Composition for improving swell of skin comprising NYPA fruticans wurmb extract and fucoidan extract

Abstract

The present invention relates to a composition for reducing skin swelling, including a Nypa fruticans extract and a fucoidan extract. Specifically, the composition exhibits excellent efficacy in maintaining homeostasis by increasing nitric oxide(NO) levels in vascular endothelial cells to an appropriate level, inhibiting the activation of an epithelial sodium channel(ENaC) to reduce sodium reabsorption, and promoting the excretion of water together with sodium from the body, thereby reducing skin swelling.

Description

COMPOSITION FOR IMPROVING SWELL OF SKIN COMPRISING NYPA FRUTICANS WURMB EXTRACT AND FUCOIDAN EXTRACT

[0001] The present invention relates to a composition including a Nypa fruticans extract and a fucoidan extract for reducing skin swelling, and more particularly, to a composition having excellent efficacy in reducing skin swelling by including a Nypa fruticans extract and a fucoidan extract as active ingredients, thereby inhibiting the activation of an epithelial sodium channel (ENaC) to lower the reabsorption rate of sodium, excreting water together with sodium from the body to improve skin swelling, and ameliorating blood circulation problems caused by a decrease in abnormal production of nitric oxide(NO).

[0002] Skin swelling occurs when excessive water or salt accumulates in the skin. This reduces skin elasticity and hydration, contributing to wrinkles and aging. Severe skin swelling may lead to skin cracking, loss of skin elasticity and radiance, and increased wrinkles.

[0003] Such skin swelling is known to be caused by a variety of factors, including dehydration, excessive salt intake, stress, increased free radicals, lack of sleep, allergic reactions, and hormonal imbalances.

[0004] In particular, when salt accumulates excessively in the body, fluid abnormally accumulates outside the blood vessels, leading to edema, a condition in which water accumulates in the interstitial tissue. Specifically, when salt levels are excessively high in the body, sodium channels(ENaC) present in the epithelial cells of the collecting ducts of the kidneys promote sodium reabsorption. Consequently, water reabsorption promotes water retention to maintain body homeostasis, resulting in skin swelling due to excessive water retention.

[0005] Furthermore, normal vascular endothelial cells continuously secrete small amounts of nitric oxide(NO), a vascular relaxant, to maintain vascular tone at a basal level. Chronically reduced NO secretion may lead to venous endothelial damage and inflammation, which may lead to valve damage and the development of varicose veins or chronic venous insufficiency. Therefore, NO, secreted from vascular endothelial cells by basal response or stimulation, properly maintains vascular tone and plays a crucial role in maintaining vascular homeostasis through mechanisms such as preventing leukocyte adhesion and platelet aggregation.

[0006] A variety of products, including functional foods and cosmetics, are being released to reduce skin swelling, but most of these products lack significant effects or may cause skin irritation due to physical pressure or chemical ingredients.

[0007] Meanwhile, with the growing demand for products made from natural ingredients that are non-irritating to the human body and do not cause skin problems, extensive research is being conducted to discover and enhance the efficacy of natural ingredients. Furthermore, research is actively underway on mixtures of different natural extracts, utilizing the functions of various natural extracts. However, mixing one extract with another may lead to problems such as precipitation, formulation instability, and reduced user experience, and the desired effects are often not fully implemented.

[0008] Therefore, the present inventors have conducted ongoing research to develop a composition effective in reducing skin swelling using a combination of natural extracts, and as a result, discovered that when a Nypa fruticans extract and a fucoidan extract are optimally combined, α-ENaC function is inhibited to reduce the reabsorption rate of sodium, discharge water from the body along with sodium, and ameliorate blood circulation problems caused by abnormally reduced NO production, exhibiting an excellent effect in reducing skin swelling, thereby completing the present invention.

[0009] An objective of the present invention is to provide a composition including a Nypa fruticans extract and a fucoidan extract as active ingredients, thereby exhibiting an excellent effect in reducing skin swelling.

[0010] The technical problems of the present invention are not limited to above-mentioned technical problems, and other technical problems that are not mentioned will be readily understood by those skilled in the art from the description below.

[0011] An embodiment of the present invention provides a composition for reducing skin swelling, including a fucoidan extract and a Nypa fruticans extract as active ingredients, and the fucoidan extract and the Nypa fruticans extract are included in a weight ratio of 1:0.05 to 1:20.

