KR102471051B1 - Composition for preventing and alleviating redness and atopy using natural materials, and cosmetics containing the same - Google Patents

Abstract

The present invention relates to a composition for preventing and relieving redness and atopy using natural materials and a cosmetic comprising the same. The present invention provides a plant extract-based composition capable of preventing and improving skin troubles caused by skin redness and atopy, and provides a functional cosmetic containing the composition, which can be used for a long time without side effects and exhibits the above-described activity. It is intended to be used as a functional product depending on the active ingredient.
A cosmetic composition for preventing or alleviating skin flushing and facial flushing according to an embodiment of the present invention is a eoseongcho extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof. In addition, the cosmetic composition for preventing or alleviating atopic dermatitis according to another embodiment of the present invention Eoseongcho extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof.

Description

Composition for preventing and alleviating redness and atopy using natural materials and cosmetics containing the same

The present invention relates to a composition for preventing and relieving redness and atopy using natural materials and a cosmetic comprising the same. In more detail, the present invention provides a plant extract-based composition capable of preventing and improving skin troubles caused by skin flushing and atopy, and provides a functional cosmetic containing the composition, which can be used for a long time without side effects and the above-mentioned It is intended to be used as a functional product according to the active ingredient showing activity.

Redness of the skin, also called skin redness or face redness, is mainly known as a kind of menopausal syndrome, but in addition, it refers to symptoms in which the skin color becomes red due to sensitivity to sunlight, heat, stress, etc.

Skin flushing (or facial flushing) is most severe in early winter when temperature changes are severe and dry, and recently, skin flushing in men as well as women in their 10s and 20s is on the rise. You don't have to worry much about skin troubles that appear and disappear for a short time due to seasonal changes, but if left untreated, they can develop into serious skin diseases such as rosacea and telangiectasis.

It is reported that skin reddening is caused by angiogenic factors. Vascular endothelial growth factor (VEGF) is a representative angiogenic factor that induces angiogenesis, and is an important factor regulating not only angiogenesis but also vasodilation and vasculogenesis in embryogenesis.

On the other hand, it is known that the expression of VEGF is increased by an external stimulus such as ultraviolet light. The secreted VEGF binds to the VEGF receptor present on the surface of vascular endothelial cells and promotes the proliferation of vascular endothelial cells and the secretion of extracellular matrix degrading enzymes, which is involved in the mechanism involved in the movement of vascular endothelial cells and the adhesion of vascular endothelial cells. It is known to do

Angiogenesis is a process in which new blood vessels are created from existing blood vessels. Including the migration of vascular endothelial cells constituting blood vessels, the invasion reaction that passes through the extracellular matrix (ECM), which is the barrier between cells, the growth of cells constituting blood vessels, and the stages of differentiation (tube-like formation) in which blood vessel shapes are created. do.

In a normal state, the equilibrium between angiogenesis inducers and angiogenesis inhibitors is maintained, and when the equilibrium is broken, angiogenesis occurs. In addition to skin flushing and facial flushing, angiogenesis is known to affect skin aging, wound healing, melasma, dark circles, and hair.

On the other hand, when the skin barrier function is damaged by external stimuli, excessive inflammation and immune responses occur, and the damaged tissues and cells secrete various cytokines, chemokines, and growth factors including VEGF. increases. Through this, various immune cells gather in the tissue through the blood and lymphatic vessels, and an inflammatory response occurs.

Angiogenesis and increased vascular permeability caused by secreted VEGF promote a process in which various immune cells gather into damaged tissues through blood vessels, and when continuously occurring, it leads to chronic inflammation. Through this mechanism, the skin appears red, and in severe cases, a phenomenon in which microvessels are visible on the skin surface appears.

Atopy occurs mainly in infancy and is also called fetal fever, and mainly occurs as the content of ceramides in the stratum corneum decreases. Although the cause of atopic dermatitis has not yet been clearly identified, it is presumed to be caused by disturbance of the immune system.

In general, atopic dermatitis is dry due to a decrease in the moisture retention function of the stratum corneum, and as the skin barrier function decreases, skin allergies and the like easily occur, resulting in severe itching, repeated recurrence, and dermatitis easily caused by various stimuli. do. In many cases, it worsens repeatedly at intervals of 1 to 2 weeks, and if proper management is not performed, it progresses to a subacute lesion of erythematous rash accompanied by exudation or a chronic lesion of lichenification thickened like leather.

