KR20050011889A - Anti-aging composition for external application to the skin containing ingredients having a protective effect for the intercellular communication capacity - Google Patents

Abstract

PURPOSE: Provided is an anti-aging composition for external application to the skin which contains ingredients having protective effect on intercellular communication capacity. Therefore, it maintains intercellular communication capacity to prevent and delay skin aging. CONSTITUTION: The anti-aging composition for external application to the skin is characterized by containing, as an active ingredient, 0.001-30.0 wt.% of one of retinoic acid, resveratrol, epigallocatechin gallate or genistein which has protective effect on intercellular communication capacity, normalizing effect on connexin expression and promoting effect on procollagen expression.

Description

Anti-aging composition for external application to the skin containing ingredients having a protective effect for the intercellular communication capacity}

The present invention relates to an anti-aging external composition for skin containing a substance having a protective action against information transfer between skin cells. More specifically, the present invention is a retinoic acid, resveratrol, epigallocatechin gallate having a protective action against information transfer ability between skin cells, normalizing expression of connectin or promoting expression of procollagen. The present invention relates to an anti-aging skin external composition comprising at least one of (epigallocatechin gallate, EGCG) or genistein.

Life is composed of cells that perform various functions, and these cells are composed of a combination of proteins, which are expressions of information stored in genes. Many physiological phenomena for life support, such as growth and differentiation, metabolism, memory and learning, emotional expression, reproduction and aging, are regulated by the organic exchange of information between cells, which are the basic units of life. maintain.

The exchange of information between cells is brought into the cell through signal transduction, in which signals from neurotransmitters such as acetylcholine, various hormones, growth factors, etc. are caused by the recognition of these substances by specific receptors on the cell membrane. By delivery and amplification. Signal transduction in cells is influenced by the three secretory forms of autocrine, paracrine, and endocrine. This is made possible by the dynamic and finely regulated networks, which can cause direct and indirect causes of diseases such as cancer, diabetes, and dementia.

In skin cells, there are many reports on the growth and differentiation of skin cells by these signal transduction processes. Among them, the junctions in the cell membrane play a big role in the signal transduction between cells. One of which is well known (Experimental Dermatology, 9, 77-96, 2000). Among these junctions, the most researched in terms of their function is gap junction, which plays a role of intercellular pathway, maintaining homeostasis, regulating cell growth, and stimulation between cells. In addition to acting as a pathway for signaling that maintains and regulates the physiological activity of cells, such as maintaining a synchronized response, it has recently been a marker of psoriasis skin (Journal of Investigative Dermatology, 111, 72-76, 1989). Or as a marker of aged skin (Experimental Gerontology, 37, 1113-1120, 2002), and its importance in cancer tissue formation is also known.

Recently, the composition and role of connectin (connexin), a protein constituting gap junctions, in keratinocytes, melanocytes and fibroblasts, which are skin cells, have been revealed. Gap junctions can maintain the homeostasis of skin cells such as gap junctions of forming cells, gap junctions of dermal and melanocytes, and gap junctions between dermal cells. Fact has been reported in several literatures (Journal of Cell Science, 113, 1535-1542, 2000).

Therefore, the present inventors can maintain the homeostasis of skin cells by activating signal transmission through gap junction from the above description, and thus can prevent and delay aging, thereby improving information transfer ability between skin cells through gap junction. A study was conducted to search for substances that have efficacy to protect and enhance. In particular, the present inventors have found that the information transfer ability between skin cells is reduced by the addition of TPA (phorbol 12-O-tetradecanoate 13-acetate) or by irradiation of ultraviolet rays and used for the search of the substance.

As a result, the present inventors have found that retinoic acid, resveratrol, epigallocatechin gallate, and genistein are proteins that constitute gap junctions that serve as a pathway for information transmission between skin cells as a function of protecting and improving information transmission ability between skin cells. It has the effect of normalizing the expression of connectin and promoting the expression of collagen precursor procollagen, so it can be used as an active ingredient of the external skin composition for protecting information transfer between skin cells. The present invention was completed by revealing that it can be applied to an anti-aging skin composition for anti-aging.

Therefore, an object of the present invention is characterized by containing as at least one selected from the group consisting of retinoic acid, resveratrol, epigallocatechin gallate and genistein as an active ingredient as a substance having a protective action against information transfer between skin cells. It is to provide a skin external preparation composition for protecting information transfer ability between skin cells.

Another object of the present invention is to provide an anti-aging external composition for skin, characterized in that it has an anti-aging action due to the protective action against information transfer between skin cells.

1 is a method for reducing information transfer between skin cells through gap junction by TPA treatment of retinoic acid, resveratrol, epigallocatechin gallate and genistein, which are substances having a protective effect on information transfer between skin cells according to the present invention. This is the result of confirming the effect.

Figure 2 is to reduce the information transfer between skin cells through gap junction by ultraviolet treatment of retinoic acid, resveratrol, epigallocatechin gallate and genistein as a substance having a protective action against intercellular skin information transfer ability according to the present invention. This is the result of confirming the effect.

Figure 3 is a result confirming the normalizing the leucine expression of retinoic acid, resveratrol, epigallocatechin gallate and genistein as a substance having a protective action against information transfer between skin cells according to the present invention.

