Composition for enhancing adrenomedullin gene expression

Abstract

To provide a composition that contains a plant component as an active ingredient and has the action of enhancing adrenomedullin gene expression.SOLUTION: One embodiment of the present invention relates to a composition for enhancing adrenomedullin gene expression, containing 6-shogaol and ethanol extract of Japanese pepper as active ingredients. Another embodiment of the present invention relates to a composition containing 6-shogaol and ethanol extract of Japanese pepper as active ingredients. Still another embodiment of the present invention relates to a composition containing 6-shogaol, and one or more selected from γ-sanshool, α-sanshool and hydroxy-α-sanshool in a total of 2-10 mol per 6-shogaol 1 mol.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a composition for enhancing adrenomedullin gene expression. The present invention also relates to a composition having the effect of enhancing the expression of the adrenomedullin gene. [Background technology]

[0002] Adrenomedullin is a circulatory regulatory peptide with vasodilatory properties. It is produced by various organs such as the circulatory system and digestive system, and has been reported to have important physiological activities such as vasodilatory properties, angiogenesis, antibacterial properties, anti-enteritis properties, gastric mucosa protection, and inhibition of thrombus formation (Patent Document 1).

[0003] Patent Document 1 describes an adrenomedullin production enhancer containing a compound derived from ginseng (= Korean ginseng) and a compound derived from dried ginger and / or a compound derived from Japanese pepper as active ingredients. Example 2 of Patent Document 1 describes that a combination of ginseng (= Korean ginseng) extract and dried ginger extract, a combination of ginseng extract and Japanese pepper extract, a combination of dried ginger extract and Japanese pepper extract, ginseng extract alone, dried ginger extract alone, and Japanese pepper extract alone were administered to rats, and VC (vascular conductance), an index of blood flow, was measured and the increase rate was calculated. The combination of ginseng extract and dried ginger extract and the combination of ginseng extract and Japanese pepper extract showed a clearly higher increase in VC than each of the single administrations, but no change in the increase rate of VC was observed in the combination of dried ginger extract and Japanese pepper extract.

[0004] Non-patent document 1 describes that Daikenchuto (containing Japanese pepper, dried ginger, and Korean ginseng), a type of herbal medicine, promotes the release of adrenomedullin from intestinal epithelial cells, thereby promoting blood flow in the intestine, and that Daikenchuto and one of its components, 6-shogaol, have the effect of promoting the release of adrenomedullin from intestinal epithelial cells, and that this effect is inhibited by an antagonist of TRPA1, a TRP channel.

[0005] Patent Document 2 describes an anti-inflammatory composition containing Japanese pepper, dried ginger, ginseng (= Korean ginseng) and glue candy. Patent Document 2 describes that this anti-inflammatory composition can treat and alleviate inflammatory symptoms in the digestive organs.

[0006] Patent Document 3 describes a gastrointestinal mucosa protecting agent containing Japanese pepper or an extract thereof as an active ingredient. Patent Document 3 also describes that the gastrointestinal mucosa protecting agent may be mixed with other medicinal active ingredients such as dried ginger or an extract thereof.

[0007] Patent Document 4 describes a hair tonic that contains five kinds of herbal extracts, namely, Chinese hawthorn, Japanese angelica, red pepper, and Chinese laurel. Patent Document 4 describes that the five kinds of herbal extracts in the hair tonic increase peripheral blood flow by dilating the local peripheral blood vessels in the hair loss area, thereby promoting the activity of hair roots. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] International Publication WO2009 / 104248 [Patent Document 2] JP 2002-145791 A [Patent Document 3] JP 2007-291008 A [Patent Document 4] Japanese Patent Application Publication No. 2-17117 [Non-patent literature]

[0009] [Non-Patent Document 1] Am J Physiol Gastrointest Liver Physiol 304:G428-G436,2013 Summary of the Invention [Problem to be solved by the invention]

