Agents that increase the expression of intercellular adhesion factors

Abstract

To provide a novel agent for increasing the expression of intercellular adhesion molecules.SOLUTION: An agent for increasing the expression of intercellular adhesion molecules comprises extract from plants belonging to Rosa, Rosaceae, as an active ingredient.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an agent for increasing the expression of intercellular adhesion factors.

Background Art

[0002] In the epithelial tissue of the skin, epithelial cells Allies form a cell layer by closely binding to each other. The adhesion of epithelial cells is composed of cell adhesion structures such as tight junctions (TJ), adherens junctions (AJ), and desmosomes (DS). Among these, tight junctions exist to function as a barrier so that epithelial cells in contact with the outside world do not allow harmful substances and the like to permeate into the cells. As factors constituting tight junctions, claudin and occludin, which are cell membrane proteins, and Zonula occludens protein that binds to these proteins are known.

[0003] Factors constituting such tight junctions are also known to be deeply involved in maintaining homeostasis by functioning as a barrier separating the inside and outside of tissues in endothelial cells such as blood vessels and lymphatic vessels. When the factors constituting tight junctions decrease, the structure of tight junctions is destroyed, causing various diseases. For example, it has been clarified that the localization of Zonula occludens protein-1 (ZO-).

[0004] In addition, the present inventors have discovered that the adhesion between vascular endothelial cells and lymphatic endothelial cells decreases in the presence of cortisol (Non-Patent Document 2). From this finding, it is considered that when cortisol increases due to stress, the adhesion between cells decreases, water leaks from blood vessels and lymphatic vessels, water accumulates inside the skin, and swelling occurs.

[0005] In recent years, efforts have been made to search for components that promote the formation of tight junctions in epithelial tissue in order to prevent or improve such diseases. For example, a tight junction formation promoter containing mandarin orange extract as an active ingredient (Patent Document 1), an epithelial tight junction formation promoter containing processed garlic extract with a high concentration of sulfur-containing garlic components as an active ingredient (Patent Document 2), and an inhibitor of decreased cell adhesion containing peach kernel (Prunus persica Batsch) extract as an active ingredient (Patent Document 3) have been disclosed. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] Japanese Patent Publication No. 2016-222558 [Patent Document 2] Japanese Patent Publication No. 2016-113368 [Patent Document 3] Japanese Patent Publication No. 2020-132546 [Non-patent literature]

[0007] [Non-Patent Document 1] Olga Tornavaca et al. J Cell Biol (2015) 208 (6): 821 838 [Non-Patent Document 2] "Stress hormones cause facial swelling," [online], June 28, 2019, POLA ORBIS HOLDINGS, Inc., [Retrieved September 14, 2021], Internet<http: / / www.pola-rm.co.jp / pdf / release_20190628.pdf> [Overview of the Initiative] [Problems that the invention aims to solve]

[0008] As mentioned above, it has become clear that a decrease in intercellular adhesion function can cause various health problems in the body. Therefore, the object of the present invention is to provide a novel agent for increasing the expression of intercellular adhesion factors. [Means for solving the problem]

[0009] The present invention, which solves the above problems, is an agent for increasing the expression of intercellular adhesion factors, in which an extract of a plant belonging to the genus Rosa of the family Rosaceae is used as an active ingredient. The cell adhesion factor expression enhancer of the present invention can promote the expression of cell adhesion factors and strengthen cell adhesion. Furthermore, the cell adhesion factor expression enhancer of the present invention is useful in preventing or improving the decline in biological functions caused by impaired cell adhesion function.

[0010] In a preferred embodiment of the present invention, the extract is an extract obtained by supercritical CO2 extraction. Extracts obtained by supercritical CO2 extraction are excellent at promoting the expression of intercellular adhesion factors and strengthening intercellular adhesion.

[0011] In a preferred embodiment of the present invention, the extract mainly consists of hydrocarbon waxes. Rose extract, which is primarily composed of hydrocarbon waxes, is excellent at promoting the expression of intercellular adhesion factors and strengthening intercellular adhesion.

[0012] In a preferred embodiment of the present invention, it is used to promote intercellular adhesion in blood vessels and / or lymphatic vessels.

[0013] In a preferred embodiment of the present invention, the present invention is used for the prevention and / or improvement of edema.

[0014] In a preferred embodiment of the present invention, it is used to improve skin condition.

[0015] In a preferred embodiment of the present invention, the intercellular adhesion factor is a tight junction-related factor.