[0012] The composition of the present invention inhibits activation of the epithelial sodium channel(ENaC) to reduce sodium reabsorption and increase the amount of urine excreted, which contains both sodium and water, thereby alleviating skin swelling caused by excessive water retention.

[0013] In addition to the effect of reducing skin swelling, the composition of the present invention also provides excellent skin-improving effects, including improved hydration, anti-inflammation, enhanced skin elasticity, and increased skin density.

[0014] The composition of the present invention utilizes natural extracts, and thus is non-irritating to the human body and may be used in a variety of products, including foods and cosmetics.

[0015] To help better understand the drawings cited in the detailed description of the present invention, a brief description of each drawing is provided.

[0016] FIG. 1 shows a schematic diagram illustrating the sodium channel protein(ENaC) structure.

[0017] FIG. 2 shows the results of measuring the α-ENaC inhibitory efficacy of Comparative Examples 1 to 3 and Examples 1 to 6, as well as the results of Western blot analysis of α-ENaC.

[0018] FIG. 3 shows the results of a human umbilical vein endothelial cell (HUVEC) cytotoxicity evaluation of a fucoidan hot water extract and a Nypa fruticans ethanol extract at various concentrations.

[0019] FIG. 4 shows the rate of NO production increase of Examples 9 to 21 compared to the normal cells.

[0020] An embodiment of the present invention provides a composition for reducing skin swelling, including a fucoidan extract and a Nypa fruticans extract as active ingredients, and the fucoidan extract and the Nypa fruticans extract are included in a weight ratio of 1:0.05 to 1:20.

[0021] In addition, a total content of the fucoidan extract and the Nypa fruticans extract may be 40% to 70% by weight based on a total weight of the composition.

[0022] In addition, the fucoidan extract and the Nypa fruticans extract may be obtained by hot water extraction.

[0023] In addition, the fucoidan extract may be obtained by hot water extraction and the Nypa fruticans extract may be obtained by ethanol extraction.

[0024] In addition, the fucoidan extract and the Nypa fruticans extract may be in dry powder form.

[0025] In addition, the composition may inhibit activation of an epithelial sodium channel(ENaC).

[0026] In addition, the composition may increase nitric oxide (NO) production to maintain a function of vascular endothelial cells.

[0027] In addition, a food composition including the above-mentioned composition is provided.

[0028] In addition, a cosmetic composition including the above-mentioned composition is provided.

[0029] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Generally, the nomenclature used herein and the experimental methods which will be described later are those well known and commonly employed in the art. Furthermore, when describing embodiments of the present invention, detailed descriptions of related known components or functions will be omitted if they are deemed to hinder understanding of the embodiments of the present invention. Furthermore, while embodiments of the present invention will be described below, the technical spirit of the present invention is not limited or restricted thereto, and can be modified and implemented in various ways by those skilled in the art.

[0030] When a part is herein said to "comprise" a certain component, unless specified otherwise, this means that it may further include other components, rather than exclude other components. The term "and / or" as used herein includes a combination of a plurality of related items or any one of a plurality of related items.

[0031] In an embodiment of the present invention, a composition for reducing skin swelling, including a Nypa fruticans extract and a fucoidan extract, is provided.

[0032] Skin swelling occurs when water is not properly excreted from the body, leading to water accumulation in the skin. Excessive sodium intake activates the sodium channel protein(ENaC), promoting sodium and water reabsorption and increasing body water levels. This increased water content increases blood osmotic pressure, thereby increasing intravascular pressure. This, in turn, causes body fluid to leak out of the blood vessels and accumulate between tissues, potentially leading to skin swelling.

[0033] The epithelial sodium channel(ENaC) is an essential membrane protein that facilitates sodium ion transport, playing a crucial role in regulating electrolyte ion balance, body fluid volume, and blood pressure. As illustrated in FIG. 1, the structure of ENaC is composed of α-ENaC, an essential factor in sodium channel activation, and -ENaC and γ-ENaC, which function to facilitate activation of ENaC at the cell surface and enhance ENaC activity.

[0034] To suppress sodium reabsorption by the ENaC, α-ENaC inhibitors such as amiloride or triamterene may be used, and to suppress cell surface activation and ENaC activity enhancement, -ENaC and γ-ENaC activation may be inhibited. Furthermore, sodium reabsorption may be suppressed by controlling SGK-1, which regulates ENaC activity, or Nedd4-2, which promotes its degradation.