There are two main causes of atopic skin. One is due to genetic causes, which are due to a congenital deficiency of γ-linoleic acid (GLA), a skin protective film intercellular bonding material. This is caused by the fact that the moisture on the skin's surface is easily lost. Another cause is an environmental cause, which is caused by a dry living environment, active oxygen, bacteria, viruses, fungi, and the like.

In addition, atopic dermatitis (atopic dermatitis) is a chronic recurrent inflammatory skin disease accompanied by pruritus (itching), dry skin, and characteristic eczema. It usually starts in infancy or childhood, but recently it often occurs in adults or older people as well. The cause of atopic dermatitis is not yet known with certainty, and it has been reported that environmental factors, genetic predisposition, immunological abnormality, abnormal skin barrier, etc. are the main causes. Topical steroids and local immunomodulators (calcineurin inhibitors) are the main treatments used as a treatment for atopic dermatitis, and antihistamines are also commonly used to suppress itching. In addition, there is a demand for multifaceted and systematic treatment to avoid allergens, irritants, and stress that aggravate or cause skin symptoms.

KR 10-2010-0120490 A KR 10-2017-0100245 A

An object of the present invention is to provide a natural product-derived composition capable of preventing and alleviating skin flushing and facial flushing.

An object of the present invention is to provide a composition derived from natural products capable of preventing and alleviating atopic dermatitis.

An object of the present invention is to provide a composition that can be used for a long period of time without problems of cytotoxicity and other side effects through the natural product-derived composition.

Another object of the present invention is to provide a cosmetic product containing the composition and the cosmetic product in various formulations.

In order to achieve the above object, the composition for preventing or alleviating skin flushing and facial flushing according to an embodiment of the present invention is Eoseongcho extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof.

The extract may be extracted with any one selected from water, alcohol having 1 to 10 carbon atoms, and mixtures thereof using a solvent.

A composition for preventing or alleviating atopic dermatitis according to another embodiment of the present invention is Eoseongcho extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof.

The extract may be extracted with any one selected from water, alcohol having 1 to 10 carbon atoms, and mixtures thereof using a solvent.

A cosmetic for preventing or alleviating facial flushing according to another embodiment of the present invention may include the above composition.

A cosmetic for preventing or alleviating atopic dermatitis according to another embodiment of the present invention may include the above composition.

Hereinafter, the present invention will be described in more detail.

A composition for preventing or alleviating skin flushing and facial flushing according to an embodiment of the present invention is a houttuynia cordata extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof.

When using the extract, it is possible to relieve skin irritation and provide a cooling effect that lowers the heat generated in the skin. In addition, by improving the blood flow, it is possible to show the effect of preventing and improving skin flushing and facial flushing.

The Eoseongcho (Houttuynia cordata) is also called medicinal wheat and was named Eoseongcho because it smells fishy from the leaves. It shows vigorous fertility like a weed, and is often eaten as a tea.

The bupyeongcho (Spirodela polyrhiza) refers to an annual floating aquatic plant belonging to the duckweed family. Donga (冬芽) generated from the mother body in autumn sinks into the water to overwinter and rises to the surface again the following year to breed, so it can be seen as a perennial, and grows wild in all regions of Korea.

Centella asiatica is a perennial plant commonly found in the mountains and fields of the southern islands of the Korean Peninsula. The main stem extends sideways, and there are two degenerated scale-like leaves near the nodes where the roots come out. , the length of the petiole is 4 to 20 cm.

Preferably, the composition may include bupyeongcho extract. In the case of a single extract, when using the bupyeongcho extract, the effect of preventing and improving skin flushing and facial flushing through blood flow improvement, cooling effect, and anti-inflammatory activity may be most excellent.

Preferably the composition is Eoseongcho extract; bupyeongcho extract; Centella asiatica extract may be mixed. In the case of mixing of each extract, a synergistic effect may appear depending on the difference in active ingredients contained in each extract and the interaction of activities.

More preferably, the composition includes bupyeongcho extract, 30 to 80 parts by weight of eoseongcho extract based on 100 parts by weight of the bupyeongcho extract; 60 to 120 parts by weight of Centella asiatica extract may be mixed. In the case of the above range, it is possible to achieve excellent skin flushing and facial flushing prevention and improvement effects through the synergistic effect according to the interaction of each extract. In addition, through a synergistic effect, it is possible to achieve an excellent effect with a smaller content, so that it can be used for a long time without problems such as cytotoxicity.

