KR101737276B1 - Composition comprising Artemisia annua essential oil having skin regeneration and anti-obesity - Google Patents

Abstract

The present invention relates to a composition comprising essential oils extracted from Sugarcane mellifera having skin regeneration, antioxidant and anti-obesity effects. The essential oil extracted from the mugwort of the present invention has activity of inducing proliferation of epidermal keratinocyte, activity of inducing type I and type IV collagen synthesis and secretion of epidermal keratinocyte, serine / threonine kinase (AKT) (ERK1 / 2), thereby regulating cell proliferation. Thus, it has an excellent effect on skin regeneration, exhibits an ABTS radical or DPPH radical scavenging activity, exhibits an excellent antioxidative effect, And has an excellent anti-obesity effect.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for skin regeneration and antiobesity,

The present invention relates to the use of essential oils extracted from dogwood mugwort with skin regeneration, antioxidant and anti-obesity effects.

Mugwort is a perennial herbaceous plant with strong vitality and fertility, belonging to Artemisia (Compositae). It is widely grown in Asia, Europe, and America, including China and Korea. About 400 species are distributed worldwide It is known that about 300 species are alive. It is known as a nutritionally superior food because of high contents of green leaf protein and essential fatty acid. Mugwort has been studied for its functional properties such as antioxidant, anticancer, immunity enhancement and availability as livestock feed, and attempts have been actively made to apply it to various foods. In Korea, it has been used in the treatment of calm, convulsions, paralysis and systemic stiffness, abdominal pain, blood clots, chronic hepatitis, anorexia and chronic gastroenteritis. Or has been used as a drinking and horticultural plant. Recently, research on the availability of mugwort as anticancer, antioxidant, antibacterial and livestock feed has been studied. In Korea, researches on myrrh, mugwort, mugwort, and mugwort are the mainstream.

On the other hand, Artemisia annua L. is a perennial herbaceous plant belonging to Asteraceae, distributed worldwide in tropical Asia, and in Korea, it grows wild in various fields and fields across the country. In the oriental medicine, it is used as antipyretic agent, hemostatic agent, dermatological treatment agent and insecticide in the name of 菁 蒿, and the antimicrobial, antiviral and antioxidant activities are known. In China, it has been used as herbal medicine for the treatment of malaria. Artemisinin, a type of terpenoids sesquiterpene lactone, has a strong anti-malarial effect and is currently used as a medicine. It is also found in mature plant fires and leaf keratoses, and it is known to cause a strong deterrent effect on herbivorous insects and mammals, as well as a deadly effect on pathogenic protozoa such as malaria. In addition, recently, it has been evaluated as a herb medicine which has been attracting worldwide attention because it has proved its anticancer activity selectively necrosis of breast cancer cells.

Arteannuin, scopoletin, coumarin and eupatin are the main components of mung bean, and these components are related to anticancer and antimicrobial activities. Studies have been done to find out, and tissue culture and genetic transformation for mass production of major components have been studied. Recently, it has been reported that the antioxidant activity of mugwort, which is one of the medicinal plants with high total antioxidant ability, has been reported by the phenol compounds contained in the sample, and studies on the physiological activity of mugwort It is true.

In addition, it is difficult to obtain cow mugwort and it is difficult to cultivate it. Therefore, it is not commercialized so that it can be consumed as a healthy food of the general public.

Korean Patent No. 1,332,214

Accordingly, the present inventors have completed the present invention by confirming the fact that essential oil extracted from Clostridium diffuseis is effective for skin regeneration, exhibits antioxidative activity, and exhibits an adipocyte differentiation inhibitory effect.

Accordingly, it is an object of the present invention to provide a skin regeneration composition containing mugwort essential oil.

Another object of the present invention is to provide an antioxidative composition comprising the mugwort essential oil.

It is a further object of the present invention to provide an anti-obesity composition comprising dogwood mugwort essential oil.

In order to accomplish the above object, the present invention provides a skin regeneration composition comprising essential oil extracted from Artemisia princeps as an active ingredient.

In one embodiment of the present invention, the essential oils are selected from the group consisting of 1,8-cineol, Germacrene D, caryophyllene, β-Farnesene, Camphor, β-Selinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, α-humulene, But are not limited to, γ-Muurolene, Santolina triene, myrcene, 4-Terpineol, Camphene, α-Copaene, Bicyclogermacrene, β-thujone, 2,6,6,9-tetramethyl-Piperitone-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, Piperitone, -Terpinene, β-Cubebene, 4-methylene-1-methyl-2- (2-methyl-1-propen- 3,7-dimethyl-, (Z) -1,3,6-Octatriene, Eugenol, α-Cedrol, Vulgarone B, 1-bornyl acetate, Borneol L borneol L), 1-Phellandrene, 1,2,3,4,4a, 5,6,8a-octahy 4-methyl-4-methylene-1- (1-methylethyl) -, (1alpha, 4aalpha, 8aalpha) -Naphthalene, terpinolene, terpineol ), 1,3-bis (1,1-dimethylethyl) -Benzene, Azunol, and Artemesia triene.