[0010] An objective of the present invention is to provide a composition which contains a naturally derived component as an active ingredient and has the effect of enhancing adrenomedullin gene expression. [Means for solving the problem]

[0011] The inventors have discovered that a composition containing 6-shogaol and an ethanol extract of Japanese pepper as active ingredients has the effect of synergistically enhancing the expression of the adrenomedullin gene, and have completed the present invention as described below. (1) A composition for enhancing adrenomedullin gene expression, comprising 6-shogaol and an ethanol extract of Japanese pepper as active ingredients. (2) The composition according to (1), wherein the ethanol extract of Japanese pepper contains one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool. (3) A composition containing 6-shogaol and an ethanol extract of Japanese pepper. (4) The composition according to (3), wherein the ethanol extract of Japanese pepper contains one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool. (5) A composition containing 6-shogaol and one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool in a total amount of 2 to 10 moles per mole of 6-shogaol. (6) The composition according to (5), comprising 2 to 10 moles in total of one or more selected from γ-sanshool and α-sanshool per mole of 6-shogaol. Effect of the Invention

[0012] According to the present invention, there is provided a composition which contains a naturally-derived component as an active ingredient and has the effect of enhancing the expression of the adrenomedullin gene. [Brief description of the drawings]

[0013] [Figure 1] Figure 1 shows the relative mRNA expression level of adrenomedullin to β-actin in rat-derived small intestinal cell lines cultured in medium supplemented with 10 μM 6-shogaol, 25 μg / mL ethanol extract of Japanese pepper, 50 μg / mL ethanol extract of Japanese pepper, a combination of 10 μM 6-shogaol and 25 μg / mL ethanol extract of Japanese pepper, or a combination of 10 μM 6-shogaol and 50 μg / mL ethanol extract of Japanese pepper, with the expression level under control conditions taken as 1. [Diagram 2] FIG. 2 shows the results of the experiments using 10 μM 6-shogaol, 10 μg / mL or 20 μg / mL Fr.3-1 (28 μM or 56 μM as HAS), a combination of 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL Fr.3-1, 10 μg / mL or 20 μg / mL Fr.3-2 (31 μM or 62 μM as AS), 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL Fr.3-3 (31 μM or 62 μM as AS), and 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL Fr.3-4 (31 μM or 62 μM as AS). The relative mRNA expression level of adrenomedullin to β-actin in rat-derived small intestinal cell lines cultured in medium supplemented with a combination of Fr.3-2, 10 μg / mL or 20 μg / mL of Fr.3-3 (28 μM or 56 μM as GS), or a combination of 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL of Fr.3-3 is shown, with the expression level under control conditions taken as 1. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] <6-Shogaol> 6-Shogaol may be derived from a plant such as ginger, or may be chemically synthesized, but is preferably derived from a plant.

[0015] 6-Shogaol may be used in the form of an extract from a plant such as ginger that contains 6-shogaol, or it may be used in the form of a purified or concentrated compound.

[0016] When 6-shogaol derived from ginger is used, the ginger may be dried ginger or shokyo, which are herbal medicines.

[0017] A plant extract containing 6-shogaol can be prepared by subjecting a plant raw material to an extraction operation using an extraction medium. As the extraction medium, an organic solvent, water, hot water, or a supercritical fluid such as supercritical carbon dioxide can be used, and a solvent is particularly preferred. As the solvent, an alcohol such as ethanol, an organic solvent such as ethyl acetate, hexane, acetone, or a mixture thereof is particularly preferred, and ethanol and / or ethyl acetate are particularly preferred. When producing an extract by solvent extraction, the plant raw material is immersed in an appropriate amount of solvent (for example, 0.5 to 20 times the amount of the plant raw material by weight), and appropriately stirred or left to elute the solvent-soluble components into the solvent. The extraction time is not particularly limited, but can be 5 to 120 minutes, for example, 15 to 30 minutes. The extraction temperature is not particularly limited, but can be 0°C to 125°C, for example, 25°C to 125°C. After extraction, the solvent fraction containing the solvent-soluble components and the solid fraction such as cell walls are separated by solid-liquid separation means (centrifugation, filtration (diatomaceous earth filtration, etc.)), and the solvent fraction is obtained as an extract. The obtained extract may be used as it is as a 6-shogaol-containing extract, or if necessary, the extract may be further subjected to further purification means such as concentration, solvent fractionation, chromatography (column chromatography, gas chromatography, high performance liquid chromatography (HPLC), supercritical fluid chromatography, etc.) and / or recrystallization, and the resulting product may be used as a 6-shogaol-containing extract. The plant raw material used for extraction may be in its original form, or may be cut to an appropriate size or shape, or may be crushed, pulverized, or ground, or may be in the form of squeezed juice.