The composition includes 1 to 10 parts by weight of peppermint extract based on 100 parts by weight of the bupyeongcho extract; 5 to 20 parts by weight of apple mint; It may further include 1 to 10 parts by weight of sage and 5 to 20 parts by weight of basil extract. In the case of using the composition, the effect of improving skin flushing and facial flushing by improving blood flow along with a skin cooling effect may be more excellent.

The extract may be extracted with any one selected from water, alcohol having 1 to 10 carbon atoms, and mixtures thereof using a solvent.

A composition for preventing or alleviating atopic dermatitis according to another embodiment of the present invention is Eoseongcho extract; bupyeongcho extract; It may be one containing any one selected from the group consisting of centella asiatica extract and mixtures thereof.

The Eoseongcho extract; bupyeongcho extract; When containing any one selected from the group consisting of centella asiatica extract and mixtures thereof, it may exhibit relief and improvement effects on atopic dermatitis.

Preferably, the composition may include bupyeongcho extract. In the case of a single extract, when using the bupyeongcho extract, the activity of alleviating atopic dermatitis and improving the atopic symptoms through antioxidant, anti-inflammatory and antihistamine activities may be high.

Preferably the composition is Eoseongcho extract; bupyeongcho extract; Centella asiatica extract may be mixed. In the case of mixing of each extract, a synergistic effect may appear depending on the difference in active ingredients contained in each extract and the interaction of activities.

More preferably, the composition includes bupyeongcho extract, 30 to 80 parts by weight of eoseongcho extract based on 100 parts by weight of the bupyeongcho extract; 60 to 120 parts by weight of Centella asiatica extract may be mixed. In the case of the above range, it is possible to achieve excellent atopic and atopic dermatitis improvement effects through synergistic effects according to the interaction of each extract. In particular, through the synergistic effect according to the interaction, it is possible to produce an excellent effect with a smaller content, so that it can be used without problems such as cytotoxicity due to long-term use.

The composition includes 1 to 10 parts by weight of peppermint extract based on 100 parts by weight of the bupyeongcho extract; 5 to 20 parts by weight of apple mint; It may further include 1 to 10 parts by weight of sage and 5 to 20 parts by weight of basil extract. In the case of the composition, it is possible to exhibit antioxidative and anti-inflammatory activities against atopic dermatitis itself, as well as an effect of improving atopy, thereby preventing and improving atopic dermatitis.

The extract may be extracted with any one selected from water, alcohol having 1 to 10 carbon atoms, and mixtures thereof using a solvent.

A cosmetic for preventing or alleviating facial flushing according to another embodiment of the present invention may include the above composition.

A cosmetic for preventing or alleviating atopic dermatitis according to another embodiment of the present invention may include the above composition.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, a solution, suspension, paste, gel, cream, lotion, powder, soap, surfactant-containing cleanser, oil , Powder foundation, emulsion foundation, wax foundation, and may be formulated as a spray, but is not limited thereto. More specifically, softening lotion, nourishing lotion, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation, oil-in-water (O/W) type or water-in-oil (W/O) type formulation or ointment.

In addition, if necessary, functional substances such as whitening agents, moisturizing agents, anti-inflammatory agents, antibacterial agents, antifungal agents, vitamins, sunscreens, antibiotics, anti-acne agents, perfumes, and dyes may be included, which are in amounts commonly used in the cosmetic field. It may be included in the cosmetic composition according to the present invention.

A cosmetic composition according to another embodiment of the present invention is prepared by including the composition for skin regeneration and skin moisturizing as an active ingredient.

Regeneration cream according to another embodiment of the present invention is prepared by including the cosmetic composition as an active ingredient.

A moisturizing cream according to another embodiment of the present invention is prepared by including the cosmetic composition as an active ingredient.

The dermatologically acceptable carrier may include purified water, oil, wax, fatty acid, fatty alcohol, fatty acid ester, surfactant, moisture absorbent, thickener, antioxidant, viscosity stabilizer, chelating agent, buffer, preservative, lower alcohol, and the like. However, it is not limited thereto, and the type and concentration are various, and include parts that can be changed by those skilled in the art within the scope of the present invention.