[0035] Furthermore, when excessive sodium intake leads to excessive production of reactive oxygen species(ROS), or when ENaC activation increases inflammatory factors(cytokines such as COX-2 and PGE2, cell adhesion molecules such as VCAM-1, interleukin, and hs-CRP), the actions of antioxidant or anti-inflammatory substances may reduce vascular permeability and alleviate swelling.

[0036] The present inventors have confirmed that when a suitable combination of a Nypa fruticans extract and a fucoidan extract is applied, the function of one of the α-ENaC, -ENaC, and γ-ENaC of the ENaC is inhibited, or the regulatory factors of the ENaC(SGK-1 or Nedd4-2) are controlled to inhibit sodium reabsorption, thereby reducing the rate of sodium reabsorption and promoting the excretion of water along with sodium, ultimately reducing skin swelling.

[0037] Furthermore, the composition of the present invention can improve the function of a tight junction, a key element in maintaining skin cell volume, thereby suppressing cell shrinkage and water loss. By reinforcing the skin cell barrier and enhancing its moisture retention capacity, the composition can reduce skin swelling.

[0038] Furthermore, the composition of the present invention exhibits antioxidant and anti-inflammatory effects that suppress the expression of excessively produced ROS or inflammatory factors(cytokines such as COX-2 and PGE2, cell adhesion molecules such as VCAM-1, interleukin, and hs-CRP) to reduce intravascular pressure, thereby reducing body swelling.

[0039] Specifically, the composition of the present invention may inhibit α-ENaC activation to suppress sodium reabsorption by the ENaC. This inhibition of α-ENaC function reduces the rate of sodium reabsorption, which in turn inhibits water reabsorption necessary for maintaining body homeostasis, resulting in increased urine output containing sodium and water. Therefore, by inhibiting α-ENaC activation, the composition of the present invention can promote the excretion of excess water accumulated in the body together with urine, thereby alleviating body edema and skin swelling.

[0040] The nipa palm(Nypa fruticans), the only species in the subfamily Nypa, a monotypic subfamily of the palm family, is distributed throughout South Asia, Southeast Asia, and Oceania. Nypa fruticans refers to a young shoot of the palm tree and is extremely high in polyphenols, which have antioxidant, blood circulation-promoting, anti-inflammatory, and cancer-preventing effects. Nypa fruticans is also rich in vitamin E and saponins, which help with cardiovascular disease, cataracts, and arthritis. Nypa fruticans is also known to have efficacy in constipation alleviation, detoxification, and hangover alleviation.

[0041] Fucoidan is a sulfated, high-molecular weight polysaccharide having a viscous structure found in trace amounts in brown algae such as wakame and kelp. Fucoidan is an ingredient that protects the algae from seawater currents and external stimuli, serving as a protective barrier against marine microorganisms.

[0042] The composition of the present invention includes both a Nypa fruticans extract and a fucoidan extract, and the composition has an excellent effect of inhibiting α-ENaC in the kidneys, thus reducing the sodium reabsorption rate and excreting sodium and water together from the body, thereby reducing skin swelling caused by excessive water intake. Furthermore, the composition has an excellent effect of regulating appropriate NO production in vascular endothelial cells, thereby maintaining normal vascular function and alleviating skin swelling through vasodilation and blood flow stasis alleviation. Better effects can be obtained when the Nypa fruticans extract and the fucoidan extract are combined, compared to when the Nypa fruticans extract and the fucoidan extract are used alone.

[0043] In the present invention, the term "extract" refers to a substance obtained by extracting components from a natural product. For example, it is a broad concept that includes substances obtained by extracting solvent-soluble components from a natural product using water or an organic solvent, and substances obtained by extracting only specific components from a natural product. The term "extract" may encompass all extracts, fractions, dilutions, concentrates, or dried products thereof obtained in each step, such as extraction, fractionation, or purification.

[0044] In the present invention, extraction may be performed by utilizing any extraction method commonly used in the art, such as hot water extraction, ethanol extraction, immersion extraction, cold maceration extraction, reflux extraction, supercritical extraction, subcritical extraction, dissolution, compression, high-temperature extraction, high-pressure extraction, or ultrasonic extraction. Specifically, hot water extraction or ethanol extraction may preferably be used.

