CN112367969A

CN112367969A - Antioxidant agent

Info

Publication number: CN112367969A
Application number: CN201980045137.XA
Authority: CN
Inventors: 菅原知宏
Original assignee: Toyobo Co Ltd
Current assignee: Toyobo Co Ltd
Priority date: 2018-09-14
Filing date: 2019-07-11
Publication date: 2021-02-12
Also published as: JP2023171681A; JPWO2020054204A1; WO2020054204A1

Abstract

The present invention provides an antioxidant for safely and effectively reducing oxidative stress of an organism, maintaining health, and preventing various diseases and symptoms caused by oxidative stress by promoting the expression of antioxidant-related enzymes and redox-related factors in the organism. An antioxidant agent comprising an extract of Albizzia julibrissin, which has an effect of promoting gene expression of an antioxidant-related enzyme and/or a redox-related factor, in particular.

Description

Antioxidant agent

Technical Field

The present invention relates to an antioxidant containing an albizzia julibrissin extract as an active ingredient. In particular, the present invention relates to an antioxidant which acts on cells to thereby increase the antioxidant ability of the cells.

Background

In recent years, various kinds of reactive oxygen species have been particularly attracting attention as a main factor for oxidizing biological components, and adverse effects on the living body have been problematic. The reactive oxygen species refer to oxygen molecules and molecules related thereto, which have improved chemical reactivity as compared with oxygen molecules in a stable state, and are generated during energy metabolism in the living body. Examples of the reactive oxygen species include superoxide (. O) as a radical species^2－) Hydroxyl radicals (. OH), hydroperoxy radial (HHO.), Nitric Oxide (NO); hydrogen peroxide (H) as a non-radical species₂O₂) Singlet oxygen (a)¹O₂) Peroxynitrite (ONOO-), lipid hydroperoxide (LOOH), and the like. These reactive oxygen species are the second messengers of the in vivo signal transmission system and are essential molecular species for controlling cell proliferation, differentiation, and vital activities such as bactericidal action in macrophages and the like. On the other hand, it is considered that an excessive amount of reactive oxygen species causes damage to cells and tissues, but is harmful.

Against the oxidative stress caused by these excessive reactive oxygen species, the living body has a mechanism of inhibiting the generation of the reactive oxygen species as an antioxidant mechanism for protecting against the oxidative stress. The mechanism for inhibiting the generation of reactive oxygen species mainly comprises antioxidative enzymes such as catalase, superoxide dismutase (SOD), Glutathione Peroxidase (GPX), etc., and inhibits the reactive oxygen species by an enzymatic reaction. The active oxygen species are changed from superoxide to hydrogen peroxide and hydroxyl radical, and in the process, superoxide is eliminated by SOD, and hydrogen peroxide is eliminated by catalase. In addition, GPX, Glutathione Reductase (GRD), Thioredoxin Reductase (TRD), and Peroxiredoxin (PRX) eliminate hydrogen peroxide and hydroxyl radicals by controlling the oxidation/reduction state of Glutathione (GSH) and Thioredoxin (TXN). By the synergistic action of these antioxidant-related enzymes and redox-related factors, the number of reactive oxygen species in the cell is maintained at an appropriate level.

As described above, it is considered that it is effective to increase the expression levels of the antioxidant-related enzyme and the redox-related factor in order to suppress cell damage caused by reactive oxygen species in the living body. These antioxidant-related enzymes and redox-related factors are known to be promoted by ultraviolet irradiation stress, but are expected to be components that act safely and efficiently on living bodies and promote expression of antioxidant-related enzymes and redox-related factors.

As such components, for example, Kaempferol (Kaempferol) and quercetin, which are flavonols, widely distributed in tea plants, have been reported to promote the expression and activity of glutathione and thioredoxin in the living body (patent document 1).

The search for a component that safely and effectively acts on an antioxidant-related enzyme and/or a redox-related factor in a living body is important not only from the viewpoint of providing a safer and more effective component to the market, but also from the viewpoint of diversification of preparations, expansion of options in formulation, and synergistic effects due to the combined use of different components. Needless to say, development of new ingredients acting on antioxidant-related enzymes and/or redox-related factors is required.

On the other hand, albizia julibrissin is a deciduous tree of leguminous plants naturally grown in japan on the forest edge of mountains in the south of the northeast region, in open fields and on well-exposed wetlands, and is also distributed and cultivated in overseas china, southeast asia and the like. The preparation which is prepared by collecting barks during the flowering period, washing the barks with water, drying the barks in the sun and cutting the barks into proper lengths is called as cortex albiziae, and is taken in folk in anticipation of arthritis, lumbago, diuresis, edema and strong body; for external use, cold-wet cloth is applied to affected part with decoction for swelling, pimple, contusion, traumatic injury, and arthralgia, or used as bathing material. It is known that bark contains triterpenoid saponins (most of them are albizzia saponins), flavonoids (gelaldone, isoocanin, luteolin, etc.), tannins, and the like (non-patent document 1).

As the effect of albizzia on living bodies, active oxygen scavenging action and aldose reductase inhibiting action have been reported which have an albizzia-derived component as an active ingredient, but the effect is focused on active oxygen scavenging action, i.e., the action thereof remains in scavenging effect, and therefore, active oxygen scavenging is not achieved by stimulating antioxidant-related enzymes and redox-related factors of living bodies (patent document 2). Further, although an effect of the albizia bark extract in inhibiting melanin production and an effect in promoting collagen production for the skin have been reported, no factor related to oxidation resistance is mentioned (patent document 3).

Documents of the prior art

Patent document

Patent document 1: japanese patent No. 5833438

Patent document 2: japanese laid-open patent publication No. 2000-128798

Patent document 3: japanese patent No. 5253862

Non-patent document

Non-patent document 1: hehan medicine No.759(2016 month 8 year)

Summary of The Invention

Problems to be solved by the invention

In view of the above background, the present invention addresses the problem of promoting the expression of antioxidant-related enzymes and/or redox-related factors in an organism, thereby safely and effectively reducing oxidative stress in the organism, maintaining health, and preventing various diseases and symptoms caused by oxidative stress.

Means for solving the problems

In view of the above circumstances, the present inventors have conducted extensive studies and as a result, found that an albizia julibrissin extract has an antioxidant effect, and thus completed the present invention. The albizzia julibrissin extract was found to promote gene expression of SOD, TXN, PRX, TRD, GRD, Glutaredoxin (GXN) among antioxidant-related enzymes and/or redox-related factors in the organism. By using the antioxidant of the present invention as an active ingredient for promoting the expression of an antioxidant-related enzyme and/or a redox-related factor in an organism, it is possible to eliminate excessive reactive oxygen species in the organism and alleviate various oxidative stresses in the organism without causing stress to the organism to which the antioxidant is applied.

