KR20110110069A - A composition containing lindera obtusiloba leaf extract for skin whitening and wrinkle treatment - Google Patents

Abstract

The present invention provides a cosmetic composition or functional food composition for skin whitening and anti-wrinkle containing ginger tree leaf extract. The whitening and anti-wrinkle effect of the present invention is to reduce melanin content, inhibit the expression of protein involved in melanogenesis, inhibit melanocyte formation in melanocytes of melanocytes induced pigmentation by UVB, melanin pigment distributed in the epidermis It is confirmed by the action of reducing the thickness of the epidermis and reducing the thickness of the epidermis. Therefore, the composition comprising the ginger tree leaf extract according to the present invention can be used as a cosmetic composition or a functional food composition having no side effects and exhibiting skin whitening effect or skin wrinkle improvement effect of excellent efficacy.

Description

A composition containing Lindera obtusiloba leaf extract for skin whitening and wrinkle treatment}

The present invention relates to a composition for skin whitening, and more particularly to a composition for skin whitening and / or anti-wrinkle comprising ginger tree leaf extract.

Substances known to be effective in skin whitening include vitamin C, lettuce extract, kojic acid, arbutin, licorice extract, and hydroquinone. Koji acid except been identified as a carcinogen, and experimental use is prohibited as a cosmetic ingredient, hydroquinone are toxic to melanocytes (melanocyte), in vivo (in As a result of the in vivo test, there is a risk of permanent discoloration, and it is also known as a carcinogen, and it is prohibited to import as a cosmetic ingredient in Korea. Known chemicals as described above, although there is a whitening effect, it is accompanied by a problem of toxicity or side effects on the skin, and its use is limited. Therefore, in recent years, researches are being actively conducted to develop skin whitening ingredients derived from natural products which can minimize side effects on the skin.

Ginger tree ( Lindera obtusiloba BL.) is a deciduous shrub of the dicotyledonous plant of the genus Trifoliaceae, which is distributed in Korea, Japan and China, and its flowers are used for ornamental purposes, and its branches, leaves, and fruits are used for food. The branches and bark are also used for medicinal purposes, and are known to be effective for bruising fish and postpartum swelling and limb pain. However, the effects associated with skin whitening or wrinkle improvement of ginger tree have not been studied or known to date.

The present inventors have conducted research to develop a material from natural products that exhibits excellent skin whitening and wrinkle improvement effects with less side effects and ensures safety, and as a result, the leaves of ginger trees, which have been in large quantities throughout Korea but have not been widely utilized, are used as raw materials. From the extract to make excellent skin whitening and wrinkle improvement efficacy was completed the present invention.

It is an object of the present invention to provide a composition for skin whitening and / or wrinkle improvement that exhibits excellent efficacy without toxicity and side effects.

The object of the present invention can be achieved by providing in the form of a cosmetic composition or functional food composition or pharmaceutical composition for skin whitening and / or anti-wrinkle including ginger tree leaf extract.

The composition comprising the ginger tree leaf extract according to the present invention can solve the problem of toxicity or side effects on the skin by using natural products as well as exhibits excellent skin whitening and wrinkle improvement effect. The whitening and anti-wrinkle effect of the present invention is to reduce melanin content, inhibit the expression of protein involved in melanogenesis, inhibit melanocyte formation in melanocytes of melanocytes induced pigmentation by UVB, melanin pigment distributed in the epidermis It is confirmed by the action of reducing the thickness of the epidermis and reducing the thickness of the epidermis. Therefore, the composition comprising the ginger tree leaf extract according to the present invention can be used as a cosmetic composition or a functional food composition having no side effects and exhibiting skin whitening effect or skin wrinkle improvement effect of excellent efficacy.

Figure 1 shows the results of confirming the melanin content after treatment with ginger leaf extract 1, 5, 10, 50 ㎍ / ㎖ in B16 melanoma F10 cell line for 72 hours.
Figure 2 shows the results of confirming the expression of proteins involved in melanin synthesis through Western blotting technique after treatment for 10 hours, 50 ㎍ / ㎖ ginger leaf extract for 72 hours to B16 melanoma F10 cell line.
Figure 3 is a photograph showing the melanocyte activity in the ear tissues of experimental animals every 3, 6, 9 weeks after oral administration of ginger tree leaf extract to the experimental animals through the waveguide staining method.
Figure 4 is a photograph showing the melanin distribution in the ear tissues of experimental animals every 3, 6, 9 weeks after oral administration of ginger tree leaf extract to the experimental animals through Fontana Mason staining method.

