HK1167345A - Expression promoter for redox-related factor - Google Patents

Description

Redox-related factor expression promoter

Technical Field

The present invention relates to an expression promoter of a redox-related factor comprising kaempferol (kaempferol), quercetin (quercetin) or a glycoside thereof, or any combination of the above kaempferol, quercetin and a glycoside thereof as an active ingredient, and an expression promoter of a redox-related factor comprising ginkgo biloba extract, kaempferia galanga extract or a combination thereof as an active ingredient.

Background

The organism is redox-regulated to maintain its survival and constant. Redox regulation in organisms relies primarily on reversible redox reactions of sulfhydryl groups on cysteine residues present in intracellular proteins, thereby controlling various cellular functions. It is known that factors involved in redox regulation (redox-related factors) maintain a reduced state in cells and exert an essential role for cell survival, while they also play an important role in scavenging active oxygen in the living body. If the above redox-related factors do not function normally, active oxygen in the organism is excessively generated, thereby suffering from excessive oxidative stress. Such excessive oxidative stress is considered to cause various diseases and/or aging by oxidizing and damaging DNA and/or biopolymers such as protein and lipid. Therefore, in recent years, research has been actively conducted on redox-related factors that function as a system for scavenging active oxygen that causes oxidative stress.

The glutathione system is known as a redox-related factor that acts as a defense mechanism against oxidative stress of an organism. Glutathione is a tripeptide reported to be present in mammals and has the following effects: utilizing the reduction of a thiol group in a molecule thereof, the action as a coenzyme, the action involved in the formation of thioether amino acid and other detoxification mechanisms, the action of protecting thiol enzyme (thiol enzyme) or other cellular components, the action of promoting excretion of harmful substances, the action of improving cholinesterase activity to prevent allergy, and the enzyme activation action. However, since glutathione precipitates in preparations and most cells cannot take up glutathione, a component for promoting intracellular glutathione production is extremely useful. As such components, γ -tocopherol or δ -tocopherol has been reported so far, and a glutathione production promoter containing these components has been disclosed (japanese patent laid-open No. 2007-284430 (patent document 1)).

Further, as other redox-related factors, thioredoxin system is known, and the thioredoxin system also functions as a defense mechanism against oxidative stress in organisms (cell engineering vol.25, No.2, 2006, p143-148 (non-patent document 1)). Furthermore, it has been reported that transgenic mice of thioredoxin become long-lived (Mitsui A, et al: Artificial Redox Signal (2002) 4: 693-696 (non-patent document 2)), and that thioredoxin is resistant to cerebral infarction, diabetes (Takagi Y, et al: Proc Natl Acad Sci USA (1999) 96: 4131-4136 (non-patent document 3), Hotta M, et al: J Exp Med (1998) 188: 1445-1451 (non-patent document 4)), and effective against allergic dermatitis (Japanese patent laid-open No. 2007-269671 (patent document 2)), and thus thioredoxin is widely used as an active oxygen scavenger (Japanese patent laid-open No. 9-157153 (patent document 3)). However, since the thioredoxin system is a high-molecular protein, these molecules cannot be taken into cells even if these compounds are directly administered to a subject. Therefore, although it is desired to promote the expression of thioredoxin system in living body, only wormwood extract or green perilla extract has been known so far as a component for promoting the expression thereof (international publication No. WO2006/033351 (patent document 4)).

As described above, even when redox-related factors such as a glutathione system and/or a thioredoxin system are directly administered to a living body, they cannot be efficiently taken into cells, and thus there is a possibility that redox-regulating functions and/or antioxidant functions cannot be exerted in the living body. Although it is known that the expression of these redox-related factors can be enhanced by ultraviolet irradiation stress, chemical substances that act safely and efficiently on living organisms have not been known so far, and the expression mechanism of redox-related factors in living organisms has not been elucidated in detail. Therefore, it is still desired to find a safe and effective component for promoting the expression of a redox-related factor in vivo.