[0045] In an embodiment, the fucoidan extract and the Nypa fruticans extract of the present invention may be obtained by hot water extraction. The active ingredients that affect the skin swelling alleviation effect of the fucoidan extract and the Nypa fruticans extract of the present invention are water-soluble substances that readily dissolve in pure water. Hot water extraction of the fucoidan extract and the Nypa fruticans extract may effectively extract these active ingredients. When the Nypa fruticans extract is obtained by hot water extraction the amount of polyphenols, which have antioxidant, blood circulation-promoting, anti-inflammatory, and cancer-preventing effects, as well as vitamin E and saponins, which have therapeutic effects on cardiovascular disease, cataracts, and arthritis and constipation relief, detoxification, and hangover relief efficacy, can be effectively increased.

[0046] In another embodiment, the fucoidan extract of the present invention may be obtained by hot water extraction, and the Nypa fruticans extract may be extracted by ethanol extraction. When the composition of the present invention is prepared by extracting the Nypa fruticans extract with ethanol, active ingredients such as polyphenols, flavonoids, and tannins can be efficiently extracted, and the risk of spoilage and contamination during the extraction process can be reduced through inhibition of microbial growth, thereby ensuring preservative stability. In addition, ethanol can be easily evaporated and removed even at low temperatures, thereby minimizing thermal denaturation of active ingredients and facilitating the drying, powdering, and concentration processes.

[0047] In an embodiment, the solvent used in the extract may be water, ethanol, or a mixture thereof, specifically purified water or ethanol. However, the present invention is not limited thereto, and various solvents applicable in the art may be used alone or in combination. For example, the extraction solvent may include water (or distilled water); lower alcohols having 1 to 4 carbon atoms, such as methanol, ethanol, propyl alcohol, and butyl alcohol; polyhydric alcohols, such as propanediol, butanediol, hexanediol, methylpropanediol, glycerin, butylene glycol, propylene glycol, pentylene glycol, and dipropylene glycol; and hydrocarbon solvents, such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; or mixtures thereof.

[0048] In an embodiment, hot water extraction may be performed using hot water at a temperature of 60 to 140 °C. Because pure water is used as an extraction medium, hot water extraction has the advantage of avoiding problems arising from the toxicity of an organic solvent.

[0049] In an embodiment, the ethanol extraction may be performed using an ethanol aqueous solution of 30% to 70%, but is not limited thereto. When ethanol of the above-mentioned concentration is used, the skin swelling-reducing effect of the composition according to the present invention may be further enhanced.

[0050] In an embodiment, the extract may be in liquid form or in dried powder form. Various drying methods commonly used in the art, such as freeze drying, vacuum drying, hot air drying, and spray drying, may be applied.

[0051] In an embodiment, the extract may be prepared or processed through various methods, such as vacuum distillation or concentration.

[0052] The composition of the present invention may include a Nypa fruticans extract and a fucoidan extract in a weight ratio of 1:0.05 to 1:20, specifically a weight ratio of 1:0.1 to 1:18, and more specifically a weight ratio of 1:0.1 to 1:15. When the Nypa fruticans extract and the fucoidan extract are included in the above-described mixing ratio, the sodium reabsorption rate due to inhibition of α-ENaC function in the kidney can be sufficiently reduced, and the NO expression level in vascular endothelial cells can be increased, resulting in an excellent skin swelling relief effect.

[0053] In an embodiment, the Nypa fruticans extract and the fucoidan extract may be included in an amount of 40% to 70% by weight based on a total weight of the composition. When the Nypa fruticans extract and the fucoidan extract are included in an amount of less than 40% by weight based on a total weight of the composition, the α-ENaC function inhibition effect and / or NO production activation effect may not be sufficient, resulting in a minimal effect on reducing skin swelling. In addition, when the Nypa fruticans extract and the fucoidan extract are included in an amount exceeding 70% by weight, the α-ENaC function inhibition effect and / or NO production activation effect may be reduced, resulting in an insufficient effect on reducing skin swelling, but the present invention is not limited thereto.

[0054] In an embodiment, the composition may inhibit the activation of the ENaC and thus reduce the reabsorption rate of sodium and water in the kidneys, thereby exhibiting excellent skin swelling reduction effects. Furthermore, in addition to the swelling reduction effect, the composition may have an excellent effect in at least one of reducing swelling, improving hydration, enhancing elasticity, and increasing skin density. Specifically, the composition of the present invention can alleviate skin swelling by improving skin hydration, enhancing elasticity and increasing density, and suppressing factors that cause skin swelling.

