KR102197883B1 - A composition comprising heat water extract of gojiberry, fig, and agastache rugosa as active ingredients - Google Patents

Abstract

The present invention relates to a composition comprising a hot water extract of Goji berry, fig and pear chow scent, and exhibits a synergistic effect in antioxidant activity, immunity enhancement, whitening, wrinkle improvement, collagen degrading enzyme inhibition, collagen production promoting activity, while cytotoxicity is It is found to be low, and is suitable for antioxidant, immunity enhancement, whitening, and skin wrinkle improvement.

Description

A composition containing hot water extracts of Goji berry, fig and pear chorion as active ingredients {A COMPOSITION COMPRISING HEAT WATER EXTRACT OF GOJIBERRY, FIG, AND AGASTACHE RUGOSA AS ACTIVE INGREDIENTS}

The present invention relates to a composition for antioxidant, immunity enhancement, whitening, or wrinkle improvement comprising hot water extracts of Goji berry, fig and pear chorion as an active ingredient.

Due to the recent development of science and technology and modern medicine, the average life span of humans is gradually increasing, and accordingly, interest for health is increasing in order to improve the quality of life. This trend is increasing regardless of generation and age. In addition, as a lot of interest in medicines, health supplements, and cosmetics containing natural products is increasing, research and development for this is increasing. In particular, the efficacy of natural medicines and the functions of health supplements and cosmetics including natural products are expanding to the basic medical scope such as antioxidant, cholesterol suppression, obesity prevention effect, immunity and disease prevention and aging suppression.

Gojiberry is also known as Chinese boxthorn or Matrimony Vine, and represents a variety of species belonging to the Solanacea family and Lycium genus. Among the various species in the genus Lycium , Lycium barbarum and Lycium chinense are most commonly used in oriental medicine. For example, Lycium, also known as Fructus lycii The barbarum fruit has long been known as a medicinal plant that helps maintain health in general, especially in Asia. In addition, recent studies report that Lycium barbarum not only promotes and regulates the immune system, but also has anti-aging properties and anti-cancer properties, reducing blood fat and sugar levels, or reducing arterial blood pressure. U.S. Patent Application Publication No. 2006/0198810 discloses the efficacy of a mixture of rose fruit, Chinese angelica extract and wolfberry extract to promote fat synthesis of skin cells, in particular. Also Lycium The properties of certain constituents of barbarum are already known. According to this, zeaxanthin dipalmitate is known for its antioxidant and anti-inflammatory properties, whereas Lycium barbarum Polysaccharides (LBP) and 2-O-(β-D-glucopyranosyl)-ascorbic acid (an analog of vitamin C) are known for their immunostimulatory and antioxidant properties.

Fig Tree ( Ficus carica L) is a deciduous shrub belonging to the Moraceae family and is native to the western Mediterranean coast of Asia. It is 2~4m in height, and the bark is grayish-white, the leaves are displaced and thick in the shape of a wide egg, the surface is rough, and there are hairs on the back side. Emulsion is known to be effective for athlete's foot, boils, and hemorrhoids, leaves for athlete's foot and neuralgia, and fruit for lowering blood pressure and constipation. Republic of Korea Patent Registration No. 10-0512285 discloses a medicinal soap composition having a fungal and antibacterial effect containing a fig fruit or leaf extract isolate.

Agastache rugosa (Fisch. et Meyer) O. Kuntze, Kwakhyang) is a perennial herb of the Lamiaceae family (Labiatae), and is used as a medicine after drying the above-ground part and outpost. The herbal name is called Gwakhyang or baechohyang (Latin name: Agastachis herba), and in China, Pogostemon cablin of Dongil is called Gwanggwakhyang (Latin name: Pogostemonis herba) and is commonly used with Kwakhyang. Kwakhyang has traditionally been used as an herbal medicine for anorexia, vomiting, dry stomach, diarrhea, and antipyretic, and research on this has been conducted by using a combination of amino acids and plant extracts such as Kwakhyang to moisturize and lubricate the stratum corneum. There are studies on external preparations (Japanese Unexamined Patent Publication No. 9-71527). Substances such as methylchavicol, rosmarinic acid, and anethole have been reported through the study of ingredients for Kwakhyang.