[0018] <Ethanol extract of Japanese pepper> The part of Japanese pepper used for preparing the ethanol extract is not particularly limited, but the peel, fruit, seeds, leaves, stems, etc. can be used, with the peel being particularly preferred.

[0019] The method for preparing the ethanol extract of Japanese pepper is not particularly limited. For example, Japanese pepper is immersed in an appropriate amount of ethanol (for example, 0.5 to 20 times the amount of Japanese pepper by weight), and appropriately stirred or left to dissolve soluble components in ethanol. The extraction time is not particularly limited, but can be 5 to 120 minutes, for example, 15 to 30 minutes. The extraction temperature is not particularly limited, but can be 60°C to 125°C, for example, 85°C to 100°C. After extraction, a solution fraction containing ethanol-soluble components and a solid fraction such as cell walls are separated by the solid-liquid separation means, and the solution fraction is obtained as an extract. The obtained extract may be used as it is as an ethanol extract, or if necessary, the extract may be further subjected to further purification means such as concentration, solvent fractionation, chromatography (column chromatography, gas chromatography, high performance liquid chromatography (HPLC), supercritical fluid chromatography, etc.) and / or recrystallization, and the resulting product may be used as an ethanol extract. The form of the Japanese pepper used for extraction may be in the original form, or cut to a suitable size or shape, or crushed, pulverized or ground form, or squeezed juice.

[0020] The Japanese pepper used for preparing the ethanol extract of Japanese pepper may be a water extraction residue obtained by performing a water extraction treatment before the above-mentioned extraction with ethanol, that is, Japanese pepper from which water-soluble components have been removed by water extraction. Here, the water extraction of Japanese pepper is, for example, performed by immersing Japanese pepper in an appropriate amount of water (for example, 0.5 to 20 times the amount of Japanese pepper by weight) and appropriately stirring or leaving it to elute the water-soluble components into the water. The extraction time is not particularly limited, but can be 5 minutes to 120 minutes, for example, 15 minutes to 30 minutes. The extraction temperature is not particularly limited, but can be 60°C to 125°C, for example, 85°C to 100°C. After the extraction, the aqueous fraction and the solid fraction are separated by the solid-liquid separation means, and the solid fraction is collected and dried as necessary to obtain a Japanese pepper water extraction residue, which can be used for preparing the ethanol extract.

[0021] The ethanol extract of Zanthoxylum japonicum may be formulated into a desired form such as powder, granules, etc. For formulation, a suitable excipient can be used.

[0022] The ethanol extract of Zanthoxylum persica preferably contains one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool.

[0023] A preferred example of the ethanol extract of Japanese pepper contains γ-sanshool at a concentration of preferably 50% by weight or more, more preferably 70% by weight or more. Another preferred example of the ethanol extract of Japanese pepper contains α-sanshool at a concentration of preferably 50% by weight or more, more preferably 70% by weight or more. Another preferred example of the ethanol extract of Japanese pepper contains hydroxy-α-sanshool at a concentration of preferably 50% by weight or more, more preferably 70% by weight or more.