That is, the gist of the present invention is as follows.

Item 1, antioxidant containing albizzia julibrissin extract.

The antioxidant according to item 1, wherein the albizzia julibrissin extract is an aqueous extract from a plant of the albizzia genus.

The antioxidant according to item 1 or 2, wherein the albizia julibrissin extract has an effect of promoting gene expression of an antioxidant-related enzyme and/or a redox-related factor.

The antioxidant according to any one of claims 1 to 3, wherein the antioxidant-related enzyme and/or redox-related factor is at least one or more selected from the group consisting of superoxide dismutase (SOD), Peroxiredoxin (PRX), Thioredoxin (TXN), Thioredoxin Reductase (TRD), Glutathione Reductase (GRD), and Glutaredoxin (GXN).

The antioxidant according to any one of claims 1 to 4, wherein the Albizzia julibrissin extract has an effect of reducing intracellular active oxygen.

The external preparation for skin according to item 6, wherein the antioxidant according to any one of items 1 to 5 is prepared.

The external preparation for skin according to item 6, wherein the albizzia julibrissin extract is contained in an amount of 0.00001 to 5.0 wt%.

Effects of the invention

By using the antioxidant of the present invention, it is expected to improve the antioxidant ability in vivo, and an external skin preparation aimed at improving the antioxidant ability can be provided.

Brief Description of Drawings

FIG. 1 is a graph showing the results of evaluation of the effect of promoting the expression of a target gene (SOD) in epidermal keratinocytes of a human normal newborn in example 1.

FIG. 2 is a graph showing the results of evaluating the promoting effect of expression of a target gene (TXN) in epidermal keratinocytes of a human normal newborn in example 1.

FIG. 3 shows the results of evaluation of the effect of promoting expression of a target Gene (GXN) in epidermal keratinocytes of a normal human newborn in example 1.

FIG. 4 is a graph showing the results of evaluation of the effect of promoting the expression of a target gene (SOD) in human normal neonatal fibroblasts in example 2.

FIG. 5 is a graph showing the results of evaluating the promoting effect of the expression of a target gene (TXN) in human normal neonatal fibroblasts in example 2.

FIG. 6 shows the results of evaluation of the effect of promoting the expression of a target Gene (GXN) in human normal neonatal fibroblasts in example 2.

FIG. 7 is a graph showing the results of evaluating the promoting effect of the expression of a target gene (PRX) in human normal neonatal fibroblasts in example 2.

FIG. 8 is a graph showing the results of evaluating the promoting effect of expression of a target gene (TRD) in human normal neonatal fibroblasts in example 2.

FIG. 9 is a graph showing the results of evaluation of the effect of promoting the expression of a target Gene (GRD) in human normal neonatal fibroblasts in example 2.

FIG. 10 is a graph showing the results of evaluating the effect of reducing reactive oxygen species in human normal neonatal fibroblasts in example 3.

FIG. 11 is a graph showing the results of evaluation of the effect of promoting the expression of antioxidant-related genes in cells obtained from a living body in example 4.

Detailed Description

Specific examples of the method for extracting an albizzia julibrissin extract used in the present invention are shown below. The plant parts in the extract are mainly bark, but bark is not limited to bark, and all plant parts can be used. In addition, it goes without saying that it is not limited to the examples described below.

In the present invention, the albizzia julibrissin extract refers to a cut product, a dried powder and an extract thereof, other than the plant body itself of the albizzia genus plant. Further, the "extract" includes any of a state containing the extraction solvent and a state from which the extraction solvent is removed, and in addition, includes those subjected to further purification treatment after removal of the extraction solvent.

The albizzia julibrissin extract can be obtained by extracting a plant body of a plant belonging to the genus albizzia, usually a branch or leaf thereof, freshly or by drying and pulverizing the plant body, and then extracting the pulverized plant body with water, preferably hot water at 70 to 100 ℃, or an organic solvent as an extraction solvent. The extraction may be carried out by combining water extraction and organic solvent extraction as necessary, but in consideration of the influence when applied to living organisms, it is preferable to carry out the extraction only by water extraction. For example, water; alcohols such as methanol, ethanol, propanol, and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate; chain and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; hydrocarbons such as hexane, cyclohexane, petroleum ether, and the like; aromatic hydrocarbons such as benzene and toluene; pyridines; among fats and oils, waxes, and other oils, water, alcohols, and water-alcohol mixtures are preferable, and it is particularly preferable to use hot water as the extraction solvent.

In the case of hot water extraction, usually, the pulverized plant material of the albizia plant is extracted under heating reflux. The extraction conditions vary depending on the solvent used, and for example, in the case of extraction with water, extraction with 1 to 100 parts by weight of water at a temperature of 4 to 100 ℃ for 10 minutes to 7 days is preferable for 1 part by weight of Albizzia julibrissin; it is more preferable that 5 to 20 parts by weight of water is used for extraction of 1 part by weight of the albizia bark at a temperature of 70 to 100 ℃ for 30 minutes to 2 hours. In addition, from the viewpoint of further improving the extraction efficiency, pressurization, stirring, ultrasonic treatment, or the like may be performed simultaneously with the extraction treatment. Further, after the extraction treatment, the residue may be separated by filtration or centrifugation.

The albizzia julibrissin extract usually gives a tarry dark brown extract when the extraction solvent is distilled off under reduced pressure, and if necessary, an excipient may be added. The extraction solvent may be used as it is in the dosage form or an extract from which the extraction solvent has been separated may be used in the dosage form, in which case the extraction solvent is non-toxic, for example, in the case where the extraction solvent is water. The extract after separation of the extraction solvent may also be redissolved in a suitable solvent, such as water, for dispensing. The extract thus obtained may be purified or the titer may be increased by a purification method such as separation extraction, partition between two solvents, separation adsorption/elution with an appropriate adsorbent, chromatography, or the like.

The albizzia julibrissin extract obtained as described above can be used as it is as an antioxidant, but it is preferably used by preparing it with an external preparation. The amount of the external preparation to be prepared is determined by the degree of absorption, the degree of action, the form of the product, the frequency of use, and the like, and is not particularly limited, but it is desirable that the concentration is in the range of 0.00001 to 5.0% by weight, preferably in the range of 0.00005 to 3.0% by weight, and particularly preferably in the range of 0.0001 to 1.0% by weight, based on the dry weight. When the content of the albizzia julibrissin extract is less than 0.00001 wt%, the sufficient effect cannot be exerted, and the effect is almost constant even if the content is 5.0 wt% or more.

