KR20100023013A - Composition of skin external application containing ginseng berry extracts - Google Patents

Abstract

PURPOSE: A skin external composition containing Panax ginseng fruit extract is provided to promote collagen synthesis and ensure effects of anti-oxidation and DNA protection. CONSTITUTION: A skin external composition for anti-aging contains 0.001-50 weight% of Panax ginseng fruits extract as an active ingredient. The extract is a water or ethanol extract. The extract is produced by drying the fruits of Panax ginseng by hot air or light and extracting with water or ethanol. The composition is used in the form of skin softener, astringent, nutrition lotion, nutrition cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, body lotion, body cream, body oil or body essence.

Description

Composition of skin external application containing ginseng berry extracts}

The present invention relates to a composition for external application for skin containing ginseng fruit extract, and more specifically, to promote collagen production in the skin by containing ginseng fruit extract having a unique ingredient and composition as the active ingredient in the ground portion of ginseng, MMP-1 inhibition The present invention relates to a skin external composition having an effect and at the same time having an anti-aging and anti-wrinkle effect through an antioxidant effect and a DNA damage protection effect. In addition, the external composition for skin according to the present invention exhibits a skin whitening effect through the effect of inhibiting melanogenesis and improving pigmentation by ultraviolet rays, and induces and maintains the differentiation of keratinocytes of the skin to maintain skin dryness and atopic symptoms. It was able to improve sebum secretion and to improve acne and skin trouble through anti-inflammatory effect. It improves blood circulation through improving peripheral blood circulation and makes skin clear and transparent.

Ginseng (Panax ginseng C.A. Meyer) is a plant belonging to the genus Ogapi ginseng and has been used in Korea, China, and Japan for more than 2,000 years, and has been used to prevent disease and extend life. Known efficacy and effects of ginseng to the central nervous system, anticarcinogenic effect, anticancer activity, immune function regulation, antidiabetic effect, liver function anti-inflammatory effect, cardiovascular disorders improvement, anti-arteriosclerosis, blood pressure control action, menopausal Disability improvement, effects on osteoporosis, antistress and anti-fatigue, antioxidant activity and anti-aging effects (Korea Ginseng 'Ingredients and Efficacy', Korea Ginseng and Tobacco Research Institute, 56-112, 1996).

Ginsenoside, a representative physiologically active ingredient of ginseng, is distributed evenly in the ground and underground parts of ginseng. Especially, the composition and composition of ginsenosides are different depending on the parts such as ginseng root (root), ginseng leaf and ginseng fruit. (Attele AS et al, Biochem Pharmacol, 58; 1685-1693, 1999). In particular, ginseng fruit has been reported to show better results than ginseng root in antidiabetic efficacy based on the components and contents of ginseng root (root) and other ginsenosides (Dey L. et al., Phytomedicine, 10; 600; -605, 2003).

Recently, as consumers' interest in natural cosmetics has increased and many cosmetics using herbal ingredients have been released, ginseng is also being studied as a plant material having important skin effects of cosmetics, but most of them are extracts or ginseng roots of ginseng root. Only the side ingredients and ginseng polysaccharides are used to inhibit skin aging and wrinkle improvement by ginseng fruit components.

In addition, ginseng fruit is considered more precious than ginseng since ancient times, and has been selected and harvested for the purpose of obtaining seeds. Ginseng fruit is harvested only once in 4 years of ginseng cultivation period. It is difficult to produce good seedlings because seed is too small when harvested in 3 years old. Seeds are faithful and good when grown in 5 years old or older. In addition to inhibiting development, the tissue is not dense, so red ginseng quality may be greatly reduced when manufacturing red ginseng. In addition, the number and frequency of red ginseng deterioration is greatly reduced when the number of crops is collected more than two times during the cultivation period.

In US Patent No. 6,524,626, ginseng fruit juice is used in a cosmetic composition by freeze-drying, it is described that by using a combination of ginseng fruit juice and other natural products they exhibit moisturizing and skin softening effect, which is according to the present invention It is clearly distinguished from cosmetic use as an anti-aging effect and an antiwrinkle effect exhibited by the extract.

Human skin is changed by a number of internal and external factors as it ages. That is, internally, the secretion of various hormones that regulate metabolism decreases, and the function of immune cells and the activity of cells decreases, thereby reducing the biosynthesis of immune proteins and constituent proteins necessary for living organisms. Due to the increase in the amount of ultraviolet rays that reach the surface of the sun's rays and intensifying environmental pollution, free radicals and free radicals increase, thereby reducing the thickness of the skin, increasing wrinkles, reducing elasticity, The color of the skin becomes dull, skin problems frequently occur, and there are various changes such as blemishes, freckles, and black mushrooms.

As aging progresses, symptoms such as changes and decreases in the content and arrangement of collagen, elastin, hyaluronic acid and glycoproteins, which are constituents of the skin, appear and are subject to oxidative stress caused by free radicals and active harmful oxygen. In addition, cyclooxygenase-2 (Cox-2, cyclooxygenase), an enzyme that produces proinflammatory cytokine, which is known to cause inflammation, in most cells constituting the skin due to aging or ultraviolet rays. Biosynthesis of the enzyme increases the biosynthesis of matrix metalloproteinase (MMP), an enzyme that breaks down skin tissues by these inflammatory factors, and the production of nitric oxide (NO) by iNOS (inducible nitric oxide synthase). It is known to increase. In other words, the biosynthesis of the substrate material is reduced by the reduction of cellular activity and micro-inflammation due to naturally occurring endogenous aging, and by external factors such as the increase of stress caused by various harmful environments and the increase of free radical species caused by sunlight. As a result, decomposition and degeneration are accelerated, and the skin matrix is destroyed and thinned, and various symptoms of skin aging appear. Therefore, a lot of research is being conducted on the active ingredient that can prevent and improve the phenomenon of aging.

There are many factors involved in determining the skin color of a person, including the activity of melanocytes (melanocytes) that make melanin pigment, the distribution of blood vessels, the thickness of the skin and the presence or absence of pigments in the body such as carotenoids and bilirubin. Are important.

Especially, the most important factor is a black pigment called melanin which is produced by the action of various enzymes such as tyrosinase in melanocytes in the human body. The formation of this melanin pigment is influenced by genetic factors, physiological factors related to hormone secretion, stress, and environmental factors such as ultraviolet irradiation.

Melanin pigment, which is produced from melanocytes of the skin of the body, is a phenolic polymer material having a complex form of black pigment and protein. It protects skin organs below the dermis by blocking ultraviolet rays from the sun and free radicals generated in the skin body. It plays a useful role in protecting proteins and genes in the skin.

As such, melanin produced by stress stimulation inside and outside the skin is a stable substance that does not disappear until it is released to the outside through skin keratinization even if the stress disappears. However, if the melanin is produced more than necessary to cause hyperpigmentation, such as blemishes, freckles, and spots, cosmetically bad results.

Today, Asian women prefer white and clean skin like white jade, and this is an important standard of beauty, increasing the demand for treatment and cosmetic problems for hyperpigmentation.