Patent document 1: japanese patent laid-open No. 2007 & 284430

Patent document 2: japanese laid-open patent publication 2007-269671

Patent document 3: japanese patent laid-open No. 9-157153

Patent document 4: international publication No. WO2006/033351

Non-patent document 1: cell engineering Vol.25, No.2, 2006, p143-148

Non-patent document 2: mitsui A, et al: artificial Redox Signal (2002) 4: 693-696

Non-patent document 3: takagi Y, et al: proc Natl Acad Sci USA (1999) 96: 4131-4136

Non-patent document 4: hotta M, et al: j Exp Med (1998) 188: 1445-1451

Disclosure of Invention

The present invention addresses the problem of promoting the expression of a redox-related factor in an organism, thereby safely and effectively reducing oxidative stress in the organism, maintaining the health of the organism, and preventing various diseases and symptoms caused by oxidative stress.

At this time, as components for enhancing the expression and activity of the glutathione system and the thioredoxin system, which are redox-related factors in the organism, the present inventors found kaempferol (kaempferol) and quercetin (quercetin) which are flavonols. Unexpectedly, the above effects are characteristic of kaempferol and quercetin in flavonols and are not observed in structurally similar myricetin. Compared with the case of directly applying the molecules of the glutathione system and/or the thioredoxin system, the application of kaempferol and/or quercetin can more effectively promote the expression and the activity of the redox-related factors such as the glutathione system and/or the thioredoxin system in the organism, so that the invention can more effectively relieve various oxidative stresses in the organism. Flavonols such as kaempferol and/or quercetin are widely distributed in tea plants, are nontoxic to organisms, and can be safely ingested. Thus, when kaempferol and quercetin are used as active ingredients for promoting the expression of redox-related factors in the living body, excess active oxygen in the living body can be eliminated without giving stress to the living body to which the composition is applied.

Accordingly, the present application encompasses the following inventions:

[1] a redox-related factor expression promoter comprising kaempferol, quercetin or a glycoside thereof, or any combination of the above-mentioned kaempferol, quercetin and a glycoside thereof as an active ingredient.

[2] The redox-related factor expression promoter according to [1], wherein the redox-related factor is 1 or more selected from thioredoxin, thioredoxin reductase, glutathione reductase and glutaredoxin.

[3] An expression promoter for a redox-related factor comprising ginkgo biloba extract, kaempferia galanga extract or a combination thereof as an effective ingredient.

[4] The redox-related factor expression promoter according to [3], wherein the redox-related factor is 1 or more selected from thioredoxin, thioredoxin reductase, glutathione reductase and glutaredoxin.

[5] Use of kaempferol, quercetin or a glycoside thereof, or any combination of the above-mentioned kaempferol, quercetin and a glycoside thereof, for the preparation of a redox-related factor expression promoter.

[6] Use of Ginkgo biloba extract, Kaempferia galanga extract or their combination for preparing redox-related factor expression promoter.

By promoting the expression of the redox-related factor in the living body, it is possible to safely and effectively reduce active oxygen in the living body and protect the living body from oxidative stress. Thereby maintaining the health of the organism and preventing various diseases and symptoms caused by oxidative stress.

Drawings

Fig. 1 is a graph showing that kaempferol enhances the expression of redox-related factors in an organism.

Fig. 2 is a graph showing the expression of redox-related factors in a kaempferol-enhanced skin equivalent model.

Fig. 3 is a graph showing a comparison of the effects of flavonoids on enhancing the expression of the thioredoxin system.

FIG. 4 is a graph showing that ginkgo biloba extract and kaempferia galanga extract enhance the expression of thioredoxin reductase.

FIG. 5 is a graph showing the effect of kaempferol and quercetin to promote the activity of thioredoxin reductase.

FIG. 6 is a graph showing the effect of kaempferol and the extract of Ginkgo biloba and Kaempferia galanga on the inhibition of the production of inflammatory cytokines (IL1 β).

Detailed Description

In 1 embodiment of the present invention, there is provided a redox-related factor expression promoter comprising kaempferol, quercetin or a glycoside thereof, or any combination of the above-mentioned kaempferol, quercetin and a glycoside thereof as an active ingredient.