Figure 4 is a result confirming the procollagen expression promoting effect of the retinoic acid, resveratrol, epigallocatechin gallate and genistein as a substance having a protective action against information transfer between skin cells according to the present invention.

In order to achieve the above object, the present invention is characterized in that it provides a retinoic acid, resveratrol, epigallocatechin gallate or genistein as a substance having a protective action against information transfer between skin cells. That is, the present invention is characterized by providing information protection function between skin cells by the action of the retinoic acid, resveratrol, epigallocatechin gallate or genistein.

More specifically, the protective function of information transmission between skin cells of the retinoic acid, resveratrol, epigallocatechin gallate or genistein according to the present invention is a function of protecting the information transmission between skin cells through gap junctions. It is due to the expression normalizing action of connectin, which is a constituting protein, and the action of promoting expression of procollagen, which is a precursor of collagen.

In addition, the present invention is characterized in that it provides an external skin composition for protecting information transfer ability between skin cells containing at least one selected from the group consisting of the retinoic acid, resveratrol, epigallocatechin gallate and genistein as an active ingredient. do.

In the present invention, when the retinoic acid, resveratrol, epigallocatechin gallate or genistein is treated, the information transfer between skin cells through the gap junction reduced by other causes, including TPA and ultraviolet light, by recovering By containing any one or more of the substances as an active ingredient, there is a feature to provide an anti-aging external composition for skin that can exhibit an excellent anti-aging action by the protection and enhancement of information transfer ability between skin cells.

In the present invention, it has been found that substances such as retinoic acid, resveratrol, epigallocatechin gallate, and genistein have a protective effect on information transmission ability between skin cells, which has not been previously known.

The substance that may be added to the external preparation composition for skin may be added in an amount of 0.001 to 30% by weight, preferably 0.01 to 20% by weight, based on the total weight of the composition, but is not limited thereto.

Substances having a protective action against information transfer between skin cells are as follows.

Retinoic acid is the body's active form of vitamin A.

Resveratrol is a compound found in a variety of plants, in particular large amounts are detected from red or red wine.

Epigallocatechin gallate is a major ingredient such as green tea.

Genistein is a protein component extracted from soybeans.

The anti-aging external composition for skin according to the present invention is used for the purpose of protecting information transfer ability between skin cells. The formulation is not particularly limited as long as it is a formulation capable of achieving the above object, and may be, for example, an external composition for skin having a formulation of supple cosmetics, nourishing cosmetics, massage cream, nourishing cream, pack, gel, or skin adhesive cosmetic Can be.

In addition, in the external preparation composition for skin of each formulation, other components other than the above components can be appropriately selected and blended by those skilled in the art without difficulty according to the formulation or purpose of use of other external preparations.

Hereinafter, the structure and the effect of the present invention will be described in more detail with reference to test examples. However, the scope of the present invention is not limited to these.

[Test Example 1] Measurement of information transfer ability between skin cells through gap junction

The intercellular information transfer ability of retinoic acid, resveratrol, epigallocatechin gallate and genistein was measured as follows.

Retinoic acid, resveratrol, epigallocatechin gallate and genistein were purchased from Sigma Corporation (St. Louis, MO, USA) and used.

The cell line used in the experiment was based on the results of Dr. Human keratinocytes HaCaT cell line distributed from Fusenig was used to dispense 1.5 × 10 ⁶ cells into 60 mm dishes and incubate for 1 day, then test samples at 1 μM concentration, namely retinoic acid and resveratrol, 2 hours before the start of the test. , Epigallocatechin gallate and genistein, respectively. Cell culture was performed at 37 ° C. and 5% CO ₂ under DMEM (Dulbecos modified eagles medium, GIBCO BRL, Life Technologues) medium containing 10% fetal bovine serum (FBS). The control group was cultured in the same medium without the test substance added. After incubation for 2 hours, the wash buffer was washed with 1 × phosphate buffer (PBS, phosphate buffered saline, Sigma, St. Louis, MO, USA).

After treatment with TPA (phorbol 12-O-tetradecanoate 13-acetate) at a dose of 10 ng / ml, the cells prepared as described above were treated with the test samples of the above concentrations and incubated for 1 hour, followed by El-Fouly's method. The information transfer ability between skin cells was confirmed by scrape-loading / dye transfer (SL / DT) analysis using Lucifer Yellow.

In the same way, the effect of the test sample on the ultraviolet A (UVA) 10J / ㎠ was confirmed, wherein the incubation time after the UV A irradiation was incubated at 37 ℃, 5% CO ₂ conditions to 2 hours.

The effect of the test sample on the intercellular skin information transfer ability of TPA treatment or UV irradiation was observed and compared visually through a fluorescent microscope. The results are shown in FIGS. 1 and 2, respectively.

In FIGS. 1 and 2, the substance observed as granules was photographed in the shape of granules through observation of a fluorescence microscope, in which fluorescent substance lucifer yellow was spread to surrounding cells through a gap junction. It can be seen that the fluorescent material did not spread well in the group treated with TPA or UV as compared to the control material spreading well.

