Humectant, skin barrier function improving agent, Anti-inflammatory agent, hyaluronic acid promoting agent, and melanin production inhibitor

Abstract

The present invention provides a humectant, a skin barrier function improving agent, an anti-inflammatory agent, a hyaluronic acid promoting agent, and a melanin production inhibitor.　Provided are a humectant, a skin barrier function improving agent, an anti-inflammatory agent, a hyaluronic acid promoting agent, and a melanin production inhibitor that contain cis-3-hexenol as an active ingredient.

Description

Moisturizing agent, skin barrier function improver, anti-inflammatory agent, hyaluronic acid increaser, and melanin production inhibitor

[0001] The present invention relates to a moisturizing agent, a skin barrier function improving agent, an anti-inflammatory agent, a hyaluronic acid increasing agent, and a melanin production inhibitor, each containing cis-3-hexenol as an active ingredient.

[0002] Cis-3-hexenol is an aromatic component found in green tea, and its aroma is known to have effects such as fatigue relief when smelled (Patent Document 1). However, this effect is exerted by smelling the aroma, and is different from the effect achieved when cis-3-hexenol is directly applied to the skin. Patent Document 2 describes that cis-3-hexenol has an inhibitory effect on melanin production by melanoma cells.

[0003] International Publication No. 2005 / 000286 Japanese Patent Application Laid-Open No. 2022-174825

[0004] The present inventors aim to obtain moisturizing agents, skin barrier function improving agents, anti-inflammatory agents, hyaluronic acid increasing agents, melanin production inhibitors, and the like.

[0005] The present inventors have discovered that applying cis-3-hexenol directly to the skin exerts beneficial effects on the skin, such as moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and inhibiting melanin production, and have thus arrived at the present invention.

[0006] The present invention relates to the following: [1] A moisturizing agent containing cis-3-hexenol as an active ingredient. [2] A skin barrier function improving agent containing cis-3-hexenol as an active ingredient. [3] An anti-inflammatory agent containing cis-3-hexenol as an active ingredient. [4] A hyaluronic acid increaser containing cis-3-hexenol as an active ingredient. [5] A melanin production inhibitor containing cis-3-hexenol as an active ingredient. [6] A FLG gene expression promoter containing cis-3-hexenol as an active ingredient. [7] A GBA gene expression promoter containing cis-3-hexenol as an active ingredient. [8] A COX2 gene expression inhibitor containing cis-3-hexenol as an active ingredient. [9] A HAS2 gene expression promoter containing cis-3-hexenol as an active ingredient.

[10] An agent for suppressing MIF gene expression, comprising cis-3-hexenol as an active ingredient.

[0007] By directly applying cis-3-hexenol, it is expected that the effects of moisturizing the skin, improving the barrier function, anti-inflammatory, increasing hyaluronic acid, and suppressing melanin production can be expected.

[0008] Cis-3-hexenol (CAS number: 928-96-1) is an unsaturated alcohol that, together with its isomer trans-2-hexenol, is also known as green leaf alcohol, and is an aroma component contained in green tea and the like.

[0009] As mentioned above, Patent Document 1 describes that the aroma of cis-3-hexenol provides an anti-fatigue effect, but this effect is exerted by smelling the aroma, and in the examples, the effect is actually demonstrated when a subject inhales cis-3-hexenol. Patent Document 2 describes that cis-3-hexenol has the effect of inhibiting melanin production by melanoma cells. The present inventors have discovered that direct application of cis-3-hexenol to the skin exerts favorable effects on the skin, such as moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and inhibiting melanin production.

[0010] Therefore, the present invention provides a moisturizer, a skin barrier function improver, an anti-inflammatory agent, a hyaluronic acid increaser, and a melanin production inhibitor (hereinafter, these may be collectively referred to as the agent of the present invention) that contain cis-3-hexenol as an active ingredient.