[0016] In a preferred embodiment of the present invention, the tight junction-related factor is tight junction protein-1 (Zonula occludens-1, ZO-1). By setting the content as described above, the effect of enhancing cell adhesion is more effectively exerted.

[0017] The present invention also relates to a topical skin agent for increasing the expression of an intercellular adhesion factor, which contains an agent for increasing the expression of the above intercellular adhesion factor. The topical skin agent of the present invention can promote the expression of an intercellular adhesion factor and strengthen intercellular adhesion.

Effects of the Invention

[0018] The agent for increasing the expression of an intercellular adhesion factor of the present invention can improve the expression of an intercellular adhesion factor. Further, the agent for increasing the expression of an intercellular adhesion factor of the present invention exhibits the effect of preventing or ameliorating the decrease in the barrier function of epithelial tissue and diseases such as swelling caused by the decrease in intercellular adhesion by strengthening intercellular adhesion.

Brief Description of the Drawings

[0019] [Figure 1] It is an immunohistochemical photograph showing the expression of ZO-1 protein in the case where no rose supercritical CO2 extract is added (a) and the case where 0.1% rose extract is added (b) to lymphatic endothelial cells in Test Example 1. The presence of ZO-1 between cells and nuclei inside the cells can be confirmed. The scale bar in the photograph is 100 μm. [Figure 2] It is a graph showing the occupancy area ratio per unit area of the stained portion of ZO-1 protein in the immunohistochemical staining image of FIG. 1. [Figure 3] It is an immunohistochemical photograph showing the expression of ZO-1 protein in the case where no rose supercritical CO2 extract is added (a) and the case where 0.1% rose supercritical CO2 extract is added (b) to vascular endothelial cells in Test Example 2. The presence of ZO-1 between cells and nuclei inside the cells can be confirmed. The scale bar in the photograph is 100 μm. [Figure 4]Figure 3 is a graph showing the area ratio of the ZO-1 protein stained region per single area in the immunohistochemical staining image. [Modes for carrying out the invention]

[0020] (1) Extracts from plants belonging to the genus Rosa of the family Rosaceae. The present invention provides an expression enhancer for intercellular adhesion factors, comprising an extract of a plant belonging to the genus Rosa of the family Rosaceae (hereinafter also simply referred to as rose extract). Examples of plants belonging to the genus Rosa of the family Rosaceae include Rosa Jurlique, Rosa damascena, Rosa multiflora, Rosa centifolia, Rosa rugosa, Rosa chinensis, Rosa moschata, Rosa alba, Rosa alpina, Rosa canina, Rosa cinnamonea, Rosa gallica, Rosa repens, Rosa rubrifolia, Rosa rubiginosa, Rosa sempervirens, Rosa spinosissima, Rosa styosa, Rosa tomentosa, and Rosa villosa, with Rosa Jurlique and Rosa gallica being particularly preferred.

[0021] The part of the plant belonging to the genus Rosa that can be extracted can be appropriately selected depending on the purpose, and examples include leaves, stems, flowers, fruits, and roots. In the present invention, it is preferable to use the flowers, especially the petals.

[0022] For extraction, it is preferable to pre-dry the extraction part of a plant belonging to the genus Rosa. In this invention, freeze-dried or hot-air-dried material is preferably used in the extraction process.

[0023] Extraction methods include immersion in an extraction solvent at room temperature, cooled, or heated, under normal pressure, pressurized, or reduced pressure; distillation methods such as steam distillation; pressing to obtain an extract; and supercritical fluid extraction. However, in the present invention, extraction by supercritical fluid extraction is preferred.

[0024] When extraction is performed using supercritical fluid extraction, propane, ethylene, water, etc., can be used as the supercritical fluid, but in this invention, carbon dioxide is preferred.

[0025] Supercritical fluid extraction using carbon dioxide (supercritical CO2 extraction) is preferably performed under conditions of a temperature of 40-90°C, a pressure of 8-60 MPa, and a CO2 flow rate of 20 times or more the weight of the petals. By performing supercritical fluid extraction under the above conditions, solid active ingredients (hereinafter also referred to as solid components) can be obtained.

[0026] The cell adhesion factor expression enhancer of the present invention can use the above-mentioned solid component as a rose extract according to the present invention.