The present invention provides a skin whitening and / or anti-wrinkle composition comprising a ginger tree leaf extract, and more specifically provides a cosmetic composition or functional food composition comprising a ginger tree leaf extract.

In the present invention, the ginger leaf extract is an extract from a polar solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof, and preferably means an extract from 70% ethanol. .

In order to obtain the ginger tree leaf extract using an extraction method commonly used in the art, for example, 20 to 100 ℃ as a solvent of 5-20 times the weight of the ginger tree leaf, preferably 10-20 times the raw material, Preferably, extraction is repeated once to three times for 1 hour to 2 days, preferably 2 hours to 1 day, at an extraction temperature of 65 to 85 ° C., and methods such as reflux extraction and ultrasonic extraction may be applied.

Ginger leaf extract according to the present invention, preferably contained in 0.001 to 50% by weight relative to the total weight of the composition.

Decreased melanin content and inhibition of expression of proteins involved in melanogenesis were observed when the composition of the present invention was treated to melanoma cell lines, and the composition of the present invention was orally administered to mice induced with pigmentation by UVB. Inhibition of melanoma production at the site, reduction of melanin pigments distributed in the epidermis, and reduction of the thickness of the epidermis were confirmed. Therefore, the composition comprising the ginger tree leaf extract according to the present invention may exhibit an excellent skin whitening and wrinkle improvement effect as a cosmetic composition or a functional food composition to be administered orally directly applied to the skin.

When the composition of the present invention is a cosmetic composition, the components included in the cosmetic composition include components commonly used in cosmetic compositions in addition to ginger leaf extract, for example, antioxidants, stabilizers, solvents, vitamins, pigments and flavorings. Conventional adjuvants such as, and carriers. In addition, the cosmetic composition may further include a skin absorption enhancer to enhance the effect.

Cosmetic compositions of the present invention can be prepared in any formulation conventionally prepared in the art, for example solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, oils, powder foundations, emulsions It may be formulated as a liquid foundation, wax foundation, spray or the like. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

Ginger leaf extract according to the present invention may be used in an amount of 0.001 to 5% by weight based on the total weight of the cosmetic, but is not limited thereto.

In addition, the composition of the present invention may be a functional food. Ingredients included in the functional food composition of the present invention includes additives commonly used in foods in addition to ginger leaf extract, for example, flavoring, acidulant, preservatives, sweeteners, antioxidants, stabilizers, colorants and nutrition enhancers, etc. It may include.

Of course, the compositions according to the invention may also be formulated in the form of drugs or pills, such as injections.

Ginger leaf extract according to the present invention can be used in an amount of 1 to 20% by weight based on the total weight of the food, but is not limited thereto.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples, which are merely illustrative of the present invention and the present invention is not limited thereto.

Example  1: Preparation of Ginger Extract

The ginger leaves were washed and dried to freeze at -80 ℃ and then lyophilized to powder. 70% ethanol was added to 100 g of the leaf in the powder state 10 times the weight of the sample and extracted three times at 80 ℃ three times. The extract was concentrated under reduced pressure at 40 ° C. and freeze dried to obtain a powder. The yield was 26.8% and the powder obtained was used for the following experiment.

However, the preparation method of the ginger tree extract described in Example 1 is only one example, and conditions such as concentration and temperature and time of ethanol may be changed, and may be based on various other known extraction methods. However, if the manufacturing conditions of the ginger tree extract is changed, the yield may vary slightly.

Experimental Example  One : Ex vivo ( In vitro ) In  Melanin content measurement

In order to confirm the melanin production inhibitory effect of the ginger leaf extract according to the present invention, the change of melanin content was measured by an in vitro experiment with B16 melanoma F10 cell line.