[0011] Moisturizing, improvement of skin barrier function, anti-inflammation, increased hyaluronic acid, and suppression of melanin production can be confirmed, for example, by measuring increases in gene expression and protein production of FLG, GBA, HAS2, etc., and decreases in gene expression and protein production of COX2, MIF, etc., in biological samples such as keratinocytes and skin fibroblasts. Gene expression levels can be measured using techniques known in the art, such as quantitative PCR and Northern blotting. For example, probes for mRNA of FLG, GBA, COX2, HAS2, and / or MIF, etc., may be used. Protein levels can be measured using techniques known in the art, such as Western blotting, immunostaining, ICM, and ELISA. However, in addition to the above-mentioned methods, moisturizing, improvement of skin barrier function, anti-inflammation, increase in hyaluronic acid, and inhibition of melanin production can be confirmed by any method, such as visual inspection, TEWL, measurement of the amount of keratinized outer membrane, measurement of specific proteins and lipids, measurement of the amount of hyaluronic acid by ELISA in skin samples such as skin models or skin cells such as fibroblasts, etc.

[0012] The present invention also provides a FLG gene expression promoter, a GBA gene expression promoter, a COX2 gene expression inhibitor, a HAS2 gene expression promoter, and a MIF gene expression inhibitor, each containing cis-3-hexenol as an active ingredient. In one aspect, the FLG gene expression promoter promotes FLG gene expression in epidermal keratinocytes. In one aspect, the GBA gene expression promoter promotes GBA gene expression in epidermal keratinocytes. In one aspect, the COX2 gene expression inhibitor inhibits COX2 gene expression in epidermal keratinocytes. In one aspect, the HAS2 gene expression promoter promotes HAS2 gene expression in skin fibroblasts. In one aspect, the MIF gene expression inhibitor inhibits MIF gene expression in skin keratinocytes.

[0013] FLG (Filaggrin) is a gene encoding filaggrin, which is involved in the skin's moisture retention function as an NMF. GBA (β-Glucocerebrosidase) is a gene encoding an enzyme that produces ceramide EOP (ceramide 1), which is involved in the skin's barrier function. COX2 (Prostaglandin-Endoperoxide Synthase 2) is a gene encoding an enzyme that produces PGE2, an inflammatory mediator involved in melanin production. HAS2 (Hyaluronan Synthase 2) is a gene encoding a hyaluronic acid synthesis enzyme that is produced in dermal cells and is involved in the skin's moisture retention. MIF (Macrophage migration inhibitory factor) is a gene that encodes an enzyme that inhibits macrophage migration, inhibits the degradation of tyrosinase protein, and enhances melanin production.

[0014] Gene expression can be measured by techniques known in the art, such as quantitative PCR and Northern blotting, as described above. In one embodiment, promotion of gene expression refers to an increase with a statistically significant difference (e.g., Student's t-test, Dunnett's test) at a significance level of 5%, and / or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more. Inhibition of gene expression refers to a decrease with a statistically significant difference (e.g., Student's t-test, Dunnett's test) at a significance level of 5%, and / or a decrease of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100%.

[0015] Protein expression can be measured by techniques known in the art, such as Western blotting, immunostaining, ICM, and ELISA, as described above. In one embodiment, promotion of protein expression refers to an increase with a statistically significant difference (e.g., Student's t-test, Dunnett's test) at a significance level of 5%, and / or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more. Inhibition of protein expression refers to a decrease with a statistically significant difference (e.g., Student's t-test, Dunnett's test) at a significance level of 5%, and / or a decrease of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, or 100%.

[0016] Hyaluronic acid can be measured by techniques known in the art, such as ELISA, for example, sandwich ELISA, etc. In one embodiment, an increase in hyaluronic acid means an increase that is statistically significant at a significance level of 5% (e.g., Student's t-test, Dunnett's test), and / or an increase of, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more.

[0017] The biological sample may be a sample capable of measuring moisturizing, improving skin barrier function, anti-inflammatory, hyaluronic acid increase, and melanin production inhibitory effects, or a cell culture capable of measuring gene expression levels, such as FLG, GBA, COX2, HAS2, and / or MIF, protein production levels corresponding to these genes, and / or hyaluronic acid production levels. For example, keratinocytes or fibroblasts are preferably used. Alternatively, a skin sample or a three-dimensionally constructed cultured skin model may be used. The cells may be derived from any animal, but from the perspective of developing cosmetics and pharmaceuticals, human-derived cells are preferred.