[0027] The rose extract according to the present invention is preferably a viscous wax. Furthermore, the rose extract preferably has a hydrocarbon wax as its main component, and more preferably has paraffin wax as its main component. In this invention, a component whose content ratio relative to the total amount of rose extract is 50% or more is considered the main component. The content ratio of the components can be expressed, for example, as an area percentage (%) measured by gas chromatography.

[0028] Paraffin wax is preferably composed mainly of linear saturated hydrocarbons (n-paraffins). The number of carbon atoms in the linear saturated hydrocarbons is preferably 20 to 35, and more preferably 23 to 33. The content of chain saturated hydrocarbons with 20 to 35 carbon atoms is preferably 50 to 80%, more preferably 60 to 70%, of the total amount of rose extract.

[0029] In this invention, the content of each component of the rose extract is determined by the component ratio (area percentage) measured by gas chromatography.

[0030] Furthermore, the paraffin wax preferably contains one or more selected from heptacosane, nonacosane, hentricontane, and pentacosane, and more preferably contains heptacosane and one or more selected from nonacosane, hentricontane, and pentacosane.

[0031] The rose extract preferably contains one or more selected from sitosterol, amilenol, and geranyl stearate. The content of each of sitosterol, amilenol, and geranyl stearate is preferably 0.5 to 6%, more preferably 1 to 5%, of the total amount of the rose extract.

[0032] In the present invention, it is preferable to dissolve the solid rose extract obtained by supercritical fluid extraction in an oil to obtain a solution-type extract (hereinafter also referred to as rose extract).

[0033] The oil used to dissolve the solid components is not particularly limited as long as it can solubilize the solid components. In the present invention, it is preferable to use a solution obtained by mixing one or more selected from dicaprylyl carbonate, olive oil, and tri(caprylic / capric acid) glyceryl, and it is more preferable to use a solution obtained by mixing equal amounts of dicaprylyl carbonate, olive oil, and tri(caprylic / capric acid) glyceryl.

[0034] The mixing of the rose extract and the oil agent described above is preferably carried out under heating, specifically to 40°C. Furthermore, it is preferable to stir and mix for 10 minutes or more.

[0035] After stirring and mixing, the resulting rose extract can be cooled and filtered using a mesh or similar to obtain the desired rose extract. The rose extract obtained in this manner, or a suitable concentration thereof, can be used as an expression enhancer for the cell adhesion-promoting factor of the present invention.

[0036] The content of rose extract relative to the total amount of the cell adhesion factor expression enhancer of the present invention is preferably 0.001 to 10% by mass, more preferably 0.01 to 10% by mass, even more preferably 0.05 to 5% by mass, and particularly preferably 0.1 to 1% by mass, based on the dry mass.

[0037] (2) Intercellular adhesion factor Cell-to-cell adhesion factors are proteins that constitute cell-to-cell adhesion. In the present invention, preferred examples of cell-to-cell adhesion factors include tight junction-related factors between cells, such as claudins and occludins, which are the backbone proteins of tight junctions, and tight junction proteins (Zonula occludens protein) that bind to these proteins and form a supporting structure.

[0038] Examples of tight junction proteins include Zonula occludens protein-1 (ZO-1), Zonula occludens protein-2 (ZO-2), and Zonula occludens protein-3 (ZO-3). The cell adhesion factor expression enhancer of the present invention is particularly excellent at increasing the expression of ZO-1.

[0039] Furthermore, the intercellular adhesion factor expression enhancer of the present invention has the effect of promoting the expression of intercellular adhesion factors in epithelial tissue. The present invention is particularly effective in improving the expression of intercellular adhesion factors in cells of blood vessels or lymphatic vessels within epithelial tissue.

[0040] (3) Dosage form The intercellular adhesion factor expression enhancer of the present invention may be used by administering the intercellular adhesion factor expression enhancer in its undiluted form into the body, or it may be used after being diluted to any desired concentration. Furthermore, the intercellular adhesion factor expression enhancer of the present invention can be formulated in the form of a topical skin preparation or an oral preparation by appropriately combining it with any component used in formulation. In the present invention, it is preferable that the present invention be in the form of a topical skin preparation containing an agent that increases the expression of intercellular adhesion factors.

[0041] Examples of topical skin preparations include cosmetics, quasi-drugs, and topical medicines. Cosmetics and quasi-drugs are more preferred because they can be used on a daily basis. Furthermore, their dosage forms are not particularly limited. When used as a cosmetic, it is preferable to use it in the form of a lotion, emulsifier, gel, cream, or ointment. More specifically, examples include lotions, emulsions, serums, creams, gels, packs, hair creams, and sprays, with lotions, emulsions, creams, or serums being particularly preferred.