The measuring method is as follows. When the B16 melanoma F10 cell line was dispensed at 1 x 10 ⁴ cells per well into 24 well-plates and adhesion was confirmed, the ginger leaf extract obtained in Example 1 was 1, 5, 10, 50 μg / ml. Each concentration was treated with DMEM (Dulcco's Modifeid Eagle Medium) and incubated for 72 hours at 37 ℃, 5% CO ₂ incubator. After 72 hours, the cells in the wells were washed twice with PBS and then lysed with 1 N NaOH. The melanin amount was measured at an absorbance of 470 nm.

As a result, as shown in Figure 1, the total melanin content of the B16 melanoma F10 cell line was reduced by more than 10% in the pre-treated group of ginger leaf extract. In particular, the amount of melanin was decreased by 22% in the 50 ㎍ / ㎖ ginger leaf extract treatment group. This is the result of a significant decrease in the total amount of melanin in the cells in the range that does not inhibit the proliferation of the cells. As a result, the ginger leaf extract according to the present invention was confirmed to exhibit a whitening effect by reducing melanin.

Experimental Example  2: measurement of protein expression involved in melanin synthesis

In order to confirm the melanin production inhibitory effect of the ginger leaf extract according to the present invention, the protein involved in melanogenesis, ERK (Extracelluar signal-regulated kinase), MITF (Microphthalmia-associated transcription factor), and tyrosinase (tyrosinase) Protein expression was measured in the subject.

ERK is an important signaling mediator involved in cell proliferation and differentiation, ERK signaling pathway is involved in melanogenesis and inhibition of ERK signaling pathway causes hyperpigmentation. Known. Microphthalmia-associated transcription factor (MITF) is involved in pigmentation, proliferation, and survival of melanocytes. MITF is also known to be an important protein strongly involved in tyrosinase and tyrosinase relate protein-1 (TRP-1). Tyrosinase is well known as the most important enzyme that is directly involved in melanin synthesis.

The measuring method is as follows. When the B16 melanoma F10 cell line was dispensed in 24 well-plates at a cell number of 1 X 10 ⁴ per well, and then adhesion was confirmed, the ginger leaf extract obtained in Example 1 was treated at a concentration of 10 and 50 ㎍ / ml. Incubated with DMEM (Dulcco's Modifeid Eagle Medium) for 72 hours in a 37 ° C., 5% CO ₂ incubator. After 72 hours, the cells in the wells were washed twice with PBS, followed by lysis buffer [composition: 62.5 mM Tris-HCl (pH 6.8), 2% SDS, 5% β-mercaptoethanol, protease inhibitors (Complete ^™ , Roche, Mannheim, Germany), 2 mM phenylmethylsulfonylfluoride, 10 mM EDTA, 50 mM NaF, and 1 mM Na ₃ VO ₄ ] to collect the cells and extract proteins.

Each 10 μg of protein was electrophoresed using SDS-polyacrylamide gel and then transferred to the PVDF membrane. The membrane was blocked with 5% dried milk and then refrigerated overnight with 1: 1000 primary antibody dilution with 5% BSA. Development was performed using a Chemiluminescence Plus Kit (Amersham International, Little Chalfont, UK).

As a result, as shown in Figure 2, the ginger leaf extract increased the expression of ERK, and inhibited the expression of MITF and tyrosinase. Therefore, the ginger leaf extract according to the present invention was confirmed to exhibit a whitening effect by reducing melanin.

Experimental Example  3: Wave  Through staining Melanosite  Active observation

In order to confirm the inhibitory effect of melanin production by oral administration of the ginger leaf extract according to the present invention, induction of pigmentation in the experimental animal by UVB irradiation and oral administration of the ginger tree leaf extract and the activity of melanocytes by dopa staining method Was observed.

For the measurement, the experimental animals were treated as follows. C57BL / 6 females (18 g, 8 weeks old) were used as experimental animals and were purchased from KOATEK (KOATEK, Incheon, Korea) for 1 week, then divided into non-UVB and UVB groups for 10 days in the morning. After irradiating UVB at 100 mJ / cm ² , pigmentation was induced, and as shown in Table 1, the non-UVB group (n = 15 in each group), only-UVB group, and ascorbic acid (70 mg / kg) ) Oral administration group, ginger tree leaf extract obtained in Example 1 orally administered three groups (40 mg / kg, 80 mg / kg, 120 mg / kg) were divided into 9 weeks breeding.