In one embodiment of the present invention, the essential oil may exhibit activity of inducing proliferation of epidermal keratinocytes, inducing type I and type IV collagen synthesis and secretion of epidermal keratinocytes.

In one embodiment of the present invention, the essential oil may regulate cell proliferation by inducing phosphorylation of serine / threonine kinase (AKT) and extracellular signal regulated kinase (ERK1 / 2).

In one embodiment of the present invention, the essential oil may be used at a concentration of 0.0001 to 1 μg / mL.

In one embodiment of the present invention, the essential oil may be one prepared from a fungus by the steam distillation method.

In addition, the present invention provides an antioxidant composition comprising an essential oil extracted from Sugarcane mugwort as an effective ingredient.

In one embodiment of the present invention, the essential oils are selected from the group consisting of 1,8-cineol, Germacrene D, caryophyllene, β-Farnesene, Camphor, β-Selinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, α-humulene, But are not limited to, γ-Muurolene, Santolina triene, myrcene, 4-Terpineol, Camphene, α-Copaene, Bicyclogermacrene, β-thujone, 2,6,6,9-tetramethyl-Piperitone-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, terpene, 1-methyl-1-propen-1-yl) -1-vinylcycloheptane, 3,7-dimethyl-, (Z) -1,3,6-Octatriene, Eugenol, α-Cedrol, Vulgarone B, 1-boronyl acetate, borneol L, 1- L-Phellandrene, alpha-terpinolene, 1,2,3,4,4a, 5,6,8a (1 alpha, 4a alpha, 8a alpha) -Naphthalene, cis- beta -terpineol, 1,3-bis (4-methyl- 1,1-dimethylethyl) -Benzene, Azunol, and Artemesia triene.

In one embodiment of the present invention, the essential oil may exhibit an ABTS radical or a DPPH radical scavenging activity.

In one embodiment of the present invention, the essential oil may be used at a concentration of 0.1 to 50 μg / mL.

In one embodiment of the present invention, the essential oil may be one prepared from a fungus by the steam distillation method.

In addition, the present invention provides an anti-obesity composition comprising essential oil extracted from Artemisia princeps as an active ingredient.

In one embodiment of the present invention, the essential oils are selected from the group consisting of 1,8-cineol, Germacrene D, caryophyllene, β-Farnesene, Camphor, β-Selinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, α-humulene, But are not limited to, γ-Muurolene, Santolina triene, myrcene, 4-Terpineol, Camphene, α-Copaene, Bicyclogermacrene, β-thujone, 2,6,6,9-tetramethyl-Piperitone-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, terpene, 1-methyl-1-propen-1-yl) -1-vinylcycloheptane, 3,7-dimethyl-, (Z) -1,3,6-Octatriene, Eugenol, α-Cedrol, Vulgarone B, 1-boronyl acetate, borneol L, 1- L-Phellandrene, alpha-terpinolene, 1,2,3,4,4a, 5,6,8a (1 alpha, 4a alpha, 8a alpha) -Naphthalene, cis- beta -terpineol, 1,3-bis (4-methyl- 1,1-dimethylethyl) -Benzene, Azunol, and Artemesia triene.

In one embodiment of the present invention, the essential oil may exhibit an activity of inhibiting adipocyte differentiation.

In one embodiment of the present invention, the essential oil may be used at a concentration of 0.05-5 / / mL.

In one embodiment of the present invention, the essential oil may be one prepared from a fungus by the steam distillation method.

According to the present invention, the essential oil extracted from the clary mugwort exhibits the activity of inducing the proliferation of epidermal keratinocytes, inducing the type I and type IV collagen synthesis and secretion of epidermal keratinocytes, activating the serine / threonine kinase (AKT) It can induce phosphorylation of extracellular signal-regulated kinase (ERK1 / 2) and regulate cell proliferation, thus showing excellent effect on skin regeneration.

In addition, the essential oil extracted from the mugwort shows excellent antioxidative activity, exhibiting an ABTS radical or DPPH radical scavenging activity, exhibiting an activity of inhibiting adipocyte differentiation, and exhibiting an excellent anti-obesity effect.