Cosmetics prepared from the cosmetic composition of the present invention can be prepared in the form of general emulsified formulations and solubilized formulations. Emulsified cosmetics include nutrition lotion, cream, essence, etc., and solubilized cosmetics include softening lotion.

The cosmetic may be prepared in the form of an adjuvant that can be applied topically or systemically, which is commonly used in the art, by containing a dermatologically acceptable medium or base in addition to the extract of the present invention.

Suitable cosmetic formulations include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), non-ionic forms. It may be provided in the form of a follicular dispersion, cream, toner, lotion, powder, ointment, spray or conceal stick. In addition, it can be prepared in the form of a foam (foam) or the form of an aerosol composition further containing a compressed propellant.

Products to which the cosmetic composition of the present invention can be added include, for example, cosmetics such as astringent lotion, softening lotion, nutrient lotion, various creams, essences, packs, foundations, etc., and cleansing, face wash, soap, treatment or beauty essence etc. Specific formulations include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrient lotion, massage cream, nutrient cream, moisture cream, hand cream, essence, nutrient essence, pack, soap, shampoo, and cleansing foam. , cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, or eye shadow.

The present invention provides a natural product-derived composition capable of preventing and alleviating skin flushing and facial flushing.

The present invention provides a natural product-derived composition capable of preventing and alleviating atopic dermatitis.

The present invention provides a composition that can be used for a long period of time without problems of cytotoxicity and other side effects through the natural product-derived composition.

The present invention can provide cosmetics containing the composition and the cosmetics in various formulations.

1 relates to cytotoxicity evaluation results according to an embodiment of the present invention.
2 relates to cytotoxicity evaluation results according to an embodiment of the present invention.
3 relates to cytotoxicity evaluation results according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Preparation Example: Preparation of extract]

1. Preparation of bupyeongcho extract (AE)

After drying the washed bupyeongcho, it was pulverized, extracted with hot water, and filtered to prepare bupyeongcho extract (AE).

2. Preparation of Eoseongcho Extract (BE) and Centella Asiatica Extract (CE)

BE and CE were prepared in the same manner as the manufacturing method of the above-mentioned bupyeongcho extract.

3. Preparation of Additional Mixed Formulations

Peppermint extract (P1) in the same manner as in the above AE; Apple mint extract (P2); Sage extract (P3) and basil extract (P4) were prepared, and in order to confirm the synergistic effect of mixing each extract, compositions of M1 to M10 were prepared by mixing in formulations as shown in [Table 1] below.

M1 M2 M3 M4 M5 M6 M7 M8 M9 AE 100 100 100 100 100 100 100 100 100 BE 10 30 50 80 100 50 50 50 50 CE 30 60 90 120 150 90 90 90 90 P1 - - - - - 0.1 One 10 20 P2 - - - - - One 5 20 30 P3 - - - - - 0.1 One 10 20 P4 - - - - - One 5 20 30

(unit: parts by weight)

4. Preparation of Ethanol Extract Formulations

Dried bupyeongcho, eoseongcho, and centella asiatica were respectively ethanol as a solvent, and immersed each bupyeongcho, eoseongcho, and centella asiatica for 48 hours to obtain ethanol extract (AEW) of bupyeongcho; Eoseongcho extract (BEW); Centella asiatica ethanol extract (CEW) was prepared. In addition, the AEW to CEW were replaced with AE to CE to prepare a mixed formulation with the composition shown in [Table 1].

[Experimental Example: Activity Evaluation for Composition]

1. Cytotoxicity evaluation

MTT analysis was performed using the MTT assay, which is an animal alternative test method proposed by the Ministry of Food and Drug Safety and a test standard presented by the Ministry of Food and Drug Safety. After separating the RBL-2H3 cells cultured in the cell culture dish with Trypsin, mix them in a new medium, count the cell lines using a counting chamber and a microscope, and then dispense into 96 well plates at a concentration of 1 to 105 cells/ml. Samples were prepared by concentration and cultured for 24 hours. The cell line RBL-2H3 cells were divided and cultured for 24 hours, and then AE, BE, and CE were treated for each concentration. After removing the medium, MTT solution was added at a concentration of 1 μg/ml and reacted at 37 ° C for 4 hours, followed by MTT. After removing the solution and dispensing 100 μl of DMSO to dissolve the reactants formed, reacting for 30 minutes, the absorbance at 540 nm was measured with an ELISA Reader. The survival rate for each cell was compared and analyzed by expressing the difference in absorbance with respect to the control group as a percentage.