The inventors of the present invention have been studying anti-oxidation, immunity enhancement, whitening, and skin aging anti-aging substances originating from natural products. Compositions containing hot water extracts of goji berry, fig and pear chow scent, antioxidant activity, immunity enhancement, whitening, wrinkle improvement, collagen The present invention was completed by confirming that the cytotoxicity was low while exhibiting a synergistic effect in inhibiting degrading enzymes and promoting collagen production.

It is an object of the present invention to provide a composition for antioxidant, immunity enhancement, whitening, collagen decomposition enzyme inhibition, collagen production promotion, and wrinkle improvement comprising hot water extracts of Goji berry, fig and pear chorion as active ingredients.

The present invention provides an antioxidant composition comprising the extract of Goji berry, fig and pear chorion as an active ingredient.

In another aspect, the present invention provides a composition for enhancing immunity comprising an extract of Goji berry, fig and pear chorion as an active ingredient. As used herein, the term "immunity enhancement" refers to increasing the immune response or activity of the immune system in vivo.

In another aspect, the present invention provides a composition for skin whitening comprising an extract of Goji berry, fig and pear chorion as an active ingredient. In the present invention, "skin whitening" refers to a result of inhibition of melanin production, and specifically, as melanin production is inhibited, symptoms resulting from an increase in melanin, such as melasma, freckles, skin aging, etc. Can be understood.

In another aspect, the present invention provides a composition for inhibiting collagen degrading enzyme comprising the extract of Goji berry, fig and baechohyang as an active ingredient.

In another aspect, the present invention provides a composition for promoting collagen production comprising the extract of Goji berry, fig and pear chorion as an active ingredient.

In another aspect, the present invention provides a composition for improving wrinkles, comprising the extract of Goji berry, fig and pear chorion as an active ingredient.

When the extract is obtained by treating an extraction solvent, various extraction solvents may be used, and preferably, a polar solvent may be used. Suitable polar solvents include purified water, alcohol (methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol, etc.), acetic acid, DMFO ( dimethyl-formamide) and DMSO (dimethyl sulfoxide), and more preferably, the extraction solvent used in the present invention is purified water, an anhydrous or hydrated lower alcohol having 1-4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), It may contain a mixed solvent of the lower alcohol and purified water. Most preferably, the extract of the present invention is a hot water extract.

As used herein, the term "extract" has the meaning commonly used as a crude extract in the art as described above, but broadly includes a fraction obtained by additionally fractionating the extract. That is, the extract according to the present invention includes not only those obtained by using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a cut-off value with a constant molecular weight, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods carried out are also included in the extract. In addition, the extract according to the present invention may be prepared in a powder state by an additional process such as re-extraction, distillation under reduced pressure, freeze drying, spray drying, or a combination thereof.

According to an embodiment of the present invention, the extract is (i) Goji berry 5 to 20 parts by weight, or 7 to 12 parts by weight, or 10 to 15 parts by weight; (ii) 5 to 20 parts by weight of figs, or 7 to 12 parts by weight, or 10 to 15 parts by weight; And (iii) 60 to 90 parts by weight of baecho flavor, or 65 to 85 parts by weight, or 70 to 80 parts by weight, and the mixture may be extracted.

According to one embodiment of the present invention, the extract may be obtained by mixing the extracts for each of Goji berry, fig and pear chorion in a weight ratio of 1:1:1, 2:1:1, or 2:1:2.

According to one embodiment of the present invention, the composition of the present invention comprises a pharmaceutically effective amount of the extracts of Goji berry, fig and pear chorion of the present invention described above; And it is a pharmaceutical composition comprising a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the above-described efficacy or activity of the extract.

When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used at the time of formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. It does not become. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components. In addition, the pharmaceutical composition of the present invention can be administered orally or parenterally, and is preferably applied by way of oral administration.

The appropriate dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration mode, patient's age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity. It can be, and the general dosage is in the range of 0.001-100 mg/kg for adults.

Another aspect of the present invention provides a food composition comprising the extract of Goji berry, fig and pear chorion of the present invention as an active ingredient.

