KR101915952B1 - Composition for anti-oxidant, anti-inflammatory and skin regeneration comprising Houttuynia cordata thumb extract as effective component - Google Patents

Abstract

The extract of the present invention is extracted by an extraction method including an enzyme treatment and an ultrahigh pressure homogenization treatment. The extraction method solubilizes the insoluble fibrous component to improve extraction yield and physiological activity, It is possible to secure competitiveness.
In addition, the antioxidant and anti-inflammatory composition comprising the extract of Rhododendron sulphate of the present invention as an active ingredient inhibits the antioxidant function and the generation of NO, which is an inflammation mediator, so that the composition for cosmetics for antioxidant and anti-inflammation and the pharmaceutical for the prophylaxis and treatment of inflammatory diseases Can be usefully used as a composition.

Description

TECHNICAL FIELD The present invention relates to a composition for antioxidant, anti-inflammatory and skin regeneration comprising an extract of Aspergillus niger as an active ingredient,

The present invention relates to a composition for antioxidant, antiinflammation and skin regeneration, which comprises a perennial herb extract as an active ingredient.

Active oxygen generated by the body's enzymes, reductive metabolites, chemicals, pollutants and environmental factors destroys cellular components such as lipids, proteins, sugars and DNA irreversibly and irreversibly, It is known to cause disease. In addition, various peroxides such as lipid peroxides, which are produced as a result of lipid peroxidation by these active oxygen, also cause oxidative destruction to cells, resulting in various dysfunctions and various diseases.

Therefore, attention has been focused on the development of antioxidants such as free radical scavengers or lipid peroxides and other peroxidation inhibitors.

BHA (butylated hydroxy anisole) and BHT (butylated hydroxy toluene) are known as synthetic antioxidants, but they are known to cause serious toxic effects on gastrointestinal mucosa, lung, kidney and circulatory system when overexcited or long-term ingestion. Development of alternative antioxidants is required.

Thus, research on natural antioxidants that are harmless to human body and have excellent antioxidant power has been carried out for a long time and it is expected that natural antioxidants harmless to human body can be developed by exploring free radical scavenging function from plant extracts.

On the other hand, the inflammatory reaction is one of biological tissue defense responses to external stimuli. It is involved in physiological and pathological processes of various diseases and is a mechanism to repair or regenerate damaged tissue by external stimuli. These inflammatory responses are characterized by physicochemical, chemical and bacteriological changes in various tissues such as skin and joint tissues.

It has been reported that inflammation can cause a variety of intractable diseases such as dementia, cardiovascular disease, cancer, obesity, arthritis, and diabetes due to persistent inflammation reaction such as fever, erythema and edema. When inflammatory reaction occurs, inflammatory mediators such as nitric oxide (NO), prostaglandin E ₂ (PGE ₂ ) and inflammatory cytokines are secreted. Nitric oxide (NO) is an indicator of inflammatory response. It is synthesized by enzyme (NOS). The formation of nitric oxide (NO) plays a role in the immune response, but nitric oxide (NO), which is overproduced by iNOS induced by lipopolysaccharide (LPS) or inflammatory cytokines, , Genetic mutations, nerve damage, and the like.

Conventional anti-inflammatory drugs are classified into steroidal and non-steroidal anti-inflammatory drugs. Most synthetic anti-inflammatory drugs are accompanied with various side effects in addition to the main action. Therefore, there is a desperate need for the development of anti- , In particular, the early reaction of inflammation through the inhibition of the generation of nitrogen oxides (NO) is a major target of development of anti-inflammatory drugs.

In addition, skin is the primary protective membrane of the human body, which protects organs in the body from external environmental influences such as temperature and humidity changes, ultraviolet rays, and pollutants, and plays an important role in the maintenance of body homeostasis such as body temperature control. The keratin, which is located at the outermost part of the skin, is a tissue formed by the change of skin cells. The stratum corneum is composed of dead cells. In the dermal layer, the dermally formed cell layer in the connective tissue is lifted to the epidermis, and the cell becomes viable and changes into a firm and regular cell layer. The nucleus and cytoplasm escape and form hard stratum corneum like bricks. This stratum corneum acts as a defense line to protect the moisture of our body from the outermost part of the skin tissue and to protect the various substances coming in from outside while firmly supporting the skin. That is, when the skin surface is scratched, the rapid regeneration ability of the epidermis promotes the recovery of the wound, thereby preventing further infection through the wound and reducing the scarring on the skin surface.