FIG. 1 is a flowchart illustrating a process of extracting essential oil from cryptomeria japonica according to an embodiment of the present invention. Referring to FIG.
FIG. 2 is a graph showing the effect of the crude oil of mugwort-mugwort essential oil according to an embodiment of the present invention on epidermal keratinocyte migration.
FIG. 3 is a graph showing the effect of the crude oil of Artemisia princeps on the epidermal keratinocyte proliferation according to an embodiment of the present invention.
FIG. 4 is a graph showing the effect of the synthetic oil of the artificial mugwort mugwort on the epidermal keratinocyte signaling process according to an embodiment of the present invention.
FIG. 5 is a graph showing the effect of P. japonica essential oil according to one embodiment of the present invention on the type I or IV collagen synthesis and secretion of epidermal keratinocytes.
FIG. 6 is a graph showing the effect of the ragweed essential oil according to an embodiment of the present invention on the ABTs radical scavenging ability.
FIG. 7 is a graph showing the effect of DPPH radical scavenging ability of P. japonica essential oil according to an embodiment of the present invention.
FIG. 8 is a graph showing the effect of the crude oil of mugwort-mugwort essential oil according to an embodiment of the present invention on differentiation of adipocytes.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description, detailed description of known techniques well known to those skilled in the art may be omitted. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear. Also, terminologies used herein are terms used to properly represent preferred embodiments of the present invention, which may vary depending on the user, intent of the operator, or custom in the field to which the present invention belongs.

Therefore, the definitions of these terms should be based on the contents throughout this specification. Throughout the specification, when an element is referred to as "comprising ", it means that it can include other elements as well, without excluding other elements unless specifically stated otherwise.

The present invention relates to a composition containing essential oils extracted from mugwort, and more particularly to a composition having skin regeneration, antioxidant or anti-obesity effect including essential oils extracted from mugwort.

According to the present invention, the dog milk mugwort essential oil can be prepared from dog wormwood using a conventional extraction method. Preferably by a steam distillation method or a steam distillation-extraction method simultaneously.

According to one embodiment of the present invention, the mugwort essential oil was prepared by washing the mugwort, distilling it with steam distillation method, cooling it at a low temperature, and separating and purifying the mugwort mugwort.

According to another embodiment of the present invention, the essential oil extracted from the crude oil of the present invention can be obtained by washing the dried milk powder, drying and pulverizing it, and subjecting the pulverized product to anhydrous or hydrous alcohol having 1 to 4 carbon atoms, acetone, glycerin, ethylene glycol , Followed by cooling or warming with one or more extraction solvents selected from the group consisting of propylene glycol, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane. For example, a first extraction using one or more of an anhydrous and a lower alcohol having 1 to 4 carbon atoms, a polar solvent containing acetone and butylene glycol, and then successively extraction with ethyl acetate, diethyl acetate, diethyl ether , And a low-polarity solvent including benzene, chloroform, and hexane. Further, the extract obtained in the above may be further concentrated in a distillation apparatus to a suitable temperature and concentrated.

The extraction method may be carried out for about 12 to 96 hours in the case of cold beating, or by aging at a temperature of 4 to 25 DEG C for 1 to 20 days in an anhydrous or hydrolyzed alcohol having 1 to 4 carbon atoms, ethyl acetate or diethyl ether as a solvent, The active ingredient can be extracted. In the case of warming, it is preferable that the heating is carried out at a temperature close to the reflux temperature of the solvent for about 5 to 24 hours although it depends on the kind and temperature of the extraction solvent.

The essential oil extracted from the mugwort according to the present invention contains various physiological activities. More specifically, the essential oil contains 1,8-cineol, Germacrene D, Caryophyllene,? -Farnesene, Camphor,? -Serinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept- 2-ene,? -Humulene,? -Muurolene, Santolina triene, myrcene, 4-terpineol, Camphene, Copa 9-tetramethyl-Piperitone-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, ene, Piperitone,? -terpinene,? -cubebene, 4-methylene-1-methyl-2- (2-methyl-1-propen- vinyl-Cycloheptane, δ-Cadinene, 3,7-dimethyl-, (Z) -1,3,6-Octatriene, Eugenol, α-Cedrol, Vulgarone B ), l-boronyl Borneol L, 1-Phellandrene, 1,2,3,4,4a, 5,6,8a-octahydro-7-methyl-4-methylene- 1- ( 1-methylethyl) -, (1 alpha, 4a alpha, 8a alpha) -Naphthalene, alpha-terpinolene, cis- beta -terpineol, 1,3-bis ) -Benzene, Azunol, and Artemesia triene, but the present invention is not limited thereto.

