Factor xiii a expression enhancer and screening method using expression of factor xiii a as index

Abstract

To provide a novel composition that enhances the expression of Factor XIII A (Coagulation Factor XIII A).SOLUTION: Provided is an agent for enhancing the expression of Factor XIII A (Coagulation Factor XIII A), containing as an active ingredient one or more extracts selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an agent for enhancing Factor XIIIA expression, an agent for promoting cell proliferation, an agent for enhancing collagen production, an agent for enhancing TGF-β2 expression, an agent for repairing damaged DNA, an agent for suppressing DNA damage, and an agent for suppressing DNA damage. The present invention also relates to a screening method using the expression of Factor XIIIA as an index. [Background technology]

[0002] Factor XIII exists in plasma as a heterotetramer (A2B2) consisting of a dimer of the A subunit (Factor XIII A) and a dimer of the B subunit (Factor XIII B), but Factor XIII A is present in the cytoplasm of megakaryocytes, platelets, monocytes, macrophages, etc. (Non-patent Document 1).

[0003] Factor XIII has traditionally been said to be a factor involved in mechanisms related to the blood and vascular systems. In recent years, it has been suggested that Factor XIIIA also contributes to wound healing processes, angiogenesis, and the like (Non-Patent Document 2). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Thrombosis and Hemostasis 2018;29(6):703-706 [Non-Patent Document 2] Physiol Rev 91: 931-972, 2011 Summary of the Invention [Problem to be solved by the invention]

[0005] In light of the above-mentioned prior art, the present inventors have discovered that expression of Factor XIIIA is significantly enhanced in cells that have fallen out of the cell cycle and are in a dormant state, i.e., in the G0 phase, and by conducting screening focusing on the expression of Factor XIIIA, they have discovered a component that further enhances the expression of Factor XIIIA whose expression is enhanced in the G0 phase.

[0006] That is, an objective of the present invention is to provide a novel composition that enhances the expression of Coagulation Factor XIII A.

[0007] Furthermore, based on the above-mentioned new discoveries, the present invention has an objective of providing a novel method for screening substances that have the effect of enhancing collagen production, substances that have the effect of enhancing TGF-β2 (Transforming Growth Factor β2) expression, and substances that have the effect of repairing damaged DNA and / or inhibiting DNA damage. [Means for solving the problem]

[0008] The present invention, which solves the above-mentioned problems, provides an agent for enhancing expression of Coagulation Factor XIII A, comprising, as an active ingredient, one or more selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Cynara in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0009] In a preferred embodiment, the present invention is an agent for enhancing expression of Coagulation Factor XIII A in cells in the G0 phase.

[0010] The present invention, which solves the above-mentioned problems, also relates to an agent for promoting cell proliferation, comprising, as an active ingredient, one or more selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Cynara in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0011] The present invention, which solves the above-mentioned problems, also relates to an agent for enhancing collagen production, comprising, as an active ingredient, one or more selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Cynara in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0012] The present invention, which solves the above-mentioned problems, also relates to an agent for enhancing expression of Transforming Growth Factor β2 (TGF-β2), comprising, as an active ingredient, one or more selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Cynara in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0013] The present invention, which solves the above-mentioned problems, also relates to an agent for repairing damaged DNA and / or suppressing DNA damage, comprising, as an active ingredient, one or more extracts selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Cynara in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0014] In a preferred embodiment, the present invention relates to an agent for promoting cell proliferation induced by enhanced expression of Coagulation Factor XIII A, an agent for enhancing collagen production, an agent for enhancing expression of Transforming Growth Factor β2 (TGF-β2), and an agent for repairing damaged DNA and / or suppressing DNA damage.

[0015] In a preferred embodiment, the present invention relates to an agent for promoting cell proliferation induced by enhanced expression of Coagulation Factor XIII A in cells in the G0 phase, an agent for enhancing collagen production, an agent for enhancing expression of Transforming Growth Factor β2 (TGF-β2), and an agent for repairing damaged DNA and / or suppressing DNA damage.

[0016] In a preferred embodiment of the present invention, the agent of the present invention contains an extract of a plant belonging to the genus Passiflora in the family Passifloraceae and an extract of a plant belonging to the genus Cynara in the family Asteraceae as active ingredients.

[0017] In a preferred embodiment of the present invention, the extract of a plant belonging to the genus Passiflora in the family Passifloraceae is Passiflora edulis peel extract, the extract of a plant belonging to the genus Cynara in the family Asteraceae is Cynara scolymus leaf extract, and the extract of a plant belonging to the genus Echinacea in the family Asteraceae is Echinacea angustifolia leaf extract.

[0018] In a preferred embodiment of the present invention, the agent according to the present invention is an external agent for the skin.

[0019] The present invention, which solves the above-mentioned problems, also relates to a screening method for screening a substance having an effect of enhancing collagen production, using the expression of Coagulation Factor XIII A in cells as an indicator.

[0020] The present invention, which solves the above-mentioned problems, also relates to a screening method for screening for a substance that has the effect of enhancing TGF-β2 (Transforming Growth Factor β2) expression, using the expression of Coagulation Factor XIII A in cells as an indicator.

[0021] The present invention, which solves the above-mentioned problems, also relates to a screening method for screening for a substance having an effect of repairing damaged DNA and / or an effect of suppressing DNA damage, using the expression of Coagulation Factor XIII A in cells as an indicator.

[0022] In a preferred embodiment of the screening method of the present invention, the cell is a fibroblast. Effect of the Invention

[0023] According to the present invention, a novel composition that enhances the expression of Coagulation Factor XIII A can be provided. Furthermore, the present invention can provide novel methods for screening substances that have the effect of enhancing collagen production, substances that have the effect of enhancing TGF-β2 (Transforming Growth Factor β2) expression, and substances that have the effect of repairing damaged DNA and / or inhibiting DNA damage. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] <Agent> The present invention relates to an agent for enhancing expression of Coagulation Factor XIII A (hereinafter referred to as "Factor XIII A") (herein referred to as a "Factor XIII A expression enhancer") comprising, as an active ingredient, one or more extracts selected from the group consisting of extracts of plants belonging to the genus Passiflora in the family Passifloraceae (hereinafter referred to as "Passifloraceae Passiflora"), extracts of plants belonging to the genus Cynara in the family Asteraceae (hereinafter referred to as "Asteraceae Cynara"), and extracts of plants belonging to the genus Echinacea in the family Asteraceae (hereinafter referred to as "Asteraceae Echinacea").