In response to this demand, ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione or derivatives thereof, or substances having tyrosinase inhibitory activity have been used in combination with cosmetics or pharmaceuticals, but they are insufficient. Its use has been limited due to its whitening effect, safety issues on the skin, formulation and stability problems when formulated in cosmetics, and the like.

The most important function of the outermost epidermis of the skin is to protect the skin and protect against various stimuli (physical and chemical stimulating factors such as chemicals, air pollutants, dry environment and ultraviolet rays) from the outside. It is a protective function that prevents excessive divergence of moisture through the body, and this protective function can be maintained only when the stratum corneum composed of keratinocytes is normally formed. The outermost stratum corneum (horney layer) of the epidermis is formed from keratinocytes and consists of the differentiated keratinocytes and the surrounding lipid layer (J. Invest. Dermatol. 1983; 80: 44- 49). Keratinocytes are characteristic cells whose basal cells, which continuously proliferate in the epidermal layer, undergo morphological and functional changes in stages and rise to the surface of the skin. Exfoliated and new keratinocytes take over their functions. This repetitive sequence of changes is called "epidermal differentiation" or "keratinization." During keratinization, keratinocytes form the stratum corneum by generating natural moisturizing factor (NMF) and intercellular fat (ceramide, cholesterol, fatty acids), making the stratum corneum firm and flexible. It has a function as).

However, the stratum corneum can easily lose its function due to excessive washing, lifestyle factors such as bathing, environmental factors such as dry air and pollutants, and endogenous diseases such as atopic or aged skin. In recent years, more and more people are complaining of dry skin symptoms and all kinds of obstacles due to various factors. Therefore, many studies have been conducted to supply moisture from the outside or to prevent water loss from the body in order to maintain proper skin moisture, and various types of moisturizers having moisture retention capabilities have been developed and mainly cosmetics. It is widely used in the field.

However, as the risk factors for humans increase and the aged population rapidly increases in the living environment, the turnover rate of the stratum corneum is slowed, the lipid synthesis ability of the keratinocytes is degraded, or the epidermis is normal in the epidermis. Due to poor cell division, maturation and differentiation, the amount of moisturizing factors and lipids in the stratum corneum is reduced, so that humans with skin with a condition that is unable to maintain normal stratum corneum function, that is, the skin barrier function is deteriorated There is a growing trend.

Due to the abnormal division and differentiation of epidermal cells, the skin causes various skin diseases such as dry skin, atopy and psoriasis, and these diseases can alleviate the symptoms slightly with a conventional moisturizer having only water retention function. However, it is difficult to expect radical healing.

In general, the sebum in the skin including the scalp and face plays a role in maintaining the moisturizing of the skin and preventing the invasion of microorganisms, but excessive secretion of sebum promotes hair loss, worsens acne, promotes the enlargement of pores by acne, and seborrheic dermatitis. There are a number of diseases that can occur.

These secretions are caused by a number of causes, the most important of which is the sebaceous gland cells are activated by the amount of dihydrotestosterone (DHT), one of the hormones involved in promoting sebum secretion in sebaceous gland activity. Is oversecreted. That is, in the case of hair loss, testosterone (T) is converted into dihydrotestosterone, one of the male hormones, by the 5α-reductase type 2 intracellularly and at the same time the receptor protein in the cytoplasm. (receptor) enters the fluid and causes hair loss, but in the skin or sebaceous glands, testosterone is converted to dihydrotestosterone by 5-a-reductase type 1, which activates sebaceous gland cells to differentiate Promoting it leads to excessive secretion of sebum in the sebaceous glands, causing acne (J. Invest Dermatol 105: 209-214 Diane etc).

Skin problems, such as acne and hair loss that occur outside of excessive sebum secretion, are further exacerbated by the skin's fine inflammatory response. If you look at the development of acne, excessive sebum accumulates in the hair follicles, which activates the acne bacteria and causes inflammation.

Acne due to sebum hypersecretion by inhibiting the production of prostaglandins, especially the production of prostaglandin E2 (PGE2) in the cellular inflammatory response, and the production of NO (nitric oxide), a pro-inflammatory factor by iNOS It can alleviate skin problems such as

However, as a natural material safe for skin, there are not many raw materials that reduce sebum secretion through a fundamental cause and at the same time relieve inflammation.

Thus, the present inventors found that the ginseng fruit extract has a skin effect due to different ingredients and compositions from general ginseng and red ginseng, while studying cosmetics having skin efficacy using ginseng fruit, which is the ground part of ginseng, and the present invention. It was completed.

Accordingly, it is an object of the present invention to prepare a ginseng fruit extract having an antioxidant effect, collagen production and MMP-1 inhibitory effect from ginseng fruit, and to provide a skin external composition for anti-aging and wrinkle improvement containing the same as an active ingredient. .

It is also an object of the present invention to provide an external composition for skin whitening by inhibiting melanin production and improving pigmentation by containing the ginseng fruit extract as an active ingredient.

In addition, the object of the present invention by containing the ginseng fruit extract as an active ingredient to the skin moisturizing effect or atopic dermatitis improving effect to induce and maintain the normal differentiation of epidermal keratinocytes and improve atopic symptoms To provide.

It is also an object of the present invention to provide an anti-inflammatory skin external composition for improving skin troubles including acne by controlling the sebum control effect and anti-inflammatory effect, that is, the production of inflammatory factors by containing the ginseng fruit extract as an active ingredient. It is.

It is another object of the present invention to provide a skin external preparation composition for improving skin color to expand capillaries of the skin, promote blood circulation, and make the skin clear and transparent by containing the ginseng fruit extract as an active ingredient.

In order to achieve the above object, the present invention provides a skin external composition for inhibiting skin aging and wrinkle improvement containing ginseng fruit extract as an active ingredient.

In another aspect, the present invention provides a skin whitening composition for skin whitening showing the skin whitening effect by inhibiting melanogenesis and improving pigmentation by containing the ginseng fruit extract as an active ingredient.

In another aspect, the present invention provides a skin moisturizing or atopic dermatitis composition for improving the skin moisturizing effect and atopic symptoms through the normal integration and maintenance of epidermal keratinocytes by containing the ginseng fruit extract as an active ingredient.

In another aspect, the present invention provides an anti-inflammatory external composition for skin having the effect of improving acne and skin troubles by controlling the sebum control effect and inflammatory response by containing the ginseng fruit extract as an active ingredient.

In another aspect, the present invention provides a skin external preparation composition for improving skin color by promoting peripheral blood circulation by improving the blood circulation by containing the ginseng fruit extract as an active ingredient.

In the present invention, by containing the ginseng fruit extract having a different ingredient and composition from the ground of ginseng as an active ingredient, inhibiting skin aging, skin wrinkle improvement, skin whitening effect, skin moisturizing and atopic symptoms, acne and skin trouble improvement An external preparation composition for skin having an effect and an effect of improving skin coloration could be provided.

1 is a graph analyzing the ginsenoside components of the ginseng fruit extract of Example 1 and the ginseng extract of Comparative Example 1.
2 is a graph showing the active oxygen removal activity of the test substance.
Figure 3 is a graph measuring the amount of collagen production of ginseng fruit extract of Example 1.
Figure 4 is a graph measuring the inhibitory effect of MMP-1 biosynthesis of ginseng fruit extract of Example 1.
Figure 5 is a graph comparing the nitrogen monoxide inhibitory effect of the ginseng fruit extract and red ginseng extract of Example 1.
Figure 6 is a graph showing the amount of NO produced by the ginseng fruit extract of Example 1.
7 is a graph showing the amount of NO produced by red ginseng extract.

