KR20050107649A - Composition containing paeoniflorin for inhibiting skin aging - Google Patents

Abstract

The present invention relates to a composition for inhibiting skin aging comprising peonyflorin, and in particular, it enhances the activity and function of skin cells and promotes the synthesis of collagen fibers to improve the toughness and elasticity of skin tissues as well as to produce lipid peroxides. It provides a composition for inhibiting natural skin aging that can be reduced.

Description

Composition for inhibiting skin aging containing peoniflorin {COMPOSITION CONTAINING PAEONIFLORIN FOR INHIBITING SKIN AGING}

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to a composition for inhibiting skin aging including peoniflorin, and more particularly, to improve the toughness and elasticity of skin tissues by enhancing the activity and function of skin cells and promoting the synthesis of collagen fibers. In addition, the present invention relates to a composition comprising peoniflorin having the effect of significantly inhibiting natural skin aging by reducing lipid peroxide production.

[Private Technology]

Peonyflorin is one of the main components of the roots of peony, such as peony or peony, and is a colorless form of substance called peony saponin. Peonyflorin is known to have vasodilation, anti-inflammatory, anti-hypersensitivity and immunostimulating effects, and is also used to treat protective spasms of the red cell membrane.

Skin aging is mainly classified into intrinsic aging and photoaging. Natural aging is due to internal factors (mainly genetic factors) in the body, which are characterized by wrinkles and skin extension. Photoaging is caused by overlapping damage caused by UV rays with the progress of natural aging, rough UV exposure area, deep wrinkles, thick, irregular pigmentation, vascular expansion, poor keratinization of epidermis, abnormal growth. In addition, deformation of elastic fibers of dermis and accumulation of degradation products occur. Ultraviolet light also accelerates the breakdown of collagen and elastic proteins in the dermis, intensifying damage to the skin and, in severe cases, developing skin cancer. In addition, ultraviolet light causes wrinkle formation, skin elasticity, skin dryness, skin glossiness, skin roughness, freckles, capillary rupture or skin color shift ( J Pathol , 1997, 180; 80-89).

In order to prevent skin photoaging, UV blockers are applied to household goods and cosmetics, and other skin light aging products such as retinoids or hydroxy acids (alpha-hydroxy acid or butylated hydroxyanisol) have been developed. It's happening. However, studies on skin aging have been focused on photo aging, and studies on skin aging delay due to natural aging have been insufficient.

In order to solve the above problems, it is an object of the present invention to provide a composition that can suppress the natural skin aging.

Another object of the present invention is to provide a composition for inhibiting skin aging comprising peonyflorin.

In order to achieve the above object, the present invention provides a composition for inhibiting skin aging comprising peonyflorin as an active ingredient.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention, while searching for a substance that can inhibit the natural skin aging, peonyflorin, known as the main ingredient of the neck and roots enhances the activity and function of skin cells and promotes the synthesis of collagen fibers, It was confirmed that not only contributes to the improvement of elasticity but also inhibits skin aging by reducing the production of lipid peroxides, and completed the present invention based on this.

Peonyflorin of the present invention can be used by purchasing a commercially available material, or by using a well-known method of separating and purifying from the roots of plants belonging to the genus Peony, such as peony or peony, or at least 50% by weight of peonyflorin Can be used in place of the root extract of the plant containing above. For example, the root extract may be prepared by the following method. The root of Paeonia lactiflora Pallas is extracted and concentrated three times with 5 times of 75% alcohol water each time, 3 times each time. The remaining alcohol of the concentrate is blown off, and 1 times of water is added to dissolve it to make a suspension. The suspension is triturated three times with 1x of ethyl ether, and the mixture is suspended and partitioned. The aqueous solution of the lower layer is taken to form a concentrate. The fractionated concentrate was poured into a 200-300 mesh silica gel column, and the fluidized phase was separated by chloroform: acetone (4: 1), concentrated, dried, and powdered.

The present invention provides a composition for inhibiting skin aging comprising a plant extract containing peonyflorin or peonyflorin as an active ingredient. The active ingredient may be included in 0.001 to 50% by weight in the composition for inhibiting skin aging, preferably 0.01 to 10% by weight. If the content thereof is less than 0.001% by weight, the effect may be insignificant, and if the content thereof exceeds 50% by weight, it may be uneconomical in terms of the effect compared to the amount used.

The composition of the present invention may further comprise conventional ingredients provided for the use of the prophylaxis and / or treatment of skin damage such as skin aging, photo damage, wrinkles, acne and psoriasis in addition to the aforementioned peonyflorin. Examples include alpha-hydroxy acids, antioxidants and tretinoin.

In addition, the composition of the present invention may further comprise a pharmaceutically acceptable carrier or excipient. Such carriers or excipients may be materials used in conventional pharmaceuticals, detergents or cosmetics, and may be, for example, diluents, emulsifiers, fungicides, disintegrants, lubricants and flavoring agents. Representative carriers or excipients include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, carbohydrates, stearic acid, magnesium, talc, silicic acids, paraffins, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrrolidone, and the like. It doesn't happen.