[0025] <Active ingredients> The agent of the present invention contains, as an active ingredient, one or more selected from extracts of plants belonging to the genus Passiflora in the family Passifloraceae, extracts of plants belonging to the genus Artemisia in the family Asteraceae, and extracts of plants belonging to the genus Echinacea in the family Asteraceae.

[0026] A preferred example of a plant belonging to the genus Passiflora in the family Passifloraceae according to the present invention is Passiflora edulis (hereinafter referred to as "passiflora edulis"). Furthermore, as a plant belonging to the genus Cynara of the Asteraceae family according to the present invention, preferred example is artichoke (Cynara scolymus) (hereinafter referred to as "artichoke"). Furthermore, as a plant of the genus Echinacea in the family Asteraceae according to the present invention, preferred examples include Echinacea angustifolia (hereinafter referred to as "Echinacea angustifolia").

[0027] Regarding the plant extract of the present invention, the whole plant may be used, or the plant body, above-ground parts, roots and stems, trunks, leaves, stems, spikes, flower buds, etc. may be used, but it is preferable to crush or chop them in advance to improve the extraction efficiency. Suitable examples of the extraction solvent include one or more polar solvents selected from water, alcohols such as ethanol, isopropyl alcohol, butanol, polyhydric alcohols such as 1,3-butylene glycol, propylene glycol, ketones such as acetone, methyl ethyl ketone, ethers such as diethyl ether, tetrahydrofuran, etc. The extraction solvent is preferably water, alcohols, or a mixture of water and alcohols.

[0028] Specific examples of extraction methods include adding 1 to 30 parts by mass of a solvent to 1 mass of the part of a plant body or the like used for extraction or its dried matter, soaking the part for several days at room temperature or several hours at a temperature near the boiling point, cooling to room temperature, and then removing insoluble matter and / or the solvent as desired, and fractionating and purifying the part by column chromatography or the like, but the extraction method is not limited to these.

[0029] The plant extract of the present invention will be described in detail below in terms of preferred embodiments.

[0030] In the case of obtaining an extract of a plant belonging to the genus Passiflora of the family Passifloraceae according to the present invention, it is preferable to obtain the extract from the pericarp of a plant belonging to the genus Passifloraceae. In addition, it is preferable to use water, 1,3-butylene glycol or a mixture thereof as an extraction solvent when obtaining an extract of a plant belonging to the genus Passifloraceae.

[0031] The extract of the plant belonging to the genus Passiflora of the family Passifloraceae according to the present invention is preferably a Passiflora edulis pericarp extract extracted with the above-mentioned extraction part and extraction solvent. In addition, the Passiflora edulis pericarp extract according to the present invention may be a commercially available product.

[0032] When obtaining an extract of a plant belonging to the genus Artichoke of the Asteraceae family according to the present invention, it is preferable to obtain the extract from the leaves of a plant belonging to the genus Artichoke of the Asteraceae family. In addition, when obtaining an extract of a plant belonging to the genus Artichoke of the Asteraceae family, it is preferable to use water, 1,3-butylene glycol, or a mixture thereof as an extraction solvent.

[0033] The extract of the plant belonging to the genus Artichoke of the Asteraceae family according to the present invention is preferably an artichoke leaf extract extracted with the above-mentioned extraction part and extraction solvent. The artichoke leaf extract according to the present invention may be a commercially available product (such as "Artichoke Leaf Extract" manufactured by Ichimaru Pharcos Co., Ltd.).

[0034] When obtaining an extract of a plant belonging to the genus Echinacea of ​​the Asteraceae family according to the present invention, it is preferable to obtain the extract from the leaves of a plant belonging to the genus Echinacea of ​​the Asteraceae family. When obtaining an extract of a plant belonging to the genus Echinacea of ​​the Asteraceae family, it is preferable to use water, ethanol or a mixture thereof as an extraction solvent.

[0035] The extract of the plant belonging to the genus Echinacea of ​​the Asteraceae family according to the present invention is preferably an echinacea leaf extract extracted with the above-mentioned extraction part and extraction solvent. In addition, the echinacea leaf extract according to the present invention may be a commercially available product (such as "echinacea leaf extract" manufactured by Ichimaru Pharcos Co., Ltd.).

[0036] ≪Applications≫ As shown in the Examples below, the extracts of the aforementioned plants have the effect of enhancing the expression of Factor XIII A. Therefore, the extracts of the aforementioned plants can be used for enhancing the expression of Factor XIII A. That is, the present invention can be in the form of a Factor XIII A expression enhancer containing the extracts of the aforementioned plants as an active ingredient. The present invention can also provide a composition for enhancing Factor XIIIA expression, which comprises the above-mentioned plant extract as an active ingredient.

[0037] Here, the Factor XIIIA is encoded by the F13A (F13A1) gene. The Factor XIIIA expression enhancer of the present invention is preferably an expression enhancer of the F13A gene.

[0038] Factor XIII A has been shown to act in the wound healing process, angiogenesis, promote fibroblast and macrophage migration, reduce apoptosis, decrease vascular permeability, form and stabilize the extracellular matrix, and increase the proliferation rate of fibroblasts (Physiol Rev 91: 931-972, 2011). Therefore, the Factor XIII A expression enhancer of the present invention can be used for the purpose of treating diseases associated with Factor XIII A. For example, the present invention can be used as a component for wound healing, angiogenesis, promotion of fibroblast and macrophage migration, reduction of apoptosis, reduction of vascular permeability, and formation and stabilization of the extracellular matrix.

[0039] Here, the method for detecting whether or not expression of Factor XIIIA in cells is enhanced is not particularly limited, and any method that can be appropriately selected by those skilled in the art can be used.

[0040] In the present invention, the type of cells in which expression of Factor XIIIA is enhanced is not particularly limited, but is preferably a fibroblast, more preferably a dermal fibroblast, such as a normal human dermal fibroblast (NHDF).

[0041] As shown in the Examples below, the present inventors have found that cell proliferation is promoted in the presence of Factor XIIIA. This suggests that cell proliferation can be promoted by enhancing the expression of Factor XIIIA. Therefore, the above-mentioned plant extracts having the effect of enhancing the expression of Factor XIIIA can be used to promote cell proliferation. That is, the present invention can be in the form of an agent for promoting cell proliferation (referred to as a "cell proliferation promoter" in this specification) containing the above-mentioned plant extract as an active ingredient. The present invention can also provide a composition for promoting cell proliferation, which contains the above-mentioned plant extract as an active ingredient.