In the present specification, the term "redox-related factor" means a factor that undergoes an oxidation or reduction reaction with a thiol group of a cysteine residue of a protein present in a living body, for example, a mammal, preferably a human body, and maintains a reduced state in a cell. It is considered that modification of protein-protein interaction by oxidation of a thiol group at a cysteine residue in a protein present in a cell plays a particularly important role in activation of the cell and/or processing of an information transduction substance, that is, a signal transduction mechanism. The oxidation/reduction (redox) of the thiol group at the cysteine residue is involved in the regulation of various life phenomena such as gene transcription, protein intracellular localization, synthesis, and degradation, and cell proliferation and/or cell death. As systems related to redox in such organisms, a glutathione system and a thioredoxin system are known, and each of them is composed of a group of molecules. These redox-related factors are also known to be involved in the scavenging of reactive oxygen species generated and present in organisms.

The thioredoxin system is composed of factors such as thioredoxin and thioredoxin reductase in particular. Thioredoxin (TRX) has been discovered as a coenzyme that supplies hydrogen to ribonucleotide reductase, an enzyme essential for DNA synthesis in Escherichia coli. TRX is a protein of about 12kDa conserved in various organisms such as prokaryotes and/or humans, has a three-dimensional structure including 4 β -sheets and 3 α -helices, and has an active site of-Cys-Gly-Pro-Cys-. TRX has an oxidized form (S-S) in which disulfide bonds are formed between 2 cysteine residues in its active site and a reduced form (-SH, SH-), which reduces the disulfide bonds of the substrate protein, thereby becoming oxidized itself. Oxidized TRX is reduced by NADPH and thioredoxin reductase, thereby regenerating reduced form. As one of the substrate proteins, a catalase family collectively called peroxiredoxin (peroxiredoxin) is known. Reduced peroxiredoxin reduction H₂O₂. Then, the redox protein that has become oxidized is reduced by thioredoxin as a substrate. Thioredoxin that has been changed to the oxidized form is reduced by thioredoxin reductase. Among such active oxygen detoxification systems, the thioredoxin system plays an important role.

The glutathione system is composed in particular of factors such as glutathione, glutathione reductase and glutaredoxin. Glutathione is a tripeptide formed from glutamic acid, cysteine and glycine, is present in a large amount in cells as a nonproteinaceous thiol component, and is known as a main antioxidant substance in hydrophilic regions in cells. Glutathione reacts with active oxygen (e.g., superoxide or hydrogen peroxide) to form a stable glutathione radical, which becomes a 2-mer (GSSG: oxidized glutathione), and then glutathione reductase transfers electrons from NADPH to GSSG, thereby generating GSH (reduced glutathione). Glutaredoxin (GRX) has an active site, Cys-Pro-Tyr-Cys-, which is homologous to thioredoxin, and is called the thioredoxin superfamily because of its homology to thioredoxin. GRX has an oxidized form (S-S) and a reduced form (-SH, SH-) in which disulfide bonds are formed between 2 cysteine residues in its active site. Like thioredoxin, the reduced form reduces the disulfide bond of the substrate protein, and changes itself to the oxidized form. There are many unclear points as to their biological activity, but are known to be associated with redox regulation and/or oxidative stress responses in organisms.

In this way, by activating redox-related factors such as thioredoxin, thioredoxin reductase, glutathione reductase and glutaredoxin or promoting their expression, it is possible to reduce excess active oxygen in the living body and protect the living body from oxidative stress.

As described above, this time, the inventors obtained the following surprising recognition: kaempferol and quercetin activate the redox-related factors and promote their expression. The flavonols comprising kaempferol and quercetin have the general formula shown below,

kaempferol: r₁＝H；R₂＝H

Quercetin: r₁＝OH；R₂＝H

Myricetin: r₁＝OH；R₂＝OH

They are 1 of the flavonoids widely distributed in plants. These flavonols are known to have antioxidant effects, and these flavonols have also been reported to have a vascular strengthening effect and/or an anti-inflammatory effect. However, it has not been known so far that flavonols can promote the expression of redox-related factors in organisms, and it is extremely unexpected that, among these flavonols whose chemical structures are extremely similar, only kaempferol and quercetin specifically promote the expression of redox-related factors. Based on the above knowledge, it was clarified that kaempferol and quercetin are extremely useful as active ingredients for promoting the expression of redox-related factors.

Further, it is known that flavonols form various glycosides, and glycosides of kaempferol and quercetin may be used as active ingredients in the redox-related factor expression promoter of the present invention. In the present invention, sugars which form glycosides with them are not limited, and examples of aldoses include glucose, mannose, galactose, fucose, rhamnose, arabinose, and xylose, and examples of ketoses include fructose.