As can be seen in Figure 1, retinoic acid, resveratrol, epigallocatechin gallate and genistein are the same as the information transfer ability in the control group for the reduction of intercellular skin information transfer through gap junctions induced by TPA treatment. It can be expected that the test sample will exhibit an anti-aging effect through maintaining skin always.

In addition, as can be seen in Figure 2, retinoic acid, resveratrol, epigallocatechin gallate and genistein are the information transfer ability in the control group for the reduction of the information transfer between skin cells through the gap junction induced by UVA treatment It shows the same degree of recovery effect, and through this, the test sample can be expected to exhibit anti-aging action through maintaining skin always.

Test Example 2 Expression Measurement of Connexin 43 by RT-PCR Analysis

Total RNA was extracted from cells cultured and tested in the same manner as in Test Example 1 using Trizol (Gibco Laboratories, USA) and then provided in one step RNAPCR kit (AMV) (TAKARA BIO INC. Japan). RT-PCR (reverse transcriptional polymerase chain reaction) analysis was performed. At this time, as a primer, the expression for leucine 43 was confirmed by using a primer for leucine 43 (Bionia, Korea). The results are shown in FIG.

As can be seen in Figure 3, retinoic acid, resveratrol, epigallocatechin gallate and genistein have the effect of restoring the expression of leucine, which is increased by TPA or UVA, to normal levels at control levels, and thus the above test. The sample was expected to exhibit anti-aging through maintenance of homeostasis, and the following Test Example 3 was performed.

Test Example 3 Anti-aging Effect of Retinoic Acid, Resveratrol, Epigallocatechin Gallate, and Genistein on Hairless Mouse Skin

About 40 weeks old hairless mice (female hairless mouse) was tested for the anti-aging effect of the test substance through histological immunohistochemistry for procollagen (procollagen) present in the skin. For use in the test, a cream consisting of each composition of Table 1 was prepared and used as a sample. As a control, a cream having a composition of Comparative Example 1 containing none of the samples was prepared and used.

ingredient Content (% by weight) Comparative Example 1 Example 1 Example 2 Example 3 Example 4 glycerin 3.0 3.0 3.0 3.0 3.0 Butylene glycol 2.5 2.5 2.5 2.5 2.5 Propylene glycol 2.0 2.0 2.0 2.0 2.0 Sukualan 5.0 5.0 5.0 5.0 5.0 Caprylic / Capric Triglycerides 3.3 3.3 3.3 3.3 3.3 Stearic acid 0.5 0.5 0.5 0.5 0.5 Carboxy Vinyl Polymer 0.3 0.3 0.3 0.3 0.3 Liquid paraffin 7.0 7.0 7.0 7.0 7.0 Beeswax 4.5 4.5 4.5 4.5 4.5 Cetostearyl alcohol 1.5 1.5 1.5 1.5 1.5 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 Glycerin Monostearate 4.5 4.5 4.5 4.5 4.5 Sorbitan monosulfate 0.05 0.05 0.05 0.05 0.05 antiseptic Quantity Quantity Quantity Quantity Quantity Retinoic acid - 0.1 - - - Resveratrol - - 0.1 - - Epigallocatechin gallate - - - 0.1 - Genistein - - - - 0.1 Purified water Remaining amount Remaining amount Remaining amount Remaining amount Remaining amount Sum 100.0 100.0 100.0 100.0 100.0

The cream consisting of the compositions of Examples 1 to 4 and Comparative Example 1 was applied to the back of the hairless mouse by using Tegaderm ^TM for 2 days, followed by one more time after the rest for 1 day. The site was biopsy and immunohistostaining for procollagen was performed. The results are shown in FIG.

As can be seen in Figure 4, retinoic acid, resveratrol, epigallocatechin gallate and genistein have the effect of increasing the expression of procollagen compared to the control group, and thus the test substances are the information transfer between skin cells Maintaining homeostasis showed anti-aging activity.

As described above, the present invention contains a retinoic acid, resveratrol, epigallocatechin gallate, and genistein, which are effective in improving information transmission between skin cells and constitute a gap junction that is a pathway for information transmission between skin cells. Providing the effect of normalizing the expression of connectin, which is a protein, and the effect of promoting the expression of procollagen, thereby providing a skin external preparation composition which can achieve an excellent anti-aging effect through maintaining information transfer ability between skin cells. Excellent effect.

Claims (4)

Intercellular skin cell information transfer ability As a substance having a protective action, it contains at least one selected from the group consisting of retinoic acid, resveratrol, epigallocatechin gallate and genistein as an active ingredient. Protective skin external preparation composition. According to claim 1, wherein the intercellular skin information transfer ability protection action is a skin external preparation composition for protecting the skin cell information transfer ability, characterized in that to protect the information transfer between the skin cells through the gap junction. The composition for external application of skin for protecting information transfer ability between skin cells according to claim 1 or 2, wherein the active ingredient is contained in an amount of 0.001 to 30.0% by weight based on the total composition. The skin external preparation composition according to any one of claims 1 to 3 is an anti-aging skin external composition for preventing or preventing skin aging due to a protective effect on information transfer between skin cells.