulfur-containing compound found in an Allium plant extract.
[6] The pharmaceutical composition for prevention or treatment of metabolic syndrome according to [5], wherein the sulfur-containing compound found in the Allium plant extract contains alliin, allicin, diallyl disulfide, diallyl trisulfide or diallyl tetrasulfide object.
[7] The pharmaceutical composition for preventing or treating metabolic syndrome according to [5] or [6], wherein the 67LR agonist comprises EGCG, oolong tea polymerized polyphenol, procyanidin C1, a derivative thereof, or a 67LR agonist antibody.

According to the present invention, it is possible to provide a composition effective for prevention, improvement or treatment of metabolic syndrome.

(A) is a graph which shows the result of having measured the transition of the body weight of the mouse | mouth of each group in Experimental example 1. FIG. (B) is a graph which shows the result of having measured transition of calorie intake of the mouse of each group in example 1 of an experiment. (A) is a graph which shows the result of having measured the mass of the white adipose tissue of the mouse | mouth of each group in Experimental example 2. FIG. (B) is a graph showing the results of measuring the mass of adipose tissue around the kidney of each group of mice in Experimental Example 2. (C) is a graph which shows the result of having measured the mass of the testicular periphery fat tissue of each group of mice in Experimental Example 2. (A) is a graph which shows the result of having measured the triglyceride in the serum of the mouse | mouth of each group in Experimental example 3. FIG. (B) is a graph showing the results of measurement of triglycerides in the liver tissue of each group of mice in Experimental Example 3. In Experimental example 4, it is a graph which shows the result of having measured the cholesterol in the serum of the mouse | mouth of each group. (A) to (h) are graphs showing the results of measuring the expression level of mRNA of a gene encoding a lipid metabolism-related protein in Experimental Example 5. (A)-(i) is a graph which shows the result of having measured the expression level of mRNA of the gene which codes a lipid metabolism related protein in Experimental example 6. FIG. (A)-(f) is a graph which shows the result of having measured the expression level of mRNA of the gene which codes a lipid metabolism related protein in Experimental example 7. FIG. (A)-(j) is a graph which shows the result of having measured the expression level of mRNA of the gene which codes a lipid metabolism related protein in Experimental example 8. FIG. (A)-(f) is a graph which shows the result of having measured the expression level of mRNA of the gene which codes a lipid metabolism related protein in Experimental example 9. FIG. In Experimental example 10, it is a graph which shows the result of having measured the glucose level (blood glucose level) in the serum of the mouse | mouth of each group. In Experimental example 11, it is a graph which shows the result of having measured the free fatty acid in the serum of the mouse | mouth of each group. (A) is a graph which shows the result of having measured the blood insulin density | concentration of the mouse | mouth of each group in Experimental example 12. FIG. (B) is a graph showing the results of calculating HOMA-IR values of mice in each group in Experimental Example 12. (A) is a graph showing the results of measuring the expression of the MCP1 gene in mesenteric adipose tissue in Experimental Example 13. (B) is a graph showing the results of measuring the expression of the MCP1 gene in the adipose tissue around the kidney in Experimental Example 13. (A) is a graph showing the results of measuring Aspartate Aminotransferase (AST) activity in the serum of mice of each group in Experimental Example 14. (B) is a graph showing the results of measurement of Alanine aminotransferase (ALT) activity in the serum of each group of mice in Experimental Example 14. (A)-(c) is a graph which shows the result of having measured the expression level of mRNA of the gene which codes a lipid metabolism related protein in Experimental example 15. FIG.

[Food Composition for Prevention or Improvement of Metabolic Syndrome]
In one embodiment, the present invention provides a food composition for preventing or improving metabolic syndrome, comprising a 67LR agonist and a sulfur-containing compound found in an onion plant extract as active ingredients.

As will be described later in the Examples, the inventors ingested a high-fat high-sucrose diet, and mice ingested the food composition of the present embodiment were able to suppress weight gain, suppress body fat accumulation, and serum. Decreased neutral fat in the middle and liver, decreased blood cholesterol, improved lipid metabolism at the gene level, suppressed blood sugar level increase, decreased serum free fatty acid, improved insulin resistance, inflammation related to white adipose tissue It was clarified that it showed significant effects such as suppression of gene expression and suppression of liver damage.

Therefore, the food composition of the present embodiment can be suitably used for the prevention or improvement of metabolic syndrome.

In addition, as will be described later in the Examples, the inventors have found that the above-described effects of preventing or improving the metabolic syndrome are observed when a 67LR agonist alone or a sulfur-containing compound alone found in an Allium plant extract is ingested. It was clarified that it was observed only when a 67LR agonist and a sulfur-containing compound found in an Allium plant extract were used in combination.

In the present specification, “containing as an active ingredient” means that the composition contains a 67LR agonist and a sulfur-containing compound found in an Allium plant extract in an amount sufficient to obtain the effects of the present invention. Meaning containing, specific content can be suitably set according to a composition (other components).