[0024] <Composition for enhancing adrenomedullin gene expression> One or more embodiments of the present invention relate to a composition for enhancing adrenomedullin gene expression, which contains 6-shogaol and an ethanol extract of Zanthoxylum japonicum as active ingredients. The composition according to this embodiment preferably does not contain one or more selected from ginseng and ingredients derived from ginseng, and more preferably does not further contain hawthorn and / or ingredients derived from hawthorn, dong qui and / or ingredients derived from dong qui, and capsicum and / or ingredients derived from capsicum.

[0025] The subject of the composition according to this embodiment is typically a human, but is not limited to humans and may be other non-human animals, for example, mammals other than humans.

[0026] Adrenomedullin is a peptide that has physiological activities such as vasodilatation and angiogenesis. By enhancing the gene expression of adrenomedullin, it is possible to increase blood flow. Furthermore, by increasing blood flow by enhancing the gene expression of adrenomedullin, it is possible to achieve effects such as antibacterial action, antienteritis, gastric mucosa protection, and inhibition of thrombus formation.

[0027] Patent Document 1 describes that a combination of a compound derived from Korean ginseng and a compound derived from dried ginger and / or a compound derived from Japanese pepper has an effect of enhancing adrenomedullin production, while it describes that a combination of dried ginger extract and Japanese pepper extract did not have a synergistic effect of increasing blood flow. Non-Patent Document 1 describes that Daikenchuto and one of its components, 6-shogaol, have the effect of promoting the release of adrenomedullin from intestinal epithelial cells. Patent Document 2 describes an anti-inflammatory composition containing Japanese pepper, dried ginger, Korean ginseng and glue candy, but does not describe the effect of a composition that does not contain Korean ginseng. Patent Document 3 describes that Japanese pepper or its extract has a gastrointestinal mucosa protecting effect, and Patent Document 4 describes that a combination of five herbal extracts (ethanol extract in the example) of Chinese ginseng, Japanese cratae, Japanese angelica, Japanese capsicum, and Chinese bell pepper has a hair growth effect by increasing peripheral blood flow. Patent Documents 1 to 4 and Non-Patent Document 1 do not disclose a combination of 6-shogaol and an ethanol extract of Japanese pepper.

[0028] The composition of this embodiment contains 6-shogaol and an ethanol extract of Japanese pepper as active ingredients, and has the effect of synergistically enhancing the expression of the adrenomedullin gene, which cannot be predicted from the information described in Patent Documents 1 to 4 and Non-Patent Document 1.

[0029] 6-shogaol and the ethanol extract of Japanese pepper are both components with strong pungent tastes. By combining the two, the expression of the adrenomedullin gene can be synergistically enhanced, so the total amount of 6-shogaol and the ethanol extract of Japanese pepper required to enhance the expression of the adrenomedullin gene can be reduced compared to the case of using each alone. Therefore, the composition for enhancing the expression of the adrenomedullin gene according to this embodiment is suitable for use as a food or drink with reduced pungent taste.

[0030] Enhanced gene expression of adrenomedullin can be confirmed by preparing cDNA from mRNA in cells taken from a subject who has ingested or administered the composition of this embodiment, or in cells cultured in the presence of the composition of this embodiment, and using the cDNA as a template, performing a nucleic acid amplification reaction using a primer set capable of specifically amplifying at least a portion of the cDNA base sequence of adrenomedullin (5' untranslated region, coding region and / or 3' untranslated region, preferably the coding region), and detecting the amount of amplified product from the reaction.

[0031] Furthermore, the enhanced expression of the adrenomedullin gene can be confirmed by detecting the amount of adrenomedullin peptide in cells taken from a subject who has taken or been administered the composition according to this embodiment, or in cells cultured in the presence of the composition according to this embodiment. When the amount of adrenomedullin peptide is increased in the cells, it can be evaluated that the expression of the adrenomedullin gene is enhanced.

[0032] The amino acid sequence of the precursor of human-derived adrenomedullin (preproadrenomedullin) is shown in SEQ ID NO: 3, and the cDNA base sequence of human-derived adrenomedullin is shown in SEQ ID NO: 4. In SEQ ID NO: 4, the coding region is from positions 179 to 736.