[0042] When the intercellular adhesion factor expression enhancer of the present invention is used as an oral preparation, it is preferable to use it in the form of a food composition containing the intercellular adhesion factor expression enhancer of the present invention as an active ingredient. More specifically, it is preferable to use it in the form of a supplement having a dosage form such as a general food, tablet, granule, or drink.

[0043] When the intercellular adhesion factor expression enhancer of the present invention is used in a topical skin preparation, the intercellular adhesion factor expression enhancer of the present invention may be contained in an amount of preferably 0.01 to 10% by mass, more preferably 0.05 to 5% by mass, and even more preferably 0.1 to 5% by mass, based on the total amount of the topical skin preparation. Furthermore, the rose extract content (dry mass) in the topical skin preparation of the present invention may be preferably 0.0001 to 0.1% by mass, more preferably 0.0005 to 0.05% by mass, and even more preferably 0.001 to 0.05% by mass, relative to the total amount of the topical skin preparation, based on the mass of the solvent-removed plant extract (dry product). If the value is above the lower limit, the effect of the topical skin preparation of the present invention will be exerted, and if it is below the upper limit, it is thought that the plateauing of the effect can be avoided.

[0044] (4) Other ingredients The following describes the ingredients that may be included in topical skin preparations such as cosmetics. For example, oily components such as hydrocarbons, esters, triglycerides, fatty acids, and higher alcohols, surfactants such as anionic surfactants, amphoteric surfactants, cationic surfactants, and nonionic surfactants, polyhydric alcohols, thickeners and gelling agents, antioxidants, UV absorbers, colorants, preservatives, powders, etc., can be added as desired. Furthermore, other active ingredients besides the intercellular adhesion factor expression promoter of the present invention may be included, as long as they do not interfere with the effect of the intercellular adhesion factor expression promoter of the present invention. Examples of active ingredients are not particularly limited, but include whitening ingredients, wrinkle-improving ingredients, anti-inflammatory ingredients, and plant and animal extracts.

[0045] The topical skin preparation of the present invention can be prepared by treating the cell adhesion factor expression promoter of the present invention with the optional components described above by a conventional method.

[0046] The present invention's agent for increasing the expression of intercellular adhesion factors can be used to prevent or improve diseases and skin conditions caused by intercellular adhesion, and is particularly suitable for treating diseases and skin conditions caused by weakening of the barrier function of epithelial or endothelial cells due to the weakening of tight junctions between cells.

[0047] Furthermore, it is thought that reduced cell adhesion in blood vessels and lymphatic vessels leads to fluid leakage from these vessels, causing fluid to accumulate inside the skin and resulting in swelling. Therefore, the cell adhesion factor expression enhancer of the present invention can be used to prevent or improve various diseases caused by leakage of body fluids due to instability of vascular structures, such as edema, by strengthening cell adhesion in blood vessels or lymphatic vessels. In other words, in a preferred embodiment of the present invention, it can be used to promote intercellular adhesion in blood vessels and / or lymphatic vessels, and can also be used to prevent and / or improve edema.

[0048] Furthermore, reduced intercellular adhesion and stagnation of blood and lymphatic flow due to edema are thought to affect fluid exchange and oxygen supply within tissues, leading to skin problems. Therefore, in a preferred embodiment of the present invention, the present invention can be used to improve skin condition. In this invention, "improvement of skin condition" refers to the suppression and / or improvement of the deterioration of skin condition, and skin condition includes skin texture, sagging, elasticity, flexibility, roughness, skin color, etc.

[0049] Furthermore, it is preferable that the topical skin preparation of the present invention be in a form that includes the indication of use "for promoting the expression of intercellular adhesion factors." Alternatively, the indication of use may include "prevention and / or improvement of edema," "improvement of skin," etc.

[0050] The aforementioned "display" includes all displays that have the function of informing consumers of the aforementioned use. In other words, any display that can evoke or infer the aforementioned use falls under the category of "display," regardless of the purpose of the display, the content of the display, or the object or medium on which it is displayed. Furthermore, the phrase "with a label attached" means that there is a labeling act that aims to associate the label with the topical skin preparation (product).