[0018] The present invention also provides a method for moisturizing a subject, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and suppressing melanin production, which comprises applying cis-3-hexenol or the agent of the present invention or a composition containing same.

[0019] Examples of subjects to which the method of the present invention can be applied include subjects with dry skin or the like who need or desire to moisturize, subjects with reduced skin barrier function who need to improve it, subjects with inflammation who need to suppress it or desire to prevent it, subjects with a hyaluronic acid deficiency who desire to prevent it, and subjects with reduced gene expression levels or protein production levels of FLG, GBA, and / or HAS2, etc., or increased gene expression levels or protein production levels of COX2 and / or MIF, etc.

[0020] The method according to the present application is for cosmetic purposes and may exclude medical procedures performed by doctors or medical professionals. The method according to the present application may also be a method for supporting cosmetic procedures of a subject.

[0021] The present invention also provides a composition for moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and inhibiting melanin production, which comprises cis-3-hexenol or the agent of the present invention. The composition of the present invention may be a composition for moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and inhibiting melanin production. The composition may be a cosmetic, pharmaceutical, or quasi-drug.

[0022] The composition can be applied via any route, such as transdermal, oral, transmucosal, nasal, intravenous, intraarterial, or subcutaneous, but is preferably applied transdermally, i.e., to the skin, from the viewpoint of acting on the skin. The skin can be applied to any part of the body, such as the face, head, neck, limbs, or trunk.

[0023] In the case of cosmetics, the composition can be incorporated into facial or body cosmetics such as lotions, emulsions, serums, creams, lotions, packs, essences, and gels, makeup cosmetics such as foundations, makeup bases, and concealers, and even bath additives. In the case of transdermal pharmaceuticals, the composition can be formulated into topical skin preparations. The form is not particularly limited as long as it is applicable to the skin, and any dosage form can be applied, for example, a solution, emulsion, solid, semi-solid, powder, powder dispersion, water-oil two-layer separation, water-oil-powder three-layer separation, ointment, gel, aerosol, mousse, stick, etc. In addition, bases and excipients commonly used in cosmetics and topical skin preparations, such as preservatives, emulsifiers, and pH adjusters, may be used.

[0024] The amount of active ingredient in the agent or composition of the present invention can be selected as desired from the viewpoint of exerting moisturizing, improving skin barrier function, anti-inflammatory, and / or hyaluronic acid-increasing effects. For example, cis-3-hexenol can be blended at 0.0005 to 100.0 mM. From the viewpoint of fully exerting its effects, it can be blended preferably at 0.05 mM or more, for example, at 0.5 mM or more. On the other hand, since cis-3-hexenol has an aroma, from the viewpoint of avoiding an overly strong odor, it can be blended preferably at 10.0 mM or less, more preferably at 5.0 mM or less. The above components may be combined in any ratio, and in such cases, it is preferable that the total amount of those components is within the above range.

[0025] The present invention also provides cis-3-hexenol for moisturizing, improving skin barrier function, anti-inflammation, and / or increasing hyaluronic acid, wherein preferably, the moisturizing, improving skin barrier function, anti-inflammation, and / or increasing hyaluronic acid is mediated by promoting gene expression of FLG, GBA, and HAS2, and / or suppressing gene expression of COX2. In one aspect, the moisturizing is epidermal moisturizing. In one embodiment of this aspect, epidermal moisturizing is achieved by promoting gene expression or protein production of FLG, GBA in keratinocytes, and / or HAS2 in dermal fibroblasts. In one aspect, the improvement of skin barrier function is epidermal skin barrier function. In one embodiment of this aspect, the improvement of epidermal skin barrier function is achieved by promoting gene expression or protein production of GBA in keratinocytes. In one aspect, the anti-inflammation is epidermal anti-inflammation. In one embodiment of this aspect, anti-inflammatory effects in the epidermis are achieved by suppressing gene expression or protein production of COX2 in keratinocytes. In one aspect, the increase in hyaluronic acid is an increase in dermal hyaluronic acid. In one embodiment of this aspect, the increase in dermal hyaluronic acid is achieved by promoting gene expression or protein production of HAS2 in skin fibroblasts. In one aspect, the inhibition of melanogenesis is an inhibition of epidermal melanogenesis, and is achieved by suppressing gene expression or protein production of MIF and / or COX2 skin in keratinocytes.