[0055] In another embodiment, the composition may alleviate skin puffiness by increasing appropriate NO production in vascular endothelial cells. NO serves as a vasodilator, promoting microcirculation and alleviating stagnant blood flow within capillaries, thereby reducing excess fluid accumulation within tissues. This process may normalize vascular permeability and improve lymphatic circulation.

[0056] In an embodiment, the composition may be a food composition.

[0057] The term "food" as used herein refers to a food manufactured and processed using raw materials or ingredients with beneficial functional properties for the human body. The term refers to a food with high medical or therapeutic effects, processed to effectively exhibit bioregulatory functions in addition to providing nutrition. The term may be used interchangeably with terms known in the art, such as functional foods. For example, the food may be manufactured as a health functional food, a health supplement, a special nutritional supplement, a functional beverage, and the like, or the composition according to the present invention may be added to natural foods, processed foods, and the like. The food may be manufactured as powder, granules, tablets, capsules, syrups, or beverages. There is no limitation on the form the food may take, and the food may encompass all foods in the conventional sense.

[0058] For example, the food may include beverages and various drinks, fruits and processed foods thereof(canned fruits, jams, etc.), fish, meat and processed foods thereof(ham, bacon, etc.), breads and noodles, cookies and snacks, dairy products(butter, cheese, etc.), and the like, and it may include all functional foods in the conventional sense. In addition, it may also include food used as animal feed.

[0059] The food composition according to the present invention may further include food additives and other suitable auxiliary ingredients commonly used in the art. For example, the composition may further contain flavoring agents, natural carbohydrates, sweeteners, vitamins, electrolytes, colorants, pectic acid, alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonating agents, and the like.

[0060] For example, as a flavoring agent, natural flavoring agents including thaumatin, stevia extract, and the like, or synthetic flavoring agents including saccharin, aspartame, and the like may be used. As a natural carbohydrate, monosaccharides such as glucose and fructose, disaccharides including maltose and sucrose, polysaccharides including dextrin and cyclodextrin, and sugar alcohols including xylitol, sorbitol, erythritol, and the like may be used. As a sweetening agent, natural sweeteners such as thaumatin, stevia extract, or synthetic sweeteners such as saccharin, aspartame, and the like may be used. The food additives, other auxiliary components, and the like are preferably added and used in a trace amount as long as the purpose for which they are added may be achieved. The trace amount, when expressed numerically, may be in the range of 0.0005% to about 0.5% by weight based on a total weight of the food composition. Those skilled in the art may easily select the blending amount of the above-mentioned components within a range that does not impair the purpose and effect of the present invention.

[0061] In an embodiment, the composition may be a cosmetic composition. In the present invention, the term "skin" refers to tissue covering the body surface of an animal, and is a broad concept encompassing not only tissue covering the body surface of the face or body, but also the scalp.

[0062] In an embodiment, when the composition of the present invention is used as a cosmetic composition, it may be manufactured into various products effective in reducing swelling, improving skin hydration, enhancing skin elasticity, and increasing skin density.

[0063] In an embodiment, the composition of the present invention may be manufactured into functional cosmetics such as nourishing creams, essences, and ampoules, or basic cosmetics such as toners and lotions. Furthermore, it may be manufactured into soaps, cleaning agents, cleansing creams, and cleansing water, or into color cosmetics such as lipsticks, lip balms, mascara, blushers, shading products, highlighters, makeup bases, foundations, pacts, concealers, and skin covers. In addition, it may be manufactured as a product that adheres to the skin, such as a pack or hydrogel slimming patch, or as a product that is sprayed onto the skin, such as a mist or aerosol.

[0064] In an embodiment, the cosmetic composition of the present invention may be provided in a formulation commonly manufactured in the art. For example, it may be provided in the form of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, an emulsion, a solid, a gel, an oil, powder, a paste, or a foam aerosol.

[0065] In an embodiment, other ingredients commonly used in the art, such as thickeners, pH adjusters, isotonic agents, surfactants, stabilizers, preservatives, antiseptics, UV absorbers, sterilizers, moisturizers, blockers, antioxidants, organic pigments, inorganic pigments, fragrances, vitamins, and the like may be further included, as long as the effects of the present invention are not impaired. Those skilled in the art may easily select the blending amounts of these ingredients, as long as they do not impair the purpose and effects of the present invention.

[0066] Hereinafter, the following examples and experimental examples are presented to further illustrate the present invention, but the present invention is not limited thereto.