Experimental group Population n = 15 Non-UVB 20% propylene glycol Control group (only-UVB) 20% propylene glycol Positive control group (ascorbic acid administration group) 70 mg / kg
Experimental group (ginger tree leaf extract administration group)
40 mg / kg 80 mg / kg 120 mg / kg

FIG. 3 is a photograph showing melanocyte activity in dorsal tissues by dopa-staining at the expense of 5 mice per experimental group every 3, 6 and 9 weeks of oral administration. Dopa staining is an immunological tissue staining method characterized by knowing the number of melanocytes in the epidermis. Irradiation at UVB 100 mJ / cm ² for 10 days may lead to pigmentation. As a result, the number of cells after UVB irradiation increased 3-4 times, the size was larger than before irradiation, and the dendrites developed significantly. Up to 20 weeks after discontinuation of UVB irradiation, the levels remained higher than before. Oral administration of Ginger leaf extract in different concentrations affects the proliferation of melanocytes, indicating a melanin formation inhibitory effect. Ginger leaf extract 40 mg / kg inhibitory effect was observed to be weak compared to other concentrations, compared with only UVB irradiation group was confirmed that the number and size of melanocytes was reduced. Ginger leaf extract 80 mg / kg and 120 mg / kg showed a similar pattern, and ascorbic acid 70 mg / kg was found to have a melanin pigment inhibitory effect is similar or slightly better. Therefore, the ginger leaf extract according to the present invention was confirmed to exhibit a whitening effect by reducing melanin even by oral administration.

Experimental Example  4: Observation of the distribution of melanin pigment by special staining method

In order to confirm the inhibitory effect of melanin production by oral administration of the ginger leaf extract according to the present invention, induction of pigmentation in the experimental animal by UVB irradiation and oral administration of ginger tree leaf extract Fontana Mason silver (Fontana-Masson silver) ) Melanocyte distribution in the entire epidermal layer was observed by staining.

Treatment of experimental animals and experimental groups were the same as in Example 3.

Figure 4 is a photograph showing the distribution of melanin pigment in the ear tissue by Fontana Masson silver staining at the expense of 5 per experimental group every 3, 6, 9 weeks oral administration. Fontana Masson silver staining is an immunological tissue staining method that can identify melanin pigments distributed throughout the epidermal layer. As a result, only UVB irradiation group showed melanin pigment distribution throughout the epidermal layer. However, when ascorbic acid or ginger leaf extract was administered to 3/6/9 weeks, the distribution of melanin was found only in a part of the epidermis, and the positive dose group of 80 mg / kg and 120 mg / kg of ginger leaf extract was positive. Compared with the control group 70 mg / kg ascorbic acid, the melanin pigment in the epidermal layer was similar or slightly less. Therefore, the ginger leaf extract according to the present invention was confirmed to exhibit a whitening effect by reducing melanin even by oral administration.

On the other hand, it was also possible to observe the thickness change of the epidermis through the Fontana and Mason silver staining method. As shown in FIG. 4, it was found that the epidermal layer was thickened by the ultraviolet rays irradiated. However, oral administration of Ginger leaf extract could identify the epidermal layer thinning over time. The thickening of the epidermal layer is due to premature aging due to photoaging. That is, as ultraviolet rays accumulate on the body, the skin layer becomes thicker, melanocytes increase, collagen formation decreases, elastin fibers harden, wrinkles appear on the skin, and elastic fiber degeneration symptoms appear. It was confirmed that this photoaging can be prevented by administering the ginger leaf extract, and thus, the ginger leaf extract according to the present invention showed an improvement in skin wrinkles.

Claims (7)

Skin whitening composition comprising a ginger tree leaf extract. The composition of claim 1, wherein the composition is a cosmetic composition. The composition of claim 1, wherein the composition is a functional food composition. The composition of claim 1, wherein the ginger tree leaf extract is extracted using water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof as a solvent. The composition of claim 4, wherein the ginger leaf extract is extracted using 70% ethanol as a solvent. The composition of claim 1, wherein the ginger leaf extract is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. The composition of claim 1, wherein the composition comprising the ginger tree leaf extract is a composition for skin whitening and wrinkle improvement.

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