[0033] The content of 6-shogaol and ethanol extract of Japanese pepper in the composition according to this embodiment is not particularly limited, and for example, the total amount of the composition may contain 0.1 to 95% by mass of 6-shogaol and ethanol extract of Japanese pepper. The blending ratio of 6-shogaol and ethanol extract of Japanese pepper in the composition according to this embodiment is not particularly limited. For example, the amount of ethanol extract of Japanese pepper may be 2 to 100 parts by mass, preferably 3 to 20 parts by mass, per part by mass of 6-shogaol.

[0034] When the composition according to the present embodiment includes the ethanol extract of Japanese pepper containing one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool, the content of one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool is preferably 2-10 mol in total, more preferably 2-7 mol per 1 mol of 6-shogaol.

[0035] When the composition according to the present embodiment contains the ethanol extract of Japanese pepper containing one or more selected from γ-sanshool and α-sanshool, the content of one or more selected from γ-sanshool and α-sanshool is preferably 2-10 mol in total, more preferably 2-7 mol per 1 mol of 6-shogaol.

[0036] When the composition according to the present embodiment contains an ethanol extract of Japanese pepper containing γ-sanshool, the content of γ-sanshool is preferably 2 to 10 moles in total, more preferably 2 to 7 moles per mole of 6-shogaol.

[0037] <Composition containing 6-shogaol and ethanol extract of Japanese pepper> Another embodiment of the present invention relates to a composition containing 6-shogaol and an ethanol extract of Zanthoxylum japonicum. The composition according to this embodiment preferably does not contain one or more selected from ginseng and ingredients derived from ginseng, and more preferably does not further contain Crataegus chinensis and / or ingredients derived from Crataegus japonicum, Angelica acutiloba and / or ingredients derived from Angelica acutiloba, and Capsicum chinensis and / or ingredients derived from Capsicum chinensis.

[0038] The subject of the composition according to this embodiment is typically a human, but is not limited to humans and may be other non-human animals, for example, mammals other than humans.

[0039] The composition according to this embodiment has an effect that could not be predicted from the matters described in Patent Documents 1 to 4 and Non-Patent Document 1, that is, it can synergistically enhance the expression of the adrenomedullin gene.

[0040] The composition according to this embodiment is suitable for use as a food or drink with reduced spiciness.

[0041] The content of 6-shogaol and ethanol extract of Japanese pepper in the composition according to this embodiment is not particularly limited, and for example, the total amount of the composition may contain 0.1 to 95% by mass of 6-shogaol and ethanol extract of Japanese pepper. The blending ratio of 6-shogaol and ethanol extract of Japanese pepper in the composition according to this embodiment is not particularly limited. For example, the amount of ethanol extract of Japanese pepper may be 2 to 100 parts by mass, preferably 3 to 20 parts by mass, per part by mass of 6-shogaol.

[0042] When the composition according to the present embodiment includes the ethanol extract of Japanese pepper containing one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool, the content of one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool is preferably 2-10 mol in total, more preferably 2-7 mol per 1 mol of 6-shogaol.

[0043] When the composition according to the present embodiment contains the ethanol extract of Japanese pepper containing one or more selected from γ-sanshool and α-sanshool, the content of one or more selected from γ-sanshool and α-sanshool is preferably 2-10 mol in total, more preferably 2-7 mol per 1 mol of 6-shogaol.

[0044] When the composition according to the present embodiment contains an ethanol extract of Japanese pepper containing γ-sanshool, the content of γ-sanshool is preferably 2 to 10 moles in total, more preferably 2 to 7 moles per mole of 6-shogaol.

[0045] <A composition containing 6-shogaol and at least one selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool in a total amount of 2 to 10 moles per mole of 6-shogaol> Yet another one or more embodiments of the present invention relate to a composition that contains 6-shogaol, and one or more selected from 2 to 10 moles in total of γ-sanshool, α-sanshool, and hydroxy-α-sanshool per mole of 6-shogaol.The composition according to this embodiment preferably does not contain one or more selected from ginseng and ginseng-derived components, and more preferably does not further contain hawthorn and / or components derived from hawthorn, angelica and / or components derived from angelica, and capsicum and / or components derived from capsicum.