[0026] Furthermore, the present invention also provides use of cis-3-hexenol in the manufacture of a medicament for moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and inhibiting melanin production.

[0027] All documents mentioned herein are incorporated by reference in their entirety.

[0028] The following examples of the present invention are for illustrative purposes only and do not limit the technical scope of the present invention. The technical scope of the present invention is limited only by the claims. The present invention may be modified, for example, by adding, deleting, or substituting components of the present invention, provided that the modifications do not depart from the spirit of the present invention.

[0029] To confirm the moisturizing, skin barrier function improvement, anti-inflammatory, hyaluronic acid-increasing, and melanin production-inhibiting effects of cis-3-hexenol, the expression of genes involved in these functions was analyzed. More specifically, the expression levels of FLG, GBA, COX2, HAS2, and MIF were measured using the methods described below.

[0030] Example 1: Effect of cis-3-hexenol on keratinocytes 1-1: Cell culture HaCaT cells were cultured in a 24-well plate (Cat No. 3526, Corning, USA) at a density of 10.0 × 10 ４ The cells were seeded at a density of 10.0% (v / v) fetal bovine serum (FBS, Cat No. SH30071.03, Hyclone, UK) and 1.0% (v / v) antifungal agent (Antibiotic-Antimycotic 100X, Cat No. 15240-062, Invitrogen, USA) in Dulbecco's Modified Eagle Medium (DMEM, Cat No. 043-30085, Wako, Japan) at a density of 10.0% (v / v) cells / well and cultured for 24 hours in a CO2 incubator (CO2 concentration 5%, 37°C). After removing the medium, the cells were replaced with medium supplemented with cis-3-hexenol (CAS: 928-96-1, Wako Pure Chemical Industries, Ltd.) to the final concentrations shown in the table below, and further cultured for 48 hours in a CO2 incubator. As a control, medium without cis-3-hexenol was used.

[0031] 1-2: RNA extraction and purification, quantification, and purity measurement. RNA was extracted and purified using PureLink™ RNA Mini Kit (Cat No. 12183018A, Invitrogen, USA). A portion of the purified RNA was placed in a UV-transparent 96-well plate and diluted 10-fold with Tris-EDTA buffer. The absorbance at 230 nm, 260 nm, and 280 nm (OD230, OD260, OD280) was measured using a microplate reader (SPARK® 10M TECAN, Switzerland). The RNA concentration was calculated using OD260, and the RNA concentration was adjusted to 10 μg / mL by dilution with TE buffer.

[0032] 1-3: Gene Expression Analysis by Real-Time PCR RNA was reverse-transcribed using SuperScript™ IV VILO™ Master Mix with ezDNase (Cat No. 11766050, Invitrogen, USA). 4 μL of SuperScript™ IV VILO™ Master Mix and 6 μL of nuclease-free water were added per well of an 8-tube array. cDNA was synthesized using a real-time PCR system (QuantStudio™ 3, Applied Biosystems, USA) by heating at 25°C for 10 minutes, 50°C for 10 minutes, and 85°C for 5 minutes. Ten microliters of TaqMan® Fast Advanced Master Mix (Cat No. 4444557, Applied Biosystems, USA), 1 μL of TaqMan Gene Expressor, 7 μL of UltraPure™ Distilled Water (Invitrogen, Cat No. 10977-015, USA), and 2 μL of cDNA were added per well to a PCR plate, and the plate was sealed. Real-time qPCR was performed using a primer for FLG (Hs00856927_g1), a primer for GBA (Hs00986836_g1), a primer for COX2 (Hs00153133_m1), or a primer for MIF (Macrophage migration inhibitory factor) (Hs00236988_g1), and a primer for GAPDH (Hs02786624_g1) as an internal standard gene, to measure the threshold of each gene, such as FLG, in a cis-3-hexenol-supplemented medium. After calculating the cycle (Ct) value, the Ct value was corrected using GAPDH to obtain a ΔCt value. Assuming that the gene amount doubles per cycle, the gene expression level in the cis-3-hexenol-supplemented medium was calculated, assuming that the gene expression level in the control was 1. For gene expression analysis, the average value of 3 wells of a 24-well plate was used for each treatment group.

