JPWO2010010786A1 - β3 adrenergic receptor expression enhancer - Google Patents

Abstract

The present invention seeks to expand the use of kuzuka by finding a new function of kuzuka, and provides a novel β3 adrenergic receptor expression enhancer. Since the β3 adrenergic receptor expression enhancer of the present invention contains a processed product of Kuzuka, it has a lipid metabolism improving action that promotes fat degradation in white adipocytes and promotes fat burning in brown adipocytes. Furthermore, lifestyle-related diseases (for example, diabetes) caused by obesity can be prevented.

Description

  The present invention relates to a β3 adrenergic receptor expression enhancer containing a processed kuzuhana product.

  Kuzu is a large vine plant of the legume family, and katsu starch collected from its roots has long been used as a raw material for Japanese confectionery. The roots and flowers of kuzu are called kuzu root and kuzuka, respectively, and are used as raw materials for Kampo medicines for symptoms such as antipyretic drugs, analgesics, antispasmodics and sweating. In particular, unlike other legumes, Kuzuka has been shown to have various effects such as an action for improving liver damage, an action for preventing hangover, and an action for improving urinary nitrogen metabolism (Patent Documents 1 to 3). ).

  Recently, it is also known that Kuzuka extract has a leptin action-improving obesity-suppressing action (Patent Document 4), an antihypertensive action (Patent Document 5), a body fat reducing action (Patent Document 6), and the like.

Japanese Patent No. 3454718 Japanese Patent Publication No. 8-32632 JP-A 64-68318 JP 2007-137861 A JP 2006-304766 A International Publication No. 2006/11245 Pamphlet

  An object of the present invention is to expand the field of use of Kuzuka by finding a new function of Kuzuka.

  The present inventor conducted intensive studies on the functionality of kuzuka and found that the kuzuka processed product has an excellent β3 adrenergic receptor expression enhancing action.

  The present invention provides a β3 adrenergic receptor expression enhancer containing a processed kuzuhana product.

  In one embodiment, the processed Kuzuka product is a Kuzuka extract.

  According to the present invention, a novel β3 adrenergic receptor expression enhancer is provided. The use of Kuzuka can be expanded by finding that it has a new function of β3 adrenergic receptor expression enhancing action of the Kuzuhana treatment product.

FIG. 1 (a) shows the amount of UCP-1 mRNA, FIG. 1 (b) shows the amount of β3 adrenergic receptor mRNA, and FIG. 1 (c) shows the amount of PGC-1α mRNA. FIG. 2 (a) shows the amount of mRNA of β3 adrenergic receptor, FIG. 2 (b) shows the amount of HSL mRNA, and FIG. ) Indicates the amount of UCP-2 mRNA. FIG. 3A is a graph showing the amount of mRNA in white adipocytes of mice in each group, FIG. 3A shows the amount of UCP-1 mRNA, FIG. 3B shows the amount of PPARγ mRNA, and FIG. ) Indicates the amount of LPL mRNA. It is a graph which shows transition of the weight of the mouse | mouth of each group. It is a graph which shows the mass of the organ / tissue which the mouse | mouth of each group extracted. It is a graph which shows the relative mass of the extracted organ and tissue with respect to the body weight of the mouse | mouth of each group.

  The β3 adrenergic receptor expression enhancer of the present invention contains a processed kuzuka product as an active ingredient. Furthermore, the β3 adrenergic receptor expression enhancer of the present invention may contain components (hereinafter sometimes referred to as “other components”) that are generally used as raw materials for foods, pharmaceuticals, and the like, as necessary.

(Katsuhana treatment product)
Kuzuka, the flower part of Kuzu, contains flavonoids, saponins, and tryptophan glycosides. In the present specification, “Kuzuhana” refers to a flower collected at the stage from the bud to the fully opened flower.

  In the present specification, the “Kuzuhana processed product” refers to a product obtained by subjecting Kuzuka to at least one of a drying process, a pulverizing process, and an extracting process.

  In the present specification, “dried kuzuka” refers to a product obtained by drying kuzuka, and “dried kuzuka” refers to a dry powder obtained by crushing or crushing kuzuka after drying. Good, does not contain the following Kuzuhana extract.

  In the present specification, in particular, “Kuzuka extract” refers to an extract of Kuzuka juice, an extract extracted from Kuzuka or Kuzuka dry product, a concentrate obtained by concentrating these juice or extract, It refers to a dry powder (extract powder) obtained by drying a juice or an extract.

  Hereinafter, a method for preparing a kuzu flower dried product, a kuzu flower dried powder, and a kuzu flower extract will be described.

  The dried kuzu flower is obtained by drying kuzu flowers by sun drying, hot air drying or the like. Preferably, it is dried until the water content is 10% by mass or less.