[0067]

[0068] Preparation Example. Preparation of extracts and mixtures

[0069] (1) Preparation of Nypa fruticans hot water extract

[0070] A 10-fold volume of distilled water was added to each gram of Nypa fruticans, and extraction was performed with the resulting mixture under hot water at 95±2 °C for six hours. The solids from the extract obtained by the hot water extraction were removed, and the remaining solution was vacuum-concentrated. After concentration, the resulting concentrate was mixed with dextrin in a 7:3 ratio and spray-dried to obtain a Nypa fruticans hot water extract.

[0071] (2) Preparation of Nypa fruticans ethanol extract

[0072] A 10-fold volume of 50% ethanol was added to each gram of Nypa fruticans, and the extraction temperature was set at 70 °C. First extraction was performed in a solvent with a 10-fold volume of the sample volume (v / w) for four hours, and second extraction was performed in a solvent with a 10-fold volume of the sample volume (v / w) for two hours. After the extraction was complete, the solids were removed, and the remaining solution was vacuum-concentrated and then freeze-dried to obtain a Nypa fruticans ethanol extract.

[0073] (3) Preparation of fucoidan hot water extract

[0074] Dried seaweed sporophyll was extracted with purified water, and the extract was concentrated after solid-liquid separation to prepare a primary concentrate. After removing alginate from the concentrate, a secondary concentration was performed. Subsequently, polysaccharides were separated by adding ethanol and steeping, followed by purification of the precipitate. After purification, the precipitate was dissolved in water and freeze-dried to prepare a fucoidan hot water extract.

[0075] (4) Preparation of composition for reducing skin swelling

[0076] The fucoidan hot water extract and the Nypa fruticans hot water extract prepared above were mixed in the ratios shown in Table 1 below to prepare example compositions.

[0077] Ratio of fucoidan hot water extract (A): Nypa fruticans hot water extract (B)Example 11 : 1Example 21 : 2Example 31 : 4Example 41 : 5Example 51 : 10Example 61 : 20Example 72 : 1Example 85 : 1

[0078] The compositions of Examples 1 to 8 in Table 1 above were prepared by adjusting the concentration of the Nypa fruticans hot water extract based on 0.5 μg / mL of the fucoidan hot water extract.

[0079] In addition, the prepared fucoidan hot water extract and Nypa fruticans ethanol extract were mixed in the ratios shown in Table 2 to prepare example compositions.

[0080] Ratio of fucoidan hot water extract (A): Nypa fruticans ethanol extract (C)Example 920 : 1Example 1010 : 1Example 115: 1Example 124 : 1Example 133 : 1Example 142 : 1Example 151 : 1Example 161 : 2Example 171 : 3Example 181 : 4Example 191 : 5Example 201 : 10Example 211 : 20

[0081] The compositions of Examples 9 to 21 of Table 2 above were prepared by adjusting the concentration of the fucoidan hot water extract based on 2.5 μg / mL of the Nypa fruticans ethanol extract.

[0082]

[0083] Experimental Example 1. Evaluation of epidermal cell(HaCaT) cytotoxicity

[0084] To evaluate the cytotoxicity of the compositions of Examples 1 to 8 on epidermal cells (HaCaT), HaCaT cells treated with 145 mM NaCl were treated with the compositions of Examples 1 to 8, and the viability of the cells was measured.

[0085] Specifically, 200 μL of the HaCaT cell line was plated in a 96-well plate at the appropriate concentration and cultured for 24 hours in an incubator at 37 °C and 5% CO2. The cultured cells were treated with the compositions of Examples 1 to 8 at various concentrations and cultured for 22 hours in an incubator at 37 °C and 5% CO2. Thereafter, 10 μL of cell counting kit-8(CCK-8) solution was added per well and incubated for two hours under the same culture conditions. Thirty minutes before the end of the reaction, 145 mM NaCl was added, and the resulting mixture was allowed to re-react. The absorbance was measured at 450 nm using a microplate reader (Thermo, UV / Vis). Cell viability was derived using Equation (1). The results are shown in Table 3 below.

[0086] Equation (1)

[0087] Example 1Example 2Example 3Example 4Example 5Example 6Example 7Example 8(A) : (B)1:11:21:41:51:101:202:15:1Cell viability9093968987759597

[0088] Referring to Table 3, it was confirmed that the compositions of Examples 1 to 8 prepared by including the fucoidan hot water extract and the Nypa fruticans hot water extract in appropriate content ratios exhibited no cytotoxicity in the HaCaT cytotoxicity evaluation.