In the present invention, the ginseng fruit extract may be mixed in an amount of 0.001 to 50% by weight based on the total weight of the composition, depending on the form of the skin external preparation composition. The ginseng fruit extract is preferably 0.001% by weight or more when considering the effect, and 50% by weight or less is preferable when considering the formulation of the external preparation for skin.

The ginseng fruit extract according to the present invention is prepared by extracting the flesh and skin of the ginseng fruit separated and separated from the seeds by sun drying or hot air drying, followed by water or ethanol extraction.

The cosmetic form according to the invention contains a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example emulsions, suspensions, microemulsions, microcapsules, microgranules, or ionic (liposomes) obtained by dispersing an oil phase in solution, gels, solids, pasty anhydrous products, aqueous phases, and It may be provided in the form of a nonionic vesicle dispersant or in the form of a cream, skin, lotion, powder, ointment, spray or cone stick. These compositions can be prepared according to conventional methods in the art. The composition according to the invention can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

Cosmetics of the present invention may be used in the preparation of fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers. Cosmetics such as fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics or It may contain adjuvants commonly used in the field of dermatology. Such adjuvants are introduced in amounts generally used in the cosmetic or dermatological arts.

The external preparation for skin containing the ginseng fruit extract of the present invention is not particularly limited in the formulation, for example, supple cosmetics, astringent cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing It may be formulated into a cosmetic such as cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil and body essence.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples and Test Examples. However, these experimental examples and examples are provided only for the purpose of illustration in order to help the understanding of the present invention is not limited to the scope and scope of the present invention.

Example 1 Preparation of Ginseng Fruit Extract

1) Ginseng fruit pretreatment: Raw ginseng fruit was harvested, seed was separated and removed, and then the ginseng fruit dried raw material was prepared by sun drying or hot air drying.

2) Preparation of ginseng fruit extract: 3 g of water was added to 1 kg of dried ginseng fruit extract to extract reflux, and then filtered and concentrated under reduced pressure at 40 to 45 ° C. to obtain 300 g of ginseng fruit extract.

Comparative Example 1 Preparation of Ginseng Root Extract

Except for using ginseng root (root) instead of ginseng fruit in Example 1 was prepared in the same manner as in Example 1.

Test Example 1 Comparison of Ingredients of Ginseng Fruit Extract

<Ginsenoside (Ginseng Saponin) Analysis of Ginseng Fruit and Ginseng Root>

In Example 1 and Comparative Example 1, respectively, ginseng fruit and ginseng root extract were prepared, and then, the extracts were treated with ether to remove fat-soluble components, followed by extracting and concentrating ginsponin with butanol (BuOH). Cenoside component analysis was performed and the results are shown in FIG. 1 and Table 1. FIG.

Example 1 Comparative Example 1 Irradiated Ponin Content (Dry Weight) 33.42% 16.70% PD / PT ratio 0.73 3.23

The ginseng fruit extract prepared in Example 1 had about twice the content of the ginseng root extract prepared in Comparative Example 1 in the content of irradiated phonynin, and ginsenosides were PD (Protopanaxadiol)-"ginsenoside Rb1, Rb2. And Rc and Rd "and PT (Protopanaxatriol)-" Ginsenoside Re, Rg1 and Rg2 "ratios were 0.73 and 3.23, respectively. .

Mineral Analysis of Ginseng Fruit Extract

In order to distinguish the ginseng fruit extract prepared in Example 1 has the characteristics as 'fruit' different from ginseng, mineral components including vitamins were analyzed, and the results are shown in Table 2 below.

ingredient content ingredient content Potassium (mg / 100g) 5865.57 Magnesium (mg / 100g) 354.38 Calcium (mg / 100g) 819.26 Zinc (mg / 100g) 178.49 Iron (mg / 100g) 59.31 Vitamin A (µg / 100g, RE) 213.11 Phosphorus (mg / 100g) 187.17 Vitamin B1 (mg / 100g) 12.29 Vitamin B2 (mg / 100g) 8.45 Vitamin B6 (mg / 100g) 10.50 Vitamin C (mg / 100g) 4.91 Vitamin E (mg / 100g, a-TE) 23.61 Vitamin K (μg / 100g) 232.12 Niacin (mg / 100g, NE) 5.76 Pantothenic Acid (mg / 100g) 5.87 Folic Acid (µg / 100g) 349.97

As described above, the ginseng fruit used in the present invention contains more ginseng saponin than the ginseng root, and at the same time, the composition of the saponin is opposite to that of the ginseng root. It was confirmed that, based on this, the following experiments on the skin efficacy was carried out.

[Test Example 2] skin aging inhibition and wrinkle improvement effect

1) Antioxidant Effect of Ginseng Fruit Extract

Antioxidant effect was investigated through experiments comparing the ability to remove reactive oxygen species (ROS) generated from cells by ultraviolet (UV) irradiation. As a positive control, trolox, which is generally used to compare antioxidant effects, was used, which was also compared with white and red ginseng extracts (FIG. 2). In addition, the group not treated with the test substance was used as a control.

In the results of Figure 2, the ginseng fruit extract of Example 1 according to the present invention significantly erases the active oxygen generated by UV compared to the control in the human HaCaT keratinocytes monolayer culture system (human HaCaT keratinocytes monolayer culture system) It showed an effect. This is a good activity similar to trolox, which is used as an activity indicator of antioxidants, and contrasted with white and red ginseng extracts without significant scavenging activity. That is, it was confirmed that the ginseng fruit extract according to Example 1 of the present invention may have an effect of preventing wrinkle formation, elasticity reduction and pigmentation by significantly eliminating active oxygen which causes skin aging.

2) Inhibitory activity against DNA oxidative damage by single cell gel electrophoresis (SCGE, Comet assay)

After culturing human fibroblasts for 24hr, the ginseng fruit extract and vitamin E of Example 1 were treated by concentration, and after 12hr, H ₂ O ₂ (final concentration 10-3M), which was a positive control, was treated by electrophoresis. The stained result was observed by fluorescence microscope. In this case, using an image analyzer KOMET 3.1 (Kinetic Imaging, England) was analyzed the length (um) of the tail (um) resulting from DNA cleavage in each of the 25 cells, the results are shown in Table 3 below. Indicated.

Inhibitory Effect of Ginseng Fruit Extract and Vitamin E on DNA Oxidative Damage Test substance Tail length (μm) % Inhibition H ₂ O ₂ 115.6 - H ₂ O ₂ + Example 1 (10 ppm) 75.8 34.4 H ₂ O ₂ + Example 1 (50 ppm) 65.7 43.2 H ₂ O ₂ + Example 1 (100 ppm) 52.1 54.9 H ₂ O ₂ + Vitamin E (10 ppm) 70.6 38.9

In the results of Table 3, the ginseng fruit extract of Example 1 by cell gel electrophoresis showed significant oxidative DNA damage inhibitory activity, whereby the ginseng fruit extract according to the present invention is a hydroxyl radical (.OH) It was confirmed that it acts as a DNA oxidative damage inhibitor that inhibits the DNA single strand break.