Thus, skin regeneration-promoting materials which are harmless to human body and excellent in formulation stability are very useful for the treatment of wounds on the skin surface and inhibition of scarring, so that research has been carried out, and by exploring the function of regenerating skin from natural substances, It is anticipated that it will be possible to develop natural skin regeneration-promoting materials which are harmless and excellent in formulation stability.

On the other hand, Houttuynia cordata Thunb. Is a medicinal herb called medicinal herb, Chinese medicine, or Chinese medicine. Horseshoe contains a variety of essential oils and is called Horseshoe because it gives off a unique odor such as fishy smell. Saururus chinensis is a perennial plant that blooms in May-July, and it grows mainly in coastal areas such as Jeonnam, Gyeongnam, Ulleungdo and Jeju Island. In Japan, it is widely distributed throughout the country and is widely used as a civilian herb medicine. It is a medicinal plant that has attracted the attention of many people in recent years in Korea and is widely cultivated. According to the Food and Agriculture Agency 's food analysis table, the content of Hwasungcho contains 12.8% of proteins in leaves and stems. The minerals are 25 times higher than rice, 10 times higher than brown rice, especially calcium is 20 times higher than rice. There are also a number of vitamin B blooms that are rare in plants. Hwasungcho is a medicinal herb used for gonorrhea, enteritis, urinary tract infection, pneumonia and bronchitis.

Korean Patent Registration No. 1070469 discloses a composition for improving atopic dermatitis comprising a rhizome extract, a rhizome extract, a Saururus chinensis extract, a Ginkgo extract, and a golden extract. Korean Patent Registration No. 1591872 discloses a composition for improving atopic dermatitis, Allergic and antioxidant functional cosmetic composition containing the extract of the present invention and the cosmetic composition of the present invention. However, the present invention provides a method for extracting the anti-inflammatory, antiallergic and antioxidant functional cosmetic composition, Antioxidant, anti-inflammatory and skin regenerating cosmetic compositions have not been disclosed.

The present invention provides a composition for antioxidant, antiinflammation and skin regeneration, comprising the extract of Aspergillus oryzae as an active ingredient, and the extract of Aspergillus oryzae, which is an effective ingredient, is subjected to enzymatic treatment and ultrahigh pressure homogenization And confirming that the herbal extract of the present invention has excellent antioxidative, anti-inflammatory and skin regenerating effects, thus completing the present invention.

In order to achieve the above object, the present invention provides an antioxidant and anti-inflammatory cosmetic comprising an extract of a perennial herb prepared by a method comprising the step of adding an enzyme to a mixture of water and a hydrolysis, Lt; / RTI >

In addition, the present invention provides a pharmaceutical composition for antiinflammation comprising the above extract of Aspergillus as an active ingredient.

In addition, the present invention provides a cosmetic composition for promoting skin regeneration comprising the above extract of Aspergillus as an active ingredient.

The present invention also provides a pharmaceutical composition for the treatment of wounds comprising the above extract of Aspergillus as an active ingredient.

The extract of the present invention is extracted by an extraction method including an enzyme treatment and an ultrahigh pressure homogenization treatment. The extraction method solubilizes the insoluble fibrous component to improve extraction yield and physiological activity, It is possible to secure competitiveness.

 In addition, the antioxidant and anti-inflammatory composition comprising the extract of Rhododendron sulphate of the present invention as an active ingredient inhibits the antioxidative function and the generation of NO, which is an inflammation mediator, and promotes the regeneration of fibroblasts, A pharmaceutical composition for the prevention and treatment of inflammatory diseases and a pharmaceutical composition for the treatment of wounds.

Fig. 1 shows the result of confirming the regeneration effect of skin cells after UV irradiation of the Hwasungcho extract.
Fig. 2 shows the results of confirming the effect of Hwasungcho extract on inhibition of NO production by LPS in macrophages.

The present invention provides a cosmetic composition for antioxidant and anti-inflammation comprising, as an active ingredient, a perennial herbal extract prepared by a method comprising the steps of:

(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;

(b) dissolving the fish cord powder prepared in the step (a) in water and adding an enzyme to hydrolyze it;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the hydrolyzate treated with ethanol in step (c).

It is preferable to use any one selected from the group consisting of outstretched outposts, leaves, flowers, stems and roots, more preferably outposts, but the present invention is not limited thereto.