(67LR agonist)
In the food composition of the present embodiment, as the 67LR agonist, any substance that can transmit a signal to the 67 kDa laminin receptor (67LR) can be used without particular limitation. The 67LR agonist is preferably one that can be added to food.

More specific 67LR agonists include, for example, EGCG, oolong tea polymerized polyphenol, procyanidin C1, and derivatives thereof.

EGCG is an ester of epigallocatechin and gallic acid and is a kind of catechin. EGCG is a catechin that is particularly abundant in tea among plants.

Examples of EGCG derivatives include compounds in which some functional groups are changed using EGCG as a basic skeleton. More specifically, methylated EGCG is mentioned, for example. Specific examples of methylated EGCG include (−)-epigallocatechin-3-O- (3-O-methyl) gallate represented by the following formula (1). The compound represented by the following formula (1) is a compound contained in “Benifuuki”, which is one of green tea varieties, and is safe to add to food.

Oolong tea polymerization polyphenol is a general term for compounds in which catechins are bound in a complex manner formed by an enzymatic reaction or a thermal polymerization reaction in a unique method of producing oolong tea called semi-fermentation. Oolong tea polymerization polyphenols include, for example, dimers of catechins and trimers of catechins. Examples of catechin dimers include oolong homobisflavans such as oolong homobisflavan A, monodesgaloyl oolong homobisflavan A, oolong homobisflavan B, oolong homobisflavan C, and the like.

Examples of the derivative of oolong tea polymerized polyphenol include compounds in which some functional groups are changed using oolong tea polymerized polyphenol as a basic skeleton.

Procyanidin C1 is a trimer of epicatechin that is abundant in plants such as apples and grapes. Examples of the derivatives of procyanidin C1 include compounds in which some functional groups are changed using procyanidin C1 as a basic skeleton.

By derivatizing EGCG, oolong tea polymerized polyphenol, procyanidin C1, etc., for example, physical properties such as water solubility and fat solubility of these compounds, biology such as 67LR agonist activity, retention in blood, biotoxicity, etc. Activity and the like can be adjusted to a desired range.

The 67LR agonist may be a solvate of EGCG, oolong tea polymerized polyphenol, procyanidin C1, or a derivative thereof, a pharmaceutically acceptable salt, or a solvate of a pharmaceutically acceptable salt. It may be.

Examples of the pharmaceutically acceptable salt include inorganic acid salts, alkali metal salts, alkaline earth metal salts, metal salts, ammonium salts, organic amine addition salts, amino acid addition salts, and the like. More specifically, for example, inorganic acid salts such as hydrochloride, sulfate, hydrobromide, nitrate, phosphate; acetate, mesylate, succinate, maleate, fumarate, Organic salts such as citrate and tartrate; alkali metal salts such as sodium salt and potassium salt; alkaline earth metal salts such as magnesium salt and calcium salt; metal salts such as aluminum salt and zinc salt; ammonium salt and tetra Examples include ammonium salts such as methylammonium salts; organic amine addition salts such as morpholine and piperidine; amino acid addition salts such as glycine, phenylalanine, lysine, aspartic acid, and glutamic acid.

In addition, solvates of EGCG, oolong tea polymerized polyphenol, procyanidin C1 or derivatives thereof, and solvates of pharmaceutically acceptable salts are particularly pharmaceutically acceptable solvates. There is no limitation, and examples thereof include hydrates and organic solvates.

The 67LR agonist may be chemically synthesized or purified from a natural product. Or you may make it contain by using the natural product containing a 67LR agonist for the raw material of a food composition. Examples of natural products containing 67LR agonist include green tea, green tea extract, oolong tea, oolong tea extract, black tea, black tea extract, apple, apple extract, grape, grape extract, grape seed, grape seed extract, Cocoa beans, cacao bean extract, apricot, apricot extract, kiwi, kiwi extract, cherry, sanranbo extract, strawberry, strawberry extract, cranberry, cranberry extract, blueberry, blueberry extract, loquat, loquat extract Product, black soybean, black soybean extract, cinnamon, cinnamon extract, pine bark, pine bark extract, peanut, peanut extract and the like.

The food composition of the present embodiment may contain one of the 67LR agonists described above alone, or may contain a mixture of two or more.

(Sulfur-containing compounds found in allium plant extracts)
In the food composition of the present embodiment, examples of the Allium plant include garlic, leeks, leeks, onions, and rakkyo.

Also, examples of sulfur-containing compounds found in the genus plant extract include alliin, allicin, diallyl disulfide, diallyl trisulfide, diallyl tetrasulfide, and the like.

Here, alliin is a sulfur-containing compound contained in, for example, about 1% by mass in garlic. Alliin is an odorless compound and is stored in the cytoplasm of bulb mesophyll storage cells. On the other hand, alliinase, an enzyme that acts on alliin, is stored in the vacuole of vascular sheath cells.

In intact garlic, no odor is detected because the enzyme and substrate are in different parts. However, when the garlic bulb tissue is damaged, the enzyme alliinase and the substrate alliin react to produce allicin. Allicin is a compound that has a characteristic smell of garlic.