3) Type I Procollagen Assay

Human fibroblasts were cultured in a 12-hole plate incubator at a concentration of 10 ⁵ and then replaced with a medium containing 1 ppm and 10 ppm of ginseng fruit extract of Example 1. Cells were harvested on the third day of culture to quantify the amount of type I procollagen produced by ELISA. The result was calculated as a comparison with the measured value of the control group that does not contain the ginseng fruit extract of Example 1 to 100, TGF-b was used as a positive control group. It was also compared with white and red ginseng extracts as ginseng root extract.

In the normal human fibroblast monolayer culture system, the ginseng fruit extract of Example 1 showed an effect of markedly promoting the production of type I procollagen as compared to the control group. That is, the ginseng fruit extract of Example 1 was able to suppress the reduction in collagen production caused by the aging of the human skin, and showed a wrinkle improvement effect (Fig. 3).

4) MMP-1 expression inhibition assay

Human fibroblasts were cultured in a 12-hole plate incubator at a concentration of 10 ⁵ and then irradiated with ultraviolet B at 40 mJ / cm 2, and then replaced with a medium containing 1 ppm and 10 ppm of ginseng fruit extract of Example 1. Cells were harvested on the second day of culture (harvest) to quantify the amount of matrix metalloproteinase I (MMP-1) produced by ELISA. The result was calculated as a comparison with the value measured using the UV control group containing no test substance 100, and the positive control group was TGF-b. It was also compared with white and red ginseng extracts as ginseng root extract.

The ginseng fruit extract of Example 1 according to the present invention in human normal fibroblast monolayer culture system significantly inhibited the expression of MMP-1 induced by UVB 40 mJ / cm 2. Therefore, the ginseng fruit extract of Example 1 according to the present invention inhibits the biosynthesis of the skin tissue degrading enzyme MMP-1 caused by internal aging or external environmental factors, inhibits skin aging and is effective in improving wrinkles. (FIG. 4).

5) Biosynthesis Inhibition Effect of Cyclooxygenase-2 (COX-2) by Ultraviolet Light

Human fibroblasts were cultured in a 12-hole flat plate incubator at a concentration of 10 ⁵ , and then irradiated with UVA at 15 J / cm 2. The ginseng fruit extract of Example 1 and the ginseng root extract of Comparative Example 1 were 0.1 ppm and 1 ppm, respectively. And medium containing 10 ppm. Cells were harvested on the second day of culture (harvest) and the amount of cyclooxygenase-2 (COX-2) produced was quantified using the Western blot method. The UV control group containing no test substance was calculated as 100 and compared with the value measured by a densitometer, and the results are shown in Table 4 below.

Test substance (concentration: ppm) COX-2 biosynthesis (%) Example 1 10 11 One 38 0.1 63 Comparative Example 1 10 72 One 91 0.1 99 Control 100

From the results of Table 4, the ginseng fruit extract of Example 1 concentration-dependently reduced the biosynthesis of cyclooxygenase-2 by ultraviolet rays, skin tissue degradation by the prostaglandin E2 (PGE2) product of COX-2 Could be prevented.

6) Inhibitory effect of UV on tumor necrosis factor alpha (TNF-a)

After culturing human keratinocytes in a 12-hole plate incubator at a concentration of 10 ^5, UV B was irradiated at 30 mJ / cm 2, and then the ginseng fruit extract of Example 1 and the ginseng root extract of Comparative Example 1 were 0.1 ppm and 1, respectively. Replace with medium containing ppm and 10 ppm. After 6-24 hours of culture, the cells were harvested (harvest) and the amount of tumor necrosis factor alpha (TNF-a) generated using the ELISA (Pharmingen 555212) method was quantified. The results calculated by comparison with the measured values are shown in Table 5 below.

Test substance (concentration: ppm) TNF-a biosynthesis (%) Example 1 10 16 One 38 0.1 72 Comparative Example 1 10 74 One 85 0.1 92 Control 100

From the results of Table 5, the ginseng fruit extract of Example 1 according to the present invention can be caused due to the biosynthesis of tumor necrosis factor alpha by reducing the concentration of the biosynthesis of tumor necrosis factor alpha (TNF-a) by ultraviolet rays It was confirmed that skin aging can be prevented beforehand.

7) Skin wrinkle improvement effect on human skin

Forty subjects with facial wrinkles aged 30 to 50 years were prepared with the nutrition cream of Example 2 and Comparative Example 2 containing the ginseng fruit extract of Example 1 having the composition shown in Table 6 below. The skin wrinkle improvement effect was compared and evaluated (unit: weight%).

ingredient Example 2 Comparative Example 2 Ginseng Fruit Extract 0.1 - Beeswax 10 10 Polysorbate 60 1.5 1.5 Sebum 60 Cured Castor Oil 2 2 Sorbitan sesquioleate 0.5 0.5 Liquid paraffin 10 10 Squalane 5 5 Caprylic / Capric Triglycerides (Estasan: Uniqema) 5 5 glycerin 5 5 Butylene glycol 3 3 Propylene glycol 3 3 Triethanolamine 0.2 0.2 Preservative, coloring, flavoring Quantity Quantity Purified water Remaining amount Remaining amount

Example 2 was used for the left side of the subject and Comparative Example 2 was used for the right side for 3 months. The skin condition was measured on both sides of the face before the use of the cream, and then three months after the use of the cream, the same area was remeasured to measure the change in skin wrinkles. In the constant temperature and humidity room of 24 ° C. and 40% RH, the wrinkles of the tail of the eyes were replicated, and skin wrinkles were measured by a Visiometer system (C + K). The amount of change in skin wrinkles was calculated according to the following equation.

In Equation 1, "Tdi" is a measurement value at D90, and "Tdo" is a measurement value at D0.

As a result of calculation according to Equation 1, the skin wrinkles at the site using Comparative Example 2 showed a decrease of 6.5 ± 4% (mean ± standard deviation), while the skin wrinkles at the site using Example 2 were 39 ± 11%. It showed a decrease of, showing an excellent skin wrinkle improvement effect.

8) Improvement of skin elasticity in human body

The skin elasticity improving effect of Example 2 and Comparative Example 2 prepared by the composition of Table 6 was measured. Forty women over 30 years old were divided into two groups, and the skin elasticity measuring device (Cutometer SEM 575, C + K Electronic) was applied to the face twice a day for 12 weeks. Co., Germany) to measure the skin elasticity, the results are shown in Table 7 below. The results in Table 7 refer to the nature of skin viscoelasticity of the skin elasticity meter.

Test substance Skin elasticity effect Example 2 0.46 ± 0.12 Comparative Example 2 0.29 ± 0.09

As shown in the results of Table 7, when the nutrition cream of Example 2 containing the ginseng fruit extract of Example 1 was used, the skin elasticity enhancing effect was about 150%.