The food composition according to the present invention includes all forms such as functional food, nutritional supplement, health food and food additives. These types can be prepared in various forms according to conventional methods known in the art. For example, as a health food, the extracts of Goji berry, fig and pear chorion of the present invention can be prepared and consumed in the form of tea, juice and drinks, or granulated, encapsulated, and powdered to be consumed. In addition, in addition to the extracts of Goji berry, fig and pear chorion of the present invention, a known substance or active ingredient known to have an immunity enhancing, whitening, collagen degrading enzyme inhibition, collagen production promotion, and wrinkle improvement effect is mixed together to form a composition. It can be manufactured in a form.

When the composition of the present invention is prepared as a food composition, it includes, as an active ingredient, the extracts of the goji berry, fig and pear chowder, as well as ingredients commonly added during food production, for example, protein, carbohydrate, fat, Contains nutrients, flavoring and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink, in addition to the extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, headworm extract, jujube extract, licorice extract, etc. I can make it.

In the food composition of the present invention, the preferred content of the extract of the goji berry, fig and pear chow flavor of the present invention is 0.01 to 100% by weight or 0.01 to 50% by weight of the finally prepared food. In order to use the extract of Goji berry, fig, and pear-cho scent of the present invention in the form of a food additive, it may be prepared and used in the form of a powder or a concentrate.

Another aspect of the present invention provides a cosmetic composition comprising the extract of Goji berry, fig and pear chorion of the present invention as an active ingredient.

The cosmetic composition according to the present invention may be prepared in any formulation conventionally prepared in the art, and in this case, it may be prepared including a carrier or the like commonly used in the formulation of the cosmetic composition. According to one embodiment, the carrier may be included in about 1% by weight to about 99.99% by weight, preferably about 5% by weight to about 99% by weight, based on the total weight of the cosmetic composition of the present invention.

According to one embodiment, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. It may be formulated, but is not limited thereto. More specifically, when the composition of the present invention is used as a cosmetic composition, a basic cosmetic formulation selected from lotions, emulsions, creams, essences, gels, packs and cleansing creams; It can be prepared in a formulation selected from the group consisting of formulations for makeup cosmetics such as foundation.

When the cosmetic composition according to an embodiment is formulated as a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or Zinc oxide or the like may be used. When the composition of the present invention is formulated as a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In particular, in the case of a spray, additionally chlorofluorohydro Propellants such as carbon, propane/butane or dimethyl ether.

When the cosmetic composition according to an embodiment is formulated as a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl Benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the cosmetic composition according to an embodiment is formulated as a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth, and the like may be used.

The composition comprising the extracts of Goji berry, fig and pear chow scent of the present invention as an active ingredient achieves excellent effects through synergy in antioxidant, immunity enhancing, whitening, collagen degrading enzyme inhibition, collagen production promotion, and wrinkle improvement effects. .

1 is a graph showing the antioxidant effect of hot water extracts of Goji berry, fig, and pear chorion.
2 is a graph showing the cell viability of hot water extracts of Goji berry, fig, and pear chorion.
3 is a graph showing the effect of hot water extracts of Goji berry, fig, and pear chorion on the secretion rate of TNF-α of macrophages.
4 is a graph showing the effect of hot water extracts of goji berry, fig, and pear chorion on melanin production.
5 is a graph showing the effect of inhibiting tyrosinase activity of hot water extracts of goji berry, fig, and pear chorion.
6 is an electrophoresis result showing the effect of inhibiting the expression of the MMP-1 gene derived from keratinocytes by hot water extracts of Goji berry, fig, and pear chorion.
7 is a graph showing the effect of promoting the production of type 1 procollagen by hot water extracts of goji berry, fig, and pear chorion.
8 is a graph showing the effect of increasing the activity of PPAR-α,δ by hot water extracts of Goji berry, fig, and pear chorion.

Since the present invention can apply various transformations and have various embodiments, specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is for illustrative purposes of one or more embodiments, and the scope of the present invention is not limited to these examples, and it is understood to include all transformations, equivalents, and substitutes included in the spirit and scope of the present invention Should be. In describing the present invention, when it is determined that a detailed description of a related known technology may obscure the subject matter of the present invention, a detailed description thereof will be omitted.