The antioxidant of the present invention contains an albizzia julibrissin extract as an active ingredient. The antioxidant activity in the present invention means an activity of acting on cells or tissues of a living body to promote the expression of antioxidant-related enzymes and/or redox-related factors in the living body, in particular, an activity of promoting the gene expression of antioxidant-related enzymes and redox-related factors, in addition to a scavenging activity by an antioxidant substance contained in an albizia julibrissin extract. These effects also act to reduce the amount of active oxygen in the cell.

In the present invention, the effect of promoting the gene expression of an antioxidant-related enzyme and a redox-related factor can be evaluated by adding the antioxidant of the present invention to cultured cells and quantifying the intracellular gene expression level by real-time PCR. In the present invention, the amount of intracellular active oxygen can be evaluated by adding a fluorescent substance that reacts with active oxygen to cultured cells, adding the antioxidant of the present invention, and quantifying the intensity of fluorescence emitted from the cells.

Examples of the antioxidant-related enzyme and/or redox-related factor include superoxide dismutase (SOD), peroxidase, catalase, and Peroxiredoxin (PRX) which eliminate reactive oxygen species; glutathione Reductase (GRD), Thioredoxin Reductase (TRD), Glutaredoxin (GXN) that control the oxidation/reduction state of Glutathione (GSH) and Thioredoxin (TXN); enzymes for reducing and eliminating free radicals; glutathione peroxidase which reduces hydroperoxides produced by lipid oxidation, α, β -hydrogenase which reduces intermediates (4-hydroxynonenal, etc.) having carbonyl groups, and the like; proteins (ferritin, transferrin, etc.) that store and transport transition metal ions in such a manner that the transition metal ions do not produce a Fenton (Fenton) reaction; proteases, lipases, nucleases, chlorophyll-degrading enzymes, pheophorbide a monooxygenase, etc., which decompose molecules damaged by oxidation; methionine sulfoxide reductase, DNA repair enzyme, etc., which repair molecules oxidized by reactive oxygen species.

It is presumed that the albizzia julibrissin extract of the present invention increases the amount of antioxidant enzymes and/or redox factors in the living body, reduces active oxygen produced in the living body, and exerts antioxidant ability by promoting the expression level of genes, particularly SOD, PRX, TXN, TRD, GRD, and GXN.

The antioxidant of the present invention may contain other raw materials in addition to the albizzia julibrissin extract within a range not impairing the effects of the present invention. Examples of such a raw material include water, an excipient, an antioxidant, a preservative, a wetting agent, a thickener, a buffer, an adsorbent, a solvent, an emulsifier, a stabilizer, a surfactant, a lubricant, a water-soluble polymer, a sweetener, a taste corrigent, an acidulant, and alcohols. The antioxidant of the present invention may contain other active ingredients within a range not impairing the effects of the present invention. Specific examples of the active ingredient include antioxidant ingredients, anti-aging ingredients, anti-inflammatory ingredients, whitening ingredients, cell activating ingredients, vitamins, blood circulation promoting ingredients, moisturizing ingredients, ingredients having a DNA damage preventing and/or repairing effect, anti-glycation ingredients, peptides or derivatives thereof, amino acids or derivatives thereof, hydroquinone glycosides and esters thereof, and the like.

The antioxidant of the present invention may be blended in combination with components or additives used in cosmetics, quasi-drugs, drinks, medicines, etc., as necessary, in addition to the albizzia julibrissin extract as an essential component, within a range in which the effects of the present invention are not impaired.

Examples of the fats and oils include avocado oil, almond oil, anise oil, perilla oil, olive oil, orange oil, new zealand red fish oil (orange roughy oil), sesame oil, cocoa butter, chamomile oil, carrot oil, cucumber oil, beef tallow (beefttail), tallow acid, tung oil, safflower oil, soybean oil, camellia oil, corn oil, rape oil, peach kernel oil, castor oil, cotton seed oil, peanut oil, turtle oil, mink oil, egg yolk oil, cocoa butter, palm oil, palm kernel oil, japan wax, coconut oil, beef tallow, lard, hydrogenated oil, and hydrogenated castor oil.

Examples of waxes include beeswax, carnauba wax, spermaceti wax, lanolin, liquid lanolin, reduced lanolin (reduced lanoline), hard lanolin, candelilla wax, montan wax, and shellac wax (shellac wax).

Examples of mineral oils include liquid paraffin, vaseline, paraffin, ozokerite (ozokerite), ceresin (ceresin), microcrystalline wax, polyethylene powder, squalene, squalane and pristane (pristine).

Examples of the fatty acid include natural fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, and wool acid; and synthetic fatty acids such as isononanoic acid, caproic acid, 2-ethylbutyric acid, isovaleric acid, 2-methylvaleric acid, 2-ethylhexanoic acid and isovaleric acid.

Examples of the alcohols include natural alcohols such as ethanol, isopropanol, lauryl alcohol, cetyl alcohol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol and phytosterol; synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol and 2-octyldodecanol; and polyhydric alcohols such as ethylene oxide, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol butyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1, 3-butanediol, glycerol, batyl alcohol, pentaerythritol, sorbitol, mannitol, glucose and sucrose.

Examples of esters include isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyldecyl dimethyloctanoate, cetyl lactate, myristyl lactate, diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, and propylene glycol dioleate.

Examples of the metal soap include aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate and zinc undecylenate.

Examples of gum substances (gum mate) and water-soluble polymer compounds include gum arabic, benzoin gum, gum sumac (gum dammar), guaiac, irish moss, karaya gum, tragacanth gum, locust bean gum (gum carob), quince seed (quince seeds), agar, casein, dextran, gelatin, pectin, starch, carrageenan, carboxyalkyl chitin, chitosan, hydroxyalkyl chitin, low molecular weight chitosan, chitosan salts, sulfated chitin, phosphated chitin, alginic acid, alginates, hyaluronic acid, hyaluronate, chondroitin sulfate, heparin, ethylcellulose, methylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, sodium carboxyethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, nitrocellulose, crystalline cellulose, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, sodium alginate, gelatin, sodium alginate, polyvinyl methacrylates, polyacrylates, polyalkylene oxides such as polyethylene oxide and polypropylene oxide, crosslinked polymers of polyalkylene oxides, carboxyvinyl polymers, and polyethyleneimines.