[0089]

[0090] Experimental Example 2. Evaluation of α-ENaC inhibitory efficacy

[0091] To evaluate the α-ENaC inhibitory efficacy of the compositions of Examples 1 to 6, the α-ENaC activation of the compositions of Examples 1 to 6 was measured in epithelial cells treated with 145 mM NaCl. For comparison, the α-ENaC activation was measured in epithelial cells without adding NaCl in Comparative Example 1, and the α-ENaC activation was measured in epithelial cells treated with 145 mM NaCl in Comparative Example 2, and the α-ENaC activation was measured when NaCl and a 10 μM solution of amiloride, which exhibits α-ENaC inhibitory efficacy, were added together to epithelial cells in Comparative Example 3.

[0092] Specifically, the HaCaT cell line was plated in a 60 mm dish at an appropriate concentration of 4 mL per well and cultured in an incubator at 37 °C and 5% CO2for 24 hours. The cultured cells were treated with the compositions of Comparative Examples 1 to 3 and Examples 1 to 6 diluted according to the concentration and cultured for an additional 24 hours. Thirty minutes before the end of the culture, the cells were treated with 145 mM NaCl and allowed to undergo an additional reaction. Thereafter, the cells were treated with PRO-PREPTM Protein Extract Solution(Intron, Inc.) and allowed to react at -20°C for 20 minutes and then centrifuged. The supernatant containing the separated protein was transferred to a tube and stored at -20 °C. The separated protein was electrophoresed using 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and transferred to a polyvinylidene fluoride(PVDF) membrane. After blocking for one hour in a Tris buffer containing 5% bovine serum albumin(BSA), the membrane was incubated with a primary antibody, followed by secondary antibody treatment and incubation with ImmobilonTMWestern chemiluminescent horseradish peroxidase(HRP) substrate(Millipore) for a specified period of time before development. The degree of inhibition of α-ENaC activity, which was enhanced under a NaCl concentration of 145 mM(high-sodium state), was measured. The results are shown in FIG. 2.

[0093] In addition, the α-ENaC inhibition efficacy of Comparative Examples 1 and 3 and Examples 1 to 6 was calculated based on the α-ENaC activity of Comparative Example 2, which included no active ingredient and treated epithelial cells with a 145 mM NaCl concentration. The results are shown in Table 4 below.

[0094] Comparative ExamplesExamples123123456NaCl(mM)-145145145145145145145145(A) : (B)---1:11:21:41:51:101:20Amiloride--O------α-ENaC inhibitory efficacy (%)20-211415582225

[0095] Referring to Table 4 and FIG. 2, Examples 1 to 6, which included the fucoidan hot water extract and the Nypa fruticans hot water extract in appropriate ratios, demonstrated α-ENaC inhibitory efficacy. In particular, the extract mixture compositions with the composition ratios of Examples 5 and 6 demonstrated similar or superior α-ENaC inhibitory efficacy compared to Comparative Example 3, which was prepared by adding amiloride, a commonly used agent for α-ENaC inhibition.

[0096] Therefore, the composition of the present invention is expected to have the effect of reducing sodium reabsorption by inhibiting the α-ENaC mechanism within renal epithelial cells, which causes swelling.

[0097]

[0098] Experimental Example 3. Evaluation of human umbilical vein endothelial cell(HUVEC) cytotoxicity

[0099] In order to determine whether the compositions of Examples 9 to 21 had toxicity to HUVEC, HUVEC treated with NaCl at a concentration of 145 mM was treated with the fucoidan hot water extract and the Nypa fruticans ethanol extract, and the viability thereof was measured.

[0100] Specifically, the HUVEC cell line was plated in a 96-well plate at an appropriate concentration of 100 μL per well and cultured for 24 hours under incubator conditions of 37 °C and 5% CO2. The cultured cells were treated with the fucoidan hot water extract and the Nypa fruticans ethanol extract at various concentrations and cultured for 23 hours in an incubator under the conditions of 37 °C and 5% CO2. Thereafter, cells were treated with 10 μL of CCK-8 solution per well and allowed to react for one hour under the same culture conditions. Thereafter, the absorbance was measured at 450 nm using a microplate reader (Thermo, UV / Vis). Cell viability was calculated in the same manner as in Equation (1) of Experimental Example 1, and the maximum concentration at which no cytotoxicity was observed is shown in Table 5 and FIG. 3 below.