[0042] In a preferred embodiment of the present invention, the cell proliferation-promoting agent of the present invention is an agent for promoting cell proliferation induced by enhanced expression of Factor XIIIA.

[0043] Here, the method for detecting whether or not cell proliferation is promoted is not particularly limited, and any method that can be appropriately selected by a person skilled in the art can be used.

[0044] The present invention can also take the form of a Factor XIIIA expression enhancer for promoting cell proliferation, which comprises the above-mentioned plant extract as an active ingredient. Furthermore, the present invention can also be in the form of a composition for enhancing Factor XIIIA expression, which comprises the above-mentioned plant extract as an active ingredient and promotes cell proliferation.

[0045] Furthermore, as shown in the Examples below, the present inventors have found that collagen production is enhanced in the presence of Factor XIIIA. This suggests that collagen production can be enhanced by enhancing the expression of Factor XIIIA. Therefore, the above-mentioned plant extracts having the effect of enhancing the expression of Factor XIIIA can be used to enhance collagen production. That is, the present invention can be in the form of an agent for enhancing collagen production (referred to as a "collagen production enhancer" in this specification) containing the above-mentioned plant extract as an active ingredient. The present invention can also provide a composition for enhancing collagen production, which contains the above-mentioned plant extract as an active ingredient.

[0046] In a preferred embodiment of the present invention, the collagen production enhancer of the present invention is an agent for enhancing collagen production induced by enhanced expression of Factor XIIIA.

[0047] In a preferred embodiment of the present invention, the collagen production enhancer is an agent for increasing the total collagen amount. The total collagen amount refers to the total amount of collagen having a triple helix structure. In a preferred embodiment, the total collagen can include one or more types selected from type I collagen, type III collagen, type IV collagen, type V collagen, and type VII collagen.

[0048] The present invention can also take the form of a Factor XIIIA expression enhancer for enhancing collagen production, which comprises the above-mentioned plant extract as an active ingredient. Furthermore, the present invention can also be in the form of a composition for enhancing expression of Factor XIIIA, which comprises an extract of the above-mentioned plant as an active ingredient, and which enhances collagen production.

[0049] The collagen production enhancer of the present invention can be used to prevent or improve a decrease in firmness and / or elasticity of the skin, or to lift the skin.

[0050] Here, the method for detecting whether collagen production in cells is enhanced is not particularly limited, and any method that can be appropriately selected by a person skilled in the art can be used. In addition, an example of a method for detecting whether total collagen production in cells is enhanced is a method of measuring the total collagen amount by the Sirius Red method.

[0051] Furthermore, as shown in the Examples below, the present inventors have found that the expression of TGF-β2 (Transforming Growth Factor β2) (hereinafter referred to as "TGF-β2") is enhanced in the presence of Factor XIIIA. This means that the expression of TGF-β2 can be enhanced by enhancing the expression of Factor XIIIA. Therefore, the extract of the above-mentioned plant having the effect of enhancing the expression of Factor XIIIA can be used to enhance the expression of TGF-β2. That is, the present invention can be in the form of an agent for enhancing the expression of TGF-β2 (referred to as a "TGF-β2 expression enhancer" in this specification) containing the extract of the above-mentioned plant as an active ingredient. The present invention can also provide a composition for enhancing TGF-β2 expression, which comprises an extract of the above-mentioned plant as an active ingredient.

[0052] In a preferred embodiment of the present invention, the TGF-β2 expression enhancer of the present invention is an agent for enhancing the expression of TGF-β2, which is induced by enhanced expression of Factor XIIIA.

[0053] Here, the TGF-β2 is encoded by the TGFB2 gene. The TGF-β2 expression enhancer of the present invention is preferably an expression enhancer of the TGFB2 gene.

[0054] The present invention can also take the form of a Factor XIIIA expression enhancer for enhancing the expression of TGF-β2, which comprises an extract of the above-mentioned plant as an active ingredient. Furthermore, the present invention can also be in the form of a composition for enhancing expression of Factor XIIIA, which comprises an extract of the above-mentioned plant as an active ingredient and is intended to enhance the expression of TGF-β2.

[0055] TGF-β2 is a multifunctional cytokine that controls cell proliferation, differentiation, cell death, etc. TGF-β is closely involved in cell differentiation, migration, and adhesion, and plays an important role in a wide range of areas, including ontogenesis, tissue reconstruction, wound healing, inflammation, immunity, and cancer invasion and metastasis. In mammals, TGF-β exists in three structurally very similar isoforms: TGF-β1, TGF-β2, and TGF-β3.

[0056] Here, the method for detecting whether or not the expression of TGF-β2 in cells is enhanced is not particularly limited, and any method that can be appropriately selected by those skilled in the art can be used.

[0057] Furthermore, as shown in the Examples below, the present inventors have found that damaged DNA is repaired and DNA damage is suppressed in the presence of Factor XIII A. This means that the expression of Factor XIII A can be enhanced to repair damaged DNA and / or suppress DNA damage. Therefore, the above-mentioned plant extract having the effect of enhancing the expression of Factor XIII A can be used for repairing damaged DNA and / or suppressing DNA damage. That is, the present invention can be in the form of an agent for repairing damaged DNA and / or suppressing DNA damage (referred to as "agent for repairing damaged DNA and / or agent for suppressing DNA damage" in this specification) containing the above-mentioned plant extract as an active ingredient. The present invention can also provide a composition for repairing damaged DNA and / or a composition for suppressing DNA damage, which comprises an extract of the above-mentioned plant as an active ingredient.

[0058] In addition, in a preferred embodiment of the present invention, the damaged DNA repair agent and / or DNA damage suppressor of the present invention is an agent for repairing damaged DNA and / or suppressing DNA damage induced by enhanced expression of Factor XIIIA.

[0059] The agent for repairing damaged DNA of the present invention is preferably an expression enhancer of one or more genes selected from LIG4 (DNA Ligase 4), NHEJ1 (Non-Homologous End Joining factor 1), PRKDC (Protein Kinase, DNA-Activated, Catalytic Subunit), XRCC4 (X-Ray Repair Cross Complementing 4), XRCC5 (X-Ray Repair Cross Complementing 5), and XRCC6 (X-Ray Repair Cross Complementing 6).