[Test Example 3] skin whitening effect

1) Inhibitory Effects of Melanin Production Using Pigment Cells

In order to determine the melanin production inhibitory effect of the ginseng fruit extract of Example 1, the rat pigment cells were used.

First, murine Mel-Ab cells derived from C57BL / 6 mice (Dooley, TP et al, Skin pharmacol, 7, pp 188-200) were placed in DMEM (Dulbeccos modified Eagles media) in 10% fetal placental serum, 100 Incubation was performed at 37 ° C. and 5% CO ₂ in a medium to which nM 2-O-tedecanocanophorbol-13-acetate and 1 nM cholera toxin were added. The cultured Mel-Ab cells were detached with 0.25% trypsin-EDTA, incubated at a concentration of 10 ⁵ cells / well in a 24-hole plate incubator, and then 1 ppm and 10 ppm for 3 consecutive days from the second day. Ginseng fruit extract was added and cultured. At this time, red ginseng extract and hydroquinone were used as a positive control. Then, the culture solution was removed, washed with PBS, the cells were dissolved with 1 N sodium hydroxide, the absorbance was measured at 400 nm, and the melanin production inhibition rate was calculated according to Equation 2 below. (Dooley's method).

Test substance Melanin production inhibition rate (%) Example 1 (1 ppm) 14.0 Example 1 (10 ppm) 34.4 Red Ginseng Extract (10 ppm) 5.3 Hydroquinone (control) 41.1

As shown in Table 8, the ginseng fruit extract of Example 1 according to the present invention showed an excellent melanin production inhibition rate than the red ginseng extract, it was confirmed that the melanin production inhibition rate similar to hydroquinone. Through this, the ginseng fruit extract of Example 1 according to the present invention was confirmed to exhibit an excellent whitening effect.

2) whitening effect on human skin

In order to determine the whitening effect of the ginseng fruit extract of Example 1 on the human skin was carried out the following experiment.

First, an opaque tape with a hole of 1.5 cm in diameter was attached to the upper arm of 12 healthy men, and then 1.5 ~ 2 times of ultraviolet light (UVB) of the minimum erythema dose of each subject. Was irradiated to induce skin blackening.

After the UV irradiation, the ginseng fruit extract and hydroquinone of Example 1, which were test substances, were respectively applied, and one place was observed for 10 weeks without applying anything. Skin color was measured on a weekly basis with a colorimeter CR2002 (Minolta, Japan).

Then, the difference in skin color (ΔL *) between the start time and the completion time of application of each test substance was calculated according to Equation 3 below, and the results are shown in Table 9 below. On the other hand, the whitening effect is determined by comparing ΔL * between the sample application site and the control site. When the value of ΔL * is about 2, the whitening effect of the deposited pigment is clear, and when about 1.5 or more, the whitening effect is determined. Can be.

Test substance Skin color brightness degree (△ L *) Example 1 1.76 ± 0.21 Hydroquinone (positive control) 1.90 ± 0.13 Vehicle (negative control) 0.50 ± 0.16

As shown in Table 9, the ginseng fruit extract of Example 1 according to the present invention was confirmed to show a similar degree of skin color brightness as hydroquinone. This is because the ginseng fruit extract brightens the skin color by improving pigmentation generated by ultraviolet rays.

[Test Example 4] Skin moisturizing and atopic symptoms improvement effect

1) Induction of differentiation of human keratinocytes

By measuring the amount of Cornified Envelop (CE) generated during the differentiation of keratinocytes and human skin cell lines (HaCaT) to determine the cell differentiation promoting effect of the ginseng fruit extract of Example 1 according to the present invention.

After primary cultured human keratinocytes were put in a culture flask and attached to the bottom, the test substances described in Table 10 were added to the culture solution by concentration, and 5 days until the cells grew to about 70 to 80% of the floor area. Incubated. The cells were harvested and washed with PBS (phosphate buffered saline), followed by 10 mM Tris-HCl (Tris) containing 2% Sodium Dodecyl Sulfate (SDS) and 20 mM Dithiothreitol (DTT). -HCl, pH 7.4) 1 ml was added, and the sonication, boiling, and centrifuged precipitates were again suspended in 1 ml of PBS to measure absorbance at 340 nm. Separately, a portion of the solution after the sonication was taken to measure the protein content, which was used as a reference for evaluating the degree of cell differentiation. Low calcium (0.03 mM) treated group and high calcium (1.2 mM) treated group as a negative / positive control, respectively, the test results performed by adding the test substance to the low calcium concentration is shown in Table 10 below.

Test substance Differentiation capacity in keratinocytes (%) Control Low calcium group (Ca ^{2 +} 0.03 mM) 100 High calcium group (Ca ^{2 +} 1.2 mM) 210 Red Ginseng Extract (100 ppm) 104 Example 1 1 ppm 115 10 ppm 120 100 ppm 124

Through the results of Table 10, the ginseng fruit extract of Example 1 according to the present invention was confirmed to promote differentiation in keratinocytes.

2) Effect of Transglutaminase Expression on Human Skin Cell Line

Human skin cell lines were put in 5 × 10 ⁴ cells in each 96-well plate incubator and attached for 24 hours. After treatment with the test substance to the attached skin cell line, two days later, the medium was removed, and stored in a -20 ℃ refrigerator. After freeze-thawing was repeated twice, the cells treated with material were destroyed, and then treated with acetone: ethanol (1: 1, v / v) stored at -20 ° C and left at 4 ° C for 30 minutes. The cells were fixed. Thereafter, the mixture was left at room temperature to allow the organic solvent to evaporate, blocking (1% bovine serum albumin), transglutaminase antibody, HRP anti-mouse antibody (secondary) were performed, and color development was OPD (o). -phennyldiamine) was added. Expression was measured by absorbance at 490 nm, calibration was performed by measuring the background (background) at 630 nm. The low calcium (0.03 mM) and high calcium (1.2 mM) treated groups were negative / positive controls, respectively, and the results obtained by adding test substances to the low calcium concentrations are shown in Table 11 below.

Test substance Transglutaminase Expression (%) Control Low calcium group (Ca ^{2 +} 0.03 mM) 100 High calcium group (Ca ^{2 +} 1.2 mM) 140 Red Ginseng Extract (100 ppm) 131 Example 1 1 ppm 135 10 ppm 175 100 ppm 210

In the results of Table 11, the ginseng fruit extract of Example 1 according to the present invention showed a two-fold increase compared to the control, it can be seen that the amount is expressed more than the control. Therefore, it was found that the ginseng fruit extract of Example 1 according to the present invention improves the expression of transglutaminase.