Manufacturing Example 1

Manufacturing Example 1-1

10 times of distilled water was added to each of 50 g of goji berries, 50 g of figs, and 50 g of pear chowder, extracted by heating at 95 degrees for 4 hours using a heating mantle equipped with a reflux cooling device, filtered, concentrated, and frozen. Dry to obtain each extract.

Preparation Example 1-2

The Goji berry extract, fig extract, and baechohyang extract of Preparation Example 1-1 were mixed in a weight ratio of 1:1:1 and 2:1:2, respectively.

Manufacturing Example 2

Manufacturing Example 2-1

After mixing 14.93 g of Goji berries, 9.29 g of figs, and 75.78 g of pear chowder, add 10 times of distilled water, and extract by heating at 95°C for 4 hours using a heating mantle equipped with a reflux cooling device, filtered, and concentrated. The extract was obtained by lyophilization.

Manufacturing Example 2-2

Goji berry, fig, and pear-cho scent were extracted, filtered, and concentrated in the same manner as in Preparation Example 2-1, and then spray-dried to obtain an extract.

Experimental Example 1: Measurement of antioxidant activity

Antioxidant activity was investigated using DPPH (2,2-diphenyl-1-picrylhydrazyl). After each sample was treated with DPPH (250 μM) solution (positive control: Vitamin C, 50 ug/mL), a reaction was induced at 37°C for 30 minutes. The antioxidant activity was calculated through the OD value obtained by measuring the absorbance at 517 nm after completion of the reaction, and the results are shown in FIG. 1.

As shown in Fig. 1, the group obtained by mixing the extracts obtained by hot water extraction of Goji berry, fig, and pear-cho scent separately at a ratio of 1:1:1, and Goji berry, fig, and pear-cho scent were mixed in 14.93, 9.29 and 75.78 parts by weight, respectively, and then mixed. As a result of measuring the antioxidant activity in the extracted and freeze-dried or spray-dried groups, it was confirmed that the antioxidant effect was highest in the experimental group extracted after mixing. 1, it was confirmed that the synergistic effect was more remarkable in terms of antioxidant efficacy in the method of extracting after mixing than the method of extracting and then mixing goji berry, fig, and pear-cho scent individually.

Experimental Example 2: Measurement of cell viability

2-1. Cell culture

RAW264.7 cells, a mouse macrophage cell line, were subjected to a density of 70-80% in a 100 mm cell culture dish using RPMI 1640 medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Incubate with.

2-2. Cell viability measurement

It was analyzed using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphinyltetrazolium bromide) assay method. The cultured mouse macrophage line RAW264.7 was adjusted to a concentration of 1-10 ⁶ cells/ml using RPMI 1640 medium, then inoculated into a 96 well plate, and pre-cultured at 5% CO2 and 37°C for 18 hours. Then, the medium was removed, and the medium containing the extracts at concentrations of 100, 200, and 400 ug/mL was treated and cultured. After 24 hours, 10 µl MTT solution (stock concentration: 5 mg/ml) was added and further reaction was induced for 3 hours. To complete the reaction and dissolve the formazan crystal, 100 µl DMSO was additionally added to each well. The cell viability was calculated through the OD value obtained by measuring the absorbance at 570 nm of the amount of MTT reduced to formazan, and the results are shown in FIG. 2.

As shown in FIG. 2, the group obtained by mixing the extracts extracted with hot water separately from Goji berry, fig, and pear chorion at a ratio of 1:1:1, and Goji berry, fig, and pear chow scent were mixed in 14.93, 9.29, and 75.78 parts by weight, respectively, and then mixed. It was confirmed that the cell viability in the group treated with the cells by extraction and lyophilization or spray drying was in the range of 80 to 120%. Based on the results, it was confirmed that the concentration range in which the extracts were tested did not affect cell viability.