The content of these components is specifically 6 to 7% by weight of 1,8-cineol, 5 to 7% by weight of Germacrene D, 3 to 7% by weight of caryophyllene, 1 to 3 wt% of β-Farnesene, 1 to 3 wt% of Camphor, 0.5 to 2 wt% of β-Selinene, 0.5 to 2 wt% of Sabinene, Mu] mulrolene, 0.1 to 1 wt% of [alpha] -humulene, 0.5 to 2 wt% of (1R) -2,6,6-trimethylbicyclo [3.1.1] hept- 0.1 to 1 wt.% Of Santolina triene, 0.1 to 1 wt.% Of myrcene, 0.1 to 1 wt.% Of terpineol, 0.1 to 1 wt.% Of camphene, 0.1 to 0.5% by weight of bicyclogermacrene, 0.1 to 0.5% by weight of b-thujone, 2, 6, 6, 9- 0.1 to 0.5% by weight of tetramethyl-Piperitone-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, 0.1 to 0.5% by weight of Piperitone, 0.1 to 0.5% 0.1 to 0.5% by weight of? -Cubebene, 4-methylene-1-methyl-2- (2-methyl (Z) -1,3-dimethyl-1,3-propan-1-yl) -1-vinylcycloheptane in an amount of 0.1 to 0.5% by weight, cadinene in an amount of 0.05 to 0.5% by weight, 0.05 to 0.5% by weight of 6-octatriene, 0.05 to 0.5% by weight of Eugenol, 0.05 to 0.5% by weight of chedrol, 0.05 to 0.5% by weight of Vulgarone B, 1-boryl acetate 0.05 to 0.3% by weight of borneol L, 0.05 to 0.3% by weight of borneol L, 0.05 to 0.3% by weight of 1-Phellandrene, 1,2,3,4,4a, 5,6,8a -octahydro-7-methyl-4-methylene-1- (1-methylethyl) -, (1 alpha, 4a alpha, 8a alpha) -Naphthalene 0.05-0.3 weight%, alpha -pterpinolene 0.05-0.3 weight 0.05 to 0.3% by weight of terpineol, 0.05 to 0.3% by weight of 1,3-bis (1,1-dimethylethyl) -Benzene, 0.01 to 0.1% by weight of Azunol, And 0.01 to 0.1% by weight of Artemesia triene.

In the present invention, the essential oil extracted from Clostridium perfringens contains the components exhibiting the physiological activity described above. The components induce the proliferation of dermal keratinocytes and exhibit a skin regeneration effect (see FIGS. 2 to 5 ), Exhibits an antioxidative effect due to a high radical scavenging activity (see FIGS. 6 and 7) and an effect for inhibiting differentiation of adipocytes (see FIG. 8).

Accordingly, the present invention provides a skin regeneration composition comprising an essential oil extracted from Sugarcane Artemis as an active ingredient.

The essential oil of the present invention has an activity of inducing the proliferation of epidermal keratinocytes, and exhibits activities of inducing type I and type IV collagen synthesis and secretion of epidermal keratinocytes. In addition, cell proliferation can be controlled by inducing phosphorylation of serine / threonine kinase (AKT) and extracellular signal regulated kinase (ERK1 / 2).

The essential oil may be used in a concentration of 0.0001 to 1 / / mL, more preferably 0.001 to 0.1 / / mL when the ragweed essential oil of the present invention is used as a skin regeneration composition. When the essential oil of dog mugwort is used at a concentration of less than 0.0001 μg / mL, the skin regeneration effect is insignificant. When the essential oil is used at a concentration of 1 μg / mL or more, it inhibits the proliferation of skin keratinocyte, It is not preferable.

The skin regeneration composition according to the present invention can be used, for example, as a pharmaceutical composition or a cosmetic composition.

The skin regenerative pharmaceutical composition may further contain a preservative, a stabilizer, a wetting agent or an emulsifying accelerator, a pharmaceutical adjuvant such as a salt and / or a buffer for controlling osmotic pressure, and other therapeutically useful substances, And can be formulated into various parenteral dosage forms accordingly. Parenteral dosage forms, and may be, for example, but not limited to lotions, ointments, gels, creams, patches or spray formulations.

The skin regenerating cosmetic composition is not particularly limited in formulation and can be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing But the present invention is not limited thereto, and may be manufactured by any one or more formulations selected from the group consisting of a foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

Meanwhile, the present invention provides an antioxidant composition comprising an essential oil extracted from Sugar Leaf Mugwort as an effective ingredient.

The essential oil of the present invention exhibits an ABTS radical or a DPPH radical scavenging activity and has an antioxidative effect.

When the essential oil of the present invention is used as an antioxidant composition, the essential oil may be used at a concentration of 0.1 to 50 μg / mL, It can be used at a concentration of 1 to 10 μg / mL. The antioxidative effect is insignificant when used at a concentration of less than 0.1 μg / mL, and when the essential oil is used at a concentration of 10 μg / mL or more, it is not preferable because it does not show a marked increase in the effect due to the increase of the content.