Cell viability (%) = [(absorbance of sample added group)/absorbance of non-added group] X 100

Referring to FIGS. 1 to 3 below, it can be seen that AE, BE and CE according to the present invention do not exhibit cytotoxicity at the highest concentration of 64 μg/ml, respectively. Through this, it can be confirmed that it can be used without problems such as cytotoxicity according to the use and mixing of each extract within the above range.

2. Evaluation of angiogenesis factor expression control ability

As an angiogenic factor following skin stimulation, in order to evaluate the expression inhibitory activity of VEGF (vascular endothelial growth factor), HDF cells were cultured, treated with UVB, stimulated, and secreted for each experimental example treated with each composition according to the ELISA method VEGF was measured. For objective evaluation, the purified water treatment group was set as a control, and the result of the control was fixed as an index of 10, and compared with the control, VEGF measured from 1 to 10 was expressed as an index, as shown in [Table 2] below. In the index below, the lower the number, the better the effect of improving flushing symptoms by suppressing the expression of VEGF.

Con AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 10 7 9 9 9 5 5 6 8 7 3 4 8

(Unit: Index)

Referring to [Table 2], it can be seen that the expression of VEGF is suppressed when the composition according to the present invention is used. In addition, in the case of a single formulation, it can be seen that the activity of inhibiting VEGF is excellent in the case of AE. On the other hand, in the case of a mixed formulation, it can be confirmed that a synergistic effect appears when the range is from M2 to M4. In addition, it can be seen that additional synergistic activity may occur in the case of M7 and M8. Through this, it can be confirmed that the effect of improving the flushing symptom can be achieved with a smaller content than in the case of the mixed formulation in the above range.

3. Evaluation of cooling activity

In order to confirm the skin cooling effect of the composition according to the above preparation example, normal human dermal fibroblast cells were cultured, the medium of the cultured cells was replaced with HBSS, and infrared rays were irradiated for 20 minutes, and each example was treated to express PGE2 synthase protein Inhibitory activity was evaluated. Through this, the alleviating activity of each composition against redness was analyzed according to the expression level of PGE2 synthase protein, which is a factor involved in the increase in skin temperature. On the other hand, purified water was used as a control, and for objective evaluation, the result of the control group was fixed at 10, and the results of each Example were evaluated on a scale of 1 to 10, and are shown in [Table 3] below. The lower the number of the index, the higher the activity of preventing and suppressing flushing through the cooling effect because the expression of PGE2 synthase protein is suppressed.

Con AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 10 8 9 9 9 6 6 7 9 7 4 4 7

(Unit: index)

Referring to [Table 3], it can be seen that the cooling effect is exhibited when the composition according to the present invention is used. In particular, in the case of AE, it can be confirmed that excellent skin cooling activity can be exhibited by suppressing the expression of PGE2 synthase protein. In addition, a synergistic effect according to the interaction can be confirmed in the mixed range of M2 to M4, and it can be confirmed that additional synergistic effects can be derived in the case of M7 and M8.

Therefore, in the case of using the composition disclosed in the present invention, it can be confirmed that skin flushing can be improved through the above-described resistance to UV stimulation and the skin cooling effect. In addition, as described later, it can be confirmed that the prevention and improvement effects of skin flushing and facial flushing can be achieved through excellent reducing power and anti-inflammatory activity.

4. Evaluation of DPPH radical scavenging ability and reducing power

By modifying the Brand-Williams test method, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability was measured for each composition. Each sample was treated with a test concentration of 16 μg/ml, mixed well for 10 seconds, reacted at room temperature for 20 minutes, and absorbance was measured at 525 nm. The DPPH radical content was expressed by converting the absorbance ratio of the sample treatment group and the control group into a % value. For objective evaluation, by fixing Control 1 using purified water to index 1 and fixing control 2 using vitamin C to index 10, the DPPH inhibitory activity according to the above examples was evaluated with an index of 1 to 10, and the following [Table 4]. In the index below, the lower the number, the better the antioxidant activity.