  The Kuzuka dry powder is obtained by pulverizing the Kuzuka dry product. The powderization is performed by a method commonly used by those skilled in the art, for example, a ball mill, a hammer mill, a roller mill or the like.

  Alternatively, the dried kuzuka powder can be obtained by crushing the collected kuzuka using a mass colloider, a slicer, a comitolol, etc. to obtain a kuzuka crushed product, and drying the kuzuka crushed product.

  Kuzuka extract is extracted by adding a solvent to Kuzuka, Kuzuka crushed product, Kuzuka dried product, or Kuzuka dry powder, heating as necessary, and performing centrifugation or filtration. It is obtained by collecting the extract. The kuzu flower extract is preferably obtained by extraction from dried kuzu flower or dried kuzu flower.

  Examples of the solvent that can be used to obtain the Kuzuhana extract include water, organic solvents, and hydrous organic solvents. Examples of the organic solvent include methanol, ethanol, n-propanol, n-butanol, acetone, hexane, cyclohexane, propylene glycol, ethyl methyl ketone, glycerin, methyl acetate, ethyl acetate, diethyl ether, dichloromethane, edible oils and fats, 1, Examples include 1,1,2-tetrafluoroethane, 1,1,2-trichloroethene and the like. Among these, a polar organic solvent is preferable, ethanol, n-butanol, methanol, acetone, propylene glycol, and ethyl acetate are more preferable, and ethanol is most preferable.

  Examples of the extraction method include a warm extraction method such as heating reflux, a supercritical extraction method, and the like. In these extraction methods, heating may be performed by applying pressure as necessary. When heating, it is necessary to prevent the solvent added to Kuzuka from volatilizing. When heating, the extraction temperature is preferably 50 ° C. or higher, more preferably 70 ° C. or higher, preferably 130 ° C. or lower, more preferably 100 ° C. or lower.

  The extraction time may be a time for sufficiently extracting soluble components from the extraction raw material, and may be set as appropriate according to the extraction temperature and the like. Preferably, it is 30 minutes to 48 hours. For example, when the extraction temperature is less than 50 ° C, it is preferably 6 hours to 48 hours, and when it is 50 ° C or more, it is preferably 30 minutes to 24 hours.

  The obtained extract may be concentrated or dried by a method such as concentration under reduced pressure or freeze-drying, if necessary, to form a liquid, a paste, or a powder (extract powder).

  The kuzu flower extract thus obtained can contain isoflavones and saponins. In the kuzu flower extract, isoflavones are preferably contained in an amount of 3% by mass or more, more preferably 5% by mass to 90% by mass with respect to the dry mass of the kuzu flower extract. The saponins are preferably contained in the Kuzuka extract at 1% by mass or more, more preferably 2% by mass to 50% by mass.

(Other ingredients)
The β3 adrenergic receptor expression enhancer of the present invention can contain components commonly used as raw materials for foods, pharmaceuticals and the like within a range not impairing the effects of the present invention, in addition to the processed Kuzuka product.

  Examples of such components include royal jelly, propolis, vitamins (A, C, D, E, K, folic acid, pantothenic acid, biotin, derivatives thereof, etc.), minerals (iron, magnesium, calcium, zinc, selenium). ), Chitin / chitosan, lecithin, polyphenols (flavonoids, derivatives thereof, etc.), carotenoids (lycopene, astaxanthin, zeaxanthin, lutein, etc.), xanthine derivatives (caffeine, etc.), fatty acids, proteins (collagen, elastin, etc.), Mucopolysaccharides (hyaluronic acid, chondroitin, dermatan, heparan, heparin, ketalan, and their salts), amino sugars (glucosamine, acetylglucosamine, galactosamine, acetylgalactosamine, neuraminic acid, acetylneuramin) , Hexosamine, salts thereof, etc., oligosaccharides (isomalto-oligosaccharides, cyclic oligosaccharides, etc.), phospholipids and their derivatives (sphingomyelin, ceramides, etc.), sulfur-containing compounds (alyne, sepaene, taurine, glutathione, methylsulfonylmethane) ), Sugar alcohols, lignans (such as sesamin), plant and animal extracts containing these, root vegetables (turmeric, ginger, etc.), green leaves of gramineous plants such as wheat powder, green leaves of cruciferous plants such as kale, Excipients, extenders, binders, thickeners, emulsifiers, colorants, fragrances, seasonings and the like can be mentioned.

(Β3 adrenergic receptor expression enhancer)
The β3 adrenergic receptor expression enhancer of the present invention contains the processed Kuzuka product as an active ingredient, and may further contain the other ingredients as necessary.

  White adipocytes and brown adipocytes have β3 adrenergic receptors, the only receptors that sense adrenergic in adipocytes. β3 adrenergic receptors are known to be involved in lipolysis in white adipocytes and fat burning in brown adipocytes.