Examples of the surfactant include anionic surfactants (carboxylate, sulfonate, sulfate and phosphate), cationic surfactants (amine salt and quaternary ammonium salt), amphoteric surfactants (carboxylic amphoteric surfactant, sulfuric amphoteric surfactant, sulfonic amphoteric surfactant and phosphoric amphoteric surfactant), nonionic surfactants (ether nonionic surfactant, ether-ester nonionic surfactant, block copolymer nonionic surfactant and nitrogen-containing nonionic surfactant), and other surfactants (natural surfactants, derivatives of protein hydrolysates, polymeric surfactants, titanium-silicon-containing surfactants and fluorocarbon surfactants).

Examples of vitamins include: retinol, retinal (vitamin a1), dehydroretinal (vitamin a2), carotene and lycopene (provitamin a) in the vitamin a group; thiamine hydrochloride, thiamine sulfate (vitamin B1), riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acid, pantothenic acid, biotin, choline, and inositol in the vitamin B group; ascorbic acid and ascorbic acid derivatives of the vitamin C group; ergocalciferol (vitamin D2), cholecalciferol (vitamin D3) and tachysterol in the vitamin D group; tocopherols, tocopherol derivatives and ubiquinones from the vitamin E group; and menadione (phytonadione) (vitamin K1), menaquinone (vitamin K2), menadione (vitamin K3) and menadione (vitamin K4) among the vitamin K group.

Examples of amino acids include: valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine, ornithine and histidine; and amino acid derivatives such as the aforementioned sulfates, phosphates, nitrates, citrates and pyrrolidone carboxylates.

Examples of the whitening agent include ascorbic acid, ascorbic acid derivatives, sulfur, placenta hydrolysate, ellagic acid derivatives, kojic acid derivatives, glucosamine derivatives, arbutin derivatives, hydroxycinnamic acid derivatives, glutathione, arnica essence, Scutellaria (Scutellaria) root essence, mulberry bark essence, bupleurum (Bupleurum) root essence, Saposhnikovia (Saposhnikovia) root essence, Ganoderma (Ganoderma) mycelium culture, Ganoderma mycelium culture extract, basswood essence, peach leaf essence, rose fruit essence, sophora root essence, Sanguisorba (Sanguisorba officinalis) essence, japanese angelica root essence, coix seed essence, oriental persimmon leaf essence, rhubarb essence, moutan bark essence, hamamelis essence, chestnut tree essence, forsythia suspensa (Hypericum erectum) essence and oil-soluble Glycyrrhiza (Glycyrrhiza) essence.

Examples of the moisturizer include hyaluronic acid, polyglutamic acid, serine, glycine, threonine, alanine, collagen, hydrolyzed collagen, skin-firming glycoprotein (hydrolction), fibronectin, keratin, elastin, royal jelly, heparin chondroitin sulfate, glycerophosphate, glyceroglycolipid, sphingomyelin, glycosphingolipid, linoleic acid, linoleate, eicosapentaenoic acid ester, pectin, Lactobacillus bifidus (Lactobacillus bifidus) fermentation product, lactic acid fermentation product, yeast extract, Ganoderma lucidum (Ganoderma lucidum) mycelium culture, Ganoderma lucidum mycelium culture extract, wheat germ oil, avocado oil, rice germ oil, jojoba oil, soybean phospholipid, gamma-oryzanol, hollyhock essence, coix seed essence, rehmannia root essence, jujube essence, shrimp essence, Aloe arborescens (Aloe arborescens) essence, Aloe vera essence, and Aloe vera essence, Burdock essence, rosemary essence, arnica essence and wheat bran.

Examples of the hair restorer include glyceryl pentadecanoate, Coleus (Coleus) essence, gentian essence, pine cone essence, royal jelly essence, Sasa veitchii (Sasa veitchii) essence, t-flavanone, 6-benzylaminopurine, swertia japonica (Swertia japonica) essence, carpronium chloride, minoxidil, finasteride, adenosine, nicotinamide, mulberry root essence, rehmannia root essence, and 5-aminolevulinic acid.