Con1 Con2 AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 10 One 7 9 8 7 6 6 6 8 6 3 3 7

(Unit: Index)

Referring to [Table 4], it can be seen that the antioxidant activity is confirmed in the case of the composition according to the present invention. In addition, in the case of a single extract, it can be confirmed that AE exhibits the best antioxidant activity. In addition, although not shown in [Table 4], in the case of each extract, it was confirmed that the higher the treatment concentration, the higher the antioxidant activity.

On the other hand, in the case of the mixed formulation of M2 to M4 and the mixed formulation of M7 and M8, it was confirmed that the synergistic activity according to the interaction of each active ingredient appeared.

5. Assessment of antihistamine activity

The degranulation phenomenon of mast cells is an important process for releasing histamine, and the released histamine is known as a mediator that induces allergic reactions such as itching and rash. did A blank test group treated with only saline was con1, and a control group treated only with compound48/80 was con2. 16ug/ml of each composition was treated at each concentration and histamine secretion content was measured in the test group treated with compound48/80 for comparative evaluation. For objective evaluation, the result of con 1 and the difference value were evaluated by an index of 1 to 10, and are shown in [Table 5] below. The lower the index, the better the antihistamine activity.

Con2 AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 10 5 9 7 6 4 3 4 6 5 2 3 6

(Unit: index)

Referring to [Table 5], it can be seen that in the case of the present invention, the amount of histamine secretion is suppressed compared to the control group in which the secretion of histamine is stimulated. In particular, in the case of AE, it can be confirmed that the histamine inhibitory activity is very excellent. In the case of some mixed formulations, it can be confirmed that histamine secretion stimulated by Compound 48/80 is suppressed and restored to normal through additional synergistic activity.

Through this, it can be confirmed that the composition according to the present invention can have an effect of suppressing an allergic reaction or improving atopy.

6. Evaluation of β-hexosaminidase secretion inhibitory activity

The RBL-2H3 cell line was dispensed into a 24-well plate so that 2 × 105 cells per well were sensitized with 200 ng/mL of anti-DNP IgE per well and cultured in a 5% CO ₂ incubator for 12 hours. . After washing the cell line twice with siraganian buffer, 16ug/ml of each composition was treated for each concentration and reacted again at 37°C for 30 min. After treatment with DNP-HAS (25 ng/mL) and inducing an allergic reaction for 30 min, the reaction was stopped in an ice bath for 10 min, followed by centrifugation at 12,000 rpm for 3 min. Only the supernatant was recovered and used for the measurement of beta-hexosaminidase. To measure the secretion inhibition of this enzyme, a reaction mixture of 30 μL of the supernatant and 30 μL of 1 mM p-NAG was reacted at 37 ° C for 1 hour, the reaction was terminated by adding 250 mL of 0.1 M carbonate buffer, and a microplate reader (Molecular Devices , USA) was used to measure the absorbance at 405 nm. For objective comparison, the β-hexosaminidase activity for each example was evaluated on a scale of 1 to 10 and is shown in [Table 6] below. The lower the index, the more β-hexosaminidase secretion is suppressed, which means that the allergy response is suppressed and the effect of normal recovery of the generated stimulus is high.

AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 6 9 7 8 4 4 5 7 4 3 2 6

(Unit: index)

Referring to [Table 6], it can be confirmed the inhibitory effect on the allergic reaction according to each preparation example. In particular, the stimulated control group, which was sensitized with anti-DNP IgE and treated with A23187 to induce an allergic reaction, increased the secretion of β-hexosaminidase by 40% compared to the blank test group, and the AE increased by 42% at the highest concentration of 16ug/ml compared to the control group. has decreased. In addition, it was confirmed that in some of the mixed formulations, β-hexosaminidase was largely inhibited and recovered almost similarly to the normal group.

7. Anti-inflammatory and anti-allergic evaluation

(1) Western blot analysis

After processing each sample and incubating for 1 hour, HT1080 cells stimulated with compound48/80 were added with elution buffer (50 mM Tris-HCl, pH 7.5, 0.4% Nonidet P-40, 120 mM NaCl, 1.5 mM MgCl2, 2 mM phenylmethylsulfonyl fluoride). , 80 μg/ml leupeptin, 3 mM NaF and 1 mM DTT) was added and treated at 4°C for 30 min. After electrophoresis of 10 μg of the cell lysate on a 10% Tris-HCl gel, the protein was electrically transferred to a nitrocellulose membrane. Then, 10% skim milk is pretreated on a nitrocellulose membrane, and primary antibodies (anti-IL-13, IL-4) for the target protein are treated, followed by secondary antibody treatment, and chemiluminescent ECL kit (Amersham Pharmacia Biotech) was used to detect the target protein. Western blot bands were observed using LAS3000® image analyzer (Fujifilm Life Science, Tokyo, Japan).