  The β3 adrenergic receptor expression enhancer of the present invention promotes fat decomposition in white adipocytes and promotes fat burning in brown adipocytes by enhancing expression of β3 adrenergic receptors. Therefore, since the β3 adrenergic receptor expression enhancer of the present invention has an action of improving lipid metabolism, it exhibits an anti-obesity effect and can further prevent lifestyle-related diseases (for example, diabetes) caused by obesity.

  The amount of the processed Kuzuka product contained in the β3 adrenergic receptor expression enhancer of the present invention is not particularly limited. For example, in the case where the processed kuzuka extract is a kuzuka extract, it may be contained so that the intake of the kuzuka extract is preferably 10 to 500 mg / day, more preferably 100 to 300 mg / day in terms of dry mass. .

  The β3 adrenergic receptor expression enhancer of the present invention can be used in the form of food or medicine. Depending on the application, it is processed into forms such as granules, tablets, capsules (hard capsules, soft capsules, etc.), pills, powders, liquids, tea bags, and baskets. The β3 adrenergic receptor expression enhancer thus processed may be eaten as it is, or may be taken by dissolving in water, hot water, milk or the like. If powder etc. are provided with the form of a tea bag, the extract obtained by being immersed in hot water etc. can be utilized as a drink.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated, this invention is not limited to these Examples.

(Experimental mouse)
Basic feed (MF feed; manufactured by Oriental Yeast Co., Ltd.) was given to 24 7-week-old male C57black / 6J mice (Nippon Charles River Co., Ltd.) for 8 days. After acclimatization, each group was divided into 3 groups of 8 animals so that the average weight of each group was almost equal.

  One group of mice is then fed a normal diet (LF group), the other group of mice is fed a high fat diet (HF group), and the remaining one group of mice is fed a test diet (high fat). (Feed which mix | blended Kuzuka extract with the feed) was ingested (PTE5% group). Table 1 shows the components of each feed. As the kuzu flower extract, “Kuzu no Hana Extract” (10% by mass of isoflavones and 1% by mass of saponins, which is obtained by hot water extraction from kuzu flower) was used.

  Using a Roden Cafe (made by Oriental Yeast Co., Ltd.) as a feeder, each group of mice was allowed to ingest feed and freely ingested deionized water. During the test period, adjustments were made so that there was no difference between the groups in the cumulative food intake of mice in each group. The intake amount was calculated from the feed amount including the feeder mass on the day before the measurement date and the remaining feed amount after intake.

  Each group of mice was ingested for 14 days and their body weight was measured twice a week. Further, the body weight was measured on the 15th day from the start of feed intake, and then the abdomen was opened under ether anesthesia, and the organ / tissue was removed.

(Example 1: Verification of gene expression in mouse brown adipocytes)
MRNA was quantified for various gene expression in isolated brown adipocytes of mice in the LF group, HF group, and PTE 5% group. Real Time PCR was used as the quantification method. The results are shown in FIG. A value shows the average value (relative value on the basis of LF group) of the amount of mRNA of the mouse | mouth of each group.

  As shown in FIG. 1, in UCP-1 (Uncoupling protein-1) (FIG. 1 (a)) involved in heat production and β3 adrenergic receptor upstream of UCP-1 (FIG. 1 (b)), PTE 5% group In the LF group and the mouse in the HF group were found to have significantly increased expression (*, **, and *** in the graph are p <0.05, p, respectively). <0.01 and p <0.001 indicate significant difference). Furthermore, in PGC-1α (Peroxisome proliferator-activated receptor g coactivator-1) upstream of UCP-1 (FIG. 1 (c)), the expression of the mouse in the PTE 5% group was significant compared to the mouse in the LF group. Even when compared with mice in the HF group, a tendency to increase was observed (p = 0.10).

  Therefore, the Kuzuhana extract promotes increased expression of β3-adrenergic receptor and PGC-1α upstream of UCP-1 in brown adipocytes, and further enhances heat production by enhancing the expression of UCP-1 itself. It was found to have an effect.

(Example 2: Verification of gene expression in mouse white adipocytes)
MRNA was quantified for the expression of various genes in white adipocytes of isolated testicular fat from mice in the LF group, HF group, and PTE 5% group. The results are shown in FIG. 2 and FIG. A value shows the average value (relative value on the basis of LF group) of the amount of mRNA of the mouse | mouth of each group.