[0060] The present invention can also take the form of a Factor XIIIA expression enhancer for repairing damaged DNA and / or suppressing DNA damage, comprising an extract of the above-mentioned plant as an active ingredient. Furthermore, the present invention can also be in the form of a composition for enhancing expression of Factor XIIIA, which comprises an extract of the above-mentioned plant as an active ingredient, for repairing damaged DNA and / or suppressing DNA damage.

[0061] Conditions and diseases caused by DNA damage include abnormalities in epidermal turnover, which can lead to impaired skin barrier function, hyperkeratosis, acne, pigmentation, and rough skin. Therefore, the agent for repairing damaged DNA and / or the agent for suppressing DNA damage of the present invention can be used to normalize epidermal turnover. Normalization of epidermal turnover includes improvement of skin barrier function, prevention or improvement of hyperkeratosis, prevention or improvement of acne, prevention or improvement of pigmentation, and prevention or improvement of rough skin.

[0062] Here, the method for detecting whether or not damaged DNA in a cell has been repaired and / or DNA damage has been suppressed is not particularly limited, and any method that can be appropriately selected by a person skilled in the art can be used.

[0063] Furthermore, as described above, the present inventors have demonstrated that expression of Factor XIIIA is enhanced in cells in the G0 phase. That is, in a preferred embodiment of the present invention, the Factor XIIIA expression enhancer of the present invention can be used to enhance the expression of Factor XIIIA in cells in the G0 phase.

[0064] In another preferred embodiment of the present invention, the cell proliferation-promoting agent of the present invention is an agent for promoting cell proliferation induced by enhanced expression of Factor XIIIA in cells in the G0 phase.

[0065] In another preferred embodiment of the present invention, the collagen production enhancer of the present invention is an agent for enhancing collagen production induced by enhanced expression of Factor XIIIA in cells in the G0 phase.

[0066] In another preferred embodiment of the present invention, the TGF-β2 expression enhancer of the present invention is an agent for enhancing the expression of TGF-β2 induced by enhanced expression of Factor XIIIA in cells in the G0 phase.

[0067] In another preferred embodiment of the present invention, the damaged DNA repair agent and / or DNA damage suppressor of the present invention is an agent for repairing damaged DNA and / or suppressing DNA damage induced by increased expression of Factor XIIIA in cells in the G0 phase.

[0068] In the present invention, cells that are out of the cell cycle and in a resting state are defined as cells in the G0 phase (resting phase). Cells that are in a phase other than the G0 phase, such as the G1 phase, S phase, G2 phase, or M phase, are referred to as cells in an active phase.

[0069] ≪Dosage / Dosage≫ In the present invention, the original extract obtained by extraction with any solvent may be used as is as the agent of the present invention, or the original extract may be diluted to any concentration and used as the agent of the present invention.

[0070] The total content of one or more types selected from the extracts of the plants of the present invention in the agent of the present invention is usually 0.00001 mass% or more, based on the dry mass, preferably 0.0001 mass% or more, more preferably 0.001 mass% or more, even more preferably 0.01 mass% or more, and even more preferably 0.1 mass% or more.

[0071] Furthermore, the total content of one or more types selected from the extracts of the plants of the present invention in the agent of the present invention is usually 80 mass % or less, based on the dry mass, preferably 50 mass % or less, more preferably 30 mass % or less, even more preferably 10 mass % or less, and even more preferably 1 mass % or less.

[0072] The agent according to the present invention is preferably in the form of an agent for external application on the skin or an oral agent, which can be produced by appropriately using known methods and devices.

[0073] Suitable examples of external skin preparations include cosmetics, quasi-drugs, and medicines, with cosmetics and quasi-drugs being more preferred since they can be used on a daily basis. Cosmetics can be processed into the form of lotions, emulsions, gels, creams, ointments, etc. Specific examples include lotions, milky lotions, creams, gels, packs, hair creams, sprays, sun care products, etc., and particularly preferred examples include lotions, milky lotions, and creams.

[0074] The total content of one or more types of extracts selected from the plant extracts of the present invention in the topical skin preparation is usually 0.00001 mass% or more, based on the dry mass, preferably 0.0001 mass% or more, and more preferably 0.0003 mass% or more.

[0075] In addition, the total content of one or more types selected from the plant extracts of the present invention in the topical skin preparation is usually 80 mass% or less, based on the dry mass, preferably 50 mass% or less, more preferably 30 mass% or less, more preferably 10 mass% or less, more preferably 1 mass% or less, more preferably 0.1 mass%, and even more preferably 0.05 mass% or less.

[0076] When the present invention is in the form of an external skin preparation, optional ingredients may be blended therein, within the scope of not impairing the effect of enhancing the expression of Factor XIIIA, such as whitening ingredients, wrinkle-reducing ingredients, anti-inflammatory ingredients, extracts derived from plants or animals other than the plant of the present invention, and ingredients commonly used in cosmetics other than the active ingredient. These optional ingredients may be commercially available and used, or may be synthesized by a known method and used. Each optional ingredient may have two or more effects (for example, a whitening effect and an anti-wrinkle effect).

[0077] When the agent according to the present invention is prepared as an oral agent, it is preferably in the form of a food or drink containing one or more kinds selected from the extracts of the plant according to the present invention as active ingredients.

[0078] When the agent of the present invention is prepared as an oral agent, the total content of one or more active ingredients selected from the extract of the plant of the present invention is usually 0.1 mg or more, preferably 1 mg or more, and more preferably 10 mg or more, per dose, as the dry mass of the extract, depending on the dosage form.

[0079] In addition, the total content of one or more active ingredients selected from the plant extract of the present invention depends on the dosage form, and the amount to be taken per dose is usually 2000 mg or less, preferably 1000 mg or less, and more preferably 500 mg or less, in terms of the dry mass of the extract.

[0080] Furthermore, when the present invention is in the form of an external skin preparation or an oral preparation, it is also preferable that the present invention be in a form labeled with the following uses: "for enhancing Factor XIII A expression," "for promoting cell proliferation," "for enhancing collagen production," "for enhancing TGF-β2 expression," "for repairing damaged DNA," and / or "for inhibiting DNA damage."