[0101] Fucoidan hot water extractNypa fruticans ethanol extractMaximum concentration (μg / mL)10025

[0102] Referring to Table 5 and FIG. 3, it was confirmed in the HUVEC toxicity evaluation that the fucoidan hot water extract exhibited no cytotoxicity at 100 μg / mL or less, and the Nypa fruticans ethanol extract exhibited no cytotoxicity at 25 μg / mL or less. Therefore, it may be inferred that the compositions of Examples 9 to 21, in which the fucoidan hot water extract and the Nypa fruticans ethanol extract were mixed at concentrations below the above-mentioned concentrations, would exhibit no cytotoxicity.

[0103]

[0104] Experimental Example 4. Evaluation of NO production increasing efficacy

[0105] To evaluate the efficacy of the compositions of Examples 9 to 21 in increasing NO production in vascular endothelial cells, HUVEC cells were treated with each of Examples 9 to 21, and the amount of nitric oxide(NO) produced in the cells was measured.

[0106] Specifically, the HUVEC cell line was plated in a 96-well plate at an appropriate concentration per well(100 μL) and cultured for 24 hours in an incubator at 37 °C and 5% CO2. The cultured cells were pretreated with each example for one hour. Thereafter, cells were treated with lipopolysaccharide (LPS) (1 μg / mL) in each well and cultured for an additional 24 hours under the same incubator conditions(37°C and 5% CO2). After the culture was completed, the supernatant from each well was collected and the increased amount of NO compared to normal cells was measured. The absorbance of the reaction solution was measured at 570 nm using a microplate reader(Thermo, UV / Vis). LPS 1 μg / mL was used as a negative control(NC), and celecoxib 5 μg / mL was used as a positive control(PC). All results were calculated as a percentage(%) compared to a control group(Blank Controls) and are shown in Table 6 and FIG. 4 below. Here, the control group(Blank Controls) may be considered as representing the NO production status of normal vascular endothelial cells that were not treated with any composition, and even when the NO production rate is lower than that of the control group(Blank Controls), cases where the NO production rate increases compared to the negative control(NC) may be interpreted that there is an appropriate level of NO production-increasing effect.

[0107] Example9101112131415161718192021(A) : (C)20:110:15:14:13:12:11:11:21:31:41:51:101:20NO production rate (%)48344544251234-33327434250

[0108] According to Table 6 and FIG. 4, a significant increase in NO production was confirmed in the examples compared to the control group (Blank Controls), with an increase of up to 50% compared to the control group(Example 21). In particular, in Examples 9 to 12 and Examples 18 to 21, the NO production significantly increased by approximately 30% or more compared to the control group, and Examples 13 and 17 also showed increases in NO production similar to or higher than the positive control group(Celecoxib).

[0109] Therefore, the composition of the present invention is expected to exhibit a skin swelling-reducing effect by increasing NO production, a key factor in maintaining vascular homeostasis in vascular endothelial cells, to an appropriate level, through regulation of vasodilation and blood flow.

[0110] While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

A composition for reducing skin swelling, comprising a fucoidan extract and a Nypa fruticans extract as active ingredients, wherein the fucoidan extract and the Nypa fruticans extract are included in a weight ratio of 1:0.05 to 1:20.The composition for reducing skin swelling of claim 1, wherein a total content of the fucoidan extract and the Nypa fruticans extract is 40% to 70% by weight based on a total weight of the composition.The composition for reducing skin swelling of claim 1, wherein the fucoidan extract and the Nypa fruticans extract are obtained by hot water extraction.The composition for reducing skin swelling of claim 1, wherein the fucoidan extract is extracted by hot water extraction and the Nypa fruticans extract is extracted by ethanol extraction.The composition for reducing skin swelling of claim 1, wherein the fucoidan extract and the Nypa fruticans extract are in dry powder form.The composition for reducing skin swelling of claim 1, wherein the composition inhibits activation of an epithelial sodium channel(ENaC).The composition for reducing skin swelling of claim 1, wherein the composition increases nitric oxide(NO) production to maintain a function of vascular endothelial cells.The composition for reducing skin swelling of claim 1, wherein the composition is a food composition.The composition for reducing skin swelling of claim 1, wherein the composition is a cosmetic composition.

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