Experimental Example 3: Immunity-enhancing effect according to extract concentration-Measurement of TNF-α production derived from macrophages

The cultured mouse macrophage line RAW264.7 was adjusted to a concentration of 1-10 ⁶ cells/ml using RPMI 1640 medium, then inoculated into a 96 well plate, and pre-cultured at 5% CO ₂ and 37°C for 18 hours. . Thereafter, the medium was removed, and the medium containing the extract and the control (positive control: LPS, 100 ng/mL) at concentrations of 100, 200, and 400 ug/mL, respectively, was treated and cultured. After 24 hours, the supernatant was transferred to another 96 well plate by 100 l each, and the TNF-α concentration was measured according to the manufacturer's instructions using an enzyme-linked immunosorbent assay (ELISA) kit of R&D systems, and the results are shown in FIG. Done.

As shown in Fig. 3, the group obtained by mixing the extracts obtained by extracting hot water separately from Goji berry, fig, and pear chorion at a ratio of 1:1:1, and Goji berry, fig, and pear chow scent were mixed in 14.93, 9.29, and 75.78 parts by weight, respectively, and then mixed. As a result of measuring TNF-α production in the extracted and freeze-dried or spray-dried groups, it was confirmed that the highest in the experimental group extracted after mixing. Through the above results, it was confirmed that the synergistic effect of extracting after mixing was remarkable than the method of extracting and then mixing goji berry, fig, and pear-cho scent, respectively.

Experimental Example 4: Measurement of melanin production

4-1. Cell culture

B16-F10 cells, a mouse melanoma cell line, were cultured at a density of 70-80% in a 100 mm cell culture dish using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Incubate with.

4-2. Melanin concentration measurement

B16-F10 cells, a mouse melanoma cell line, were adjusted to a concentration of 1×10 ⁵ cells/ml using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Then, it was inoculated into a 96 well plate, and pre-incubated at 5% CO ₂ and 37°C for 18 hours. Subsequently, the medium was removed and cultured by simultaneously treating the medium containing each extract and control (positive control: PTU (Phenylthiourea), 10 uM) at a concentration of 1600 ug/mL and alpha-Melanocyte-stimulating hormone (α-MSH). After 24 hours, the medium was removed and 1N NaOH was added to dissolve melanin. The melanin content was calculated by measuring the absorbance at 405 nm, and the results are shown in FIG. 4.

As shown in FIG. 4, the group obtained by mixing the extract obtained by extracting hot water with each of Goji berry, fig, and pear chorion alone in a ratio of 1:1:1, and the Goji berry, fig, and pear chow scent were mixed in a ratio of 2:1:2 and then extracted. And as a result of measuring melanin production in the freeze-dried or spray-dried group, it was confirmed that melanin production was reduced to a similar degree to that of the positive control.

Experimental Example 5: Tyrosinase enzyme activity measurement

B16-F10 cells, a mouse melanoma cell line, were adjusted to a concentration of 1×10 ⁵ cells/ml using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Then, it was inoculated into a 96 well plate, and pre-incubated at 5% CO ₂ and 37°C for 18 hours. Subsequently, the medium was removed and cultured by simultaneously treating the medium containing each extract and control at a concentration of 1600 ug/mL and alpha-Melanocyte-stimulating hormone (α-MSH). After 10 minutes, the cells were broken using RIPA buffer and then centrifuged at 14000 rpm for 20 minutes at 4° C. to separate the supernatant. Thereafter, 0.1% L-DOPA (L-3,4-dihydroxyphenylalanine) was added and incubated at 37° C. for 3 hours. Tyrosinase enzyme activity was calculated by measuring absorbance at 475 nm, and the results are shown in FIG. 5.

As shown in FIG. 5, the group obtained by mixing the extracts obtained by hot water extraction of Goji berry, fig, and pear chorion alone at a ratio of 1:1:1, and Goji berry, fig, and pear chow scent were mixed in 14.93, 9.29, and 75.78 parts by weight, respectively, and then mixed. As a result of measuring the Tyrosinase enzyme activity in the extracted, freeze-dried or spray-dried group, it was confirmed that the enzyme activity was reduced to a similar degree to that of the control group.

Experimental Example 6: Measurement of expression of MMP-1 gene derived from keratinocytes

6-1. Cell culture

HaCaT cells, a human keratinocyte line, are cultured at a density of 70-80% in a 100 mm cell culture dish using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. .