Because allicin is highly reactive, it reacts with other compounds and allicin itself. As a result, diallyl disulfide and the like are produced from allicin. Further, diallyl disulfide changes to diallyl trisulfide, diallyl tetrasulfide and the like having different numbers of sulfur. These sulfur-containing compounds derived from allicin also have a odor characteristic of garlic.

In the present specification, the term “sulfur-containing compound found in an onion plant extract” is not limited to a sulfur-containing compound derived from an allium plant, but in an extract obtained by crushing an onion plant. Means a sulfur-containing compound present in any one of its origins. The “sulfur-containing compound found in the genus Leek plant extract” may be, for example, a sulfur-containing compound extracted from a genus Leek plant, or a sulfur-containing compound extracted from a natural product other than a genus Leek plant. It may be alliin, allicin, diallyl disulfide, diallyl trisulfide, diallyl tetrasulfide and the like which are chemically synthesized.

The sulfur-containing compound found in the Allium plant extract may contain a purified compound, or may be contained by using the above-mentioned crushed material of the Allium plant as a raw material of the food composition. . The food composition of the present embodiment may contain one kind of sulfur-containing compound found in the Allium plant extract alone, or may contain a mixture of two or more kinds.

The food composition of the present embodiment seems to ingest a sulfur-containing compound found in 0.1 to 100 mg / kg body weight of 67LR agonist and 0.05 to 50 mg / kg body weight of an onion plant extract per day. May be used. Further, the food composition may be used so as to be taken once a day or divided into about 2 to 4 times a day.

The food composition of this embodiment may be, for example, in the form of a supplement, in the form of a beverage, in the form of a solid, semi-solid or gel food, It may be in the form of any cooked food. Examples of supplement shapes include capsule shapes.

The food composition of the present embodiment may be a functional display food. “Functional labeling food” means food that has been reported to the Consumer Affairs Agency as indicating the functionality on the product package based on scientific evidence. As the display, for example, “prevent or improve metabolic syndrome”, “suppress body weight gain”, “suppress body fat accumulation”, “decrease triglyceride”, “decrease blood cholesterol” , “Improving lipid metabolism”, “suppressing an increase in blood glucose level”, “decreasing free fatty acid”, “improving insulin resistance”, etc., but are not limited thereto. Moreover, even if it is a food composition without a functional display, it is also conceivable that the functionality is described in a flyer, an advertisement, etc., or displayed by voice or the like to be manufactured and sold.

The food composition of the present embodiment may be a special purpose food. Special-purpose food means food that is labeled for special use that is suitable for the growth, maintenance and recovery of health of infants, toddlers, pregnant women, sick people, etc. with the permission of the government. The food composition of the present embodiment may be a food for a sick person among special-purpose foods. Alternatively, the food composition of the present embodiment may be a food for specified health use among special purpose foods. Specified health food means food that has been recognized on the basis of scientific evidence to be useful for maintaining and improving health and is allowed to display its effects. The government will review the displayed effects and safety, and each food will be approved by the Commissioner of the Consumer Affairs Agency.

[Prevention or improvement of metabolic syndrome]
In one embodiment, the present invention provides a method for preventing or ameliorating metabolic syndrome (excluding medical practice for humans), comprising the step of causing a human or animal to take the food composition described above. In the method for preventing or improving metabolic syndrome of the present embodiment, the medical action means an action in which a doctor (including a person who receives an instruction from a doctor) performs a treatment on a human.

In the method for preventing or improving metabolic syndrome of the present embodiment, the amount of the food composition to be ingested by humans or animals is 0.1 to 100 mg / kg body weight of 67LR agonist and 0.05 to 50 mg / kg per day. The amount of the sulfur-containing compound found in the Allium plant extract of body weight can be mentioned. The food composition may be taken once a day or divided into about 2 to 4 times a day.

[Pharmaceutical composition for prevention or treatment of metabolic syndrome]
In one embodiment, the present invention provides a pharmaceutical composition for preventing or treating metabolic syndrome, which comprises a 67LR agonist and a sulfur-containing compound found in an Allium plant extract as active ingredients.

In the pharmaceutical composition of the present embodiment, examples of the 67LR agonist include EGCG, oolong tea polymerized polyphenol, procyanidin C1, EGCG derivative, oolong tea polymerized polyphenol derivative, procyanidin C1 derivative, for example, anti-67LR antibody (67LR agonist antibody), A compound represented by the following formula (2), which is methylated EGCG synthesized chemically, can be used.

Further, as the sulfur-containing compound found in the genus plant extract, the same compounds as described above can be used, and may be alliin, allicin, diallyl disulfide, diallyl trisulfide, diallyl tetrasulfide and the like.

The pharmaceutical composition of the present embodiment is administered orally, for example, in the form of tablets, capsules, elixirs, microcapsules, etc., or parenterally in the form of injections, suppositories, skin external preparations, etc. can do. More specifically, examples of the external preparation for skin include dosage forms such as ointments and patches.