On the other hand, the present invention provides an anti-obesity composition comprising an essential oil extracted from Sugar Beetle as an active ingredient.

The essential oil of the present invention exhibits excellent fat cell differentiation inhibiting activity.

When the essential oil of the present invention is used as an anti-obesity composition, the essential oil may be used at a concentration of 0.05 to 5 / / mL, 0.5 to 1 / / mL. When the concentration of the essential oil is less than 0.05 μg / mL, the effect of inhibiting adipocyte differentiation is insignificant. When the essential oil is used at a concentration of 1 μg / mL or more, It is not preferable.

The antioxidant composition or anti-obesity composition containing the essential oil extracted from the synthetic mugwort according to the present invention may be used as a pharmaceutical composition. The antioxidant composition or the anti-obesity composition containing the essential oil extracted from the mugwort- A pharmaceutically acceptable carrier, excipient or diluent.

The "pharmaceutically effective amount" as used herein refers to an amount sufficient for the physiologically active component to be administered to an animal or a human to exhibit a desired physiological or pharmacological activity. However, the pharmaceutically effective amount may be appropriately changed depending on the severity of symptoms, the age, body weight, health condition, sex, administration route and treatment period of the patient.

The term "pharmaceutically acceptable" as used herein means physiologically acceptable and does not generally cause an allergic reaction such as gastrointestinal disorder, dizziness, or the like when administered to a human. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent.

The compositions of the present invention may also be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal, and may be formulated as a variety of compositions for oral or parenteral administration . ≪ / RTI > The formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.

The composition according to the present invention may be administered through various routes including oral, transdermal, subcutaneous, intravenous or muscular, and the dose of the active ingredient may be varied depending on various factors such as route of administration, age, sex, And the like. In addition, the composition of the present invention may be administered in combination with a known compound capable of raising the desired effect.

Furthermore, the antioxidant or anti-obesity composition according to the present invention can be used not only as a pharmaceutical composition as described above, but also as a health functional food. For example, it can be easily used as a raw material for food, an ingredient, a food additive, a functional food or a beverage.

The term " food " means a natural product or a processed product containing one or more nutrients, preferably a state of being able to be eaten directly through a certain degree of processing, , Food additives, functional foods and beverages.

Examples of foods to which the food composition can be added include various foods, beverages, gums, tea, vitamin complexes, and functional foods. In addition, it is also possible to use special nutritive foods (eg crude oil, milk, infant food, etc.), meat products, fish products, tofu, jelly, noodles (eg ramen, (For example, soy sauce), candy, chocolate, gum, ice cream, milk products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods (Eg, fruit juices, vegetable beverages, beverages, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.). The food, beverage or food additive may be prepared by a conventional production method.

The above-mentioned " functional food " refers to a food group which is imparted with added value to function and express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, Refers to a food prepared by processing a body so as to sufficiently express the body's control function on the body, such as recovery, and the like. Specifically, it may be a health functional food. The functional food may include a food-acceptable food-aid additive, and may further comprise suitable carriers, excipients and diluents conventionally used in the production of functional foods.

The health supplements may be in the form of powders, granules, tablets, capsules or drinks, although not limited thereto.

Hereinafter, an embodiment of the present invention will be described in order to facilitate understanding of the present invention. It should be understood, however, that the following examples are for the purpose of promoting understanding of the present invention, but the present invention is not limited by the following examples.

≪ Example 1 >

Manufacture of mugwort essential oil

In the present invention, distilled water of distilled mugwort was subjected to steam distillation to remove water component, thereby preparing mugwort essential oil.

First, after collecting the collected mugwort, 10 ℓ of the mugwort distillate was obtained through steam distillation by 20 kg of raw milk, cooled at a low temperature, separated and purified, and the aqueous solution was removed from the obtained distillate to obtain an essential oil 100 ml was obtained.

≪ Example 2 >

Analysis of components of mugwort essential oil

The analysis of the components of the mugwort essential oil was commissioned by KBSI (korea basic science institute, seoul center). The GC / MS column was DB5-MS (30m × 50μm, 0.25μm), and the carrier gas (GC / MS) ) Was analyzed by flowing it at 1 mL / min using He. The oven temperature was maintained at 40 ° C - 2 ° C / min - 230 ° C - 5 ° C / min - 300 ° C (5 minutes) and the injection temperature was analyzed at 280 ° C. The interface temperature was 300 ℃, the ion source temperature was 230 ℃, the analyzer temperature was 150 ℃ and the mass range was 40 ~ 800 m / z.