3) Increased epidermal lipid (total ceramide) production in human skin

In order to evaluate the effect of the ginseng fruit extract of Example 1 on the increase in lipid synthesis in real human skin, 100 was used as Carbopol ETD 2020 0.5%, TEA 0.45%, test substance 5%, and deionized water. Twelve adult men and women were marked with the diameter of a 50 ml polypropylene cornical tube on the forearm, followed by 20 μl of the control and the ginseng fruit extract of Example 1 twice daily. Was divided into two groups, and the amount of lipid produced on day 3 and day 11 was analyzed by extracting lipids as follows. Wash the forearm with tap water, tape stripping once with Scotch 810 Magic tape, and cut 50 ml polypropylene conical tubes as a reservoir using a 1 ml cycle. Add hexane / ethanol (4: 1), mix for about 1 minute, transfer it to a new tube, and again treat 1 ml of cyclohexane / ethanol (1: 1) in the same manner as above. Collected on. Thereafter, the mixture was dried using nitrogen gas at 50 ° C., and then dissolved in 500 μl of chloroform / methanol (2: 1), and each test substance was silica gel using CAMAG's Automated TLC sampler-4. Dropped onto a TLC (Thin Layer chromatography) plate, and developed using a development solvent of the composition ratio as shown in Table 12 using AMD (Automated Multiple Development).

chloroform Methanol water Acetic acid Hexane Diethyl ether Petroleum ether Solvent movement distance (cm) Primary 40 10 One - - - - 3.0 Secondary 190 9 - One - - - 6.5 3rd - - - 2 12 3 - 7.5 4th - - - - - - 100 Top

After the TLC plate was developed, the size of each band was measured at a wavelength of 570 nm using a TLC scanner-II (CAMAG: densitometer). The ceramide production amount of each treatment group in which the ceramide production amount of the untreated group was converted to 100% is shown in Table 13 below.

Test substance After 3 days processing (%) After 11 days processing (%) Control No treatment group 100 100 Vehicle 103 98 Glycerin (5%) 102 94 Example 1 (5%) 108 116

As shown in Table 13, after 3 days of treatment with ginseng fruit extract of Example 1 showed an increase of about 5 to 8% compared to the negative control group (non-treated group / vehicle) and 6% of the positive control group (glycerine) Showed an increase. After 11 days, lipid production was about 16% better than the negative control and 22% better than the positive control. This demonstrates the increase in ceramide by the ginseng fruit extract of Example 1 according to the present invention.

4) Increased skin moisture in the human body

Fifty adult men and women in their fifties and sixties with skin dryness were divided into two groups, and Example 2 and Comparative Example 2 were applied to the face twice daily for four weeks in each group. Before application start, after 1 week, 2 weeks, 4 weeks after application, and after 2 weeks (total 6 weeks) after application was stopped, constant temperature, constant humidity (24 ° C, 40% relative humidity) was used as a comonometer. Skin moisture was measured, and the results are shown in Table 14 below. The test result is a percentage of the increase of the measured value after a certain period of treatment based on the Koniometer value measured immediately before the test was started.

Test substance Moisture Growth Rate (%) 1 week past 2 weeks past 4 weeks past After 6 weeks Example 2 34 41 42 33 Comparative Example 2 30 34 34 15

A conniometer is a skin moisture meter that measures the amount of moisture present on the skin by measuring the conductivity of the epidermis. As shown in Table 14, the skin moisture content was increased in the group coated with the material of Example 2 containing the ginseng fruit extract of Example 1 compared to the control group applied to Comparative Example 2. Also, two weeks after the application of the nourishing cream of Example 2 (total of 6 weeks), the value of the measured skin moisture was similar to that after 1 to 2 weeks. During the period it was found that the skin moisture by the application of the ginseng fruit extract according to the present invention is continuously maintained.

5) Atopic dermatitis improvement effect

The effect of improving atopic symptoms was measured as follows. 50 patients who were diagnosed with atopic symptoms (itch, erosion or severe dryness) or who had atopic-like symptoms by visual observation were divided into two groups, and provided in Example 2 and Comparative Example 2, respectively. The atopic dermatitis was applied twice a day for 12 weeks. Efficacy assessments were conducted after 12 weeks of treatment and subjectively judged the improvement. In the questionnaire, the symptoms of atopic dermatitis are divided into "itch", "dry", "keratogenesis", "dandruff", "erythema", "swelling", "skin cracking", and "dust and eczema". In this case, after evaluating the degree of improvement for each symptom, the average of the atopy symptom improvement effect was evaluated for each individual, and the results are shown in Table 15 below.

Test substance Number of respondents Markedly improved Improving No change worse Example 2 2 4 14 0 Comparative Example 2 6 8 6 0

As shown in Table 15, it can be seen that the treatment group to which the formulation containing the ginseng fruit extract of Example 1 according to the present invention was further improved than the control group.

[Test Example 5] Acne and skin trouble relief

1) sebum control effect: inhibitory effect of 5α-reductase activity

Mature Sproque-Dawley male rats (7-8 weeks of age) were killed with diethyl ether, followed by detachment of the abdominal prostate to remove connective tissue and buffer (0.32 M sucrose, 0.1 mm dithiothreitol). And 20 mM sodium acetate), finely chopped, and suspended in a grinder. The suspension was centrifuged to obtain supernatant, partially purified 5α-reductase, and the activity inhibition experiment of this enzyme was performed.

A portion of the supernatant obtained above was taken and placed in a buffer containing 0.2 M monobasic acid and 0.2 M dibasic acid, and then reacted with a substrate (testosterone) having ³ H as an isotope to measure the amount of dihydrotestosterone produced. It was.

The reaction solution is 1 mM dithiothreitol, 40 mM sodium phosphate (pH 6.5), 50 μM NADPH , [1,2,6,7- 3 H] testosterone / testosterone (2.2 × 109 mol) and enzyme suspension (0.8㎎ ) 565 μl of protein.

The ginseng fruit extract of Example 1 was dissolved in 10% ethanol and added to 10 μl of 100 μg / 10% ethanol per reaction. The same volume of solvent was used as a control and riboflavin was used as a positive control.

The reaction was started at the same time as adding the enzyme suspension and proceeded at 37 ° C. for 30 minutes, followed by extraction by adding 1 ml of ethyl acetate, and 100 µl of ethyl acetate was extracted from silica plastic sheet kieselgel 60 (254). 60 F254), the developing solvent system was developed with acetate-cyclohexane (1: 1).

After air drying the plastic sheet, the bath system was used to measure the isotope amount. The isotopic amount of testosterone and dihydrotestosterone remaining in the film after one week by placing the dried plastic sheet and the x-ray film together in the bath cassette. Was measured, and the results for each inhibition rate are shown in Table 16 below.

Test substance T (dpm) DHT (dpm) % Conversion % Inhibition Example 1 6878 2355 25.5 39.9 Control 7520 5536 42.4 - Positive control 6300 2530 28.7 32.4 (1) T: 3H radio activity in the testosterone region (2) DHT: 3H radio activity in the dihydrotestosterone region (3) Conversion rate: radio activity / total radio activity in the DHT region (4) Inhibition rate: 100 x (control rate vs. sample rate) / control rate

From Table 16, the ginseng fruit extract was found to have a 5α-reductase inhibitory effect.

2) Lipogenesis Inhibitory Effect

The degree of lipid synthesis in sebaceous glands was evaluated by quantifying the uptake of carbon elements necessary for lipid synthesis and assessing the degree of inhibition of lipid synthesis.

In other words, the hamster ear tissue with a lot of sebaceous glands was biopsyed for 6 hours in a C14-labeled medium, and the control group and the test substance were compared and quantified to evaluate the degree of inhibition. Test substances were used by adding 0.01% test substance solution to the medium using hamster right ear tissue, and the control group was used by adding 0.01% physiological saline to the medium using hamster left ear tissue, and the results are shown in Table 17 below. It was.