The pharmaceutical composition of this embodiment may contain a pharmaceutically acceptable carrier. As the pharmaceutically acceptable carrier, those usually used for the preparation of pharmaceutical compositions can be used without particular limitation. More specifically, for example, binders such as gelatin, corn starch, gum tragacanth and gum arabic; excipients such as starch and crystalline cellulose; swelling agents such as alginic acid; solvents for injections such as water, ethanol and glycerin; Examples thereof include adhesives such as rubber adhesives and silicone adhesives.

The pharmaceutical composition may contain an additive. Additives include lubricants such as calcium stearate and magnesium stearate; sweeteners such as sucrose, lactose, saccharin and maltitol; flavoring agents such as peppermint and red mono oil; stabilizers such as benzyl alcohol and phenol; phosphoric acid Buffers such as salts and sodium acetate; Solubilizing agents such as benzyl benzoate and benzyl alcohol; Antioxidants; Preservatives and the like.

The pharmaceutical composition is required for generally accepted pharmaceutical practice by appropriately combining the 67LR agonist described above, the sulfur-containing compound found in the Allium plant extract, the pharmaceutically acceptable carriers and additives described above. It can be formulated by mixing in unit dosage form.

The dosage of the pharmaceutical composition varies depending on the patient's symptoms, body weight, age, sex, etc., and cannot be generally determined, but in the case of oral administration, for example, 0.1 to 100 mg / kg body weight per dosage unit form And a sulfur-containing compound found in an onion extract of 0.05 to 50 mg / kg body weight may be administered. In the case of injections, for example, per unit dosage form, a sulfur-containing compound found in a 0.1 to 100 mg / kg body weight 67LR agonist and 0.05 to 50 mg / kg body weight onion plant extract. What is necessary is just to administer.

In addition, the daily dose of the pharmaceutical composition varies depending on the patient's symptoms, body weight, age, sex, etc., and cannot be determined unconditionally. For example, 0.1-100 mg / kg body weight per adult day The amount to be a sulfur-containing compound found in the 67LR agonist and 0.05 to 50 mg / kg body weight of the onion plant extract may be administered once a day or divided into about 2 to 4 times a day.

[Other Embodiments]
In one embodiment, the present invention provides a method for preventing or treating metabolic syndrome comprising administering to a patient in need of treatment an effective amount of a 67LR agonist and a sulfur-containing compound found in an Allium plant extract. provide. Here, the 67LR agonist and the sulfur-containing compound found in the Allium plant extract are the same as those described above for the pharmaceutical composition.

In one embodiment, the present invention provides a composition for preventing or treating metabolic syndrome, comprising a 67LR agonist and a sulfur-containing compound found in an Allium plant extract as active ingredients. . The composition of this embodiment may be a pharmaceutical composition. Here, the 67LR agonist and the sulfur-containing compound found in the Allium plant extract are the same as those described above for the pharmaceutical composition.

In one embodiment, the present invention provides the use of a 67LR agonist and a sulfur-containing compound found in an Allium plant extract for the manufacture of a preventive or therapeutic agent for metabolic syndrome. Here, the 67LR agonist and the sulfur-containing compound found in the Allium plant extract are the same as those described above for the pharmaceutical composition.

Next, the present invention will be described in more detail with reference to experimental examples, but the present invention is not limited to the following experimental examples.

[Experimental Example 1]
(Anti-obesity effect by combined use of green tea extract and diallyl disulfide)
Mice were ingested in combination with a high-fat, high-sucrose diet, green tea extract, and diallyl disulfide (hereinafter sometimes referred to as “DADS”), and the effect on obesity was examined.

Specifically, after a 12-week-old male C57BL / 6J mouse is preliminarily raised for 1 week, a normal diet (AIN-93G compliant diet) is fed so that the average body weight is equal. "HF / HS group" that ingests a high sucrose (High Fat High Sugar, HF / HS) diet, "green tea extract group" that ingests a high fat, high sucrose diet and green tea extract, high fat, high sucrose The food was divided into 5 groups: a “DADS group” for taking a combination of food and DADS, and a “green tea extract + DADS group” for taking a high fat and high sucrose diet, a green tea extract and DADS together. Subsequently, each group of mice was given each feed at 4 g / day and was reared for 10 weeks with free drinking water.

Thereafter, the mice were killed by blood sampling of the abdominal aorta under isoflurane anesthesia after fasting for 10 hours. Body weight and food intake were measured every other week until sacrifice.

Green tea extract was prepared as follows. First, dried Yabukita tea leaves obtained by steaming dried Yabukita tea leaves were pulverized to about 5 mm with a mill. Subsequently, 20 times (w / w) of 70 ° C. warm water was added to the ground tea leaves. After stirring at 70 ± 2 ° C. for 10 minutes, solid-liquid separation was performed with a 100-mesh wire mesh to obtain an extract. Subsequently, the extract was centrifuged at 700 × g for 10 minutes, and the supernatant was collected. Subsequently, the recovered supernatant was treated with a vacuum freeze dryer (model “FZ-12CS”, manufactured by LABCONCO) and freeze-dried to obtain a powdery green tea extract. The powdered green tea extract contained 11.9% by weight of EGCG. Table 1 below shows the results of component analysis of the prepared green tea extract (powder).