Examples of extracts and essences from animals, plants, and crude drugs include those from: uncaria gambir (Uncaria gambir) (gambir), Angelica keiskei (Angelica keiskei), acerola (acerola), Althea (Althaa), Arnica montana (arnica), avocado, Hydrangea macrophylla (hydrange macrophylla Sering var. thunbergii) (Hydrangea Makino), aloe (aloe), Aloe vera (aloe vera), nettle (nettle), Ginkgo biloba (Ginko ba biolo) (Ginkgo biloba and Ginkgo biloba), fennel, Curcuma longa (Asarum sieboldii) (Asarum root), Japanese apricot (Japanese aprica), Quercus salicina (Quercus salicina), Berberis bearberry (Arstaphylos uva-ursi), Scutellaria baicalensis (Rosaceae), Scutellaria baicalensis (bark of Scutellaria), Scutellaria baicalensis (bark of Rosaceae) (Scutellaria), Scutellaria baicalensis (root (bark of Rosa), Scutellaria (bark of Rosa grandiflora), Scutellaria (Rosa), Scutellaria sinensis (bark (Rosa), Scutellaria grandiflorum) of Rosa), Scutellaria sinensis (bark of Rosa), Scutellaria sinensis (Rosa) of Rosa), Scutellaria sinensis (bark of Rosa), Scutellaria sinensis (bark of Rosa (Rosa), Rosa sinensis, Hypericum erectum (Hypericum erectum), Laminaria sessilifolia (Lamium album), watercress (Nasturtium officinale), citrus (orange), Polygala tenuifolia (Polygala tenuifolia root), Prunella vulgaris (Prunella vulgaris), Polygonum multiflorum (Polygonum multiflorum) (Polygonum multiflorum root), Sophora (Sophora flower), Artemisia vulgaris (Artemisia Princeps) (Artemisia argyi), Curcuma zedoaria (Curcumaria zedoaria) (Curcumae rhizoma), Pueraria lobata (Pueraria lobata), Valeriana officinalis (Valeriana officinalis root), Chrysanthemum morifolium (Anthemis nobilis), Trichosanthes kirilowii (Trichosanthes kirilowii) (Trichosanthemum root), Artemisia capillaris (Artemisia pinnatifida) (Artemisia vulgaris), Glycyrrhiza glabra (Glycyrrhiza glabra), Rhynchosia typha calis japonica (Rhynchophylla), Chrysanthemum morinda officinalis (Chrysanthemum flower), Platycodon grandiflorum (Rotatum grandiflorum (Roxb.) and unripe (Chrysanthemum flower), Platycodon grandiflorum (Thunb), Platycodon grandiflorum (Roxb. grandiflorum) and Rutaceae (Rutaceae), Citrus (Rutaceae) flower, Rutaceae (Rutaceae, Rutaceae, Citrus reticulata (Citrus reticulata) (Citrus peel), cucumber, angelicae gigantis (Aralia cordifolia) and notopterygium (notopterygium root and Aralia root), almond oil (Prunus armeniaca) (almond), Lycium barbarum (Lycium chinensis) (Lycium root bark, Lycium barbarum fruit and Lycium barbarum leaf), Sophora flavescens (Sophora flavescens) (root of kuraria japonica), Cinnamomum camphora (Cinnamomum camphora), mangosteen (Sasa veitchii), grapefruit (Citrus paradisii) fruit, Cinnamomum zeylanicum (Cinnamomum zeylanicum) (cinnamon bark), Schizonepeta tenuifolia) (Schizonepeta spike), Cassia tora (cassiabarks obtusifolia) (Cassia seed), morning glory (ipoea purpurea purrea) and morning glory (Ipomoea nitida) (safflower Carthamus), Magnolia officinalis (mangostera officinalis) (mangnolia), evodia officinalis (mangosteen), gardenia officinalis (ipomea), Magnolia officinalis (acacia mangnolia officinalis) (mangnolia officinalis), gardenia officinalis (safflower Carthamus officinalis) (mangosteen), mangosteen officinalis (mangosteen officinalis) (mangosteen), myrtacea officinalis (mangosteen), myrcia) fruit (mangosteen), myrtacea officinalis (mangosteen), mangosteen (mangosteen), mangoste, Burdock, schisandra (Schizandra chinensis) (shizandra berry fruit), rice bran, wheat, Bupleurum chinense (Bupleurum scorzonerifolium) (Bupleurum root), saffron, Saponaria officinalis (Saponaria officinalis), Crataegus cuneata (Crataegus cuneata) (Crataegus fruit), Zanthoxylum bungeanum (Zanthoxylum piperitum) (Zanthoxylum bungeanum fruit), sage, Panax notoginseng (Panax notogiensis) (Panax notoginseng root), shiitake (Lentinus edodes) (shiitake), Rehmannia glutinosa (Rehmannia glutinosa) (Rehmannia root), Quisqualis indica (Quisqualis indica) (Quisqualis seed), Lithospermum erythrorhizomes (Lithospermum erythrorhizomes) (shikonishii root), Perilla frutescens (caryophylli aestivum occidentalis) (Perilla leaf and Perilla seed), oriental persimmon (persimmon powder), paeonia lactiflora (paeonia), gynura japonica (Plantago asiatica), Plantago asiatica (Plantago), calamus (Plantago), calamus (calamus), calamus (calamus) root, calamus (calamus) and calamus (calamus) root (calamus) of fruticosus (mangium) s (mangium), calamus (mangium) of fruticosus) of frutescens (mangium), calamus (mangium) and calamus (mangium) including (, Betula pendula (Betula alba), honeysuckle (Lonicera japonica) (honeysuckle and honeysuckle leaves and stems), English periwinkle, yarrow, elderberry, red bean (red bean), elderberry (Sambucus sieboldiana)), mallow, Ligusticum wallichii (Ligusticum wallichii), swertia japonica (swertia davidi), mulberry (mulberry bark and leaf), jujube, soybean, macular pigment (Aralia elata), Panax japonicus (Panaxjaponica) (rhizome of Panax japonicus), Anemarrhena asphodeloides (Anemarrhena asphodeloides) (rhizome of Anemarrhena asphodeloides), sanguisorba officinalis (sanguisorba officinalis), Houttuynia cordata (Houttuynia cordifolia), Cordyceps sinensis (sinsis) (rhizome of Chinese Angelica sinensis (plant)), pepper, Phyllanthus annua (Phyllanthus annua) (root), thyme, green tea, black tea, clove, Camellia japonica (mountain tea), Phyllanthus japonica (Citrus sinensis), Angelica sinensis (Chinese), Capsicum annuum, Capsicum annuum, black tea, Eugenia (Achillea japonica (Achillea sinensis (Acronychnum officinale) and Capsicum annuum (Acronatum) including black tea, Capsicum officinale (Acronatum annuum officinale (Acronychnum), Calendula (calndiula officinalis), Citrus aurantium (Citrus aurantium), sanguisorba officinalis, corn (corn milk), Eucommia ulmoides (Eucommia ulmoides) (bark and leaf of Eucommia ulmoides), tomato, Nandina domestica (Nandina domestica), garlic, barley (malt), dictamnus (root bark of dictamnus), Ophiopogon japonicus (Ophiopogon poponicus) (tuber of Ophiopogon japonicus), parsley, sweet potato, mint (Mentha arvensis) (mint herb), witch hazel, rose, loquat (eriobotrya japonica) leaf (loquat leaf (loguat leaf)), poria (porifera cocos) (kernel of poria), grape leaf, muskmelon, linden, peony (tree bark), lupin, sasanqua (Rosa rugosa) (yellow sage), peach seed (marjoram), honey peach seed (honey palm), honey palm, honey locust (honey bee), honey bee sprout (honey bee) and honey bee pollen (honey bee pollen), honey bee pollen, honey bee flower, honey bee pollen, honey bee sprout, honey bee pollen, honey bee flower, honey bee, Areca catechu (Areca catechu), leonurus japonicus (Leymus sibiricus), cornflower, saxifrage (striwberry geranium), Myrica rubra (myrica rubra), Alnus japonica (Alnus firma), Coix lacryma-jobi var. ma-yuen (Coix lacryma-yuen), Artemisia mongolica (Artemisia mongolica), Artemisia desertorum (Artemisia montana), lavender, apple fruit, Ganoderma lucidum (Ganoderma lucidum) (Leys ex. Fr.) Karst, lemon fruit, forsythia suspensa (Forsythia fruit), astragalus, geranium herbs, scopolia (root of scopolia), cockscomb (comb of cock), cow/human placenta extract, pig/cow stomach/duodenum/intestine extract/decomposed extract, water-soluble collagen derivatives, hydrolyzed collagen, elastin, hydrolyzed elastin, water-soluble elastin derivatives, silk protein decomposition products, and bovine blood cell protein decomposition products.

Examples of the microorganism culture metabolites include yeast essence, zinc-containing yeast essence, germanium-containing yeast essence, selenium-containing yeast essence, magnesium-containing yeast essence, fermented glutinous rice essence, gymnema extract, and lactic acid fermentation product of non-fat dry milk.