[0081] The above-mentioned "representation" includes all representations that have the function of informing consumers of the above-mentioned uses. In other words, any representation that can recall or infer the above-mentioned uses falls under the above-mentioned "representation", regardless of the purpose of the representation, the content of the representation, the object or medium on which it is displayed, etc. In addition, the phrase "labeled" means that there is a labeling action intended to cause the viewer to associate the label with a skin topical agent, etc. (product).

[0082] The act of displaying the intended use is preferably one that enables consumers to directly recognize the intended use.Specific examples include the act of describing the intended use on the product or its packaging according to the present invention, and the act of describing the intended use in advertisements for the product, price lists, or transaction documents (including those provided by electromagnetic means).

[0083] <Screening method> The present invention is a screening method for screening for substances having one or more of the following effects: promoting cell proliferation, enhancing collagen production, enhancing TGF-β2 (Transforming Growth Factor β2) expression, repairing damaged DNA, and suppressing DNA damage, using the expression of Coagulation Factor XIII A in cells as an indicator.

[0084] This screening method is an invention based on the discovery by the present inventors that the presence of Factor XIII A promotes fibroblast proliferation, enhances collagen production in the fibroblasts, and induces enhanced expression of the TGFB2 gene in the fibroblasts, repairs damaged DNA, and suppresses DNA damage.

[0085] The substances to be screened in the present invention have one or more of the following effects: promoting cell proliferation, enhancing collagen production, enhancing TGF-β2 expression, repairing damaged DNA, and inhibiting DNA damage; they may have two or more of these effects, or they may have all of these effects. Here, the substances to be screened in the present invention may include candidate substances for the active ingredients having the above-mentioned actions.

[0086] When screening for a substance having a cell proliferation-promoting effect, the present invention preferably provides a screening method comprising the steps of culturing cells in the presence of a test substance (culturing step), measuring the expression level of Factor XIII A in the cells (measuring step), and, if the expression level is higher than the expression level of Factor XIII A in cells cultured in the absence of the test substance, selecting the test substance as a substance having a cell proliferation-promoting effect (selecting step).

[0087] When screening for a substance having the effect of enhancing collagen production, it is preferable to select the test substance as a substance having the effect of enhancing collagen production in the selection step when the expression level of Factor XIII A in cells cultured in the presence of the test substance is higher than the expression level of Factor XIII A in cells cultured in the absence of the test substance.

[0088] When screening for a substance having the effect of enhancing TGF-β2 expression, it is preferable to select the test substance as a substance having the effect of enhancing TGF-β2 expression in the selection step when the expression level of Factor XIII A in cells cultured in the presence of the test substance is higher than the expression level of Factor XIII A in cells cultured in the absence of the test substance.

[0089] When screening for a substance having an effect of repairing damaged DNA and / or suppressing DNA damage, it is preferable to select the test substance as a substance having an effect of repairing damaged DNA and / or suppressing DNA damage in the selection step when the expression level of Factor XIII A in cells cultured in the presence of the test substance is higher than the expression level of Factor XIII A in cells cultured in the absence of the test substance.

[0090] The screening method of the present invention will be described below. The screening method includes a culture step, a measurement step, and a selection step. Each step will be described in detail below.

[0091] (i)Culture process The culturing step is a step of culturing cells in the presence of a test substance. The method of cell culture is not particularly limited, and adherent culture or suspension culture can be appropriately adopted depending on the properties of the cells used.

[0092] In addition, the cells used in the present invention are preferably cells of skin tissue. From the viewpoint of evaluating a substance acting mainly on the dermal tissue layer of skin tissue using the expression of Factor XIIIA as an indicator, it is preferable to use fibroblasts, and among them, dermal fibroblasts are preferable, and for example, normal human dermal fibroblasts (NHDF) can be preferably used. Fibroblasts present in subcutaneous tissue can also be used.

[0093] In the present invention, skin collected from a living body or a three-dimensional cultured skin model may be used. The skin is preferably a skin fragment collected from a living body, from which body hair has been removed as desired, and which is composed of the granular layer of the epidermis, the stratum spinosum, the basal layer of the epidermis, the dermis, and subcutaneous tissue (including adipose tissue).

[0094] A preferred three-dimensional cultured skin model is one in which cells taken from human skin (normal human epidermal keratinocytes (NHEK), normal human dermal fibroblasts (NHDF), etc.) are cultured to construct a three-dimensional structure that mimics the structure of skin, and a commercially available product in this form can also be purchased and used. A preferred example of a commercially available product is "EFT-400" sold by Kurabo Industries Co., Ltd.

[0095] In addition, in the present invention, it is preferable to use cells induced to the G0 phase. The method for inducing cells into the G0 phase can be an existing method, for example, a method of culturing cells in a serum-depleted medium for a certain period of time to obtain a cell population rich in cells in the G0 phase of the cell cycle can be preferably exemplified. Another example is a method in which cells cultured in a normal manner are labeled with Ki67 antibody, which is a cell marker for the active phase, and Ki67 (MKI67)-negative cells are separated as G0-phase cells by flow cytometry. The type of antibody is not limited, and any antibody that can label G0-phase cells can be used. In addition, it may be confirmed that cells have been induced to the G0 phase by confirming a decrease in expression of the MKI67 gene that codes for Ki67. Another example is a method in which drugs such as ubiquitin ligase Fbxw7 or cyclin C inhibitors that arrest the cell cycle are added to cells cultured in a normal manner.

[0096] The culture medium is not particularly limited, and any known medium can be used. Suitable examples of the medium include HuMedia KG2 and DMEM (Dulbecco's Modified Eagle Medium).

[0097] The period of the culture step is not particularly limited, and is preferably 3 hours or more, more preferably 6 hours or more, even more preferably 12 hours or more, and particularly preferably 1 day or more. The upper limit of the culture period is also not particularly limited, and as a guideline, it is preferably 1 month or less, more preferably 3 weeks or less, even more preferably 2 weeks or less, and particularly preferably 1 week or less.

[0098] In the culturing step, culturing is performed in the presence of a test substance. Specifically, an embodiment in which a test substance is added to a medium and culturing is performed can be mentioned. The type of test substance is not limited, and low molecular weight compounds, high molecular weight compounds, proteins, peptides, nucleic acids, etc. can be applied without limit. The test substance may be a pure compound or a mixture containing two or more components, such as a plant or animal extract.

[0099] In the culture step, one type of test substance may be used, or two or more types of test substances may be used. When evaluating a synergistic effect due to a combination of test substances, it is preferable to add two or more types of test substances.