On the other hand, the inhibition rate in Table 17 is calculated according to the following equation 4, the value is the average value for the four hamsters.

Test substance % Inhibition Example 1 47 Positive control (retinoic acid) 52

From the results of Table 17, it can be seen that the ginseng fruit extract of Example 1 according to the present invention has a lipid biosynthesis inhibitory effect.

3) Cotton cloth reduction effect

The ginseng fruit extract of Example 1 used in the present invention was dissolved in 1,3-butylene glycol to make a 1% solution, and then used as a rabbit clinical test substance in order to confirm the reduction effect of the cotton.

In the morning of the rabbit rabbit, 50% oleic acid / paraffinic oil (0.2%) was applied in the morning and rubbed with a cotton swab. It was applied twice a day in the morning (50% oleic acid / paraffin oil), in the afternoon (material) and for a month during the total experiment period. Cotton cloth is generated from about 10 days, 0.2 g each of the ginseng fruit extract of Example 1 was applied in the afternoon as a result of gradually reducing the number and size of cotton cloth from two weeks later.

Each tissue was biopsied and exfoliated, and the scrim size was measured under a microscope, and the results are shown in Table 18 below.

Test substance Average scrim size measurement (0.1 mm) N = 3 Control group (untreated group) 1.78 Oleic acid (OA) 3.5 Oleic acid (OA) + Example 1 2.3

From the results in Table 18, it can be seen that the cotton cloth of the rabbit treated with the ginseng fruit extract of Example 1 according to the present invention was significantly reduced. That is, sebum reduction was shown by the ginseng fruit extract of Example 1, and as a result, it can be estimated that the skin scleroderma was reduced by reducing the skin keratinization.

4) sebum secretion inhibitory effect

To measure the effects of oily skin, 20 to 45-year-old females with oily skin had 10 cases of Example 2 (containing ginseng fruit extract) on the forehead, and 10 others of Comparative Example 2 in the morning and evening. After applying twice a day to use, after 1 week, 2 weeks, 3 weeks and 4 weeks sebum secretion of the skin was measured using a skin oil meter (Sebumeter SM810). The experiment was carried out at 20 ℃, 20% relative humidity and the measurement results are shown in Table 19. The results are expressed as the average value of 10 subjects. For oily skin the measured value is 220 or more, for normal skin it is between 100 and 220.

Sebum secretion inhibitory effect Test substance Before application 1 week 2 weeks 3 weeks 4 Weeks Comparative Example 2 245 240 239 235 230 Example 2 245 236 231 218 209

In the results of Table 19, it was confirmed that the nutrition cream of Example 2 containing the ginseng fruit extract of Example 1 according to the present invention exhibits the effect of inhibiting excessive sebum secretion, has a sebum control effect.

5) Anti-inflammatory effect: inhibitory effect of prostaglandin E2 (PGE2) production

In order to investigate the inhibitory effect of the ginseng fruit extract according to the present invention prostaglandin E2 production was tested in the following manner (Ref .: Jeffrey, KH, Anglea, SW, Zoe, S., and Rhia CM Intracellular Measurement of Prostaglandin E2: Effect of Anti-inflammatory Drugs on Cyclooxygenase Activity and Prostanoid Expression, Analytical Biochemistry, 1999, 271, 18-28).

First, the fibroblasts isolated from human epidermal tissue were incubated for 24 hours in a CO ₂ incubator, then replaced with medium containing no FBS (fetal bovine serum) and cultured for 2 hours, and then the ginseng fruit of Example 1 Distilled water was mixed with the extract, and prepared and treated at concentrations of 1 μg / mL, 10 μg / mL and 100 μg / mL, respectively, and then incubated in 37 ° C. and 5% (v / v) CO ₂ incubator for 2 hours. .

Next, after treatment with calcium ionophore A23187 and arachidonic acid at a concentration of 50 μm for 5 minutes as a prostaglandin E2 stimulator, the cells were hemolyzed and absorbance was measured at 450 nm of ELISA reader to quantify prostaglandin E2. And the results are shown in Table 20 below. At this time, the inhibition rate shown in Table 20 is calculated based on the difference in the production of prostaglandin E2 of the skin fibroblasts not treated with the ginseng fruit extract of Example 1 and the fibroblasts treated with the ginseng fruit extract of Example 1 when arachidonic acid treatment The average value was repeated five times.

Test substance Concentration (㎍ / mL) Prostaglandin E2 (pg / 10 ⁵ cells) % Inhibition Negative Control - 25.2 ± 4.4 - Arachidonic acid - 385.1 ± 9.1 - Arachidonic Acid + Example 1 One 289.6 ± 6.5 24.8 10 246.4 ± 8.6 36.0 100 195.6 ± 14.3 49.2

In the results of Table 20, it was confirmed that the ginseng fruit extract of Example 1 according to the present invention can inhibit the production of prostaglandin E2 by the stimulation source.

6) Anti-inflammatory effect: Inhibition of biosynthesis of cyclooxygenase-2 (COX-2) induced by LPS (Lipopolysaccharide)

Human fibroblasts were cultured in a 12-hole plate incubator at a concentration of 10 ⁵ , and after LPS treatment, the ginseng fruit extract of Example 1 was replaced with a medium containing 1 ppm, 10 ppm and 100 ppm. Cells were harvested on day 2 and cultured to quantify the amount of COX-2 produced using the Western blot method. A comparison with the value measured with a densitometer using the control group as 100 is shown in Table 21 below. At this time, the control group is an untreated group cultured without treating the ginseng fruit extract of Example 1.

Test substance (concentration: ppm) COX-2 biosynthesis (%) Example 1 100 15 10 35 One 71 Control 100

As can be seen from the results of Table 21, the ginseng fruit extract of Example 1 according to the present invention by inhibiting the inflammatory response by reducing the biosynthesis of cyclooxygenase-2 induced by the inflammatory response triggering factor LPS Efficacy in alleviating skin troubles has been shown.

7) Anti-inflammatory effect: Inhibitory effect of LPS (Lipopolysaccharide) -induced tumor necrosis factor alpha (TNF-a) on biosynthesis

Human keratinocytes were cultured in a 12-hole plate incubator at a concentration of 10 ⁵ , and after treatment with LPS, the ginseng fruit extract of Example 1 was replaced with a medium containing 1 ppm, 10 ppm and 100 ppm. Cells were harvested for 6-24 hours and then harvested (harvest) to quantify the amount of tumor necrosis factor alpha (TNF-a) produced using ELISA (Pharmingen 555212) method. Table 22 shows a comparison with the value measured using the control group as 100. At this time, the control group is an untreated group cultured without treating the ginseng fruit extract of Example 1.

Test substance (concentration: ppm) TNF-a biosynthesis (%)  Example 1 100 14 10 41 One 69 Control 100

As can be seen from the results in Table 22, the ginseng fruit extract of Example 1 according to the present invention concentration-dependently reduce the biosynthesis of tumor necrosis factor alpha (TNF-a) caused by the inflammation-inducing factor LPS may be caused by this Inhibition of the inflammatory response could contribute to skin problems.