Examples of alpha-hydroxy acids include glycolic acid, citric acid, malic acid, tartaric acid, and lactic acid.

Examples of the inorganic pigments include anhydrous silicic acid, magnesium silicate, talc, kaolin, bentonite, mica, titanium mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, red iron oxide, black iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, carbon black, and calamine.

Examples of the ultraviolet absorber include p-aminobenzoic acid derivatives, salicylic acid derivatives, anthranilic acid derivatives, coumarin derivatives, amino acid compounds, benzotriazole derivatives, tetrazole derivatives, imidazoline derivatives, pyrimidine derivatives, dioxane derivatives, camphor derivatives, furan derivatives, pyridone derivatives, nucleic acid derivatives, allantoin derivatives, nicotinic acid derivatives, vitamin B6 derivatives, oxybenzone, benzophenone, guaiazulene, shikonin, baicalin, baicalein, and berberine.

Examples of astringents include lactic acid, tartaric acid, succinic acid, citric acid, allantoin, zinc chloride, zinc sulfate, zinc oxide, calamine, zinc p-phenolsulfonate, potassium aluminum sulfate, resorcinol, ferric chloride, and tannic acid.

Examples of antioxidants include ascorbic acid, ascorbate, stearate, tocopherol ester derivatives, nordihydroguaiaretic acid, Butylated Hydroxytoluene (BHT), Butylated Hydroxyanisole (BHA), p-hydroxyanisole, propyl gallate, sesamol, sesamolin, and gossypol.

Examples of anti-inflammatory agents include ichthyol, indomethacin (indomethacin), kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d/dl camphor, hydrocortisone, guaiazulene, chamazulene, chlorphenamine maleate, glycyrrhizic acid, glycyrrhetate, glycyrrhetinic acid, and glycyrrhetinate.

Examples of germicidal disinfectants include rivanol, sulphur, benzalkonium chloride, benzethonium chloride, methylrosaniline chloride, cresol, calcium gluconate, chlorhexidine gluconate, sulfonamides, mercurochrome, lactoferrin, and hydrolyzed lactoferrin.

Hair conditioners include selenium disulfide, alkylisoquinolinium bromide (alkyl isoquinoline bromide) solution, zinc pyrithione (zinc pyrithione), phencyclamate (biphenamine), dithiane (thiantol), castoreum tincture, tincture of ginger, tincture of capsicum, quinine hydrochloride, strong ammonia, potassium bromate, sodium bromate and thioglycolic acid.

Examples of perfumes include natural animal perfumes such as musk, civet, castoreum and ambergris; plant perfumes such as anise (anisie) essential oil, angelica essential oil, ylang essential oil, orris essential oil, fennel essential oil, citrus essential oil, ylang essential oil, caraway essential oil, cardamom essential oil, guaiazuki essential oil, fennel essential oil, piper spicatum essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam (copaibabalsam) essential oil, coriander essential oil, perilla essential oil, cedar essential oil, citronella essential oil, jasmine essential oil, zingiber essential oil, cedar essential oil, spearmint essential oil, mint essential oil, anise essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, gaultheria essential oil, tuunk essential oil, patchouli essential oil, rose essential oil, cypress essential oil, thujopsis essential oil, sandalwood essential oil, bitter orange leaf essential oil, bay essential oil, vetiver essential oil, fingered citron essential oil, pery essential oil, peruvian ebony essential oil, rosewood essential oil, santal, Citrus essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, galois essential oil, lemon grass essential oil, lemon essential oil, rosemary essential oil, and japanese mint essential oil; and other synthetic flavors.

Examples of the pigments/colorants include cabbage red pigment, red rice pigment, madder pigment, annatto pigment, sepia pigment, curcumin pigment, sophora pigment, krill pigment, persimmon pigment, caramel, gold, silver, gardenia pigment, corn pigment, onion pigment, tamarind pigment, spirulina pigment, buckwheat whole plant pigment, cherry pigment, laver pigment, hibiscus pigment, grape juice pigment, calendula pigment, purple sweet potato pigment, taro pigment, shellac pigment and rutin.

Examples of sweeteners include sugar, sweet hydrangea leaf, fructose, arabinose, galactose, xylose, mannose, maltose, honey, glucose, miraculin and monellin (monellin).

Examples of nutritional additives include calcined calcium, cyanocobalamin, yeast, wheat germ, soybean germ, egg yolk powder, hemicellulose and oxymyl red.

In addition, there are hormones, metal-ion blocking agents, pH adjusting agents, chelating agents, antiseptics/antifungals, refrigerants, stabilizers, emulsifiers, animal/plant proteins, decomposed animal/plant proteins, animal/plant polysaccharides, decomposed animal/plant polysaccharides, animal/plant glycoproteins, decomposed animal/plant glycoproteins, blood flow increasing agents, anti-inflammatory agents, anti-allergic agents, cell activators, keratolytic agents, wound healing agents, foaming agents, thickening agents, oral care agents, deodorants, bittering agents, seasonings, enzymes, and the like.

The dosage form of the present invention may be used in any desired dosage form and may be formulated to provide quasi-drugs, skin/hair care cosmetics, bath agents, foods, drinks or medicines in the form of ampoules, capsules, powders, granules, pills, tablets, solids, liquids, gels, foams, creams, ointments, wafers or mousses, etc.

Specific examples of cosmetic and quasi-drug products include internal/external pharmaceutical preparations; basic cosmetics, face cleansers and skin cleansers such as lotions, creams, ointments, lotions, oils and masks; hair cosmetics such as shampoo, rinse (rinse), hair care, hair cream, hair wax, hair gel, hair styling agent (hairdressing), permanent solution, hair tonic, hair dye, and hair tonic; makeup cosmetics (make-up cosmetics) such as foundation cream, white powder (white powder), face powder (face powder), lipstick, blush, eye shadow, eyeliner (liquid), mascara, eyebrow pencil, and false eyelashes; cosmetic preparations such as manicure; a perfume; bath agents, dentifrices; breath fresheners/mouthwashes; liquid odor inhibitors/deodorants; a sanitary article; sanitary napkins and wet wipes.