6-2. MMP-1 gene expression measurement

In order to investigate the expression level of MMP-1 at the transcription level, each sample and control (positive control: Vitamin C, 50 ug/mL) were treated for a certain time, irradiated with UVB ultraviolet rays, and total RNA was extracted using a Trizol reagent. . The extracted total RNA was cDNA prepared using First strand cDNA synthesis kit (Thermo scientific), and then the same amount of cDNA was amplified by PCR. The sense and antisense primer sequences used at this time were prepared by referring to existing literature, and GAPDH was used as a control gene. For PCR amplification, cDNA and MMP-1 primers for each experimental group, and GAPDH primers for the control group were used for PCR amplification using the Hipi PCR kit (Elpis biotech), including dNTP 250 uM, Tris-HCL (pH 8.3) 10 mM, KCl 50 mM, NgCl ₂ 1.5 mM. It was performed with 20ul of Hipi PCR kit. PCR was performed under conditions of denaturing at 95°C for 45 seconds, annealing at 60°C for 45 seconds, and extension at 72°C for 1 minute, and a total of 30 cycles were performed. The DNA amplified by PCR was electrophoresed on a 1.5% agarose gel, and the intensity of the fractionated DNA band was measured, and the results are shown in FIG. 6.

-MMP-1 forward 5'-ATT CTA CTG ATA TCG GGG CTT TGA-3', MMP-1 reverse 5'-ATG TCC TTG GGG TAT CCG TGT AG-3'

-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) forward 5'-CCA GCC GAG CCA CAT CGC TC-3', GAPDH reverse 5'-TGA CCT TGG CCA GGG GTG CT-3'

As shown in Fig. 6, the group obtained by mixing the extract obtained by extracting hot water separately from goji berry, fig, and pear chorion at a ratio of 1:1:1, and the goji berry, fig, and pear chow scent were mixed in 14.93, 9.29, and 75.78 parts by weight, respectively. As a result of measuring the expression of the MMP-1 gene in the extracted, freeze-dried or spray-dried group, it was confirmed that the gene expression was reduced.

Experimental Example 7: Measurement of fibroblast-derived collagen production ability

7-1. Cell culture

HS68 cells, a human fibroblast line, are cultured at a density of 70-80% in 100 mm cell culture dishes using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. .

7-2. Type 1 procollagen measurement

After the cultured human fibroblast line HS68 cells were adjusted to a concentration of 1×10 ⁶ cells/ml using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. , Inoculated on a 96 well plate, and pre-incubated for 18 hours at 5% CO ₂ and 37 ℃. Thereafter, the medium was removed and each extract-containing medium was simultaneously treated and cultured. After 24 hours, 100 μl of the supernatant was transferred to another 96 well plate, and the procollagen concentration was measured according to the manufacturer's instructions using an enzyme-linked immunosorbent assay (ELISA) kit from Abcam, and the results are shown in FIG. 7.

As shown in FIG. 7, the group obtained by mixing the extract obtained by extracting hot water separately from Goji berry, fig, and pear chorion at a ratio of 1:1:1, and Goji berry, fig, and pear chow scent were mixed in 14.93, 9.29, and 75.78 parts by weight, respectively, and then mixed. As a result of measuring the production of type 1 procollagen in the extracted and freeze-dried or spray-dried group, it was confirmed that the production amount was increased.

Experimental Example 8: Measurement of PPAR-α,δ activity derived from monkey kidney cells

8-1. Cell culture

Cos-7 cells, a monkey kidney cell line, were prepared in a 100 mm cell culture dish at a density of 70-80% using DMEM medium containing penicillin (100 IU/ml) and streptomycin (100 μg/ml) and 10% FBS. Incubate.