[0046] The subject of the composition according to this embodiment is typically a human, but is not limited to humans and may be other non-human animals, for example, mammals other than humans.

[0047] The composition according to this embodiment has an effect that could not be predicted from the matters described in Patent Documents 1 to 4 and Non-Patent Document 1, that is, it can synergistically enhance the expression of the adrenomedullin gene.

[0048] The composition according to this embodiment is suitable for use as a food or drink with reduced spiciness. In the composition according to the present embodiment, the content of one or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool is more preferably 2 to 7 moles in total per mole of 6-shogaol.

[0049] More preferably, the composition according to the present embodiment comprises one or more selected from γ-sanshool and α-sanshool, and the content of one or more selected from γ-sanshool and α-sanshool is preferably 2-10 mol in total, more preferably 2-7 mol per 1 mol of 6-shogaol.

[0050] The composition according to the present embodiment more preferably contains γ-sanshool, and the content of γ-sanshool is preferably 2 to 10 moles in total, more preferably 2 to 7 moles per mole of 6-shogaol.

[0051] In the composition of the present embodiment, one or more selected from gamma-sanshool, alpha-sanshool and hydroxy-alpha-sanshool can be derived from plants such as Japanese pepper, or can be chemically synthesized, but preferably is derived from plants.

[0052] One or more selected from γ-sanshool, α-sanshool, and hydroxy-α-sanshool may be used in the form of an extract from a plant such as Japanese pepper that contains it, or in the form of a purified or concentrated compound.

[0053] <Preferable Examples of Compositions> More preferred examples of the compositions according to the above-described embodiments disclosed in this specification (hereinafter referred to as "the compositions disclosed in this specification") will be described below.

[0054] The subject of the composition disclosed herein is preferably a subject who needs or desires to enhance gene expression of adrenomedullin. The composition disclosed herein is also effective for healthy subjects.

[0055] The composition disclosed in the present specification may be in the form of a pharmaceutical, food or drink, cosmetic, etc. Food or drink includes those in the form of functional food, food for specified health use, nutritional supplement, etc.

[0056] The compositions disclosed herein are preferably compositions that are ingested or administered orally or nasally, and more preferably compositions that are ingested or administered orally.

[0057] The composition disclosed herein may also be a composition used to enhance adrenomedullin gene expression in cells in vitro by coexisting with the cells. In this case, the composition disclosed herein is used by adding it to a medium for culturing the cells.

[0058] As used herein, the term "amount used per day" refers to the total amount of the composition disclosed herein that is ingested, administered or used in one day, and preferably refers to the total amount of the composition disclosed herein that is ingested, administered or used in one day by one human, particularly one adult. A specific example of the "amount used per day" is 0.1 g to 500 g of the composition disclosed herein.

[0059] The compositions disclosed herein may be taken, administered or used continuously or on an as-needed basis.

[0060] The form of the composition disclosed herein is not particularly limited, and may be, for example, any form such as liquid, fluid, gel, semi-solid, or solid.

[0061] The composition disclosed herein may further contain at least one other component in addition to the components described in each embodiment. The at least one other component that may be contained in the composition disclosed herein is not particularly limited, but is preferably a component that is acceptable in the final form of a pharmaceutical product, food or beverage, cosmetic product, etc.

[0062] Examples of such other components include sweeteners, acidulants, vitamins, minerals, thickeners, emulsifiers, antioxidants, water, etc. Furthermore, colorants, flavors, preservatives, antiseptics, fungicides, and further physiologically active substances may be added as necessary.