[0051] The act of labeling is preferably one that allows consumers to directly recognize the aforementioned use. Specifically, examples include the act of describing the aforementioned use on the product or packaging of the cell preparation of the present invention, and the act of describing the aforementioned use in advertisements, price lists, or transaction documents (including those provided by electronic means) relating to the product. [Examples]

[0052] The present invention will be specifically described below with reference to examples, but these are merely illustrative examples of the present invention, and the scope of the present invention is not limited thereto.

[0053] <Manufacturing Example 1> Production of rose supercritical CO2 extract (1) Extraction of active ingredients 15 kg of dried rose petals (Rosa Jurlique) were placed in a supercritical CO2 apparatus and pressurized to 45 MPa. Subsequently, extract was carried out under conditions of 80°C, 45 MPa, and a CO2 flow rate 60 times the weight of the petals. Next, the supercritical CO2 receiving tank pressure was reduced to 5.5 MPa, and the piping was evacuated to 0 MPa. After depressurization, the lid of the layered section of the separation tank was opened, and the wax components (solid extracts) remaining at the bottom of the separation tank were recovered using a scraping tool.

[0054] (2) Extraction of the extract The oils shown in Table 1 below were weighed individually, mixed, and homogenized to obtain a mixed oil.

[0055] [Table 1]

[0056] A mixed oil was added to the extract obtained by supercritical CO2 extraction. After addition, the mixture was heated to 40°C and stirred for 10 minutes.

[0057] The obtained solution was refrigerated at 5°C for 24 hours. Then, the solution was filtered using a 1000-mesh filter, and the filtered extract (hereinafter referred to as rose extract) was collected.

[0058] (3) Analysis of the components of the extract The wax components obtained in (1) above were subjected to component analysis using gas chromatography. Subsequently, the peak area of ​​each component was calculated, and the area percentage (%) for each component was determined. As a result, the wax components contained a total of 67.55% n-paraffins of C23 to C33. Furthermore, the n-paraffins included C23, C25, C27 to C29, C31, and C33. Furthermore, the wax components contained 3.6% sitosterol, 2.56% amilenol, and 2.07% geranyl stearate.

[0059] <Test Example 1> Analysis of tight junction-related factor expression in lymphatic endothelial cells using rose supercritical CO2 extract Using the rose extract obtained in Production Example 1, we evaluated the effect of rose extract on cell-cell adhesion in lymphatic endothelial cells.

[0060] (1) Preparation of cells Human lymphatic endothelial cells (HLECs) (ScienCell Research Laboratories) were seeded in flasks containing endothelial cell medium (ScienCell Research Laboratories) and cultured at 37°C and 5% CO2. Next, insert the HLEC into the 4-well slide chamber. 4.0 x 10 4 Seeds were seeded to a cell / well ratio and cultured for 48 hours.

[0061] A new endothelial cell culture medium was mixed with rose extract to a concentration of 0.1% (0.001% by dry weight). After 48 hours of incubation, the HLEC medium was replaced with the medium containing the extract and incubated for 5 days. In the control group, the following solution was added to the HLEC instead of rose supercritical CO2 extract and incubated for 5 days. Control solution: A mixture of 33% by mass of dicaprylyl carbonate, 33% by mass of caprylic / capric triglyceride, and 33% by mass of olive fruit oil.

[0062] (2) Cell fixation and immunohistochemistry The harvested lymphatic endothelial cells were washed with PBS and fixed with 4% paraformaldehyde for 10 minutes. After washing with PBS, permeabilization was performed by adding PBS containing 0.5% TritonX-100 and culturing for 10 minutes, followed by blocking with 10% Block Ace for 30 minutes.

[0063] Next, immunohistochemical staining was performed using the following antibodies. Lymphatic endothelial cells were washed with PBS, and the primary antibody was added and cultured overnight at 4°C. After washing again with PBS, the secondary antibody was added and cultured at room temperature for 30 minutes. Subsequently, nuclear staining and slide mounting were performed using Dapi-Fluoromount-G® (Southern Biotechnology Associates, Inc.). (antibody) • Primary antibody: Rabbit anti ZO-1 (200-fold dilution with 10% Block Ace) (Invitrogen) Secondary antibody: Alexa Fluor 488 (250-fold dilution with 10% Block Ace) (Invitrogen)

[0064] The obtained lymphatic endothelial cells were observed using a fluorescence microscope (KEYENCE BZ-X800), and images were acquired for analysis. The results are shown in Figure 1.