The prepared green tea extract was added to the feed of “green tea extract group” and “green tea extract + DADS group” so as to be 0.1% by mass. In addition, DADS was added to the feed of “DADS group” and “green tea extract + DADS group” so as to be 0.0189 g / kg feed. Table 2 below shows the composition of the feed of each group.

FIG. 1 (a) is a graph showing the results of measuring the change in body weight of mice in each group. In this experimental example and the following experimental examples, Tukey's test was used for statistical processing of the experimental results, and a p value of less than 0.05 was considered significant.

As a result, in the green tea extract group and the DADS group, no significant effect on the weight increase induced by the HF / HS diet was observed. On the other hand, it was revealed that the increase in body weight induced by the HF / HS diet was significantly suppressed in the green tea extract + DADS group ingested in combination with the green tea extract and DADS.

FIG. 1 (b) is a graph showing the results of measuring changes in caloric intake of mice in each group. As a result, it was clarified that the caloric intake was similar in all groups of mice. The reason why the calorie intake of the control group is low is that a normal diet was given.

From the above results, it was clarified that anti-obesity action was exhibited by ingesting the green tea extract and DADS together.

[Experiment 2]
(Inhibition of body fat accumulation by combined use of green tea extract and DADS)
From each group of mice sacrificed in Experimental Example 1, white adipose tissue, kidney peripheral adipose tissue, and testicular periphery adipose tissue were extracted and their masses were measured. Fig.2 (a) is a graph which shows the result of having measured the mass of the white adipose tissue. FIG.2 (b) is a graph which shows the result of having measured the mass of the kidney periphery fat tissue. FIG.2 (c) is a graph which shows the result of having measured the mass of the testicular periphery fat tissue.

As a result, the green tea extract group and the DADS group showed no significant effect on the accumulation of body fat induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the accumulation of body fat induced by the HF / HS diet was significantly suppressed.

[Experiment 3]
(Triglyceride lowering effect by combined use of green tea extract and DADS)
Blood collected from each group of mice sacrificed in Experimental Example 1 was allowed to stand at 37 ° C. for 2 hours to clot, and then centrifuged at 4 ° C. and 2,000 × g for 15 minutes to collect serum. Subsequently, the neutral fat in the obtained serum was measured using a commercially available triglyceride measurement kit (manufactured by Wako Pure Chemical Industries, Ltd.). Further, the neutral fat in the liver tissue collected from each group of mice in Experimental Example 1 was measured using a commercially available triglyceride measurement kit (manufactured by Wako Pure Chemical Industries, Ltd.).

FIG. 3 (a) is a graph showing the measurement results of triglycerides in the serum of each group of mice. Moreover, FIG.3 (b) is a graph which shows the measurement result of the triglyceride in the liver tissue of the mouse | mouth of each group.

As a result, in the green tea extract group and the DADS group, there was no significant effect on the increase in neutral fat induced by the HF / HS diet. On the other hand, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the increase in neutral fat induced by the HF / HS diet was significantly suppressed.

[Experimental Example 4]
(Serum cholesterol lowering effect by combined use of green tea extract and DADS)
Total cholesterol in the serum of each group of mice prepared in Experimental Example 3 was measured using a commercially available cholesterol measurement kit (Wako Pure Chemical Industries, Ltd.).

FIG. 4 is a graph showing the measurement results of cholesterol in the serum of each group of mice. As a result, in the green tea extract group and the DADS group, there was no significant effect on the increase in serum cholesterol induced by the HF / HS diet. In contrast, it was revealed that the increase in serum cholesterol induced by the HF / HS diet was significantly suppressed in the green tea extract + DADS group ingested in combination with the green tea extract and DADS.

[Experimental Example 5]
(Effects of combined use of green tea extract and DADS on expression of genes related to lipid metabolism)
CDNA was prepared from liver tissue collected from each group of mice sacrificed in Experimental Example 1, and the expression level of mRNA of each gene encoding each lipid metabolism-related protein shown in Table 3 was measured by real-time PCR. Table 3 also shows the sequence numbers of the base sequences of the primers used for PCR of each gene.

Here, SREBP-1c is a protein involved in fatty acid control. ACC1 is an enzyme involved in the control of fatty acid synthesis and fatty acid β oxidation. FAS is a multi-active domain enzyme that synthesizes fatty acids from malonyl CoA and acetyl CoA. SCD1 is an enzyme that converts saturated fatty acids into unsaturated fatty acids. ACOX1 is a fatty acid β-oxidizing enzyme. RXRα, RXRβ, and RXRγ each form a heterodimer with peroxisome proliferator-activated receptor alpha (PPAR), which is a transcription factor that regulates various target genes involved in sugar / lipid metabolism. It is a nuclear receptor that functions as an inducible transcription factor.

