JP5545938B2 - Stem cell growth factor (SCF) mRNA expression promoter - Google Patents

Description

  The present invention relates to a stem cell growth factor (SCF) mRNA expression promoter.

  Stem Cell Factor (SCF), also called Must Cell Growth Factor, C-Kit Ligand, Steel Factor, etc., is a protein produced from keratinocytes, fibroblasts, vascular endothelial cells, bone marrow stromal cells, etc. It is. SCF is known to have an action of promoting proliferation and differentiation of pluripotent hematopoietic stem cells, germ cells, mast cells, megakaryocyte precursor cells, granulocyte / macrophage precursor cells, pigment cells and the like. Moreover, it is known that the expression of SCF is enhanced by a spot site, ultraviolet irradiation or the like (see Non-Patent Document 1).

  As SCF, a membrane-bound SCF composed of 273 amino acid residues and a secreted SCF that is cleaved by the action of a proteolytic enzyme and released from the membrane are known. Membrane-bound SCF binds to the SCF receptor of the pigment cell while bound to keratinocytes and promotes the proliferation of the pigment cell. Secreted SCF is cleaved at the binding site, released from the cell membrane, and bound to the SCF receptor of the pigment cell, thereby promoting proliferation of the pigment cell. That is, it is expected that by promoting the production of SCF in living cells, the proliferation of pigment cells can be promoted, and as a result, the production of melanin can be promoted.

  The color tone of the skin and hair depends on the amount of melanin contained in the skin and hair. Melanin is converted from tyrosine to dopa and from dopa to dopaquinone by the action of the enzyme tyrosinase, which is biosynthesized in pigment cells existing in the skin and hair, and then formed through intermediates such as 5,6-dihydroxyindophenol. It is supposed to be done. By activating or growing the pigment cells, melanin is synthesized. As a result, the hair becomes black and the skin becomes brown (black).

  Whitening, which is one of the aging phenomena, is considered to be caused by a decrease in the amount of melanin produced in the hair papilla cells. Therefore, if SCF production in hair papilla cells can be promoted, melanin production can be promoted through it, and as a result, graying can be prevented or improved.

  Moreover, brown skin is being accepted as a part of fashion as a healthy skin color regardless of gender. This tendency is particularly strong in Europe and the United States, and brown skin is a kind of status symbol. Therefore, there is a demand for cosmetics that can turn the skin brown (black) without exposure to ultraviolet rays that can cause stains, buckwheat, skin cancer, and the like. Therefore, if SCF production in epidermal fibroblasts can be promoted, melanin production can be promoted through it, and as a result, it is considered that the skin can be browned (black).

  Based on such an idea, for example, an extract of edible chrysanthemum (see Patent Document 1), an extract of Antrodia camphorata (see Patent Document 2), a plant belonging to the genus Camphoraceae, The extract (refer patent document 3) etc. are known.

JP 2007-197352 A JP 2007-071119 A JP 2005-247736 A

Hachiya A et al., J. Invest. Dermatol., No. 116, 2001, p. 578-586

  An object of the present invention is to provide a stem cell growth factor (SCF) mRNA expression promoting agent that has a stem cell growth factor (SCF) mRNA expression promoting action and uses it as an active ingredient.

  In order to solve the above-mentioned problems, the stem cell growth factor (SCF) mRNA expression promoter of the present invention is an active ingredient comprising an extract from one or more plants selected from the group consisting of ashitaba, nettle and auren. It is characterized by containing as.

  According to the present invention, a stem cell growth factor (SCF) mRNA expression promoter containing a highly safe natural extract as an active ingredient can be provided.

The present invention will be described below.
The SCF mRNA expression promoter of the present invention contains, as an active ingredient, an extract from one or more plants selected from the group consisting of ashitaba, nettle and auren.

  In the present invention, the “extract” refers to an extract obtained using the above plant as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude product thereof. Both purified products and purified products are included.

  The extraction raw material used in the present invention is ashitaba (scientific name: Angelica keiskei Koidzumi), nettle (scientific name: Urtica thubergiana) or auren (scientific name: Coptis japonica Makino).

  Ashitaba (Angelica keiskei Koidzumi) is a perennial plant belonging to the genus Sisiudo that grows naturally in the Boso Peninsula, Izu Peninsula, Izu Islands, etc., and can be easily obtained from these areas. Examples of the constituent parts of Ashitaba that can be used as an extraction raw material include leaf parts, stem parts, flower parts, cocoon parts, fruit parts, fruit skin parts, fruit core parts, above-ground parts, or a mixture thereof. The leaf portion is preferred.