Identification of essential oil components was confirmed by comparing the retention indices (RI) of volatile components with the retention times of standard materials or by comparing mass spectra of volatiles with mass spectra of Wiley 7Nist 05 library. RI was determined under the same conditions as the samples using C ₆ -C ₂₄ (n-alkane). As a result, a total of 37 components were identified and listed in order of higher contents such as 1,8-Cineol, Germacrene D, and Caryophyllene.

≪ Example 3 >

Effect of mugwort essential oil on epidermal keratinocyte migration

In order to investigate the effect of mugwort essential oil on skin epidermal keratinocyte migration, the following experiment was conducted.

Human skin-derived epidermal keratinocytes (HaCat) were cultured in DMEM medium containing 10% fetal bovine serum. In order to investigate the migration rate of epidermal keratinocytes in a 48 well chemotactic boyden chamber, firstly, the lower chamber was treated with 0.0001-1 μg / mL of mugwort essential oil, and the 8 μm pore poly- carbonate membrane. Then, epidermal keratinocytes were treated with 5 × 10 ⁴ cells / 30 μL per well in the upper chamber. The chamber was incubated in a 37 ° C incubator for 90 minutes and the membrane was stained and fixed with a Diff-Quick solution. The number of cells migrated through the membrane was counted and converted to a percentage, thus securing the results of skin epidermal keratinocyte migration of dog mugwort essential oil. The negative control group did not utilize the mugwort essential oil and the positive control group was the epithermal growth factor (EGF 5 ng / mL) used for conventional wound healing and / or skin regeneration.

As a result, as shown in FIG. 2, in the positive control group, migration of keratinocytes was induced by the epidermal growth factor (EGF), whereas the epidermal keratinocyte essential oil of the present invention showed migration And it was confirmed that it did not affect.

<Example 4>

Effect of mugwort essential oil on epidermal keratinocyte proliferation

In order to investigate the effect of mugwort essential oil on skin epidermal keratinocyte proliferation, the following experiment was conducted.

Human skin-derived epidermal keratinocytes (HaCat) were cultured in DMEM medium containing 10% fetal bovine serum. Epidermal keratinocytes may form cells cells per well in ^{96 well microtiter plate 5 X 10 3} / 100μL / well to injection so that the adhesion was from 37 ℃, 5% CO ₂ incubator. 100 μL of mugwort essential oil was added to each well at a concentration of 0.0001 to 1 μg / mL and cultured for 24 hours. Ez-cytox and DMEM were mixed at a ratio of 1: 1, treated with 20 μL of each well, and incubated for 30 minutes in an incubator. The absorbance at 450 nm was measured using an ELISA plate reader (Synerge2, BioTek, USA), and the results were obtained in terms of percentages of skin epidermal keratinocyte proliferation of mugwort essential oil. The negative control group did not utilize the mugwort essential oil, and the positive control group used epithermal growth factor (EGF 50 ng / mL), which has been conventionally used for wound healing and / or skin regeneration.

As a result, as shown in Fig. 3, it was shown that the shit mugwort essential oil induces the proliferation of epidermal keratinocytes in a concentration-dependent manner. In other words, the proliferation of epidermal keratinocytes by EGF (50 ng / mL), well known as epidermal growth factor, was induced by 122.96 ± 1.87%. In comparison with the positive control, 2.28%, and at the concentration of 0.1 μg / mL, the growth induction rate was 147.82 ± 2.48%. These results indicate that the proliferation inducing effect is about 25% or more as compared with that of EGF, effectively inducing the proliferation of keratinocytes.

&Lt; Example 5 >

Effect of Mugwort Essential Oil on Signal Transduction in Epidermal Keratinocyte

To determine the effect of the oil on the phosphorylation of serine / threonine kinase (Akt) and extracellular signal-regulated kinase (ERK1 / 2), which are known to be the main signaling pathways regulating the proliferation and migration of epidermal keratinocytes (P-Akt, Akt, P-ERK1 / 2, and ERK1 / 2) were used for immuno-blotting. The negative control group did not utilize the mugwort essential oil and the positive control group was the epithermal growth factor (EGF 50 ng / mL) used for conventional wound healing and / or skin regeneration.

As a result, as shown in Fig. 4, the shit mugwort essential oil is a signal transduction system that regulates the proliferation and migration of keratinocyte cells. The epinephrine keratinocyte serine / threonine kinase (AKT) and the extracellular signal regulating kinase ERK1 / 2) in a concentration-dependent manner. In particular, it was confirmed that the concentration of 0.1 μg / ml of mugwort essential oil induces phosphorylation of AKT and ERK 1/2 more effectively than that of the known epidermal growth factor, thereby regulating cell proliferation.

&Lt; Example 6 >

Effect of Mugwort Essential Oil on the Form I or IV Collagen Synthesis and Secretion of Epidermal Keratinocytes

In order to evaluate the effects of the mugwort essential oil on the type I or IV collagen synthesis and secretion of epidermal keratinocytes, the following experiment was conducted.

