JP5988617B2 - Profilagrin gene expression promoter - Google Patents

Description

  The present invention relates to a profilagulin gene expression promoter.

  The epidermis has a thickness of about 0.2 mm, and most of the constituent cells are epidermal keratinocytes. The epidermal keratinocytes divide in the basal layer at the lowest layer of the epidermis, and move to the upper layer to the spiny layer, the granular layer, and the horny layer as they mature. This process is called keratinization.

  The stratum corneum covers the living body and functions to retain moisture and protect against intruders from the outside. Such stratum corneum is composed of various substances such as keratin, filaggrin and lipid produced by epidermal keratinocytes. Keratin forms a tonofilament as the cytoskeleton of epidermal cells and is essential for maintaining the morphology of epidermal cells.

  Filaggrin is present in a large amount in the state of profilagrin in the keratohyalin granules in the granule layer, and profilagrin is decomposed into filaggrin by dephosphorylation and the action of peptidylarginine deiminase protease during the keratinization process. Is done. It is believed that filaggrin released from profilaggrin aggregates keratin fibers compactly within the cells of keratinocytes and flattens the entire cell. Furthermore, filaggrin is decomposed into amino acids in the upper layer of the stratum corneum, and the amino acids and their derivatives provide moisture to the stratum corneum as natural moisturizing factors, so that the stratum corneum becomes flexible and an enzyme involved in stratum corneum desquamation. It also contributes to the activation of the stratum corneum and affects the normal turnover of the stratum corneum. As described above, filaggrin plays a very important role in forming a healthy stratum corneum.

  On the other hand, filaggrin decreases in synthetic power depending on the dry state, the amount of amino acids in the stratum corneum decreases (see Non-Patent Document 1), and when the natural moisturizing factor (amino acid) decreases, the molecular mobility of keratin decreases. It has been reported that the flexibility of the stratum corneum is lost (see Non-Patent Document 2). In addition, ichthyosis vulgaris (a hereditary disease characterized by dryness and desquamation of the skin and a fish scale-like appearance on the trunk and extremities) due to an abnormality in the profilagrin gene (see Non-Patent Document 3) It has been reported that the abnormality is also involved in the development of atopic dermatitis (see Non-Patent Document 4). Furthermore, it has been reported that abnormalities in the profilagrin gene are related not only to atopic dermatitis but also to bronchial asthma patients (see Non-Patent Document 5).

  Therefore, against such a decrease in stratum corneum flexibility and skin diseases, externally moisturizing natural moisturizing factors such as amino acids, polyhydric alcohols such as glycerin, water-soluble polymers such as hyaluronic acid, lipids such as ceramide, etc. Although a method of replenishing as an agent has been tried, the stratum corneum is a tissue that is created as a complex structure that is not a living organism, unlike living tissues and cells, so it is healthy if only the above moisturizer is applied directly. It is difficult to return to the stratum corneum, and a sustained effect cannot be expected. Moreover, although application | coating of a steroid agent is also considered, there was a concern of a side effect in the steroid agent.

Arch. Dermatol. Res., 288, 442-446, 1996 Skin and Beauty, 36, 4, 210-228, 2004 Nature Genetics, 38, 337-342, 2006 Nature Genetics, 38, 441-446, 2006 J. Allergy Clin. Immunol., 120, 64-68, 2007

  The present invention has been made under such circumstances, and can provide a safe and highly effective profilagrin gene expression promoter that can form a healthy stratum corneum by promoting the production of filaggrin. Objective.

In view of such circumstances, the present inventors have investigated prophylagulin gene expression promoting effects on extracts of various plants that have been used as food for many years and are highly safe. It was found to have a high profilaggrin gene expression promoting effect at a high concentration. That is, the present invention is a profilaggrin gene expression promoter comprising an extract obtained by extracting leaves of oyster family oysters with a solvent of water / 1,3-butylene glycol = 100/0 to 50/50 (mass ratio) .

  The profilaggrin gene expression promoter of the present invention consists of an extract of oyster leaves that has been used for many years as a food and has high safety and low toxicity. Therefore, the profilagrin gene expression promoter is highly safe and has an excellent effect of promoting human profilaggrin gene expression. Yes. Therefore, the profilagrin gene expression promoter of the present invention is useful for forming a healthy stratum corneum by promoting the production of filaggrin.

Hereinafter, embodiments of the present invention will be described.
The profilaggrin gene expression promoter of the present invention comprises an extract of oysters of the oyster family Oyster. The oyster used in the present invention is a plant belonging to the genus Oysteraceae, and has a scientific name of Diospyros kaki Thunb., And is also referred to as an oyster (oak tree). Oysters are usually cultivated widely to make fruits edible, and are cultivated in a wide range of regions such as East Asia and Europe and America. In addition, it has been cultivated in Japan for a long time. In addition to edible fruits, kakinoha is used in kashiwanoha sushi and kashiwanoha tea. Therefore, the oyster leaf used in the present invention is a material that has been used for many years as a food and has high safety and low toxicity.