  Nettle (Urtica thubergiana) is a perennial herb belonging to the genus Nettle, which is distributed in Honshu, Shikoku, Kyushu, the Korean Peninsula, etc. in Japan, and can be easily obtained from these areas. Examples of the constituent parts of nettle that can be used as an extraction raw material include leaf parts, stem parts, flower parts, root parts, root bark parts, above-ground parts, whole grasses, or a mixture thereof. It is.

  Ouren (Coptis japonica Makino) is a perennial plant belonging to the genus Ourenaceae that is distributed in the mountains of Hokkaido, Honshu, and Shikoku, and can be easily obtained from these regions. Examples of constituent parts of auren that can be used as an extraction raw material include, for example, leaf parts, branch parts, bark parts, trunk parts, stem parts, fruit parts, seed parts, flower parts and the like, rhizome parts, or a mixture of these parts. The rhizome is preferred.

  Although the details of the substance having SCF mRNA expression promoting action contained in the extract from one or more plants selected from the group consisting of Ashitaba, nettle and auren are unknown, they are generally used for plant extraction. The extract having this action can be obtained from the above plant by the extraction method.

  For example, after drying the plant, it is pulverized as it is or using a crusher, and subjected to extraction with an extraction solvent, whereby an extract having an SCF mRNA expression promoting action can be obtained. Drying may be performed in the sun or using a commonly used dryer. Moreover, after performing pretreatment, such as degreasing, with a nonpolar solvent such as hexane, it may be used as an extraction raw material. By performing pretreatment such as degreasing, extraction treatment with the polar solvent of the plant can be efficiently performed.

  As the extraction solvent, it is preferable to use a polar solvent, and examples thereof include water and hydrophilic organic solvents. These are used alone or in combination of two or more at room temperature or a temperature below the boiling point of the solvent. It is preferable.

  Examples of water that can be used as the extraction solvent include pure water, tap water, well water, mineral spring water, mineral water, hot spring water, spring water, fresh water, and the like, and those subjected to various treatments. Examples of the treatment applied to water include purification, heating, sterilization, filtration, ion exchange, osmotic pressure adjustment, buffering, and the like. Therefore, the water that can be used as the extraction solvent in the present invention includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones such as acetone and methyl ethyl ketone; 1,3-butylene. Examples thereof include polyhydric alcohols having 2 to 5 carbon atoms such as glycol, propylene glycol and glycerin.

  When using the liquid mixture of 2 or more types of polar solvents as an extraction solvent, the mixing ratio can be adjusted suitably. For example, when using a mixed solution of water and a lower aliphatic alcohol, it is preferable to mix 1 to 90 parts by volume of a lower aliphatic alcohol with respect to 10 parts by volume of water. When using a mixed solution, it is preferable to mix 1 to 40 parts by volume of a lower aliphatic ketone with 10 parts by volume of water, and when using a mixed solution of water and a polyhydric alcohol, water 10 It is preferable to mix 10 to 90 parts by volume of a polyhydric alcohol with respect to the volume part.

  The extraction treatment is not particularly limited as long as the soluble component contained in the extraction raw material can be eluted in the extraction solvent, and can be performed according to a conventional method. For example, the extraction raw material is immersed in an extraction solvent 5 to 15 times (mass ratio) of the extraction raw material, the soluble components are extracted at room temperature or under reflux, and then filtered to remove the extraction residue. A liquid can be obtained. The obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method in order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or a purified product thereof. Processing may be performed.

  Purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, or the like. The obtained extract can be used as it is as an active ingredient of the SCF mRNA expression promoter, but a concentrate or a dried product is easier to use.

  Since the extract from one or more plants selected from the group consisting of Ashitaba, nettle and auren obtained as described above has an SCF mRNA expression promoting action, the action is utilized. It can be used as an active ingredient of an SCF mRNA expression promoter.

  The SCF mRNA expression promoter of the present invention may consist only of an extract from one or more plants selected from the group consisting of ashitaba, nettle and auren, or a formulation of the plant extract. But you can.

  Extracts from one or more plants selected from the group consisting of ashitaba, nettle and auren can be extracted using a pharmaceutically acceptable carrier such as dextrin, cyclodextrin, etc. and other optional auxiliaries. Accordingly, it can be formulated into an arbitrary dosage form such as powder, granule, tablet, and liquid. In this case, as an auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring agent and the like can be used. Extracts from one or more plants selected from the group consisting of Ashitaba, Nettle and Ouren can be used by blending with other compositions (for example, hair cosmetics, skin cosmetics, etc.). In addition, it can be used as an ointment, a liquid for external use, a patch and the like.