[0100] In the culturing step, in parallel with culturing in the presence of the test substance, cell culture in the absence of the test substance may be carried out as a control. In this case, it is preferable to keep the culture conditions the same except for the presence or absence of the test substance.

[0101] (ii) Measurement process The measuring step is a step of measuring the expression of Factor XIII A in the cells that have been subjected to the culturing step. Preferably, in the measuring step, the expression level of Factor XIII A is measured. The method for measuring the expression level of Factor XIIIA is not particularly limited, and examples thereof include quantitative PCR and Northern blotting in which mRNA transcribed from a gene encoding Factor XIIIA is the measurement target, and immunological techniques such as immunostaining, Western blotting, and ELISA in which Factor XIIIA is used as an indicator substance. From the viewpoints of high quantitativeness and simplicity, quantitative PCR is particularly preferred. Specific embodiments of these measurement means are not particularly limited, and they can be performed by conventional methods.

[0102] Factor XIIIA is encoded by the F13A (F13A1) gene. In the measuring step, it is preferable to measure the expression of the F13A (F13A1) gene.

[0103] The sequence of the human gene encoding Factor XIIIA has been published, and when performing quantitative PCR to measure mRNA, those skilled in the art can design appropriate primers and subject them to PCR. Commercially available primers may also be used.

[0104] In addition, when the evaluation is performed using cells induced to the G0 phase, a preferable example of the measurement step is to calculate the increase rate of expression of Factor XIIIA in cells cultured in the presence of a test substance. This includes a step of measuring the expression level of Factor XIIIA in the cells and calculating, based on the expression level, the increase rate of expression of Factor XIIIA in the cells relative to the expression level of Factor XIIIA in cells cultured in the absence of the test substance. Specifically, when the expression level of Factor XIIIA in cells induced to the G0 phase and cultured in the absence of a test substance is taken as a reference value (1.00), the change in the ratio of the expression level of Factor XIIIA in cells induced to the G0 phase and cultured in the presence of a test substance can be regarded as the rate of increase in expression of Factor XIIIA. The expression level ratio is preferably 1.10 or more relative to the reference value (1.00), and more preferably 1.25 or more.

[0105] (iii) Selection step The selection step is a step of selecting a substance having a cell proliferation promoting effect, a collagen production enhancing effect, a TGF-β2 expression enhancing effect, and / or a damaged DNA repair effect and / or a DNA damage suppressing effect based on the expression of Factor XIII A measured in the measurement step. More specifically, when the expression level of Factor XIII A in cells cultured in the presence of a test substance is higher than the expression level of Factor XIII A in cells cultured in the absence of the test substance, the test substance is selected as a substance having a cell proliferation promoting effect, a collagen production enhancing effect, a TGF-β2 expression enhancing effect, a damaged DNA repair effect and / or a DNA damage suppressing effect.

[0106] In the present invention, "enhanced collagen production" preferably refers to enhanced production of total collagen.

[0107] In the present invention, "enhanced expression of TGF-β2" preferably refers to enhanced expression of the TGFB2 gene.

[0108] Furthermore, in the present invention, "repair of damaged DNA" preferably refers to enhanced expression of DNA damage repair-related genes, and more preferably refers to enhanced expression of one or more genes selected from LIG4 (DNA Ligase 4), NHEJ1 (Non-Homologous End Joining factor 1), PRKDC (Protein Kinase, DNA-Activated, Catalytic Subunit), XRCC4 (X-Ray Repair Cross Complementing 4), XRCC5 (X-Ray Repair Cross Complementing 5), and XRCC6 (X-Ray Repair Cross Complementing 6).

[0109] Furthermore, in the present invention, "inhibition of DNA damage" preferably refers to inhibition of the production of phosphorylated H2AX (γH2AX).

[0110] Here, the test substance selected in the selection step may be determined as an active ingredient as it is, or may be determined as a candidate active ingredient for use in a secondary screening.

[0111] Even when evaluation is performed using cells induced to the G0 phase, as described above, by comparing the expression levels of Factor XIII A in cells cultured in the absence of the test substance and in cells cultured in the presence of the test substance, a substance having the effect of promoting cell proliferation, enhancing collagen production, enhancing TGF-β2 expression, repairing damaged DNA, and / or suppressing DNA damage can be selected.

[0112] By screening using cells induced to the G0 phase, production of Factor XIII A is enhanced even in cells that have fallen out of the cell cycle and are in a dormant state, making it possible to screen for substances that can promote cell proliferation, enhance collagen production, enhance TGF-β2 expression, and induce repair of damaged DNA and / or suppression of DNA damage.

[0113] Furthermore, when the evaluation is performed using cells induced to the G0 phase, a preferred example is a mode in which a substance having a cell proliferation promoting effect, a collagen production enhancing effect, a TGF-β2 expression enhancing effect, a damaged DNA repair effect, and / or a DNA damage suppression effect is selected based on the "expression increase rate of Factor XIIIA in cells cultured in the presence of the test substance" calculated in the (ii) measurement step. In this embodiment, when the expression increase rate is higher than the standard value, the test substance is selected as a substance having a cell proliferation promoting effect, a collagen production enhancing effect, a TGF-β2 expression enhancing effect, a damaged DNA repair effect, and / or a DNA damage suppression effect. According to this embodiment, it is possible to screen for active ingredients based on the expression of Factor XIIIA increased by induction into the G0 phase, and therefore screening can be performed with high accuracy.

[0114] Here, the reference value is preferably the expression level of Factor XIIIA in cells induced to the G0 phase and cultured in the absence of a test substance.

[0115] The substance selected by the above-mentioned screening method using the expression of Factor XIIIA as an index can be incorporated into a composition by any preparation method. That is, the above-mentioned screening method using the expression of Factor XIIIA as an index can be suitably used for designing a composition. Suitable examples of such compositions include pharmaceuticals, quasi-drugs, cosmetics and other skin care compositions, foods and beverages, and the like. EXAMPLES

[0116] The present invention will be described in more detail below with reference to examples, but it goes without saying that the present invention is not limited to these examples.

[0117] In the examples, the plant extracts used were Passiflora edulis peel extract, artichoke leaf extract, and echinacea leaf extract, respectively, as the plant extracts of plants belonging to the Passiflora genus of the Passifloraceae family, the Artichoke genus of the Asteraceae family, and the Echinacea genus of the Asteraceae family.