(2) Immunofluorescence assay

RBL-2H3 cells cultured on a slide chamber were treated with anti-DNP-IgE for 24 hours. After processing the sample at the test concentration and incubating for 1 hour, DNP-BSA is treated for 6 hours. Cells were fixed for 15 minutes at room temperature by treatment with 10% formalin fixative, and cell membrane permeability was increased with PBS containing 0.5% tween 20. After pre-treatment with Donkey normal serum, primary antibodies (anti-IL-13, IL-4) against the target protein were dissolved in PBS containing 5% donkey normal serum and 0.1% tween 20, followed by treatment for 24 hours. Next, the remaining primary antibody was washed twice with PBS containing 0.1% tween 20, and a fluorescently labeled secondary antibody (donkey anti-rabbit conjugated CY3) was treated for 1 hour in the same manner as the primary antibody. Finally, after mounting the slide on the fluorescence shield containing DAPI, the target protein was observed using confocal microscopy.

(3) Evaluation

For each example, the anti-inflammatory activity was evaluated by observing the effect on the expression of a target protein that causes inflammatory response and itching. For objective comparison of each composition, TNF-α, IL-13, IL-4, and IL-6 expressions were comparatively evaluated, and each composition was evaluated on a scale of 1 to 10, and is shown in [Table 7] below. In the index below, the higher the number, the better the anti-inflammatory and skin soothing effect.

Con AE BE CE M1 M2 M3 M4 M5 M6 M7 M8 M9 One 6 8 8 6 5 4 5 7 4 3 3 6

(Unit: Index)

Referring to [Table 7], it can be confirmed that the composition according to the present invention can alleviate skin troubles, itchiness, etc. caused by inflammation or allergy with a skin soothing effect along with anti-inflammatory activity.

8. Evaluation of repetition effect according to ethanol extract

In the AE to CE and mixed formulations according to [Table 1], AE to CE were replaced with AEW to CEW to prepare single extracts and complex extracts, and the same evaluation was performed.

As a result, cytotoxicity was not observed at the highest concentration of 64 μg/ml, similar to that of the hot water extract. In addition, in the evaluation of angiogenesis factor expression control ability and cooling activity evaluation, AEW was the best in the case of single formulation, and synergistic effect was confirmed when treatment was performed in the mixed range of M2 to M4 in the case of combined formulation, and additional increase in the case of M7 and M8 effect could be confirmed. In addition, the DPPH radical scavenging ability, reducing power, antihistamine activity, and β-hexosaminidase secretion inhibitory activity were also excellent in AEW in the case of a single composition and in the mixing range of M2 to M4 in the case of a mixed composition, confirming that it was the same as in the case of hot water extraction. . In addition, in the case of M7 and M8, additional synergistic effects were also confirmed. Through this, it can be seen that in the case of the composition, the same trend is confirmed regardless of hot water extraction or ethanol extraction.

This can be understood from the fact that the desired active ingredient contained in the plant and its activity can be obtained using hot water or ethanol as a solvent.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concept of the present invention defined in the following claims are also made according to the present invention. falls within the scope of the rights of

Claims (5)

Eoseongcho extract; Including bupyeongcho extract and centella asiatica extract,
Further comprising peppermint extract, apple mint extract, sage extract and basil extract
A composition for preventing or alleviating skin flushing and facial flushing. According to claim 1,
The extract is extracted with a solvent selected from water, alcohol having 1 to 10 carbon atoms, and mixtures thereof.
A composition for preventing or alleviating skin flushing and facial flushing. Eoseongcho extract; Including bupyeongcho extract and centella asiatica extract,
Further comprising peppermint extract, apple mint extract, sage extract and basil extract
A composition for preventing or alleviating atopic dermatitis. A cosmetic for preventing or alleviating skin flushing and facial flushing, comprising the composition according to claim 1 or 2. A cosmetic for preventing or alleviating atopic dermatitis comprising the composition according to claim 3.

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