  As shown in FIG. 2, HSL (Hormone sensitive lipase) (FIG. 2 (b)) that promotes the degradation of neutral fat and UCP-2 (Uncoupling protein-2) (FIG. 2 ( In c)), it can be seen that the expression of the mice in the PTE 5% group was significantly increased compared to the mice in the LF group and the mice in the HF group (*, **, and *** in the graph are: P <0.05, p <0.01, and p <0.001, respectively, indicating significant differences). Furthermore, in the β3 adrenergic receptor upstream of HSL and UCP-2 (FIG. 2 (a)), it can be seen that the expression of the mice in the PTE 5% group was significantly increased as compared with the mice in the HF group. As shown in FIG. 3, PPARγ (Peroxisome proliferator-activated receptor γ) involved in adipocyte differentiation (FIG. 3 (b)), LPL (Lipo protein lipase) involved in fat uptake into cells (FIG. 3 (c) )), And UCP-1 (Fig. 3 (a)) involved in heat production, no significant difference was observed between mice in each group.

  Therefore, it was found that the Kuzuhana extract has a lipolysis enhancement action and a heat production enhancement action with increased expression of β3 adrenergic receptor in white adipocytes of testicular fat.

(Example 3: Transition of body weight of mouse)
Each group of mice was fed a diet for 14 days, and the average body weight of each group of mice was measured. The results are shown in FIG. As shown in FIG. 4, on the 15th day from the start of feed intake, the increase in body weight of the mice in the PTE 5% group and the mice in the LF group was significantly suppressed as compared with the mice in the HF group.

  Therefore, it was found that the Kuzuka extract has an anti-obesity effect even in a short period of 14 days. Furthermore, the weight transition of the mice in the PTE 5% group was almost the same as the weight transition of the mice in the LF group, suggesting that they are growing healthy.

(Example 4: Mouse organ mass)
All mice were dissected and the liver, testicular fat, mesenteric fat, perirenal fat, and retroperitoneal fat removed. FIG. 5 shows the mass of the extracted tissue, and FIG. 6 shows the relative mass with respect to the body weight of the extracted tissue.

  As shown in FIGS. 5 and 6, both the tissue mass and the relative mass of HF in both the testicular fat, the mesenteric fat, the perirenal fat, the retroperitoneal fat, and the sum of the white fat added together. The group of mice showed significantly higher values than the LF group of mice. On the other hand, the mice in the PTE 5% group showed significantly lower values than the mice in the HF group.

  Therefore, it was suggested that the suppression of the increase in body weight in Example 3 was caused by the mass of fat. The liver mass was not significantly different between the mice in each group.

(Example 5: Verification of fat accumulation in mouse liver)
The extracted liver was examined for fat accumulation. The results of the oil red staining test are shown in Table 2, and the results of Sudan III staining test are shown in Table 3. In Tables 2 and 3, 101 to 108 represent 8 mice in the LF group, 201 to 208 represent 8 mice in the HF group, and 301 to 308 represent 8 mice in the PTE 5% group. Show.

  As shown in Tables 2 and 3, in both oil red staining and Zudan III staining, the mice in the LF group and the mice in the HF group have fatty liver, steatosis, and marginal fat accumulation from the leaflet center. There was no difference. On the other hand, it was found that the mice in the PTE 5% group were remarkably improved in all of fatty liver, steatosis, and accumulation of limbic fat from the lobule center. Therefore, it was found that the Kuzuka extract has a liver fat accumulation inhibitory action.

(Example 6: Verification of whether or not Kuzuka extract acts as an agonist for β3 adrenergic receptor)
The effect of Kuzuhana extract on receptor activity was examined. Using the recombinant cells expressing human-derived β1-β3 adrenergic receptors shown in Table 4, the binding inhibition rate of the radiolabeled ligand to β1-β3 adrenergic receptors was measured. The Kuzuhana extract was dissolved in dimethyl sulfoxide (DMSO), added to the assay system shown in Table 4 so that the final concentration was 50 μg / mL, and incubated, and the inhibition rate was measured.

  Kuzuka extract did not inhibit the binding of radiolabeled ligand to β1 adrenergic receptor. Furthermore, Kuzuka extract inhibited binding of radiolabeled ligand to β2 adrenergic receptor only 12% and binding to β3 adrenergic receptor only 1%. Therefore, it was found that the Kuzuhana extract does not act as an agonist for β1 adrenergic receptor, β2 adrenergic receptor, and β3 adrenergic receptor.

  According to the present invention, a novel β3 adrenergic receptor expression enhancer containing a processed kuzuka product is provided. The β3 adrenergic receptor expression enhancer of the present invention promotes fat decomposition in white adipocytes and promotes fat burning in brown adipocytes by enhancing expression of β3 adrenergic receptors. Therefore, since the β3 adrenergic receptor expression enhancer of the present invention has an action of improving lipid metabolism, it can exert an anti-obesity effect and can further prevent lifestyle-related diseases (for example, diabetes) caused by obesity and the like. It can be used as a metabolic enhancer, anti-obesity agent, hypoglycemic agent and the like.

Claims (2)

  A β3 adrenergic receptor expression enhancer containing a processed kuzuhana product.   The β3 adrenergic receptor expression enhancer according to claim 1, wherein the processed kuzuhana product is a kuzuhana extract.

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