As described above, since kaempferol, quercetin and their glycosides are components widely distributed in plants, plant extracts containing these components can also be used to promote the expression of redox-related factors in organisms. The present inventors have found that, among plant extracts, particularly, ginkgo biloba extract and kaempferia galanga extract significantly promote the expression of redox-related factors in the organism. Accordingly, in other embodiments of the present invention, there is provided a redox-related factor expression promoter comprising ginkgo biloba extract, kaempferia galanga extract or a combination thereof as an effective ingredient.

The amount of kaempferol, quercetin or their glycosides, or any combination of the above-mentioned kaempferol, quercetin and their glycosides, which is used as an active ingredient in the redox-related factor expression promoter of the present invention, is not particularly limited, and may be about 0.00001 to 0.5 mass%, preferably about 0.0001 to 0.001 mass%, and most preferably about 0.0001 to 0.0005 mass%. The amount of the ginkgo biloba extract, the kaempferia galanga extract, or the combination thereof used as an active ingredient in the redox-related factor expression promoter of the present invention is, for example, about 0.00001 to 0.5 mass%, preferably about 0.0001 to 0.1 mass%, and most preferably about 0.0001 to 0.01 mass% in terms of the dry residue of the extract.

In the present invention, the above-mentioned active ingredients may be administered orally or parenterally in the form of an expression promoter for a redox-related factor containing these active ingredients, and they may be formulated into, for example, an oral composition or an external composition. In addition, the redox-related factor expression promoter of the present invention can maintain the health of an organism and prevent various diseases and symptoms caused by oxidative stress. As such diseases and symptoms, there are mentioned, without limitation, cancers, diabetes, autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, antiphospholipid antibody syndrome, dermatomyositis, systemic scleroderma, sjogren's syndrome, Guillain-Barre syndrome, chronic atrophic gastritis, aortic inflammation syndrome, goodpasture's syndrome, rapidly progressive glomerulonephritis, exophthalmic goiter, chronic discoid lupus erythematosus or habitual abortion, ischemic diseases, such as angina pectoris, myocardial infarction, cerebral infarction or arteriosclerosis obliterans, allergic diseases, such as bronchial asthma, allergic rhinitis, pollinosis, allergic gastroenteritis, urticaria, contact dermatitis or atopic dermatitis, skin aging, pigmentation, chapping, seborrhea, solarization, burn, acne, skin relaxation, obesity, hypertension, metabolic syndrome, anemia. Therefore, the use of the redox-related factor expression promoter of the present invention is unlimited in possibility, and can be used as a raw material or an additive for foods, beverages, cosmetics, drugs, quasi drugs, and the like in therapy or cosmetology.

Examples of the form of the composition for oral administration include general foods, functional foods, nutritional supplements, special health foods, reconstituted milks for premature babies, reconstituted milks for babies, infant foods, foods for pregnant and lying-in women, foods for the elderly, beverages, functional foods, and medicines, and the composition for oral administration may be in the form of processed forms such as tablets, powders, granules, tablets, oral liquids, suspensions, emulsions, and syrups.

The promoter for expression of a redox-related factor of the present invention may contain, in addition to the active ingredient of the present invention, various carriers generally used in foods, drinks, drugs or quasi drugs, and/or additives such as antioxidants. For example, the expression promoter for a redox-related factor of the present invention may contain, as a carrier, various carriers, extenders, diluents, extenders, dispersants, excipients, binder solvents (e.g., water, ethanol, vegetable oil), solubilizers, buffers, dissolution promoters, gelling agents, suspending agents, wheat flour, rice flour, starch, corn starch, polysaccharides, milk protein, collagen, rice bran oil, and lecithin, in addition to the active ingredient of the present invention. Examples of the additives include, but are not limited to, vitamins, sweeteners, organic acids, colorants, flavors, moisture-proofing agents, fibers, electrolytes, minerals, nutrients, antioxidants, preservatives, fragrances, humectants, natural food extracts, and vegetable extracts. Examples of the antioxidant include natural antioxidants such as tocopherols, flavone derivatives, phyllodulcin, kojic acid, gallic acid derivatives, catechins, フキ acid (fukiic acid), gossypol, pyrazine derivatives, sesamol, guaiol (guaiaol), guaiaconic acid (guaiac acid), p-coumaric acid, nordihydroguaiaretic acid (nor-dihydroguaiaretic acid), sterols, terpenes, nucleobases, carotenoids, lignans, and the like, and synthetic antioxidants including ascorbyl palmitate, ascorbyl stearate, Butyl Hydroxyanisole (BHA), Butyl Hydroxytoluene (BHT), mono-tert-butylhydroquinone (TBHQ), 4-hydroxymethyl-2, 6-di-tert-butylphenol (HMBP).