[0063] Examples of sweeteners include monosaccharides and disaccharides such as glucose, fructose, sucrose, lactose, maltose, palatinose, trehalose, and xylose, isomerized sugars (glucose-fructose liquid sugar, fructose-glucose liquid sugar, mixed isomerized sugar, etc.), sugar alcohols (erythritol, xylitol, lactitol, palatinite, sorbitol, reduced starch syrup, etc.), honey, and high-intensity sweeteners (sucralose, acesulfame potassium, thaumatin, stevia, aspartame, etc.).

[0064] The acidulant may be citric acid, malic acid, gluconic acid, tartaric acid, lactic acid, phosphoric acid, or a salt thereof, and one or more of these may be used.

[0065] Examples of vitamins include vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin E, niacin, and inositol.

[0066] Examples of minerals include calcium, magnesium, zinc, and iron.

[0067] Examples of thickening agents include carrageenan, gellan gum, xanthan gum, gum arabic, tamarind gum, guar gum, locust bean gum, karaya gum, agar, gelatin, pectin, soybean polysaccharides, and carboxymethylcellulose (CMC).

[0068] Examples of the emulsifier include glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, lecithin, plant sterols, and saponin.

[0069] Antioxidants include vitamin C, tocopherol (vitamin E), enzyme-treated rutin, and the like.

[0070] The other ingredients can be appropriately blended in amounts within the ranges that are normally used by those skilled in the art in compositions such as foods, beverages, and pharmaceuticals.

[0071] The composition disclosed herein may contain only the components described in each embodiment as active ingredients involved in enhancing the gene expression of adrenomedullin. In this case, the composition disclosed herein may contain other ingredients as described above that are not involved in enhancing the gene expression of adrenomedullin. EXAMPLES

[0072] <6-Shogaol> A commercially available reagent (Fujifilm Wako Pure Chemical Industries, Ltd.) was used as 6-shogaol (6S).

[0073] <Japanese pepper ethanol extract> A certain amount of ethanol was added to the residue of the hot water extraction of crushed Japanese pepper, and the mixture was extracted for more than 60 minutes with stirring. The extract recovered by solid-liquid separation was used as the Japanese pepper ethanol extract.

[0074] <Fractions of Japanese pepper ethanol extract> The ethanol extract of Japanese pepper was fractionated by medium pressure reversed phase chromatography (column: ODS, mobile phase: water / methanol, wavelength: 270 nm). The obtained eluates were dried using an evaporator to obtain fractions Fr.3-1, Fr.3-2, and Fr.3-3. Fr.3-1 contains 74% by weight of hydroxy-α-sanshool (HAS). Fr.3-2 contains 78% by weight of α-sanshool (AS). Fr.3-3 contains about 75% by weight of γ-sanshool (GS).

[0075] <Adrenomedullin gene expression enhancement 1> Rat-derived small intestinal epithelial cell line (RIKEN) was used at 1 × 10 per well. 5 The cells were suspended in DMEM (1 g / L glucose, 5% FBS, 4 μg / mL insulin) medium, the suspension was seeded onto a 24-well plate, and cultured at 37°C under 5% CO2 conditions.

[0076] 24 hours after the start of culture, the medium was replaced with DMEM (1 g / L glucose, 3% inactivated FBS) containing 10 μM 6-shogaol, 25 μg / mL Japanese pepper ethanol extract, 50 μg / mL Japanese pepper ethanol extract, a combination of 10 μM 6-shogaol and 25 μg / mL Japanese pepper ethanol extract, or a combination of 10 μM 6-shogaol and 50 μg / mL Japanese pepper ethanol extract, and cultured for another 6 hours. As a control, a small intestinal epithelial cell line was cultured in the same manner, except that DMEM (1 g / L glucose, 3% inactivated FBS) medium containing neither 6-shogaol nor the ethanol extract of Japanese pepper was used.

[0077] After the culture, the cells were washed with phosphate buffered saline (PBS), and the cells were harvested and RNA was extracted from the cells. The cells were harvested and RNA was extracted using an RNeasy Mini Kit (QIAGEN) according to the attached procedure.