The enzyme of step (b) is preferably a carbohydrate hydrolase or a pectin hydrolyzing enzyme, but is not limited thereto.

The carbohydrate hydrolase is any one or more enzymes selected from the group consisting of arabinase, cellulase, beta-glucanase, hemicellulase and xylanase But is not limited thereto.

The pectin hydrolyzing enzyme is preferably one or more enzymes selected from polygalacturonase and pectin esterase, but is not limited thereto.

In the cosmetic composition according to one embodiment of the present invention, it was possible to improve the yield of Houttuynia cordata extract by subjecting Houttuynia cordata to high pressure homogenization after treatment with carbohydrate hydrolase or pectin hydrolase.

In addition, in the method for producing the Hodogacus koreanum extract of the present invention, the step (b) is preferably a step of adding 10 to 14 times (w / v) water adjusted to pH 3.5 to 5.5 to the Hodgkin's powder, After adding pectinolytic enzyme, hydrolysis can be carried out at 30 to 60 ° C for 1 to 12 hours, more preferably 12 times (w / v) of water adjusted to pH 4.5 in the fish starch powder is added to the mixture After enzymatic or pectin degrading enzyme is added, it can be hydrolyzed at 50 ° C for 8 to 12 hours.

The hydrolysis of Hwasungcho powder under the above conditions could improve the extraction yield by improving the extraction yield and improving the elution of the active material by solubilizing insoluble matter of Hwasungcho by low molecular weight or molecular rearrangement.

The ethanol treatment in the step (c) is preferably performed by adding 6 to 8 times (v / v) 100% ethanol to the hydrolyzate at 20 to 30 ° C for 1 to 3 hours, 100% ethanol is added to the hydrolyzate 7 times (v / v) and the mixture is stirred at 20 to 30 ° C for 2 hours at 180 rpm.

In addition, in the method for producing an adult herbal extract of the present invention, the ethanol-treated hydrolyzate may further include centrifuging and recovering the supernatant.

In addition, in the method for producing the Houttuynia cordata extract of the present invention, the ultra-high pressure homogenization of step (d) can be homogenized at 100-120 MPa pressure, and more preferably at 110 MPa pressure.

In the present invention, it is possible to improve the extraction yield of the active material by making the fibers in the Horseshoe extract into a uniformly dispersed phase by enzymatic treatment and ultrahigh pressure homogenization treatment, and by lowering the molecular weight and solubilizing the insoluble dietary fiber.

In the cosmetic composition according to one embodiment of the present invention, the extract of Horseshoe chinensis was confirmed to have an antioxidative effect, a regenerating effect on skin cells and an inhibitory effect on NO production.

The cosmetic composition is preferably, but not limited to, at least one of inflammation selected from atopic dermatitis, acne, and inflammation caused by insect bites.

The method for producing the extract of Rhododendron sulphate according to the present invention is more preferably

(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;

(b) adding Viscozyme or Pectinex to a mixture obtained by adding 10 to 14 times (v / w) of water adjusted to pH 3.5 to 5.5 to the prepared adult flower powder of step (a) Followed by hydrolysis at 30 to 60 ° C for 1 to 12 hours;

(c) adding 6 to 8 times (v / v) 100% ethanol to the hydrolyzate of step (b) and treating the mixture at 20 to 30 ° C for 1 to 3 hours; And

(d) homogenizing the ethanol-treated hydrolyzate of step (c) under ultra-high pressure at a pressure of 100 to 120 MPa.

The viscozyme can be obtained from Aspergillus aculeatus- derived arabinase, cellulase, beta-glucanase, hemicellulase, xylanase, (xylanase).

The pectinex is a complex enzyme comprising a polygalacturonase derived from Aspergillus aculeatus .

The cosmetic composition of the present invention may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, It can have one formulation, but is not limited thereto. The cosmetic composition comprising each of these formulations may contain various bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the cosmetic composition of the present invention is a paste, a cream or a gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Etc. may be used.

When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, Propellants such as carbon, propane-butane or dimethyl ether.

When the formulation of the cosmetic composition of the present invention is a solution or an emulsion, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the cosmetic composition of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is a cleansing agent containing an interfacial active agent, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, acethionate, imidazolinium derivative, methyltaurate, sarcosinate , Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative or ethoxylated glycerol fatty acid ester.

The cosmetic composition of the present invention may further contain an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a moisturizer, a fragrance, a fragrance carrier, a protein, a solubilizer, a sugar derivative, a sunscreen, a vitamin, .