  In addition, the SCF mRNA expression promoter of the present invention is a natural extract or the like having SCF mRNA expression promoting action, if necessary, from one or more kinds of plants selected from the group consisting of Ashitaba, nettle and auren. It can be used together with the extract as an active ingredient.

  Examples of the method of administering the SCF mRNA expression promoter of the present invention to a patient include subcutaneous tissue administration, intramuscular administration, intravenous administration, oral administration, transdermal administration, and the like. A method suitable for treatment or the like may be selected as appropriate. In addition, the dose of the SCF mRNA expression promoter of the present invention may be appropriately increased or decreased depending on the type of disease, severity, individual differences among patients, administration method, administration period, and the like.

  The SCF mRNA expression promoter of the present invention can promote the expression of SCF mRNA through an SCF mRNA expression promoting action possessed by an extract from one or more plants selected from the group consisting of Ashitaba, nettle and auren. As a result, the proliferation of pigment cells and the production of melanin can be promoted to prevent or ameliorate gray hair, baldness, etc., and the skin can be browned (black). However, the SCF mRNA expression promoter of the present invention can be used for all purposes that are meaningful for exerting the SCF mRNA expression promoting action in addition to these uses.

  The SCF mRNA expression promoter of the present invention is suitably applied to humans, but can also be applied to animals other than humans as long as each effect is exhibited.

  Hereinafter, although a test example is shown and this invention is demonstrated concretely, this invention is not restrict | limited to each following example at all. In the following test examples, Ashitaba extract (manufactured by Maruzen Pharmaceutical Co., Ltd., trade name: Ashitaba extract, sample 1) and nettle extract (manufactured by Maruzen Pharmaceutical Co., Ltd., trade name: nettle extract, sample 2) are used as samples. And the oren extract (manufactured by Maruzen Pharmaceutical Co., Ltd., trade name: Ouren extract, sample 3) was used.

[Test Example 1] SCF mRNA expression promoting action test Ashitaba extract (sample 1), nettle extract (sample 2) and auren extract (sample 3) were tested for SCF mRNA expression promoting action as follows.

Normal human epidermal keratinocyte (NHEK) in normal culture medium for epidermal keratinocyte long-term culture (EpiLife-KG2) in an 80 cm ² flask at 37 ° C., 5% CO ₂ -95% air The cells were precultured under the above conditions and cells were collected by trypsin treatment.

EpiLife-KG2 was used to seed 40 × 10 ⁴ cells / 2 mL / dish in 35 mm dishes (FALCON) and cultured overnight at 37 ° C. and 5% CO ₂ -95% air. After 24 hours, the culture solution was discarded, and 2 mL of each test sample (samples 1 to 3, sample concentration shown in Table 1 below) dissolved in EpiLife-KG2 was added to each dish, and the sample was incubated at 37 ° C. and 5% CO _2. The cells were cultured for 24 hours under the condition of -95% air. After culturing, the culture solution is discarded, and total RNA is extracted with ISOGEN (NIPPON GENE, Cat. No. 311-02501), and the amount of each RNA is measured with a spectrophotometer so that it becomes 200 ng / μL. Total RNA was prepared.

  Using this total RNA as a template, the expression level of mRNA of SCF and GAPDH as an internal standard was measured. The detection was performed by real-time 2 Step RT-PCR reaction using TaKaRa SYBR PrimeScript RT-PCR Kit (Perfect Real Time, code No. RR063A) using a real-time PCR device Smart Cycler (Cepheid). The expression level of SCF mRNA was determined based on the total RNA preparation prepared from the cells cultured with and without the sample added, and a correction value was calculated with the value of GAPDH. The correction value of the sample addition at that time was calculated. From the obtained results, the SCF mRNA expression promotion rate (%) was calculated by the following formula.

SCF mRNA expression promotion rate (%) = A / B × 100
In the formula, A represents “correction value when sample is added” and B represents “correction value when sample is not added”.
The results are shown in Table 1.

  As shown in Table 1, it was found that the Ashitaba extract, the nettle extract, and the Ouren extract all have an excellent SCF mRNA expression promoting action.

  The stem cell growth factor (SCF) mRNA expression promoter of the present invention can greatly contribute to prevention / improvement of graying and promotion of browning (black) of the skin.

Claims (1)

A stem cell growth factor (SCF) mRNA expression promoter characterized by containing an extract from the leaves of Ashitaba as an active ingredient.