The composition for external use may be formulated into various forms such as ointment, cream, pack, lotion, milky lotion, bath agent, shampoo, hair tonic, hair liquid, shampoo, hair conditioner, etc., and may be appropriately blended with ingredients generally used as an external preparation, for example, whitening agent, moisturizer, oily ingredient, ultraviolet absorber, surfactant, thickener, alcohol, powder ingredient, colorant, aqueous ingredient, water, various skin nutrients, etc., as required. In addition, the redox-related factor expression promoter of the present invention may also be appropriately blended with conventional adjuvants for external preparations, for example, metal chelating agents such as disodium ethylenediaminetetraacetate, trisodium ethylenediaminetetraacetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc., agents such as caffeine, tannin, verapamil (verapamil), tranexamic acid and its derivatives, licorice extract, glabridin, hot water extract of papaya fruit, various crude drugs, tocopherol acetate, glycyrrhizic acid and its derivatives or its salts, etc., whitening agents such as vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid, etc., saccharides such as glucose, fructose, mannose, sucrose, trehalose, etc., and vitamin a compounds such as retinoic acid, retinol acetate, retinol palmitate, etc.

Representative formulations of the redox-related factor expression promoter administered in the present invention are shown below.

Protectant A

Protecting agent B

2-layer cream

Ointment

Patch preparation

Ion-introducing agent

Candy A

Tablet C

Soft capsule B

Particles A-B

Beverage A

Biscuit

Slowly adding granulated sugar while stirring butter, adding egg, folium Ginkgo extract, rhizoma Kaempferiae extract, kaempferol, quercetin and spice, and stirring. After the mixture was sufficiently mixed, the uniformly shaken weak flour was added, and the mixture was put in a refrigerator in the form of a block under low-speed stirring. Then, the mixture is molded and baked at 170 ℃ for 15 minutes to make biscuits.

Examples

Example 1 search for Agents that increase the expression of the thioredoxin System

Normal human epidermal keratinocytes (KK-400)9, KURABO) in DefinedKeratinocyte-SFM Medium (10744-₂5% and 37 ℃ under the conditions of culture. The cells at a final density of 100% were replaced with EpiLife adjusted to a calcium concentration of 1.5mM^TMCalciuM-Free Phenol Red Free medium (M-EPICF/PRF-500, KURABO). After 24 hours, kaempferol (K0018, Tokyo chemical Co., Ltd.), vitamin E (95240, FULKA), and lipoic acid ((+/-) -alpha-lipoic acid) (T1395, SIGMA) were added thereto to give a final concentration of 10. mu.M, respectively.

24 hours after the addition of the drug, RNA was isolated using QIAZOL lysine Reagent (79306, QIAGEN) and purified using RNeasy kit (74106, QIAGEN). Then, the amounts of mRNA of thioredoxin reductase, thioredoxin, glutathione reductase, glutaredoxin, ribosomal protein as an internal standard were quantified by LightCycler software (Roche) using QuantiTect SYBR Green RT-PCR Kit (204243, QIAGEN). The sequences of the primers used are as follows.