[0033] The results are shown in Table 1 below.

[0034] As shown in Tables 1 to 4, the addition of cis-3-hexenol significantly increased the expression of FLG and GBA and significantly decreased the expression of COX2 and MIF. These results suggest that promoting the expression of FLG and GBA and suppressing the expression of COX2 and MIF may contribute to moisturizing, improving skin barrier function, anti-inflammation, increasing hyaluronic acid, and / or suppressing melanin production.

[0035] Example 2: Effect of cis-3-hexenol on fibroblasts In this example, cis-3-hexenol, which was found to be effective in keratinocytes in Example 1, was used. In place of keratinocytes, human neonatal dermal fibroblast cell line NB1RGB cells (RIKEN BRC, Japan) were used in this example, and the expression level of the HAS2 gene was measured using the same materials and method as in Example 1, except that a primer (Hs00193435_m1) for HAS2 (Human Hyaluronan Synthase 2), which encodes a hyaluronic acid synthase, was used.

[0036] The results are shown in Table 5 below.

[0037] The results in Table 5 indicate that cis-3-hexenol significantly increased HAS2 expression in dermal fibroblasts. These results suggest that cis-3-hexenol promotes the synthesis of dermal hyaluronic acid by promoting HAS2 expression, which may contribute to moisturizing.

[0038] Example 3: Effect of cis-3-hexenol on hyaluronic acid content Cis-3-hexenol, which was found to have an effect of promoting HAS2 gene expression in dermal fibroblasts in Example 2, was examined in this Example to see whether it also had the effect of increasing hyaluronic acid content in dermal fibroblasts. 3-1: Cell culture The same NB1RGB cells as in Example 2 were cultured in a 24-well plate (Cat No. 3526, Corning, USA) at 10.0 x 10 ４The cells were seeded at a density of 10.0% (v / v) fetal bovine serum (FBS, Cat No. SH30071.03, Hyclone, UK) and 1.0% (v / v) antifungal agent (Antibiotic-Antimycotic 100X, Cat No. 15240-062, Invitrogen, USA) in Eagle's Minimal Essential Medium (EMEM, Cat No. 051-07615, Wako, Japan) at a density of 10.0% (v / v) cells / well and cultured for 24 hours in a CO2 incubator (CO2 concentration 5%, 37°C). After 24 hours, the medium was removed and replaced with medium supplemented with cis-3-hexenol (CAS: 928-96-1, Wako Pure Chemical Industries, Ltd.) to the final concentrations shown in Table 6 below, and the cells were further cultured for 48 hours in a CO2 incubator. As a control, medium without cis-3-hexenol was used.

[0039] 3-2: Evaluation of Cell Activation Effect The effect of cis-3-hexenol on the proliferation of dermal fibroblasts was evaluated using the following method. The medium was removed from a 96-well plate, which was then washed with 100 μL of PBS(-). Then, 100 μL of 0.5 mg / mL 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, CAS No. 298-93-1, Sigma-Aldrich, USA) solution was added, and the plate was incubated for 2 hours in a CO2 incubator. After removing the MTT solution and washing with 100 μL of PBS(-), 200 μL of 2-propanol (CAS No. 67-63-0, Wako, Japan) was added to dissolve the insoluble formazan. After uniformly dispersing the dye in the 96-well plate, the absorbance at 570 nm (OD570) was measured using a microplate reader (SPARK (registered trademark) 10M, TECAN, Switzerland). The OD570 of the control group was set at 100%, and the OD570 of the cis-3-hexenol-added group, i.e., the effect of cell proliferation, was calculated as the cell activation effect (%) of cis-3-hexenol. The average value of 3 wells of a 96-well plate was used for each treatment group to analyze the cell activation effect.