FIGS. 5 (a) to 5 (h) are graphs showing the results of measuring the expression levels of mRNAs of genes encoding the lipid metabolism-related proteins shown in Table 3, respectively. As a result, in the green tea extract group and the DADS group, no significant effect on changes in lipid metabolism-related gene expression induced by the HF / HS diet was observed. On the other hand, it was revealed that the change in lipid metabolism-related gene expression induced by the HF / HS diet was significantly suppressed in the green tea extract + DADS group ingested in combination with the green tea extract and DADS. That is, it became clear that the expression variation of the lipid metabolism-related gene whose expression is increased by the HF / HS diet is suppressed, and the expression variation of the lipid metabolism-related gene whose expression is decreased by the HF / HS diet is suppressed.

From this result, it was clarified that, when ingested in combination with green tea extract and DADS, the expression change of lipid metabolism-related genes induced by HF / HS diet is suppressed, and lipid metabolism is improved.

[Experimental Example 6]
(Effects of combined use of green tea extract and DADS on expression of lipid metabolism-related genes in white adipose tissue)
CDNA was prepared from white adipose tissue collected from each group of mice sacrificed in Experimental Example 1, and the expression level of mRNA of each gene encoding each lipid metabolism-related protein shown in Table 4 was measured by real-time PCR. Table 4 also shows the SEQ ID NOs of the base sequences of the primers used for PCR of each gene.

Here, PGC1α is a transcriptional regulator of PPAR. It promotes the expression of various genes involved in energy metabolism and mitochondrial biogenesis, and is also involved in lipolysis. PPARγ and PPARδ are transcription factors that regulate various target genes involved in sugar / lipid metabolism. UCP1 and UCP2 are proteins encoded by PPAR target genes. Uncouples oxidative phosphorylation in the inner mitochondrial membrane and dissipates energy as heat. UCP1 is abundant in brown adipocytes, and UCP2 is present in white adipocytes, skeletal muscle, spleen, small intestine and the like. CPT1α is an enzyme involved in lipolysis. RXRα, RXRβ, and RXRγ are as described above.

6 (a) to 6 (i) are graphs showing the results of measuring the expression level of the mRNA of the gene encoding each lipid metabolism-related protein shown in Table 4, respectively. As a result, in the green tea extract group and the DADS group, there was no significant effect on the decrease in lipid metabolism-related gene expression in white adipose tissue induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, the decrease in lipid metabolism-related gene expression in white adipose tissue induced by the HF / HS diet can be significantly suppressed. It became clear.

In particular, UCP1 is a protein directly involved in fat burning, and UCP2 has a similar function. For this reason, the combined use of green tea extract and DADS significantly suppresses the decrease in the expression of these proteins indicates that the combined use of green tea extract and DADS directly improves lipid metabolism.

[Experimental Example 7]
(Effects of combined use of green tea extract and DADS on expression of lipid metabolism-related genes in adipose tissue around the kidney)
CDNA was prepared from the perirenal adipose tissue collected from each group of mice sacrificed in Experimental Example 1, and the expression level of mRNA of a gene encoding each lipid metabolism-related protein shown in Table 5 below was measured by real-time PCR. Table 5 also shows the SEQ ID NOs of the base sequences of the primers used for PCR of each gene.

Here, PPARγ, PPARδ, PGC1α, UCP1, and UCP2 are as described above. UCP3 is a protein encoded by the target gene of PPAR, like UCP1 and UCP2, and has a function of uncoupling oxidative phosphorylation in the inner mitochondrial membrane and dissipating energy as heat. UCP3 is mainly present in muscle tissues such as skeletal muscle and heart.

FIGS. 7 (a) to (f) are graphs showing the results of measuring the mRNA expression levels of the genes encoding the lipid metabolism-related proteins shown in Table 5, respectively. As a result, in the green tea extract group and the DADS group, there was no significant effect on the decrease in the expression of genes related to lipid metabolism in the perirenal adipose tissue induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, the decrease in the expression of lipid metabolism-related genes in the adipose tissue around the kidney induced by the HF / HS diet is significantly suppressed. Became clear.

[Experimental Example 8]
(Effects of combined use of green tea extract and DADS on expression of lipid metabolism-related genes in brown adipose tissue)
CDNA was prepared from the brown adipose tissue collected from each group of mice sacrificed in Experimental Example 1, and the expression level of mRNA of the gene encoding each lipid metabolism-related protein shown in Table 6 below was measured by real-time PCR. Table 6 also shows the sequence numbers of the base sequences of the primers used for PCR of each gene.

Here, UCP1, UCP2, UCP3, PGC1α, PPARγ, PPARδ, CPT1α, RXRα and RXRβ are as described above. CD36 is a membrane glycoprotein present in platelets, mononuclear phagocytes, adipocytes, hepatocytes, muscle cells and some epithelia, and is involved in fatty acid transport.

8 (a) to 8 (j) are graphs showing the results of measuring the expression level of the mRNA of the gene encoding each lipid metabolism-related protein shown in Table 6, respectively. As a result, in the green tea extract group and the DADS group, there was no significant effect on the decrease in lipid metabolism-related gene expression in brown adipose tissue induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, there is a tendency to significantly suppress the decrease in lipid metabolism-related gene expression in brown adipose tissue induced by the HF / HS diet. It became clear that there was.