Thioredoxin reductase forward primer:

TGCTGGCAATAGGAAGAGATGGCTTGCAC

thioredoxin reductase reverse primer:

GCAATCTTCCTGCCTGCCTGGATTGCAACTGG

thioredoxin forward primer:

TCGGTCCTTACAGCCGCTCGTCAGACTCCA

thioredoxin reverse primer:

AGGCCCACACCACGTGGCTGAGAAGTCAAC

glutathione reductase forward primer:

GATCCTGTCAGCCCTGGGTTCTAAGACATC

glutathione reductase reverse primer:

TAACCATGCTGACTTCCAAGCCCGACAA

glutaredoxin forward primer:

ATCACAGCCACCAACCACACTAACGAGA

glutaredoxin reverse primer:

GTTACTGCAGAGCTCCAATCTGCTTTAGCC

ribosomal protein forward primer:

ACAGAGGAAACTCTGCATTCTCGCTTCCTG

ribosomal protein reverse primer:

CACAGACAAGGCCAGGACTCGTTTGTACC

the mRNA expression level was determined as the result of dividing the mRNA levels of thioredoxin reductase, thioredoxin, glutathione reductase and glutaredoxin by the mRNA level of ribosomal protein.

As a result, kaempferol significantly enhanced the expression of redox-related factors compared to vitamin E and lipoic acid, which are known to have antioxidant effects (fig. 1).

Example 2 Effect of Kaempferol on the expression of Redox-related factors in skin equivalent models

Skin equivalent models were prepared from normal human, normal human keratinocytes (Invitrogen), normal human fibroblasts (ATCC) and collagen I (Sigma), followed by high humidity, CO, in Dulbecco's modified eagle's Medium (D-MEM) (Invitrogen)₂5% and 37 ℃ under the conditions of culture. 2mM or 10mM of kaempferol was added to the culture medium, and then the model was cultured for 24 hours. Then, RNA was isolated from the epithelial layer of the skin equivalent model using QIAGEN mRNA isolation kit. Next, thioredoxin reductase and glutathione reductase, and mRNA of 36B4 as an internal standard were quantified by q-PCR (quantitative PCR using Bio-Rad MyiQ monochrome Real-time PCR detection system) using the Real-time PCR kit (Bio-Rad). Primers were purchased from QIAGEN (TXNRD 2: # QT 00070371; GSR; # QT 000383; QT 000383)25). The amount of thioredoxin reductase and the amount of glutathione reductase were divided by the amount of mRNA of 36B4 to obtain the mRNA expression level.

As a result, kaempferol significantly enhanced the expression of redox-related factors in the skin equivalent model (fig. 2).

Example 3 search for Agents that increase the expression of the thioredoxin System-Flavonoids

Normal human epidermal keratinocytes (KK-4009, KURABO) were cultured in DefinedKeratinocyte-SFM medium (10744-₂5% and 37 ℃ under the conditions of culture. The cells at a final density of 100% were replaced with EpiLife adjusted to a calcium concentration of 1.5mM^TMCalciuM-Free Phenol Red Free medium (M-EPICF/PRF-500, KURABO). After 24 hours, the following agents were added to a final concentration of 10. mu.M.

Camphanol (K0018, Tokyo Kasei Kogyo)

Quercetin (173-00403, Wako pure chemical industries, Ltd.)

Myricetin (70050, FLUKA)

Epigallocatechin gallate (E4143, CIAL)

Hesperidin (086-07342, Wako pure chemical industries, Ltd.)

5, 7, 4' -Trihydroxyflavone (012- & 18913, Wako pure chemical industries, Ltd.)

5, 7, 4' -Trihydroxyisoflavone (070-04681, Wako pure chemical industries, Ltd.)

24 hours after the addition of the drug, the amount of mRNA expression of the redox-related factor was determined by the method described in example 1.

As a result, of the above flavonoids, only kaempferol and quercetin significantly enhanced the expression of the thioredoxin system (fig. 3).

Example 4 search for agent for enhancing thioredoxin expression in the System-search for plant extract

Normal human epidermal keratinocytes (KK-4009, KURABO) were cultured in DefinedKeratinocyte-SFM medium (10744-₂5% and 37 ℃ under the conditions of culture. Cells at a final density of 100% were replaced with EpiLife adjusted to a calcium concentration of 1.5mM^TMCalciuM-Free Phenol Red Free medium (M-EPICF/PRF-500, KURABO). After 24 hours, the following agents were added to a final concentration of 0.1%.