8-2. Type 1 procollagen measurement

Cos-7 cells, a cell line derived from monkey kidney, were adjusted to a concentration of 4×10 ⁶ cells/ml using DMEM medium, inoculated into a 24 well plate, and precultured at 5% CO ₂ and 37°C for 24 hours. . Then, using Thermo Fisher's Lipofectamine, pFR-Luciferase (10 μg/ml) and pFR-PPAR (1.5 μg/ml) were introduced into the cells and cultured for 24 hours. Thereafter, the medium was removed, and the phenol free DMEM medium containing each extract was simultaneously treated and cultured for 24 hours. After incubation, the cells were lysed using a lysis buffer and reacted 1:1 with the substrate. Immediately after the reaction, the absorbance was measured with a luminometer, and the results are shown in FIG. 8.

As shown in FIG. 8, in the group extracted and spray-dried, PPAR-α and PPAR- were mixed in 14.93, 9.29, and 75.78 parts by weight of the extract of Goji berry, fig, and pear-cho scent extracted with hot water, respectively, respectively. As a result of measuring δ activity, it was confirmed that the activity was increased in the mixed extract and the mixed spray-dried group. This shows a synergistic effect in the whitening and skin epidermal cell regeneration effects in the complex extract rather than the efficacy for each single extract.

As can be seen from the results of the above experimental examples, the composition comprising the hot water extract of Goji berry, fig and pear chorion according to the present invention is effective in antioxidant, immunity enhancement, whitening, collagen degrading enzyme inhibition, collagen production promotion, and wrinkle improvement effect. It exhibits an equal effect to the positive control group, which is due to a synergistic effect between the extracts of the goji berry, fig and pearchohyang herbal medicine.

The above description is merely illustrative of the technical idea of the present invention, and those of ordinary skill in the technical field to which the present invention pertains can make various modifications and variations without departing from the essential characteristics of the present invention. Further, the embodiments disclosed in the present invention are not intended to limit the technical idea of the present invention, but to explain the technical idea, and the scope of the technical idea of the present invention is not limited by these embodiments. The scope of protection of the present invention should be interpreted by the following claims, and all technical ideas within the scope equivalent thereto should be interpreted as being included in the scope of the present invention.

Claims (8)

A functional food composition for antioxidant comprising an extract of Goji berry, fig and pear spice mixture or a mixture of Goji berry, fig and pear spice extract,
The extract of the mixture is hot water extraction by mixing 5 to 20 parts by weight of goji berry, 5 to 20 parts by weight of figs, and 60 to 90 parts by weight of pear flavor,
The mixture of the extracts is a mixture of Goji berry, fig and pear chow flavor each in hot water and then mixed in a weight ratio of 1:1:1, 2:1:1, or 2:1:2. Functional food composition for antioxidant.
A functional food composition for enhancing immunity comprising an extract of a mixture of Goji berry, fig and pear chow flavor or a mixture of Goji berry, fig and pear chow flavor extract,
The extract of the mixture is hot water extraction by mixing 5 to 20 parts by weight of goji berry, 5 to 20 parts by weight of figs, and 60 to 90 parts by weight of pear flavor,
The mixture of the extract is that after hot water extraction of Goji berry, fig and pear chow flavor, respectively, and then mixing in a weight ratio of 1:1:1, 2:1:1, or 2:1:2, functional food composition for enhancing immunity.
A functional food composition for whitening comprising an extract of Goji berry, fig and pear spice mixture or a mixture of Goji berry, fig and pear spice extract,
The extract of the mixture is hot water extraction by mixing 5 to 20 parts by weight of goji berry, 5 to 20 parts by weight of figs, and 60 to 90 parts by weight of pear flavor,
The mixture of the extract is a mixture of Goji berry, fig, and pear-cho flavor after hot water extraction, respectively, in a weight ratio of 1:1:1, 2:1:1, or 2:1:2, and a functional food composition for whitening.
A functional food composition for improving wrinkles comprising an extract of Goji berry, fig and pear vinegar flavor mixture, or a mixture of Goji berry, fig and pear vinegar flavor extract,
The extract of the mixture is hot water extraction by mixing 5 to 20 parts by weight of goji berry, 5 to 20 parts by weight of figs, and 60 to 90 parts by weight of pear flavor,
The mixture of the extracts is a mixture of Goji berry, fig and pear chow scent after hot water extraction, respectively, in a weight ratio of 1:1:1, 2:1:1, or 2:1:2, and a functional food composition for improving wrinkles.
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