Examples of the food and drink products include drink products such as refreshing beverages, carbonated beverages, nutritional supplement beverages, fruit juice beverages, and lactic acid beverages; freezing points such as ice cream, sherbet and water ice; noodles such as buckwheat noodles, wheat noodles, vermicelli, dumpling (dumpling and steamed dumpling) skin, Chinese noodles and instant noodles; confectionery (confectioneries) such as fruit hard candies (drop), candies (confections), chewing gums, chocolates, tableted candies, snacks, biscuits, jellies, jams, creams, baked goods and breads; aquatic products such as crab, salmon, mackerel, tuna, sardine, shrimp, bonito, mackerel, whale, oyster, saury, squid, red shellfish, scallop, abalone, sea urchin, salmon caviar, and abalone shell; fishery/livestock processed foods such as fish sausage, ham and sausage; dairy products such as milk powder, processed milk and fermented milk; oils/fats and processed oils/fats foods such as salad oil, frying oil, margarine, mayonnaise, shortening, whipped cream and sauces; seasonings such as soy sauce and sauce (dip); steaming and boiling bag food such as curry, stew, grilled chicken and egg rice, porridge containing vegetables or fish, fried mixed rice, fried pork chop rice, fried fish or vegetable rice, grilled eel rice, meat sauce rice, Japanese mixed rice (Kanto cooking), spicy bean curd, seasoned beef rice, meat soup, egg soup, omelet seasoned rice, dumpling (dumpling and shaomai), hamburger and meat ball; various forms of health/nutritional supplement foods; promoting health food; a tablet; capsules; nutritional supplement drinks and lozenges.

The antioxidant of the present invention can be suitably used for humans, and can also be suitably used for animals other than humans, and respective effects can be expected as much as possible.

The embodiment of the antioxidant of the present invention is not particularly limited, and examples thereof include: face washes, cleansers, lotions (e.g., whitening lotions), creams (e.g., vanishing creams, cold creams), lotions, gels, beauty lotions, face masks (e.g., jelly-like frostings, pasty wiping-off, powdered rinse-off), sheet masks, makeup removers, foundations, lipsticks, lip gloss, lip pencils, blush, pre-makeup creams, shaving creams, sun creams, after-sun lotions, deodorizing lotions, body lotions (including hand creams, foot creams), body oils, and the like.

Hereinafter, the present invention will be described in more detail based on examples. In addition, the present invention is not particularly limited to these examples.

Examples

The water extract of the bark of Albizzia julibrissin used in the following examples used the extract prepared by the method disclosed in Japanese unexamined patent publication No. 2018-58783.

Example 1: evaluation of Effect of promoting expression of target Gene in epidermal keratinocytes of human Normal newborn

The effect of promoting expression of the target gene was evaluated by the following test method using the aforementioned albizzia julibrissin extract as a test sample. The target genes in this example refer to SOD, PRX, TXN, TRD, GRD and GXN.

Human normal neonatal epidermal keratinocytes (NHEK) were inoculated into a T75 flask coated with Collagen Coating Solution (Collagen Coating Solution, manufactured by Toyo Kasei Co., Ltd.), and a medium (Humedia-KG2) for proliferating normal human epidermal keratinocytes was added thereto at 37 ℃ with 10% CO₂The culture is carried out. The culture medium is properly replaced, and the culture is continued until more than 80% confluence is formed. After removing the medium and rinsing with PBS, trypsin treatment was performed and the cells were recovered. Cells were seeded at 1X 10 in 96-well plates⁴Cell/well at 37 ℃ 10% CO₂The culture was carried out for 1 day.

The medium was removed and replaced with medium containing the sample to be tested at a given concentration, 10% CO at 37 deg.C₂The culture was conditioned for 72 hours. After the culture, the cells were collected and Cell Lysis was performed using SuperPrep (registered trademark)&RT qPCR kit (Toyo Boseki Co., Ltd.) was used to prepare cDNA from cells.

Based on the obtained cDNA, real-time PCR analysis was performed using THUNDERBIRD SYBR (registered trademark) qPCR Mix (manufactured by Toyo Boseki Co., Ltd.). The composition of the reaction solution was in accordance with the attached document of the kit. The reaction solution volume was set at 20. mu.l/well and the cDNA volume was set at 3. mu.l. The reaction cycle conditions were carried out at 95 ℃ for 1 minute → (95 ℃, 15 seconds → 60 ℃, 45 seconds) × 40 → 95 ℃, 15 seconds → 60 ℃,1 minute → 95 ℃, 15 seconds. The primers used were the base sequences shown in SEQ ID Nos. 1 to 6. The machine used a 7500 rapid real-time PCR system (applied biosystems). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control, and the expression level of the target gene was normalized (SEQ ID NO: 13-14). The normalized expression level of each target gene was divided by the expression level of the gene of the negative control not containing the test sample, and a relative value was calculated.

The results obtained are shown in FIGS. 1 to 3. As can be seen from fig. 1 to 3, in NHEK, it was confirmed that the gene expression level of antioxidant-related enzymes and/or redox-related factors was increased as compared with the negative control in which no albizia extract was added. From these results, it was found that the albizia julibrissin extract has the effects of promoting the gene expression of antioxidant enzymes and/or redox factors in epidermal cells and improving the antioxidant ability in vivo.

Example 2: evaluation of expression promoting action of target Gene in fibroblast of human Normal newborn

Human normal neonatal fibroblasts (NHDF) were inoculated into a T75 flask, and D-MEM (+ 10% FBS) was added thereto at 37 ℃ with 5% CO₂The culture is carried out. The culture medium is properly replaced, and the culture is continued until more than 80% confluence is formed. After removing the medium and rinsing with PBS, trypsin treatment was performed and the cells were recovered. Cells were seeded at 1X 10 in 96-well plates⁴Individual cells/well, 5% CO at 37 ℃₂The culture was carried out for 1 day.

The medium was removed and replaced with medium containing the sample to be tested at a given concentration, 5% CO at 37 deg.C₂The culture was conditioned for 72 hours. After the culture, the cells were collected and Cell Lysis was performed using SuperPrep (registered trademark)&RT qPCR kit (Toyo Boseki Co., Ltd.) was used to prepare cDNA from cells.

Based on the obtained cDNA, real-time PCR analysis was performed using THUNDERBIRD SYBR (registered trademark) qPCR Mix (manufactured by Toyo Boseki Co., Ltd.). The composition of the reaction solution was in accordance with the attached document of the kit. The reaction solution volume was set at 20. mu.l/well and the cDNA volume was set at 3. mu.l. The reaction cycle conditions were carried out at 95 ℃ for 1 minute → (95 ℃, 15 seconds → 60 ℃, 45 seconds) × 40 → 95 ℃, 15 seconds → 60 ℃,1 minute → 95 ℃, 15 seconds. The primers used were the base sequences shown in SEQ ID Nos. 1 to 12. The machine used a 7500 fast real-time PCR system (Applied Biosystems). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control, and the expression level of the target gene was normalized (SEQ ID NO: 13-14). The normalized expression level of each target gene was divided by the expression level of the gene of the negative control not containing the test sample, and a relative value was calculated.