In particular, it was revealed that the combined use of green tea extract and DADS significantly increased the expression of UCP1 directly involved in fat burning. This result indicates that the combined use of green tea extract and DADS directly improves lipid metabolism.

[Experimental Example 9]
(Effects of combined use of green tea extract and DADS on expression of lipid metabolism-related genes in skeletal muscle)
CDNA was prepared from the skeletal muscle collected from each group of mice sacrificed in Experimental Example 1, and the expression level of mRNA of the gene encoding each lipid metabolism-related protein shown in Table 7 below was measured by real-time PCR. Table 7 also shows the sequence numbers of the base sequences of the primers used for PCR of each gene.

Here, UCP2, UCP3, PGC1α and CD36 are as described above. LPL is an enzyme that is synthesized and secreted by adipose tissue or the like and is present on the surface of vascular endothelial cells of capillaries. LPL has a function of degrading neutral fat in blood outside the cell into free fatty acid and glycerol and taking the free fatty acid into the cell. PDK4 is an enzyme that converts the intracellular energy source from carbohydrates to lipids by inhibiting the synthesis of acetyl-CoA from pyruvic acid.

FIGS. 9 (a) to 9 (f) are graphs showing the results of measuring the expression level of mRNA of genes encoding the lipid metabolism-related proteins shown in Table 7, respectively. As a result, in the green tea extract group and the DADS group, there was no significant effect on changes in lipid metabolism-related gene expression in skeletal muscle. On the other hand, it was revealed that the expression of lipid metabolism-related genes in skeletal muscle was significantly increased in the green tea extract + DADS group ingested in combination with the green tea extract and DADS.

[Experimental Example 10]
(Inhibition of blood glucose level increase by combined use of green tea extract and DADS)
The glucose concentration (blood glucose level) in the serum of each group of mice prepared in Experimental Example 3 was measured using a commercially available glucose measurement kit (Wako Pure Chemical Industries, Ltd.).

FIG. 10 is a graph showing the measurement results of the glucose concentration (blood glucose level) in the serum of each group of mice. As a result, in the green tea extract group and the DADS group, no significant effect on the increase in blood glucose level induced by the HF / HS diet was observed. On the other hand, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the increase in blood glucose level induced by the HF / HS diet was significantly suppressed.

[Experimental Example 11]
(Serum free fatty acid lowering effect by combined use of green tea extract and DADS)
The free fatty acid concentration in the serum of each group of mice prepared in Experimental Example 3 was measured using a commercially available free fatty acid measurement kit (Wako Pure Chemical Industries, Ltd.).

FIG. 11 is a graph showing the measurement results of free fatty acids in the serum of each group of mice. As a result, in the green tea extract group and the DADS group, no significant effect on the free fatty acid concentration in serum was observed. On the other hand, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the free fatty acid concentration in the serum was significantly reduced.

[Experimental example 12]
(Insulin resistance improvement effect by combined use of green tea extract and DADS)
The insulin concentration in the serum of each group of mice prepared in Experimental Example 3 was measured by ELISA. Further, based on the measured insulin concentration and the blood glucose level measured in Experimental Example 10, a HOMA-IR value that is an index of insulin resistance was calculated. The HOMA-IR value was calculated according to the following formula (F1).
HOMA-IR value = (fasting blood insulin concentration (μU / mL) × fasting blood glucose level (mg / dL)) / 405 (F1)

FIG. 12 (a) is a graph showing the measurement results of blood insulin concentration in each group of mice. As a result, in the green tea extract group and the DADS group, there was no significant effect on the increase in blood insulin concentration induced by the HF / HS diet. On the other hand, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the increase in blood insulin concentration induced by the HF / HS diet was significantly suppressed. The increase in blood insulin concentration is caused by a decrease in insulin sensitivity.

FIG. 12 (b) is a graph showing the HOMA-IR value of each group of mice. A high HOMA-IR value indicates that insulin resistance is high, that is, insulin sensitivity is low.

As a result, in the green tea extract group and the DADS group, there was no significant effect on the increase in insulin resistance due to the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the increase in insulin resistance due to the HF / HS diet was significantly improved.

[Experimental Example 13]
(Inhibition of expression of inflammation-related genes in white adipose tissue by combined use of green tea extract and DADS)
Chronic inflammation in adipose tissue is said to contribute to obesity. Therefore, cDNA was prepared from white adipose tissue collected from each group of mice sacrificed in Experimental Example 1, and the mRNA expression level of the gene encoding monocytic chemical protein 1 (MCP1), which is an inflammation-related protein, was determined by real-time PCR. It was measured. As the primer, a sense primer (SEQ ID NO: 37) and an antisense primer (SEQ ID NO: 38) were used. MCP1 is a protein involved in tissue infiltration of monocytes and T cells in various inflammatory diseases.

FIG. 13 (a) is a graph showing the results of measuring the expression of the MCP1 gene in mesenteric adipose tissue. FIG. 13 (b) is a graph showing the results of measuring the expression of the MCP1 gene in the adipose tissue around the kidney.