Monoclonal anti-collagen I or IV Ab (2 μg / well / PBS) was injected into a 96-well microtiter plate and coated at RT for 12 hours. After blocking for 1 hour with blocking buffer, samples were treated with 100 μL per well and reacted at RT for 90 minutes. The samples were cultured in medium (CM) of epidermal keratinocyte (CM) treated with culture medium (CM), cell lysate, and milk jelly essential oil of 0.1 μg / mL and epidermal keratinocyte The lysate was processed. Then, polyclonal anti-collagen I or IV Ab was added to each well and reacted at RT for 90 minutes. Then, streptavidin-POD was treated in each well and reacted for 60 minutes at RT. Supersinal ELISA pico chemiluminescent substrate was injected at 100 μL per well and reacted for 10 min. The luminescence was measured using an ELISA plate reader. The type I or IV collagen synthesis and secretion of epidermal keratinocyte of epidermal keratinocyte The results were obtained.

As a result, as shown in Fig. 5, the shit mugwort essential oil significantly promoted the synthesis and secretion of epidermal keratinocyte type I collagen (Fig. 5A) and type IV collagen (Fig. 5B) . In particular, the type I collagen synthesis was induced 164.61 ± 0.34% in the culture medium of epidermal keratinocytes (CM), 154.80 ± 1.19% in type IV collagen synthesis, and the type I collagen And IV collagen synthesis and secretion.

&Lt; Example 7 >

Shag Essential  Antioxidant effect of oil

To confirm the antioxidative effect of mugwort essential oil, scavenging activities of ABTs radicals and DPPH radicals were confirmed.

<7-1> Effect of mugwort essential oil on ABTs radical scavenging ability

To determine the antioxidant activity of ABTs radical scavenging activity, we measured the following.

Reaction reagents were mixed with 7 mM ABTs and 2.45 mM potassium persulfate, and then reacted at room temperature for at least 12 hours. 100 μL of ABTs was reacted with 50 μL of the diluted sample at a concentration of 0.1 to 40 μg / mL for 5 minutes at room temperature, and the absorbance was measured at 734 nm. In the control group, the essential oil was dissolved in place of the sample, and the value was calculated by the following Equation 1.

<Formula 1>

ABTs activity (%) = [1- (absorbance of sample addition group) / (absorbance of no addition group)] x 100

As a result, as shown in Fig. 6, it was compared with ascorbic acid, which is a typical antioxidant. At the concentration of 10 μg / mL, the activity was 1.04 ± 0.68% in the case of ascorbic acid, Was 93.87 ± 0.34%. The antioxidant activity of essential oils was significantly higher than that of ascorbic acid.

<7-2> Effects of DPPH Radical Scavenging Ability on Mugwort Mugwort Essential Oils

The DPPH radical scavenging activity of the mugwort essential oil was measured as follows.

The hydrogen donation effect of DPPH (2-2-Diphenyl-1-picrylhydrazyl radicals) of the mugwort essential oil was measured. 0.2 mM DPPH reagent was diluted with 99% ethanol and used. 100 μL of each diluted sample was taken, and 50 μL of 0.2 mM DPPH reagent was mixed well. After leaving for 15 minutes, the absorbance was measured at 517 nm. In the control group, essential oil was dissolved instead of the sample, and the value was calculated by the following Equation 2.

<Formula 2>

DPPH activity (%) = [1- (absorbance of sample addition group) / (absorbance of no-addition group)] x 100

As a result, as shown in FIG. 7, the activity of ascorbic acid (5 μg / mL), which is a representative antioxidant, was 4.80 ± 2.46%, while the activity of the mugwort essential oil was 16.81 ± 4.81% It was shown to be excellent.

&Lt; Example 9 >

The effect of mugwort essential oil on the differentiation of pre-adipocytes

The mouse embryo 3T3-L1 precursor fat cell line was suspended in Dulbecco's modified Eagle's medium supplemented with 10% calf serum (Dulbecco's modified Eagle's medium) purchased from KCLB (Korean Cell Line Bank) at a concentration of 2 × 10 ⁵ cells / And cultured for 48 hours at 37 ° C in a CO ₂ incubator so as to be in a fusion state. The cells were cultured for 2 days in a different medium (10% FBS, 5 μg / mL insulin, 1 μM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine) and cultured at 0.5 and 1 μg / Respectively. After 2 days of incubation, DMEM medium containing 5 μg / mL insulin and mugwort essential oil were treated at 0.5 and 1 μg / mL concentration to promote differentiation into adipocytes. After culturing for 2 days, the treated dogs were treated with 0.5 and 1 μg / mL of mugwort essential oil and treated with the same dog milk mugwort essential oil after 2 days.