  The oyster leaf extract is obtained through an extraction process in which dried oyster leaves are immersed in a specific extraction solvent. Examples of the solvent used for extraction include hydrocarbons, esters, ketones, ethers, halogenated hydrocarbons, water-soluble alcohols and water. Among these, one or more of water, lower alcohol, and polyhydric alcohol is preferable, and one or more of water, ethanol, and 1,3-butylene glycol are more preferable. Examples of the extraction method include a method in which an extraction solvent is added to oyster leaves, immersed for 1 hour or longer, and filtered. The temperature at the time of extraction is a temperature not higher than the boiling point of the extraction solvent to be used. For example, when the extraction solvent is water, it is preferably 40 to 90 ° C., more preferably 50 to 80 ° C., and the extraction solvent is 50 mass. In the case of a 1% aqueous solution of 1,3-butylene glycol (BG), the temperature is preferably 30 to 90 ° C, more preferably 40 to 80 ° C. In the extraction, it is preferable to use oyster leaves which are chopped or pulverized as they are or after they are dried. The obtained extract may be used as it is, or may be concentrated by distilling off the solvent, or may be purified by treatment such as column chromatography or solvent fractionation. The obtained extract is allowed to stand at 0 to 10 ° C., preferably 2 to 6 ° C. for 3 to 10 days, preferably 5 to 8 days, and then filtered to obtain the extract. Also good.

  Since the profilaggrin gene expression promoter of the present invention is safe and low in toxicity, it can be administered by any of the administration methods including external administration, internal use, and pharmaceuticals, quasi drugs, cosmetics (for example, cosmetics) Water, emulsion, cream, gel, cosmetic liquid, pack, oil, ointment, spray, patch, etc.) and the like. The content of the oyster leaf extract (profilagulin gene expression promoter of the present invention) when blended in the preparation is 0.00005-5 mass%, preferably 0.0001-2. 5% by mass. When the content is less than 0.00005% by mass, it is difficult to exert the effect of promoting profilaggrin gene expression, and when it exceeds 5% by mass, problems are likely to occur in the stability of the preparation.

  Hereinafter, the present invention will be described more specifically with reference to examples.

[Example 1]
50 g of dried and shredded oyster leaves collected in June in Nara Prefecture were immersed in 1000 g of water as an extraction solvent at 80 ° C. for 5 hours, and then filtered using filter paper (Advantech Toyo Co., Ltd., 5C). A filtrate was obtained. The filtrate was allowed to stand at 5 ° C. for 7 days and then filtered using a filter paper to obtain an oyster leaf water extract filtrate. The solid content excluding the extraction solvent (water) in this oyster leaf water extract was 0.8% by mass.

[Example 2]
50 g of dried and shredded oyster leaves collected in June in Nara Prefecture were immersed in 1000 g of 50 mass% 1,3-butylene glycol (BG) aqueous solution as an extraction solvent at 50 ° C. for 5 hours, and then filter paper Filtration was performed using (Advantech Toyo Co., Ltd., 5C) to obtain a filtrate. The filtrate was allowed to stand at 5 ° C. for 7 days and then filtered using a filter paper to obtain a filtrate of oyster leaf BG extract. The solid content excluding the extraction solvent (BG aqueous solution) in the oyster leaf BG extract was 1.1% by mass.

  Using the oyster leaf extract obtained in Examples 1 and 2, the measurement test of the profilagrin gene expression level shown below was performed.

[Profilaggrin gene expression level measurement test]
Human foreskin-derived normal epidermal keratinocytes were used, and the culture medium was cultured using epidermal keratinocyte growth medium (Kurabo). The cells were seeded on a culture plate and cultured until subconfluent, and then various samples (extracts) were added so as to achieve the predetermined concentrations shown in Table 1. After further incubation for 6 hours, RNA was extracted from the cells. The case where no sample was added was used as a control.

  The extracted RNA was reverse transcribed to prepare cDNA, and profilaggrin mRNA was quantified by quantitative real-time PCR expression analysis. Primers specific for profilagrin were prepared based on a report by Presland R.B. et al. (J. Biol. Chem., 267, 23773-23781, 1992). GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) was used as an internal standard. The profilagrin gene expression rate was determined by using the GAPDH mRNA expression level as a correction value and the profilagrin mRNA expression level as a ratio (percentage) to the mRNA expression level in the control cells.

  As is clear from the results in Table 1, it was confirmed that the oyster leaf water extract (Example 1) and the oyster leaf BG extract (Example 2) have an excellent profilaggrin gene expression promoting effect. Therefore, the profilagrin gene expression promoter of the present invention consisting of oyster leaf extract promotes the expression of human profilagrin gene in epidermal keratinocytes, and promotes the production of filaggrin to form a healthy stratum corneum. The effect of forming can be expected.

Claims (1)

A profilagulin gene expression promoter comprising an extract obtained by extracting leaves of oysters with water / 1,3-butylene glycol = 100/0 to 50/50 (mass ratio) .