[0078] The obtained RNA was used to measure the mRNA expression levels of adrenomedullin and the internal standard β-actin by real-time PCR. The One Step TB Green PrimeScript RT-PCR Kit II (Takara Bio) was used, and the procedure was carried out according to the attached protocol. The primer sequences are as shown in the following table.

[0079] [Table 1]

[0080] The expression level of adrenomedullin mRNA was calculated as a relative value to the expression level of β-actin mRNA. Furthermore, the calculated expression level of adrenomedullin mRNA under each condition was expressed as a relative value to the calculated expression level of adrenomedullin mRNA under the control condition.

[0081] The measurements were performed for each condition (n=1). The results are shown in Figure 1.

[0082] The expression level of adrenomedullin in the small intestinal epithelial cell line was 4.17 when 10 μM 6-shogaol alone was added, 1.52 when 25 μg / mL of Japanese pepper ethanol extract alone was added, and 11.39 when 50 μg / mL of Japanese pepper ethanol extract alone was added, whereas the expression level was 14.83 when 10 μM 6-shogaol and 25 μg / mL of Japanese pepper ethanol extract were added in combination, and 25.63 when 10 μM 6-shogaol and 50 μg / mL of Japanese pepper ethanol extract were added in combination. In all cases, a remarkable synergistic effect was confirmed that far exceeded the combined effect of each component.

[0083] <Enhancement of adrenomedullin gene expression 2> A rat-derived small intestinal epithelial cell line (RIKEN) was cultured in the same manner as in the above experiment, and 24 hours after the start of culture, 10 μM 6-shogaol, 10 μg / mL or 20 μg / mL Fr.3-1 (28 μM or 56 μM as HAS), a combination of 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL Fr.3-1, 10 μg / mL or 20 μg / mL Fr The medium was then replaced with DMEM (1 g / L glucose, 3% inactivated FBS) containing Fr.3-2 (31 μM or 62 μM as AS), a combination of 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL of Fr.3-2, 10 μg / mL or 20 μg / mL of Fr.3-3 (28 μM or 56 μM as GS), or a combination of 10 μM 6-shogaol and 10 μg / mL or 20 μg / mL of Fr.3-3, and cultured for an additional 6 hours.

[0084] As a control, a small intestinal epithelial cell line was cultured in the same manner except that DMEM (1 g / L glucose, 3% inactivated FBS) medium containing no 6-shogaol, Fr.3-1, Fr.3-2 or Fr.3-3 was used.

[0085] The amount of adrenomedullin mRNA expression in the cultured cells was measured in the same manner as in the above experiment. The amount of adrenomedullin mRNA expression under each condition was expressed as a relative value to the amount of adrenomedullin mRNA expression under the control condition.

[0086] Measurements were performed for each condition (n=2 or 3), and the average value and standard deviation were calculated. The results are shown in Figure 2.

[0087] The adrenomedullin expression level in small intestinal epithelial cell lines cultured in a medium containing a combination of 10μM 6-shogaol and 10μg / mL or 20μg / mL Fr.3-1, Fr.3-2, or Fr.3-3 was significantly higher than the sum of the adrenomedullin expression level when only 10μM 6-shogaol was added and the adrenomedullin expression level when only Fr.3-1, Fr.3-2, or Fr.3-3 was added at each concentration, confirming a synergistic effect. The effect of synergistically improving the adrenomedullin expression level by combination with 6-shogaol was highest in F.3-3 (high GS content), followed by F.3-2 (high AS content), and then F.3-1 (high HAS content).

Claims

1. A composition for enhancing adrenomedullin gene expression, comprising 6-shogaol and one or more selected from γ-sanshool and α-sanshool as active ingredients (with the proviso that the composition does not contain ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, ginsenoside Rg2 and / or ginsenoside Rh1).

2. The composition according to claim 1, comprising 6-shogaol and at least one selected from 2 to 10 moles of γ-sanshool and α-sanshool per mole of 6-shogaol.

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