[0118] <Test Example 1> Enhanced expression of Factor XIII A in cells induced to the G0 phase Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using medium (10% FBS + DMEM). After the cells attached, they were cultured in medium (10% FBS + DMEM, no serum depletion (control)) or medium (0% FBS + DMEM, serum depletion), and RNA was extracted from the cells.

[0119] The collected RNA was analyzed by real-time PCR to evaluate the expression of F13A gene and, at the same time, to confirm that the cells were induced into the G0 phase, the decrease in the expression of MKI67 gene was confirmed (n=4).

[0120] [Table 1]

[0121] As shown in Table 1, a significant decrease in MKI67 gene expression was confirmed under serum deprivation conditions compared to non-serum deprivation conditions, which indicates that cells were induced to the G0 phase under serum deprivation conditions. Furthermore, it was confirmed that F13A gene expression was enhanced under serum deprivation conditions in which induction to the G0 phase was confirmed.

[0122] It was also shown that when screening for substances using cells induced to the G0 phase, the expression of Factor XIII A can be used as an index. Furthermore, since the expression of Factor XIII A is higher in the G0 phase than in the active phase, it can be said that when screening is performed using the expression of Factor XIII A as an index, more accurate screening can be performed by using cells induced to the G0 phase.

[0123] <Test Example 2> Screening of extracts that enhance the expression of Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using a medium (10% FBS + DMEM). After the cells attached, the extracts (plant extracts) listed in Table 2 below were added to the medium (0% FBS + DMEM) to a final concentration of 0.1% (dry mass: artichoke leaf extract 0.0008 mass%, Passiflora edulis peel extract 0.005 mass%, echinacea leaf extract 0.0031 mass%) to prepare an extract-containing medium. The cells were cultured in the medium, and RNA was collected from the cells. The control (NC) was subjected to the same conditions as above except that the extract was not added.

[0124] The collected RNA was analyzed using real-time PCR to evaluate the expression of the F13A gene. Specifically, the gene expression level in serum-starved cells without extract was used as the standard, and the gene expression level after extract addition in serum-starved cells was calculated. Table 2 shows the ratio of gene expression level after extract addition, when the gene expression level in serum-starved cells without extract addition is set to "1.00".

[0125] [Table 2]

[0126] As shown in Table 2, the expression level of the F13A gene was increased when 0.1% Passiflora edulis peel extract, 0.1% artichoke leaf extract, or 0.1% echinacea leaf extract was added. This indicates that plant extracts of plants belonging to the genus Passiflora in the family Passifloraceae, the genus Artichoke in the family Asteraceae, and the genus Echinacea in the family Asteraceae are capable of increasing the expression of Factor XIII A.

[0127] Furthermore, in Test Example 2, screening was performed using cells induced to the G0 phase and in which expression of the F13A gene was increased. Therefore, it can be said that the plant extracts of plants belonging to the genus Passiflora in the family Passifloraceae, the genus Artichoke in the family Asteraceae, and the genus Echinacea in the family Asteraceae are capable of further enhancing the expression of Factor XIII A in cells induced to the G0 phase.

[0128] Furthermore, when the two extracts were combined, the combination of 0.1% Passiflora edulis peel extract and 0.1% artichoke leaf extract was found to have a greater effect of enhancing Factor XIII A expression in cells in the G0 phase than when 0.1% Passiflora edulis peel extract or 0.1% artichoke leaf extract was added alone.

[0129] <Test Example 3> Cell proliferation promoting effect of Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using a medium (10% FBS + DMEM). After the cells attached, the medium (0% FBS + DMEM) was cultured for 24 hours without or with Factor XIII A, and then the medium was replaced with a medium (10% FBS + DMEM) and cell proliferation was evaluated 24 hours and 96 hours later. The cell proliferation rate was calculated by counting the number of cells 24 hours after the medium was replaced with a medium (10% FBS + DMEM) and comparing the number of cells after 96 hours. Table 3 shows the ratio of the cell proliferation rate in the Factor XIII A-added group to the cell proliferation rate in the Factor XIII A-free group (control) taken as "1.00" (n = 5). The control was prepared under the same conditions as above, except that Factor XIIIA was not added.

[0130] [Table 3]

[0131] As shown in Table 3, cell proliferation was promoted under the condition where Factor XIII A was added. This indicates that cell proliferation can be promoted by enhancing the expression of Factor XIII A. In other words, this result indicates that the agent containing the active ingredient according to the present invention can be used in the form of a cell proliferation promoter.

[0132] Furthermore, since cell proliferation is promoted by the expression of Factor XIIIA, it was demonstrated that substances having a cell proliferation-promoting effect can be screened using the expression of Factor XIIIA as an indicator.

[0133] <Test Example 4> Collagen (production) enhancement effect by Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using medium (10% FBS + DMEM). After the cells attached, they were cultured for 72 hours in medium (2% FBS + DMEM) with or without Factor XIII A, after which the cells and culture supernatant were collected.

[0134] The collected cells and culture supernatant were analyzed using the Sirius Red method to evaluate the total collagen content. Table 4 shows the ratio of the total collagen content in the Factor XIII A-added group to the total collagen content in the Factor XIII A-free group (control) taken as 1.00 (n=4). The control was prepared under the same conditions as above, except that Factor XIIIA was not added.

[0135] [Table 4]

[0136] As shown in Table 4, total collagen production was enhanced under the condition where Factor XIIIA was added. This indicates that collagen production can be enhanced by enhancing the expression of Factor XIIIA. In other words, this result indicates that the agent containing the active ingredient according to the present invention can be used in the form of a collagen production enhancer.

[0137] Furthermore, since collagen production is enhanced with the expression of Factor XIIIA, it has been shown that substances having collagen production enhancing effects can be screened using the expression of Factor XIIIA as an indicator.

[0138] <Test Example 5> Enhancement of TGF-β2 expression by Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using medium (10% FBS + DMEM). After the cells attached, they were cultured for 24 hours in medium (2% FBS + DMEM) with or without Factor XIII A, and then RNA was collected from the cells.

[0139] The collected RNA was analyzed using a DNA microarray method to evaluate genes whose expression was altered by the addition of Factor XIII A. Table 5 shows the ratio of gene expression levels in the Factor XIII A-added group to the gene expression level in the Factor XIII A-free group (control) set at "1.00" (n=3). The control was prepared under the same conditions as above, except that Factor XIIIA was not added.