The results obtained are shown in FIGS. 4 to 9. As can be seen from fig. 4 to 9, in NHDF, it was confirmed that the gene expression levels of antioxidant-related enzymes and redox-related factors were increased as compared with the negative control in which no albizia julibrissin extract was added. From these results, it was found that the albizia julibrissin extract has the effects of promoting the gene expression of antioxidant enzymes and/or redox factors in epidermal cells and improving the antioxidant ability in vivo.

Example 3: evaluation of active oxygen reduction in human Normal neonatal fibroblasts

Human normal neonatal fibroblasts (NHDF) were inoculated into a T75 flask, and D-MEM (+ 10% FBS) was added thereto at 37 ℃ with 5% CO₂The culture is carried out. The culture medium is properly replaced, and the culture is continued until more than 80% confluence is formed. After removing the medium and rinsing with PBS, trypsin treatment was performed and the cells were recovered. Cells were seeded in 96-well plates at 3X 10⁴Individual cells/well, 5% CO at 37 ℃₂The culture was carried out for 1 day.

The medium was removed and washed with HBSS, and then changed to 20. mu. M H₂DCFDA (2 ', 7' -dichlorodihydrofluorescein diacetate) at 37 ℃ with 5% CO₂Incubate (in the dark) for 30 minutes under conditions. Removal of H₂DCFDA was washed with HBSS, replaced with HBSS containing a predetermined concentration of a sample to be tested and hydrogen peroxide, and then washed with 5% CO at 37 deg.C₂The cells were cultured under the conditions of (dark place) and, after 5 hours, the fluorescence intensity was measured at an Ex/Em of 485nm/535nm using a fluorometer (ARVO MX).

The obtained results are shown in fig. 10. As can be seen from fig. 10, in NHDF, the effect of reducing intracellular reactive oxygen species induced by oxidative stress due to hydrogen peroxide was confirmed. This is considered to be due to the fact that the albizia julibrissin extract has an antioxidant effect and also promotes the gene expression of antioxidant-related enzymes and/or redox-related factors, thereby improving the active oxygen eliminating ability of the organism itself and reducing the active oxygen in the organism.

Example 4: evaluation of expression promoting action of antioxidant-related genes in cells obtained from organism

The expression promoting effect on the target gene was evaluated by the following test method using an aqueous solution containing 2.0 wt% of the aforementioned albizzia julibrissin extract as a sample to be tested. The target gene in this example is referred to as PRX.

The test sample was applied to the face of the subject, and cells were taken from the lower-jaw beard in the applied area and evaluated. Specifically, the lower jaw beard in the application area was removed from the subject before and 3 hours after the application of the test sample, and hair follicle cells were collected. The cells were immersed in RNAlater (Qiagen), and stored at 4 ℃ until RNA extraction. After completion of collection of hair follicle cells, total RNA was extracted from hair follicle cells using RNeasyplus mini kit (Qiagen) according to the attached protocol. cDNA was prepared for the extracted total RNA using ReverTra Ace (registered trademark) qPCR RT Master Mix (manufactured by Toyo Boseki Co., Ltd.) according to the attached protocol. Based on the obtained cDNA, real-time PCR analysis was performed using THUNDERBIRD SYBR (registered trademark) qPCRMix (manufactured by Toyo Boseki Co., Ltd.). The composition of the reaction solution was in accordance with the attached document of the kit. The reaction solution volume was 20. mu.l/well and the cDNA volume was 2. mu.l. The reaction cycle conditions were carried out at 95 ℃ for 1 minute → (95 ℃, 15 seconds → 60 ℃, 45 seconds) × 40 → 95 ℃, 15 seconds → 60 ℃,1 minute → 95 ℃, 15 seconds. The primers used were the base sequences shown in SEQ ID Nos. 7 to 8. The machine used a 7500 fast real-time PCR system (Applied Biosystems). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control, and the expression level of the target gene was normalized (SEQ ID NO: 13-14). The normalized expression level of each target gene was divided by the expression level of the gene of the negative control before the test sample was applied, and a relative value was calculated.

The obtained results are shown in fig. 11. As can be seen from fig. 11, it was confirmed that the cDNA obtained from hair follicle cells had an increased gene expression level of PRX 3 hours after application, as compared with the negative control before application of the albizia julibrissin extract. From this result, it was found that the albizia julibrissin extract also has the effects of promoting the gene expression of antioxidant-related enzymes and/or redox-related factors and improving the antioxidant ability in vivo in cells collected from an organism.

Example 5: prescription of skin external preparation

Hereinafter, examples of the prescription as the external preparation for skin of the present invention are shown. Any of these preparations is expected to have an effect of promoting the expression of an antioxidant-related enzyme gene by an albizzia julibrissin extract, and is effective as an antioxidant.

Toning lotion

A cosmetic lotion was prepared according to a conventional method, according to the following composition.

Gel

A gel was prepared according to a conventional method according to the following composition.

Cream

A cream was prepared according to a conventional method with the following composition.

Industrial applicability

According to the present invention, by remarkably promoting the expression of an antioxidant-related enzyme and/or a redox-related factor in a living body, it is possible to safely and effectively reduce oxidative stress in the living body, maintain health, and prevent various diseases and symptoms caused by oxidative stress, and the present invention is useful as a cosmetic or quasi-drug to be applied to a skin external preparation in particular.

Claims

1. An antioxidant contains Albizzia julibrissin extract.

2. The antioxidant agent as claimed in claim 1, wherein the albizzia julibrissin extract is an aqueous extract from a plant body of a plant belonging to the genus albizzia.

3. The antioxidant according to claim 1 or 2, wherein the albizia julibrissin extract has an effect of promoting gene expression of an antioxidant-related enzyme and/or a redox-related factor.

4. The antioxidant according to any one of claims 1 to 3, wherein the antioxidant-related enzyme and/or redox-related factor is at least one or more selected from the group consisting of superoxide dismutase (SOD), Peroxiredoxin (PRX), Thioredoxin (TXN), Thioredoxin Reductase (TRD), Glutathione Reductase (GRD) and Glutaredoxin (GXN).

5. The antioxidant according to any one of claims 1 to 4, wherein the Albizzia julibrissin extract has an effect of reducing intracellular active oxygen.

6. An external preparation for skin, wherein the antioxidant according to any one of claims 1 to 5 is formulated.

7. The external preparation for skin according to claim 6, wherein the albizia julibrissin extract is contained in a concentration of 0.00001 to 5.0 wt%.