The composition for inhibiting skin aging of the present invention can be administered orally or parenterally, and can be used as a pharmaceutical, cosmetic, food additive and cleaning agent.

The composition for inhibiting skin aging of the present invention may be prepared in various forms according to the purpose and method of use, and in the case of a medicament, for example, PLASTERS, GRANULES, lotion, LPTIONS, LINIMENTS, etc. , LIMONADES, POWDERS, Syrups, SYRUPS, LIQUIDS AND SOLUTIONS, AEROSOLS, ELIXIRS, OINTMENTS, FLUIDEXTRACTS, Emulsions (EMULSIONS), SUSPENSIONS, DECOCTIONS, AFFECTIONS, TABLETS, TABLETS, INJECTIONS, SPIRITS, CATAPLSMA, CASCELLAS, CREAMS (CREAMS), TROCHES, TINCTURES, PASTES, PILLS. In the case of cosmetics, it can be prepared in emulsion, liquid, syrup or gel form and can be used as a lotion, astringent lotion, flexible lotion, cream or pack. In the case of detergents, the formulation may be a warning agent, emulsion, liquid or powder, and may be used as a human detergent.

The amount of the composition for inhibiting skin aging is appropriately adjusted according to the absorbency, inactivation rate of the active ingredient in the body, the age, sex and condition of the patient, the type and severity of the disease, and divided once to five times a day. Can be used.

 Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention, but the protection scope of the present invention is not limited to the following examples.

Example 1 Evaluation of the Effect of Peonyflorin on Fibroblasts by MTT Analysis in vitro )

In this experiment, the effect on the cell activity of peonyflorin was confirmed.

Keratinocytes and fibroblasts are largely herpes on the dermis and epidermis, and are involved in the main functions of the epidermis and dermis. Thus, keratinocytes and fibroblasts are used as test cells in cell physiology and anti-aging studies.

Skin was taken from red skin mice within 3 days of age to remove subcutaneous tissue and treated with 0.25% trypsin for 12 hours. The skin was then separated into epidermis and dermis, respectively. The epidermis was incubated at 75% confluence in an epidermal incubator (K-SFM), and dermis obtained only fibroblast after 2 hours of trypsin treatment at 37 ° C. and 75% in DMEM medium containing 10% serum. Incubated with the confluence of.

After dissolving peonyflorin (manufactured by Shanghai Food and Process Biotechnology Co., Ltd.) in the culture medium to 0.0004 M in keratinocytes and fibroblasts, MTT ((3- (4,5-dimethylthiazol-2-yl)) -2,5-diphenyltetrazolium bromide) assay was used to determine cell activity, and the control group was also treated with the untreated peoniflorin group and vitamin C 0.002 M group. The impact is shown in Table 1 below.

Table 1

Cell activity enhancement (%) No treatment group 0 Peonyflorin 0.0004 M 25.7 Vitamin C 0.002 M 14.4

From the results of Table 1, it can be seen that peonyflorin significantly promotes cell proliferation of fibroblasts.

Example 2: Evaluation of the Effects of Peonyflorin on Skin Cell Death and Aging

In this experiment, the cell division state was confirmed through specific staining of cell DNA.

β-Gal is a substance that specifically expresses skin aging cells, and is specifically stained only in senescent cells, rather than in pre-aging and stationary cells. Each of the cultured keratinocytes and fibroblasts was treated with a minimum amount of X-gal to cover the cells, followed by staining at 37 ° C. for 24 hours. Skin aging positive cells were indigo blue at 2-4 hours and showed optimal results at 12-16 hours (Proc Natl Acad Sci USA, 1995,92; 9363-7). Then, in the flow cytometer, the number of stained cells (aging cells), the number of viable cells and the number of dead cells were measured, respectively. The results are shown in Table 2 below.

Table 2

Cell types Treatment （M） Cell growth rate Cell death rate Cell aging rate Keratinocytes Blank － － － Peonyflorin 0.0005 M 55.3% 12.3% 24.7% Vitamin E 0.0004 M 23.3% 10.5% 20.1% Fibroblast Balnk － － － Peonyflorin 0.0005 M 18.4% 11.5% 17.2% Vitamin E 0.0004 M 9.96% 15.4% 22.8%

In Table 2, peonyflorin was found to inhibit the cell death rate and cell aging rate of keratinocytes and fibroblasts. In particular, peonyflorin was found to improve the cell growth rate compared to vitamin E, it was confirmed that the skin cell aging delay effect of peonyflorin is superior to vitamin E.

Example 3: Evaluation of skin aging delay effect through animal experiment

Lipid peroxide production and collagen content change in the skin of animals were measured to evaluate the effect of peoniflorin on skin aging.