[0140] [Table 5]

[0141] As shown in Table 5, the expression of TGFB2 gene was enhanced under the condition where Factor XIIIA was added. This indicates that it is possible to enhance the expression of TGF-β2 by enhancing the expression of Factor XIIIA. In other words, this result indicates that the agent containing the active ingredient according to the present invention can be used in the form of a TGF-β2 expression enhancer.

[0142] Furthermore, because the expression of Factor XIII A is accompanied by an increase in TGF-β2 expression, it has been shown that substances that have the effect of increasing TGF-β2 expression can be screened using the expression of Factor XIII A as an indicator.

[0143] <Test Example 6> Repair effect of damaged DNA by Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using medium (10% FBS + DMEM). After the cells attached, they were cultured for 24 hours in medium (2% FBS + DMEM) with or without Factor XIII A, and then RNA was collected from the cells.

[0144] The collected RNA was analyzed by real-time PCR to evaluate genes whose expression was altered by the addition of Factor XIII A. Table 6 shows the ratio of gene expression levels in the Factor XIII A-added groups, when the gene expression level in the Factor XIII A-free group (control) was set to "1.00" (n=5). The control was prepared under the same conditions as above, except that Factor XIIIA was not added.

[0145] [Table 6]

[0146] As shown in Table 6, when Factor XIII A was added, gene expression of LIG4 (DNA Ligase 4), NHEJ1 (Non-Homologous End Joining factor 1), PRKDC (Protein Kinase, DNA-Activated, Catalytic Subunit), XRCC4 (X-Ray Repair Cross Complementing 4), XRCC5 (X-Ray Repair Cross Complementing 5), and XRCC6 (X-Ray Repair Cross Complementing 6), which act in non-homologous end joining, one of the DNA damage repair mechanisms, was enhanced. This result indicates that damaged DNA is repaired under the condition of adding Factor XIII A (F13A).

[0147] From the above, it can be said that the expression of LIG4, NHEJ1, PRKDC, XRCC4, XRCC5, and XRCC6 can be enhanced by enhancing the expression of factor XIIIA. It can also be said that damaged DNA can be repaired by enhancing the expression of factor XIIIA. In other words, this result shows that the agent containing the active ingredient according to the present invention can be used as a form of a repair agent for damaged DNA.

[0148] Furthermore, because damaged DNA is repaired in association with the expression of Factor XIIIA, it has been shown that substances that have the effect of repairing damaged DNA can be screened using the expression of Factor XIIIA as an indicator.

[0149] <Test Example 7> Inhibitory effect of DNA damage caused by Factor XIII A Normal human dermal fibroblasts (NHDFs) were seeded on a culture plate to be semi-confluent using medium (10% FBS + DMEM). After the cells attached, they were cultured for 24 hours in medium (0% FBS + DMEM) without or with Factor XIII A, and then the medium was replaced with 10% FBS + DMEM. After 24 hours, the cells were evaluated by immunostaining for γH2AX (phosphorylated H2A.X variant histone), which is known as a DNA double-strand break marker. Table 7 shows the amount of DNA damage as the ratio of the fluorescence intensity derived from γH2AX in the Factor XIII A-added group to the fluorescence intensity derived from γH2AX in the Factor XIII A-free group (control) as "1.00" (n = 5). The control was prepared under the same conditions as above, except that Factor XIIIA was not added.

[0150] [Table 7]

[0151] As shown in Table 7, the fluorescence intensity derived from γH2AX decreased under the condition where Factor XIII A was added. This result indicates that the level of DNA damage was reduced under the condition where Factor XIII A was added.

[0152] From the above, it can be said that it is possible to suppress DNA damage by enhancing the expression of Factor XIII A. In other words, this result indicates that the agent containing the active ingredient of the present invention can be used in the form of an agent for suppressing DNA damage.

[0153] Furthermore, because DNA damage was suppressed with the expression of Factor XIII A, it was demonstrated that substances that have the effect of suppressing DNA damage can be screened using the expression of Factor XIII A as an indicator. [Industrial Applicability]

[0154] According to the present invention, a composition capable of enhancing the expression of Factor XIIIA can be provided.

Claims

1. An agent for enhancing expression of Coagulation Factor XIII A, comprising as an active ingredient one or more selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.

2. The agent according to claim 1 for enhancing expression of Coagulation Factor XIII A in cells in the G0 phase.

3. An agent for promoting cell proliferation, comprising as an active ingredient one or more selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.

4. An agent for enhancing collagen production, comprising as an active ingredient one or more selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.

5. An agent for enhancing expression of TGF-β2 (Transforming Growth Factor β2), comprising, as an active ingredient, one or more selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.

6. An agent for repairing damaged DNA and / or suppressing DNA damage, comprising as an active ingredient one or more selected from an extract of a plant belonging to the genus Passiflora in the family Passifloraceae, an extract of a plant belonging to the genus Cynara in the family Asteraceae, and an extract of a plant belonging to the genus Echinacea in the family Asteraceae.

7. The agent according to any one of claims 3 to 6, which is induced by enhanced expression of Coagulation Factor XIII A.

8. The agent according to any one of claims 3 to 6, which is induced by enhanced expression of Coagulation Factor XIII A in cells in the G0 phase.

9. The agent according to any one of claims 1 to 6, comprising an extract of a plant belonging to the genus Passiflora in the family Passifloraceae and an extract of a plant belonging to the genus Cynara in the family Asteraceae as active ingredients.

10. The extract of a plant belonging to the genus Passiflora in the family Passifloraceae is a fruit peel extract of Passiflora edulis, The extract of a plant belonging to the genus Cynara in the family Asteraceae is an artichoke (Cynara scolymus) leaf extract, The agent according to any one of claims 1 to 6, wherein the extract of a plant belonging to the genus Echinacea in the family Asteraceae is an extract of Echinacea angustifolia leaves.

11. The agent according to any one of claims 1 to 6, which is an external skin agent.

12. A screening method for screening a substance having an effect of enhancing collagen production, using the expression of Coagulation Factor XIII A in cells as an indicator.

13. A screening method for screening a substance having an effect of enhancing expression of TGF-β2 (Transforming Growth Factor β2) using the expression of Coagulation Factor XIII A in cells as an index.

14. A screening method for screening a substance having an activity of repairing damaged DNA and / or an activity of suppressing DNA damage, using the expression of Coagulation Factor XIII A in cells as an indicator.

15. The method of any one of claims 12 to 14, wherein the cell is a fibroblast.

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