The present invention provides a pharmaceutical composition for antiinflammation comprising an extract of Rhododendron koreanum extract prepared by a method comprising the steps of:

(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;

(b) hydrolyzing the mixture obtained by adding the enzyme to the mixture obtained by adding the water to the prepared whitening powder of step (a);

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the ethanol-treated hydrolyzate of step (c) by ultra-high pressure.

The extract of Hwasungcho, which is an active ingredient of the pharmaceutical composition of the present invention, is as described above.

The extract of Hwasungcho, which is an active ingredient of the pharmaceutical composition of the present invention, was able to confirm the inhibitory effect on NO production.

The pharmaceutical composition is preferably, but not limited to, to ameliorate or treat inflammation caused by at least one selected from atopic dermatitis, wounds, lacerations, acne, and insect bites.

The pharmaceutical composition of the present invention is preferably, but not limited to, a carrier, an excipient and a diluent.

The pharmaceutical composition comprising the extract of Aspergillus oryzae as an active ingredient of the present invention may be various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and it is preferable to select the intravenous, intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine or intracerebral injection methods during parenteral administration, But is not limited to.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the pharmaceutical composition of the present invention may be varied depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease.

The present invention provides a cosmetic composition for accelerating skin regeneration damaged by ultraviolet rays, which comprises, as an active ingredient, a perennial herb extract prepared by a method comprising the steps of:

(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;

(b) dissolving the adult flower powder prepared in step (a) with water and adding an enzyme to hydrolyze the mixture;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the hydrolyzate treated with ethanol in step (c).

The extract of Hwasungcho, which is an effective component of the cosmetic composition for promoting skin regeneration of the present invention, is as described above.

As used herein, skin refers to skin tissue including epidermis, dermis, and subcutaneous fat.

In the present specification, skin cells refer to cells constituting the skin (epidermis, dermis, subcutaneous fat layer), and are not particularly limited. Examples of the skin cells include keratinocytes, melanocytes, and fibroblasts which are present in the epidermis and are responsible for biosynthesis of collagen and elastin.

In this specification, skin regeneration is a broad concept that includes the ability to generate new fibroblasts in response to overall skin aging and scars caused by skin epidermis and the like.

According to one embodiment of the present invention, after treatment with carbohydrate hydrolase or pectin hydrolyzing enzyme, it is possible to increase the yield of Hodgus suis extract and to regenerate damaged fibroblasts by homogenization under high pressure.

The cosmetic composition for accelerating skin regeneration comprising the extract of Aspergillus oryzae as an active ingredient of the present invention can be produced according to a manufacturing method of a person skilled in the art, as described above.

The present invention provides a pharmaceutical composition for the treatment of skin wounds induced by ultraviolet rays, which comprises, as an active ingredient, a perlite extract prepared by a method comprising the steps of:

(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;

(b) dissolving the adult flower powder prepared in step (a) with water and adding an enzyme to hydrolyze the mixture;

(c) treating the hydrolyzate of step (b) with 100% ethanol; And

(d) homogenizing the hydrolyzate treated with ethanol in step (c).

The extracts of Hwasungcho, which is an active ingredient of the pharmaceutical composition for wound healing of the present invention, are as described above.

As used herein, a wound refers to damage to epithelial tissues and connective tissue caused by intrinsic and extrinsic factors, preferably damage to the skin, and particularly preferably includes skin photoaging by burn or ultraviolet stimulation .

According to one embodiment of the present invention, after treatment with carbohydrate hydrolase or pectin hydrolyzing enzyme, it is possible to increase the yield of Hodgus suis extract and to regenerate damaged fibroblasts by homogenization under high pressure.

The pharmaceutical composition for wound healing using the extract of Aspergillus oryzae as an active ingredient of the present invention can be prepared according to the manufacturing method of the person skilled in the art, as described above.

Hereinafter, the present invention will be described in more detail with reference to Preparation Examples and Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Manufacturing example  1. Manufacture of Hwasungcho extract

In order to observe the extraction yield and composition of the extract according to the type of enzyme treatment and high - pressure homogenization, High pressure homogenization; And enzyme treatment and high pressure homogenization parallel treatment were prepared as follows.