As a result, in the green tea extract group and the DADS group, there was no significant effect on the increase in expression of the MCP1 gene induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that the increase in the expression of the MCP1 gene induced by the HF / HS diet was significantly suppressed or reduced. .

[Experimental Example 14]
(Evaluation of liver damage by combined use of green tea extract and DADS)
Aspartate Aminotransferase (AST) activity and Alanine aminotransferase (ALT) activity in the serum of each group of mice prepared in Experimental Example 3 were measured using a commercially available kit (manufactured by Wako Pure Chemical Industries, Ltd.). AST activity and ALT activity are indicators of liver damage.

FIG. 14 (a) is a graph showing the measurement results of AST activity in the serum of each group of mice. Moreover, FIG.14 (b) is a graph which shows the measurement result of the ALT activity in the serum of the mouse | mouth of each group.

As a result, in the green tea extract group and the DADS group, there was no significant effect on liver damage induced by the HF / HS diet. In contrast, in the green tea extract + DADS group ingested in combination with the green tea extract and DADS, it was revealed that liver damage induced by the HF / HS diet was significantly suppressed.

[Experimental Example 15]
(Effects of combined treatment of EGCG and DADS on expression of lipid metabolism-related genes in adipocytes)
First, the preadipocyte cell line was differentiated into mature adipocytes. Specifically, 3T3-L1 cells, a mouse preadipocyte cell line, were seeded in a 2 mL dish at 1 × 10 ⁵ cells / mL and cultured at 37 ° C. under 5% CO ₂ with water vapor saturation. . Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS) was used as the medium. Subsequently, 48 hours after the cells became confluent, dexamethasone (1 μM), insulin (10 μg / mL), and 3-isobutyl-1-methylxanthine (IBMX, 0.5 μM) were added to the medium. Replaced with% FCS-DMEM. After further incubation for 48 hours, the medium was replaced with 10% FCS-DMEM supplemented with insulin (10 mg / mL). Thereafter, the medium was replaced with 10% FCS-DMEM twice every 48 hours to differentiate the cells into mature adipocytes.

Subsequently, EGCG (5 μM), DADS (10 μM), or EGCG (5 μM) and DADS (10 μM) were added to 3T3-L1 cells differentiated into mature adipocytes, respectively, and cultured for 24 hours. In addition, cells to which neither EGCG nor DADS were added were used as controls. Thereafter, each cell was recovered and RNA was extracted. After synthesizing cDNA, the expression level of mRNA of a gene encoding each lipid metabolism-related protein shown in Table 8 below was measured by real-time PCR. Table 8 also shows the sequence numbers of the base sequences of the primers used for PCR of each gene.

Here, PGC1α, PPARγ, and LPL are as described above. 15 (a) to 15 (c) are graphs showing the results of measuring the mRNA expression levels of the genes encoding the lipid metabolism-related proteins shown in Table 8, respectively.

In FIG. 15, “EGCG” indicates the result of the EGCG group to which EGCG was added, “DADS” represents the result of the DADS group to which DADS was added, and “EGCG + DADS” represents the result of the EGCG + DADS group to which EGCG and DADS were added in combination. Results are shown.

As a result, in the EGCG group and the DADS group, no significant effect on the change in the expression of genes related to lipid metabolism in adipocytes was observed. In contrast, in the EGCG + DADS group to which EGCG and DADS were added in combination, it became clear that the expression of lipid metabolism-related genes in adipocytes was significantly increased.

This result further supports that the combined use of EGCG and DADS directly improves lipid metabolism.

According to the present invention, it is possible to provide a composition effective for prevention, improvement or treatment of metabolic syndrome.

Claims (7)

1. A food composition for preventing or improving metabolic syndrome, comprising as an active ingredient a 67 kDa laminin receptor (67LR) agonist and a sulfur-containing compound found in an extract of the genus Allium.
2. The food composition for preventing or ameliorating metabolic syndrome according to claim 1, wherein the sulfur-containing compound found in the Allium plant extract contains alliin, allicin, diallyl disulfide, diallyl trisulfide or diallyl tetrasulfide.
3. The food composition for preventing or improving metabolic syndrome according to claim 1 or 2, wherein the 67LR agonist comprises epigallocatechin gallate (EGCG), oolong tea polymerized polyphenol, procyanidin C1, or a derivative thereof.
4. A method for preventing or ameliorating metabolic syndrome, comprising a step of allowing a human or animal to take the food composition according to any one of claims 1 to 3 (however, excluding medical practice for humans).
5. A pharmaceutical composition for preventing or treating metabolic syndrome, comprising a 67LR agonist and a sulfur-containing compound found in an onion plant extract as active ingredients.
6. 6. The pharmaceutical composition for prevention or treatment of metabolic syndrome according to claim 5, wherein the sulfur-containing compound found in the Allium plant extract contains alliin, allicin, diallyl disulfide, diallyl trisulfide or diallyl tetrasulfide.
7. The pharmaceutical composition for preventing or treating metabolic syndrome according to claim 5 or 6, wherein the 67LR agonist comprises EGCG, oolong tea polymerized polyphenol, procyanidin C1, a derivative thereof or a 67LR agonist antibody.

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