[0040] 3-3: Measurement of hyaluronic acid The supernatant of the culture cultured in 3-1 was dispensed into a new 96-well plate and frozen (-80 ° C), and the amount of hyaluronic acid in this culture supernatant was measured by sandwich ELISA, and the hyaluronic acid production promoting effect of cis-3-hexenol was evaluated. 100 μL of Hyaluronan Binding Protein (HABP, Cat No. BC40, Hokudo, Japan, 1:5500) solution prepared with PBS was added to a highly adsorbent 96-well plate and incubated overnight at 4 ° C. The solidified HABP solution was removed, washed with 200 μL of PBS-T solution, and then 150 μL of 1% BSA solution was added and incubated at room temperature for 1 hour. The BSA solution was removed, and the cells were washed with 200 μL of PBS-T. Then, 100 μL of culture supernatant diluted 100-fold with PBS(-) was added and incubated at room temperature for 1 hour. Sodium hyaluronate (Cat No. 087-04511, Wako, Japan) was used as a standard. The culture supernatant was removed, and the cells were washed with 200 μL of PBS-T. Then, 100 μL of biotin-labeled HABP (Cat No. BC41, Hokudo, Japan, 1:2000) solution prepared in 0.5% BSA-containing PBS(-) was added, and the cells were left to stand overnight at 4°C. The biotin-labeled HABP solution was removed, and the plate was washed with 200 μL of PBS-T. 100 μL of Streptavidin-HRP solution (1:10,000) prepared in 0.5% BSA-containing PBS(-) was added, and the plate was left to stand at room temperature for 30 minutes. The Streptavidin-HRP solution was removed, and after washing with 200 μL of PBS-T, 100 μL of ABTS solution was added, and color development was confirmed. After homogenizing the dye in the 96-well plate, the absorbance at 405 nm (OD405) was measured using a microplate reader. The hyaluronic acid production rate of cis-3-hexenol was calculated, with the OD405 of the control group set at 100%. The OD405 of the control and cis-3-hexenol-added samples was divided by the OD570 measured in 3-2 to calculate the hyaluronic acid production rate per cell. The hyaluronic acid production rate per cell due to cis-3-hexenol was calculated by setting the hyaluronic acid production rate per cell in the control group at 100%. The average value of 3 wells of a 96-well plate was used for the analysis of hyaluronic acid production rate per treatment group.

[0041] The results of 3-2 showed that there was no significant increase or decrease in fibroblast proliferation, confirming that cis-3-hexenol had no cell-activating effect or toxicity. The results of 3-3 are shown in Table 6 as the hyaluronic acid production rate by cis-3-hexenol, when the hyaluronic acid production rate of the control was set at 100%.

[0042] The addition of cis-3-hexenol significantly increased the hyaluronic acid production rate per cell. These results confirmed that cis-3-hexenol contributes to the increase in hyaluronic acid in fibroblasts. Furthermore, taking into account the results of Example 2, it is suggested that the hyaluronic acid-increasing effect of cis-3-hexenol may be due to the promotion of HAS2 expression.

[0043] The above results indicate that cis-3-hexenol promotes gene expression of FLG, GBA, and HAS2, suppresses gene expression of COX2 and MIF, and increases hyaluronic acid. Furthermore, it is expected to have favorable effects on the skin, such as moisturizing, improving skin barrier function, anti-inflammation, and suppressing melanin production.

Claims

1. A hyaluronic acid increaser containing cis-3-hexenol as an active ingredient.

2. A moisturizer containing cis-3-hexenol as an active ingredient.

3. A skin barrier function improver containing cis-3-hexenol as an active ingredient.

4. An anti-inflammatory agent containing cis-3-hexenol as an active ingredient.

5. A FLG gene expression promoter containing cis-3-hexenol as an active ingredient.

6. A GBA gene expression promoter containing cis-3-hexenol as an active ingredient.

7. A COX2 gene expression inhibitor containing cis-3-hexenol as an active ingredient.

8. A HAS2 gene expression promoter containing cis-3-hexenol as an active ingredient.