(1) Enzyme treatment

30 ml of distilled water adjusted to pH 4.5 in 2.5 g of pulverized cord blood and enzymes (Viscozyme and Pectinex) were added at 50 unit / ml and 500 unit / ml, respectively. lt; / RTI > The enzyme was treated at the optimum time (10 hours and 2 hours), and then 70 ml of 100% ethanol was added. After extraction at 180 rpm for 2 hours at room temperature, the mixture was centrifuged at 12,000 rpm for 15 minutes and then filtered.

(2) Ultra high pressure homogenization treatment

30 ml of distilled water adjusted to pH 4.5 and 70 ml of 100% ethanol were added to 2.5 g of pulverized adult fish and then stirred. The mixture was centrifuged at 12,000 rpm for 15 minutes and then filtered to obtain a filtrate. It was passed through an orifice tube twice.

(3) Enzyme treatment and high pressure homogenization parallel treatment

30 ml of distilled water adjusted to pH 4.5 in 2.5 g of pulverized cord blood and enzymes (Viscozyme and Pectinex) were added at 50 unit / ml and 500 unit / ml, respectively. lt; / RTI > The enzymes were treated at optimal times (10 hours and 2 hours), and 70 ml of 100% ethanol was added. The mixture was extracted at 180 rpm for 2 hours at room temperature (25 ° C), centrifuged at 12,000 rpm for 15 minutes, And then passed through an orifice tube twice under a pressure of 110 MPa.

The viscozyme can be obtained from Aspergillus aculeatus- derived arabinase, cellulase, beta-glucanase, hemicellulase, xylanase, (xylanase), and Pectinex is a complex enzyme comprising polygalacturonase derived from Aspergillus aculeatus . The term " polygalacturonase "

Example  1. Identification of extraction yield

Changes in the extraction yields of various enzymes and ultrahigh pressure treatments were measured for the functional material using the extracts of. As a result, the extraction yield was improved in the enzyme treatment and the extraction yield was improved to 143.9% in the ultrahigh pressure homogenization. In particular, when the enzyme treatment and the ultrahigh pressure homogenization treatment were performed in parallel, the yield enhancement effect was superior to that of the oriental herb powder, and the highest extraction yield was obtained by increasing the ultrahigh pressure homogenization treatment by 194.9% after treatment with Viscozyme (Table 1 ).

Extraction Yield (%) of Hibiscus japonica by Enzymatic Treatment and Ultrahigh Pressure Homogeneity Treatment Treatment conditions Extraction yield Growth rate compared to untreated group (%) Untreated group 15.77 Enzyme V (viscozyme) 24.22 53.58 Enzyme P (pectinex) 30.42 92.90 Ultrahigh pressure homogeneity 38.47 143.9 Enzyme V + Ultra High Pressure Homogeneity 46.50 194.9 Enzyme P + ultrahigh pressure homogeneity 32.06 103.3

In Table 1, the untreated group is a mixture of 30 g of distilled water adjusted to pH 4.5 to 2.5 g of the fish powder.

Example  2. Antioxidant activity of Hwasungcho extract

1. ABTS radical scavenging ability

(ABTS) and 2-potassium persulfate (K ₂ S ₂ O ₈ ) were prepared in the same manner as in Example 1, except that ABTS: potassium sulfate = 2 : 1 in volume ratio to prepare a stock solution, followed by shading for 12 to 16 hours. The stock solution was diluted so that the absorbance value measured at 734 nm was 0.7 ± 0.02, and 50 μl of the extract of Hwasungcho extract was added to 100 μl of the diluted ABTS, followed by reaction at room temperature for 6 minutes. The sample was transferred to a 96-well plate and absorbance was measured at 734 nm.

The IC ₅₀ value represents the concentration value required to inhibit the radical to 50%, and the inhibition rate is calculated as shown in Equation 1 below. In the formula 1, the control group was prepared by adding distilled water instead of the Houttuynia cordata extract.

[Formula 1]

Inhibition rate (%) = {(control O.D value-sample O.D value) / control O.D value} x 100

2. DPPH radical scavenging ability

DPPH (1,1-diphenyl-2-picryl-hydrazyl) solution was added to the Horseradish Kidney Extract and the standard material in the same manner as in Table 2, and the mixture was thoroughly mixed for 30 minutes at room temperature. Absorbance at 517 nm was measured. The inhibition rate was calculated as shown in Equation 2 below.