The shape of the cells was observed using an optical microscope (Olympus, Japan). The degree of differentiation of 3T3-L1 cells was stained with Oil-red O. In order to observe the differentiated cells, the cell surface was washed twice with cold PBS, fixed with 4% formalin (in PBS) for 60 minutes, washed once with 60% isopropyl alcohol, washed with water, and sufficiently dried . The dried cells were stained with 300 μL of Oil-red O dye for 1 hour at room temperature, and the cells were washed with water and observed under a microscope. Photographs were taken. After photographing, 100% isopropyl alcohol was used to discolor the stained Oil-red O, and the absorbance was measured at 520 nm and counted.

As a result, as shown in FIG. 8, the cytotoxicity of dog milk mugwort essential oil in 3T3-L1 adipocytes was not found to be cytotoxic at a concentration of 1 μg / mL (A).

In addition, in order to examine the effect of the addition of the essential oil of dog mugwort on lipid peroxidation of 3T3-L1 adipocytes, it was stained with Oil red-o and observed with a microscope (B) In the MDI group, DMEM supplemented with MDI, which is a promoter of lipid differentiation, was treated to induce the differentiation of 3T3-L1 cells in the control group treated with only DMEM without adding the factor MDI to the culture medium. Respectively. In the case of 0.5 μg / mL, the inhibitory activity of adipocyte differentiation was 39.29 ± 7.05%, especially 78.46 ± 0.55% at the concentration of 1 μg / mL. It was confirmed that the differentiation was inhibited in a concentration-dependent manner.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (8)

1,8-cineol, Germacrene D, caryophyllene, beta-Farnesene, Camphor, beta-celllinene β-Selinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, alpha-humulene, gamma- Muurolene, Santolina triene, beta -myrcene, 4-terpineol, Camphene, alpha-Copaene, Bicyclogermacrene, β-thujone, 2,6,6,9-tetramethyl-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, Piperitone, Alpha-terpinene, 1 (S) -6,6-dimethyl-2-methylene-Bicyclo [3.1.1.] Heptane, beta-Cubebene, methyl-2- (2-methyl-1-propen-1-yl) -1-vinyl-cycloheptane, delta-Cadinene, 3,7-dimethyl-, -Octatriene, Eugenol, alpha-Cedrol, Vulgarone B, 1-Bornyl acetate, Borneol L, 1-Phellandrene, 1,2,3,4,4a, 5,6,8a-octahydro-7-methyl-4-methylene-1- (1-methylethyl ) - (1 alpha, 4a alpha, 8a alpha) -Naphthalene, alpha -terpinolene, cis- beta -terpineol, 1,3-bis -dimethylethyl) -Benzene, Azunol, and Artemesia triene in an amount of 0.05 to 5 占 퐂 / mL as an active ingredient. . The method according to claim 1,
Wherein the essential oil is prepared from dogwood by steam distillation. The method according to claim 1,
Wherein the essential oil exhibits an adipocyte differentiation-inhibiting activity. delete 1,8-cineol, Germacrene D, caryophyllene, beta-Farnesene, Camphor, beta-celllinene β-Selinene, Sabinene, (1R) -2,6,6-Trimethylbicyclo [3.1.1] hept-2-ene, alpha-humulene, gamma- Muurolene, Santolina triene, beta -myrcene, 4-terpineol, Camphene, alpha-Copaene, Bicyclogermacrene, β-thujone, 2,6,6,9-tetramethyl-Tricyclo [5.4.0.0 (2,8)] undec-9-ene, Piperitone, Alpha-terpinene, 1 (S) -6,6-dimethyl-2-methylene-Bicyclo [3.1.1.] Heptane, beta-Cubebene, methyl-2- (2-methyl-1-propen-1-yl) -1-vinyl-cycloheptane, delta-Cadinene, 3,7-dimethyl-, -Octatriene, Eugenol, alpha-Cedrol, Vulgarone B, 1-Bornyl acetate, Borneol L, 1-Phellandrene, 1,2,3,4,4a, 5,6,8a-octahydro-7-methyl-4-methylene-1- (1-methylethyl ) - (1 alpha, 4a alpha, 8a alpha) -Naphthalene, alpha -terpinolene, cis- beta -terpineol, 1,3-bis -dimethylethyl) -Benzene, Azunol, and Artemesia triene in an amount of 0.05 to 5 占 퐂 / mL as an active ingredient. . 6. The method of claim 5,
Wherein the essential oil exhibits an adipocyte differentiation inhibiting activity. delete 6. The method of claim 5,
Wherein the essential oil is prepared from dogwood by a steam distillation method.

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