[0065] (3) Expression analysis of tight junction-related factors Using the image editing software ImageJ (National Institutes of Health), the ratio of the area occupied by the ZO-1 protein-stained region to the total area of ​​three randomly selected locations from images taken with a fluorescence microscope was calculated as the cell-cell adhesion region (%). The results are shown in Figure 2.

[0066] (4) Results Figures 1 and 2 show that when the rose supercritical CO2 extract was added, the expression of ZO-1 protein increased compared to when the extract was not added. Therefore, it was revealed that the rose supercritical CO2 extract has an effect of strengthening intercellular adhesion of lymphatic cells, due to the increased expression of tight junction-related factors.

[0067] <Test Example 2> Analysis of tight junction-related factor expression in vascular endothelial cells using rose supercritical CO2 extract Using the rose extract obtained in Production Example 1, we evaluated the effect of rose extract on cell-cell adhesion in vascular endothelial cells.

[0068] (1) Preparation of cells Normal human umbilical vein endothelial cells (HUVECs) were seeded in flasks containing low-serum liquid medium for normal human vascular endothelial cell proliferation (HuMedia-EG2) (manufactured by Kurabo Industries Ltd.) and cultured at 37°C and 5% CO2. Next, insert the HUVEC into the 4-well slide chamber in 3.0 x 10 4 Seeds were seeded to a cell / well ratio and cultured for 48 hours.

[0069] A new low-serum liquid medium for normal human vascular endothelial cell proliferation was mixed with rose extract to a concentration of 0.1% (0.001% by dry weight). After 48 hours of incubation, the medium of the HUVEC cells was replaced with the medium containing the extract, and the cells were incubated for 5 days. For the control group, the same control solution as in Test Example 1 was added to the HUVEC cells, and the cells were incubated for 5 days.

[0070] (2) Cell fixation and immunohistochemistry Immunohistochemical staining of vascular endothelial cells was performed using the same procedure as in the above example 1(2). The results of the fluorescence images are shown in Figure 3.

[0071] (3) Expression analysis of tight junction-related factors Similar to the above example 1(3), the area of ​​the ZO-1 protein-stained region in the image was calculated using ImageJ based on the image in Figure 3, and expressed as the stained area (%) of cell adhesion. The results are shown in Figure 4.

[0072] (4) Results Figures 3 and 4 show that when the rose supercritical CO2 extract was added, the expression of the ZO-1 protein increased compared to when the extract was not added. Therefore, it was revealed that rose supercritical CO2 extract exerts an effect of strengthening intercellular adhesion in vascular endothelial cells, resulting in increased expression of tight junction-related factors.

[0073] <Manufacturing Example 2> Beauty serum containing rose supercritical CO2 extract as the active ingredient A topical serum was prepared according to the following formulation. Specifically, the formulation ingredients were heated to 80°C, stirred and solubilized, and then stirred and cooled to obtain the serum. The rose extract used was obtained according to Production Example 1. The dry mass of the rose extract (dry mass of the wax component) relative to the total amount of serum was 0.001%.

[0074] [Table 2]

[0075] The serum produced by the above method can be used to enhance cell-cell adhesion in blood vessels and lymphatic vessels, and to prevent and / or improve swelling, as well as to improve skin condition. [Industrial applicability]

[0076] This invention can be applied to pharmaceuticals, cosmetics, and the like.

Claims

1. The active ingredient is an extract from the petals of plants belonging to the genus Rosa in the family Rosaceae. The aforementioned extract is an extract obtained by supercritical CO2 extraction. An expression enhancer for tight junction protein-1 (Zonula occludens-1, ZO-1).

2. The extract is a ZO-1 expression enhancer according to claim 1, with hydrocarbon wax as the main component.

3. The extract comprises paraffin wax, The ZO-1 expression enhancer according to claim 1 or 2, wherein the content of chain saturated hydrocarbons having 20 to 35 carbon atoms is 50 to 80% of the total amount of the extract.

4. An expression enhancer for ZO-1 according to any one of claims 1 to 3, used to promote cell-cell adhesion in blood vessels and / or lymphatic vessels.

5. An agent for increasing ZO-1 expression according to any one of claims 1 to 4, used for the prevention and / or improvement of edema.

6. An agent for increasing the expression of ZO-1 according to any one of claims 1 to 5, used for improving skin condition.

7. A ZO-1 expression enhancer according to any one of claims 1 to 6, which is a topical skin preparation.

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