The reaction solution used for measuring DPPH radical scavenging ability DPPH sample MeOH Sample solvent A 50 μl 100 μl B 100 μl 50 μl C 50 μl 100 μl D 50 μl 100 μl

[Formula 2]

(%) = {[(C-D) - (A-B)] / (C-D)} 100

A: Absorbance of the sample

B: Blank absorbance of the sample

C: Absorbance of distilled water

D: Blank absorbance of distilled water

The results of ABTS analysis and DPPH analysis are shown in Table 3 below.

Antioxidant activity of Hwasungcho extract Treatment conditions
Radical cancellation capacity (%) ABTS DPPH Untreated group 22.13 + 0.96 86.97 + 0.44 Enzyme V (viscozyme) 40.58 ± 1.59 90.26 + - 2.48 Enzyme P (pectinex) 50.48 ± 3.65 92.15 + - 3.15 Ultrahigh pressure homogeneity 40.48 + - 0.54 88.63 + - 1.15 Enzyme V + Ultra High Pressure Homogeneity 55.59 + - 2.68 89.15 ± 3.65 Enzyme P + ultrahigh pressure homogeneity 60.18 ± 1.59 88.16 +/- 1.25

As shown in the above Table 3, the antioxidant activity of the present invention showed better antioxidative effect than that of the casein powder. In particular, the antioxidative effect of ABTS was more than twice as high as that of the untreated group (Table 3) when the enzyme treatment and the ultrahigh pressure homogenization treatment were combined.

Example  3. Measurement of cell regeneration ability of Hwasungcho extract

The regeneration rate of skin cells after UV irradiation was measured in order to investigate the skin regeneration ability by the Hwasungcho extract.

Hs68 cells were cultured at 1 x 10 < ⁵ > cells / mL And then cultured in a CO ₂ incubator for 24 hours. Then, the medium was removed, 500 μl of PBS was added, and the plate was covered with UV for 1 minute and 40 seconds. After UV irradiation, fresh DMEM medium was added, and the Houttuynia cordata extract was treated at a concentration of 100 μg / ml and cultured for 48 hours.

After incubation for 48 hours, the MTT reagent was added and allowed to stand for 3 hours. After removing the supernatant, the formed formazan was dissolved by adding 100 μL of dimethyl sulfoxide (DMSO), and the absorbance was measured at 540 nm.

As a result, the extract of Hwasung extract of the present invention prepared by the manufacturing method including the enzyme treatment and the ultra high pressure homogenization treatment step showed the best skin cell regeneration effect. In addition, it was confirmed that the extract of Hwasungsus korea prepared by the method comprising the enzyme treatment or the ultra-high pressure homogenization step was further regenerated from the skin cells (Fig. 1).

Example  4. In the macrophage of Houttuynia cordata extract LPS  On the production of nitrogen oxides (NO)

The mouse macrophage RAW264.7 cell line was dispensed into a 96-well plate to a final concentration of 1 x 10 ⁶ cells / ml, and then cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours. Then, 400 μg / ml And then treated with LPS (1 μg / ml) for 1 hour and cultured for 24 hours. 100 μl of the cell culture solution and 100 μl of the grease reagent were mixed and reacted at room temperature for 15 minutes. The absorbance was measured at 540 nm using a microplate reader, and a standard curve was prepared using sodium nitrate to calculate NO production amount .

Experimental results showed that NO production induced by LPS decreased during enzyme treatment and decreased NO production even under ultrahigh pressure homogenization treatment.

In particular, when the enzyme treatment and the ultrahigh pressure homogenization treatment were performed in parallel, it was confirmed that LPS-induced NO production was further suppressed (FIG. 2).

Claims (11)

delete delete delete delete delete delete delete delete delete A cosmetic composition for promoting regeneration of skin damaged by ultraviolet rays comprising an extract of Rhododendron koreanum extract prepared by a method comprising the steps of:
(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;
(b) dissolving the Horseshoe powder prepared in step (a) with water and adding biscozymes or pectinex to hydrolyze the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the hydrolyzate treated with ethanol in the step (c) at a pressure of 100 to 120 MPa. A pharmaceutical composition for the treatment of skin wounds induced by ultraviolet rays comprising an extract of Aspergillus niger produced by a method comprising the steps of:
(a) drying the crustacea and crushing the crustacea to produce a crustacea powder;
(b) dissolving the Horseshoe powder prepared in step (a) with water and adding biscozymes or pectinex to hydrolyze the mixture;
(c) treating the hydrolyzate of step (b) with 100% ethanol; And
(d) homogenizing the hydrolyzate treated with ethanol in the step (c) at a pressure of 100 to 120 MPa.

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