Aging skin is accompanied by significant changes. In other words, aging skin has a flat epidermal and epidermal connection, a thin layer of stratum granulosum and stratum spinosum, a decrease in collagen and elastin content, a disruption of cell layer arrangement, and black pigment cells and langerhans' cells. cells and mast cells. This phenomenon can be confirmed by histology observation (optical microscope OM, electron microscope EM, Aging skin, edited by JL Leveque & PG Agache, 1993, NY).

In addition, aging skin has a constant change in all aspects such as biochemistry and enzyme histology. In particular, skin metabolic enzyme activity is lowered, oxidoreductase system activity is weakened (SOD, GSH-Px), oxidative damage products (MDA) , Free periods, etc.) are increased. This is the basis of aging theories such as Error catastrophe theory and free-radical theory.

3-1. Sample manufacturing

10 g of stearic acid (C ₁₈ H ₃₆ O ₂ ), 70 mL of water and 20 mL of glycerol were each heated to 80 ° C. and uniformly mixed. After stirring for about 1 minute, peonyflorin or vitamin E (positive control) was added at 3 (w / v)%, methylparaben was added at 0.1 (w / v)%, and stirred for 1 minute. To this was slowly added 1 mL of KOH, after emulsification remarkably, it was stirred to about 50 ° C and packaged.

3-2. Experiment

Guinea pigs were randomly divided into groups, and each group was set to 6 animals, half male and half female. The hair on the dorsal side of the guinea pig was removed at 3 × 3 cm ² . In addition, in order to set its own control group for each guinea pig, the left side was the sample processing side prepared above, and the right side was the processing side of the negative control and the positive control group. It was applied once at 8 o'clock and 16 o'clock every day, and about 5 mg / cm ² each time for 30 days. On the 31st day of the experiment, the skin of the back of the animal was taken, the subcutaneous fat tissue was removed therefrom, and the collagen and lipid peroxidation of the partial skin were measured by electron microscope and light microscope.

3-3. result

1. Visual observation

In the group to which the sample was applied, the appearance improvement effect such as softening and shiny skin was remarkable. This was remarkable compared to the group treated with vitamin E as a positive control group. On the other hand, no apparent improvement in skin was observed in the blank control and the negative control without the peonyflorin.

2. Histological observation-Collagen content

The dorsal skin tissue was paraffin-impregnated and HE (hematoxylin and eosin) stained, followed by skin tissue changes. In addition, skin collagen was measured by adding 6 N hydrochloric acid to the skin and then measuring the content of HYP (hydroxyproline) by chloramine-T oxidation.

As a result, no significant change was observed in the epidermis of the group treated with peonyflorin, but an increase in the number of collagenous fiber bundles was observed in the dermis. Table 3 below shows the effect on the dermal collagen fiber bundle of peonyflorin, it can be seen that peonyflorin improves skin elasticity by increasing the collagen content in the dermis.

Table 3

sample Collagen content increase rate of dermis (%) Collagen fiber bundle growth rate (%) blank － － Peony florin 35.92 * 18.1 * Vitamin E 29.86 * 10.4 * * Comparison with positive control P <0.05

In addition, RER (rough endoplasmie reticulum) expansion and increase in mitochondrial number were observed in the dermis, indicating that peonyflorin increased the activity and function of skin cells.

3. Histological observation-Determination of lipid peroxide content

Superoxide dismutase (SOD) and total glutathione peroxidase (GSH-Px) are the major antioxidant damaging enzymes in organisms, which play an important role in gas oxidation and antioxidant equilibrium. Since they exhibit an antioxidant effect by removing oxygen free groups, SOD and GSH-Px activity can be interpreted as a result showing strength of oxygen free group removal ability in the body. In addition, MDA (Malondialdehyde) represents a lipid peroxide.

MDA detection was performed using thiobarboturic acid, and GSH-Px and SOD were measured by DNTB (5-5'-dithiobis-2-nitrobenzoic acid) and nitrite reduction, respectively. The results are shown in Table 4 below.

(Mathematical formula)

Concentration = (absorbance of treatment group-absorbance of blank) / absorbance of blank x 100

Table 4

sample MDA SOD GSH-Px blank － － － Peony florin 29.57 * 8.58 * 29.51 * Vitamin E 26.71 * 7.17 * 28.60 *

In the results of Table 4, it can be seen that pheoliflorin increases the SOD and GSH-Px activity of the skin tissue and inhibits lipid peroxide production.

 As mentioned above, the composition for inhibiting skin aging comprising the peonyflorin of the present invention enhances the activity and function of skin cells and promotes the synthesis of collagen fibers, thereby contributing to the improvement of toughness and elasticity of skin tissues as well as lipids. By reducing the peroxide production has the effect of significantly inhibiting natural skin aging.

Claims (4)

A composition for inhibiting skin aging comprising peonyflorin as an active ingredient. The composition of claim 1, wherein the content of peonyflorin is 0.001 to 50% by weight. The composition of claim 1, wherein the skin aging is natural aging.  The composition of claim 1, wherein the composition is a medicament, cosmetic, food additive or detergent.