8) LPS-induced NO (nitric oxide) inhibitory effect on macrophage

In order to examine the anti-inflammatory effect of the ginseng fruit extract according to the present invention was carried out LPS-induced NO (nitric oxide) inhibition experiments on macrophage (Macrophage).

Macrophages (Raw 264.7 cells) were incubated under 5% CO ₂ in medium with 10% serum. After growing at a concentration of 2 × 10 ⁵ in a 96-hole flat plate incubator and stimulating with LPS (1 ug / ml), the ginseng fruit extract and red ginseng extract of Example 1 were treated and stored at 37 ° C. for 1 hour, and then NO. The degree of production was measured to compare the effect of NO inhibition induced by LPS. The measurement of NO production was carried out using the Griess reaction method (Minghetti, L. et al., 1991, Glia 19. 152-160), the results are shown in FIG.

As shown in the results of Figure 5, the ginseng fruit extract of Example 1 according to the present invention effectively inhibited the NO production as a cytotoxic factor (Cytokine), the effect was much better than the red ginseng extract. Therefore, the ginseng fruit extract of Example 1 according to the present invention is expected to contribute to the effect of healing wounds and inhibiting wrinkles caused by skin problems through the control of the inflammatory response.

9) Acne and skin trouble improvement effect

The participants in the experiment were 10 women with severe skin problems, including acne. After washing the face with soap in a constant temperature and humidity room for 30 minutes, the skin trouble was measured by comparing the number of acne and the number with inflammation. .

The test subjects were allowed to use the nutrition creams of Example 2 and Comparative Example 2 for 4 weeks, and then the degree of acne and skin troubles was compared, and the results are shown in Table 23 below.

Example 2 (5 people) Comparative Example 2 (5 People) Skin trouble improvement rate 80% 20%

As shown in the results of Table 23, the application of Example 2 including the ginseng fruit extract of Example 1 according to the present invention for 4 weeks showed a reduction in the number of existing acne and improvement of the accompanying inflammation.

Test Example 6 Skin Color Improvement Effect

1) NO Production of Ginseng Fruit Extract in Human Uumbilical Vein Endothelial Cells

Endothelial nitric oxide synthase (eNOS) is present in human vascular endothelial cells, and its activity is increased to produce NO (nitric oxide) to expand blood vessels and promote blood circulation. After culturing human vascular endothelial cells, the amount of NO produced by treating ginseng fruit extract (FIG. 6) and general red ginseng extract (FIG. 7) in the medium was compared. In other words, vascular endothelial cells were attached at a density of 2.5 × 10 ⁴ cells / well in a gelatin-coated 24-hole plate incubator and then incubated for 12 hours in a growth medium. The vascular endothelial cells were pretreated with the ginseng fruit extract and red ginseng extract of Example 1 and the untreated group for 12 hours. Endothelial cells were treated with 10 μmol / L DAF-FM diacetate (Molecular Probe, OR) for 30 minutes in M199 medium without FBS at 37 ° C. Vascular endothelial cells were washed three times with M199 medium without FBS, placed in a parallel plate flow chamber, and stimulated with light separated from a mercury lamp. The excitation wavelength is 488 nm and NO-bound DAFs fluoresce at 515 nm. Photos were taken with a confocal laser microscope (Atto Bioscience, USA), and the brightness of the fluorescence was analyzed with Image-Pro Plus v4.5 software (Media Cybernetics, San Diego, CA, USA) to analyze NO as the intensity of fluorescence compared to the untreated group. The degree of production was compared.

As can be seen in the results of Figure 6 and Figure 7, the ginseng fruit extract of Example 1 according to the present invention showed a degree of generation of 500 to 1300% of the non-treated group, the general red ginseng extract is 100 to 200% NO The degree of production was shown (same concentration comparison 50-100 μg / ml). Therefore, the ginseng fruit extract of Example 1 according to the present invention showed an excellent ability to produce NO in vascular endothelial cells as compared to the red ginseng extract. The excellent NO (nitrogen oxide) production ability of the ginseng fruit extract of Example 1, as shown above, may eventually expand capillaries and promote blood circulation, thereby smoothly supplying nutrients to skin, inhibiting skin aging, and improving skin coloration. Will contribute to the efficacy.

2) Color improvement effect on human skin (face)

In order to evaluate the blood circulation promoting effect on the skin of the composition containing the ginseng fruit extract of the present invention, the degree of blood circulation in the skin was measured using a LDPI (Laser Doppler Perfusion Imager). LDPI is a well-known and still used device for measuring blood circulation in the skin. It is a very sensitive device that can measure not only the speed and amount of blood in the capillaries of the skin, but also the flow in the small arteries and veins. .

After washing the face with soap in a thermo-hygrostat room for 30 minutes, the initial values were measured using an IRPI (Infrad) camera, an LDPI and skin temperature measuring instrument. The participants were 20 women with cold hands and feet. LDPI was used to measure the initial blood flow in the lower part of the forehead, and the IR temperature was measured for the forehead, under eye and cheek skin.

After allowing Example 2 to be used by the subjects for one week, blood flow and skin temperature were compared with the initial measured values, and the results are shown in Tables 24 and 25 below.

LDPI results before and after use Face Before use After 1 week medium 0.952 1.169

IR results before and after use forehead Under the eyes cheek Before use 32.26 32.02 30.44 After 1 week 33.87 33.69 33.57

As described above, as a result of using Example 2 containing the ginseng fruit extract according to the present invention, it was confirmed that the color is improved due to the peripheral blood circulation improvement effect in the face of the test subjects, which ultimately according to the present invention Containing fruit extract suggests that it can contribute to the effective delivery of skin nutrients, to inhibit and retard skin aging.

Formulation Example 1 Flexible Cosmetic (Skin Lotion)

Ingredient Content (% by weight) Purified water Remaining amount Ginseng Berry Extract 0.1 Butylene glycol 2.0 Propylene glycol 2.0 Carboxy Vinyl Polymer 0.1 Fiji-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, flavoring Quantity

Formulation Example 2 Nutritious Longevity (Milk Lotion)

Ingredient Content (% by weight) Purified water Remaining amount Ginseng Berry Extract 0.1 Beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Montana 202 (Manufacturer: Seppic) 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxy Vinyl Polymer 0.1 Triethanolamine 0.2 Preservative, coloring, flavoring Quantity

Formulation Example 3 Massage Cream

Ingredient Content (% by weight) Purified water Remaining amount Ginseng Berry Extract 0.1 Beeswax 10.0 Polysorbate 60 1.5 Sebum 60 Cured Castor Oil 2.0 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Montana 202 (Manufacturer: Seppic) 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, coloring, flavoring Quantity

[Formulation Example 4] Pack

Ingredient Content (% by weight) Purified water Remaining amount Ginseng Berry Extract 0.1 Polyvinyl alcohol 13.0 Sodium Carboxymethyl Cellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 Fiji-12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservative, coloring, flavoring Quantity

Claims (2)

Anti-aging skin external composition comprising ginseng fruit extract as an active ingredient. The method of claim 1,
The ginseng fruit extract is an anti-aging skin external composition for water or ethanol extract.

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