Kaempferia galanga extracting solution: extracting rhizome of Kaempferia galanga (Kaempferia galanga) with 50% 1, 3-butanediol to obtain solution;

extracting the ginkgo leaves: a solution obtained by dissolving 50% ethanol extract of leaves of Ginkgo biloba (Ginkgo biloba L.) in 1, 3-butanediol;

sweet tea extract: a solution of hot aqueous extract of sweet tea (Rubus Suavissimus Shugan Lee. (Rosasea)) in 50% 1, 3-butanediol.

24 hours after the addition of the drug, the amount of mRNA expression of the redox-related factor was determined by the method of example 1.

As a result, it was confirmed that, among plant extracts known to contain kaempferol, quercetin and glycosides thereof, the ginkgo leaf extract or kaempferia galanga extract particularly enhanced the expression of the thioredoxin system (fig. 4).

Example 5 thioredoxin reductase Activity promoting Effect

Normal human epidermal keratinocytes (KK-4009, KURABO) were cultured in DefinedKeratinocyte-SFM medium (10744-₂5% and 37 ℃ under the conditions of culture. Cells at a final density of 100% were replaced with EpiLife adjusted to a calcium concentration of 1.5mM^TMCalciuM-Free Phenol Red Free medium (M-EPICF/PRF-500, KURABO). After 24 hours, kaempferol (K0018, Tokyo Kasei Kogyo Co., Ltd.) or quercetin (173-Pure drug industries, Ltd.) to a final concentration of 10. mu.M. After 24 hours of addition of the drug, the drug was dissolved in a buffer, and insoluble components were removed by centrifugation at 14,000rpm for 15 minutes at 4 ℃. The total protein mass IN the soluble fraction was quantified and the activity was determined according to the thioredoxin reductase activity assay of Arne Holmgren et al (METHODS IN ENZYMOLOGY, 1995, 252, 199-).

As a result, it was confirmed that kaempferol and quercetin promoted thioredoxin reductase activity (fig. 5).

Example 6 effects of kaempferol, Kaempferia galanga extract, and Ginkgo biloba extract on inhibition of inflammatory cytokine (IL 1. beta.) expression

Normal human epidermal keratinocytes (KK-4009, KURABO) were cultured in DefinedKeratinocyte-SFM medium (10744-₂Culturing was carried out at 5% and 37 ℃. Cells at a final density of 100% were replaced with EpiLife adjusted to a calcium concentration of 1.5mM^TMCalciuM-Free Phenol Red Free medium (M-EPICF/PRF-500, KURABO). After 24 hours, kaempferol was added to a final concentration of 10. mu.M, or the following plant extracts were added to a final concentration of 0.1%.

Camphanol (K0018, Tokyo Kasei Kogyo)

Kaempferia galanga extracting solution: extracting rhizome of Kaempferia galanga (Kaempferia galanga) with 50% 1, 3-butanediol to obtain solution

Extracting the ginkgo leaves: dissolving 50% ethanol extract of folium Ginkgo (Ginkgo biloba L.) in 1, 3-butanediol

After 24 hours of addition of the drug, RNA was extracted by the method of example 1, and the mRNA expression level of the inflammation-related factor (IL 1. beta.) was quantified. The primers used are as follows.

IL1 β · forward primer:

GGCCATGGACAAGCTGAGGAAGATGCTG

IL1 β reverse primer:

TGCATCGTGCACATAAGCCTCGTTATCCC

the mRNA expression level was determined as the result of dividing the mRNA level of IL 1. beta. by the mRNA level of ribosomal protein.

As a result, it was confirmed that kaempferol, kaempferia galanga extract and ginkgo biloba extract inhibited the production of IL1 β, which is an inflammatory cytokine, and thus had anti-inflammatory effects (fig. 6).

Claims (4)

1. A redox-related factor expression promoter comprising kaempferol, quercetin or a glycoside thereof, or any combination of the above-mentioned kaempferol, quercetin and a glycoside thereof as an active ingredient.

2. The redox-related factor expression promoter according to claim 1, wherein the redox-related factor is 1 or more selected from thioredoxin, thioredoxin reductase, glutathione reductase and glutaredoxin.

3. An expression promoter for a redox-related factor comprising ginkgo biloba extract, kaempferia galanga extract or a combination thereof as an effective ingredient.

4. The redox-related factor expression promoter according to claim 3, wherein the redox-related factor is 1 or more selected from thioredoxin, thioredoxin reductase, glutathione reductase and glutaredoxin.