JP2021127323A - HAS3 mRNA expression promoter - Google Patents
HAS3 mRNA expression promoter Download PDFInfo
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- JP2021127323A JP2021127323A JP2020023560A JP2020023560A JP2021127323A JP 2021127323 A JP2021127323 A JP 2021127323A JP 2020023560 A JP2020023560 A JP 2020023560A JP 2020023560 A JP2020023560 A JP 2020023560A JP 2021127323 A JP2021127323 A JP 2021127323A
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- mrna expression
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- saccharomyces
- has3
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Abstract
Description
本発明は、ＨＡＳ３ ｍＲＮＡ発現促進剤に関するものである。 The present invention relates to a HAS3 mRNA expression promoter.
ヒアルロン酸は、ムコ多糖の一種であり、細胞間の間隙に充填されることにより細胞を保持する機能を有し、さらに細胞間隙への水分の保持、組織への潤滑性や柔軟性の付与、機械的障害等の外力に対する抵抗等、数多くの機能を有している。表皮ヒアルロン酸の合成促進に関与するヒアルロン酸合成酵素３（ＨＡＳ３）の発現を促進することでヒアルロン酸の産生を促進することができれば、皮膚の荒れ、しわ、くすみ、きめの変化、弾力性の低下及び保湿機能の低下等といった皮膚の老化症状を予防、治療または改善できると考えられる。 Hyaluronic acid is a type of mucopolysaccharide that has the function of retaining cells by filling the intercellular spaces, and further retains water in the intercellular spaces and imparts lubricity and flexibility to tissues. It has many functions such as resistance to external forces such as mechanical obstacles. If the production of hyaluronic acid can be promoted by promoting the expression of hyaluronic acid synthase 3 (HAS3), which is involved in promoting the synthesis of epidermal hyaluronic acid, the skin becomes rough, wrinkled, dull, textured, and elastic. It is considered that skin aging symptoms such as deterioration and deterioration of moisturizing function can be prevented, treated or ameliorated.
現在までにヒアルロン酸合成酵素３（ＨＡＳ３）ｍＲＮＡ発現促進作用を有するものとして、甘草葉部抽出物（特許文献１参照）等が知られている。 To date, licorice leaf extract (see Patent Document 1) and the like are known to have an action of promoting hyaluronic acid synthase 3 (HAS3) mRNA expression.
本発明は、作用効果に優れたＨＡＳ３ ｍＲＮＡ発現促進剤を提供することを目的とする。 An object of the present invention is to provide a HAS3 mRNA expression promoter having excellent action and effect.
上記課題を解決するために、本発明のＨＡＳ３ ｍＲＮＡ発現促進剤は、コメのサッカロミセス・ヴェローナによる発酵物を有効成分とすることを特徴とする。 In order to solve the above problems, the HAS3 mRNA expression promoter of the present invention is characterized by containing a fermented product of rice by Saccharomyces verona as an active ingredient.
本発明によれば、コメのサッカロミセス・ヴェローナによる発酵物を有効成分とすることにより、作用効果に優れたＨＡＳ３ ｍＲＮＡ発現促進剤を提供することができる。 According to the present invention, it is possible to provide a HAS3 mRNA expression promoter having excellent action and effect by using a fermented product of rice saccharomyces verona as an active ingredient.
〔ＨＡＳ３ ｍＲＮＡ発現促進剤〕
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、コメのサッカロミセス・ヴェローナによる発酵物を有効成分として含有する。本実施形態に係るコメのサッカロミセス・ヴェローナによる発酵物は、コメをサッカロミセス・ヴェローナにより発酵して得られるものである。
[HAS3 mRNA expression promoter]
The HAS3 mRNA expression promoter of the present embodiment contains a fermented product of rice by Saccharomyces verona as an active ingredient. The fermented product of rice produced by Saccharomyces verona according to the present embodiment is obtained by fermenting rice with Saccharomyces verona.
ここで、本明細書における「コメのサッカロミセス・ヴェローナによる発酵物」には、別段の記載がある場合を除き、コメをサッカロミセス・ヴェローナにより発酵させて得られる発酵液、当該発酵液の希釈液もしくは濃縮液、当該発酵液を乾燥して得られる乾燥物、またはこれらの粗精製物もしくは精製物のいずれもが含まれる。 Here, unless otherwise specified, the term "fermented product of rice by Saccharomyces verona" in the present specification refers to a fermented liquid obtained by fermenting rice with Saccharomyces verona, a diluted solution of the fermented liquid, or a diluted solution of the fermented liquid. It includes a concentrate, a dried product obtained by drying the fermented liquid, or a crude product or a purified product thereof.
本実施形態に係るコメのサッカロミセス・ヴェローナによる発酵物において、発酵原料となるコメは、イネ科の植物イネ（学名Oryza sativa）から得られるものである。イネは日本各地で栽培されており、これらの地域から容易に入手可能である。発酵原料としては特に制限はなく、目的に応じて適宜選定することができるが、例えば、もみ、玄米、精白米またはこれらの混合物が挙げられ、これらの中でも精白米が好ましい。 In the fermented product of rice according to the present embodiment by Saccharomyces verona, the rice used as a fermentation raw material is obtained from the plant rice of the Gramineae family (scientific name: Oryza sativa). Rice is cultivated in various parts of Japan and is easily available from these areas. The fermentation raw material is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include fir, brown rice, polished rice or a mixture thereof, and among these, polished rice is preferable.
本実施形態に係るコメのサッカロミセス・ヴェローナによる発酵物は、コメをサッカロミセス・ヴェローナにより発酵させることにより得られる。発酵処理は、例えば、コメを水で分散し、発酵を行う微生物を接種することにより行うことができる。 The fermented product of rice produced by Saccharomyces verona according to the present embodiment is obtained by fermenting rice with Saccharomyces verona. The fermentation treatment can be carried out, for example, by dispersing rice in water and inoculating a microorganism for fermentation.
サッカロミセス・ヴェローナを接種する前に、発酵効率を高める観点から、コメに含まれるデンプンの分解処理および／またはコメの細胞壁の分解処理を行うことが好ましい。 Before inoculating Saccharomyces verona, it is preferable to decompose the starch contained in the rice and / or the cell wall of the rice from the viewpoint of increasing the fermentation efficiency.
デンプンの分解処理は、例えば、酵素処理、麹菌の接種等が挙げられ、特に酵素処理を行うことが好ましい。デンプン分解酵素としては、α−アミラーゼ、β−アミラーゼ、グルコアミラーゼ、イソアミラーゼ等のアミラーゼが挙げられるが、発酵効率を高める観点から、特にα−アミラーゼ、グルコアミラーゼが好ましく、さらにはグルコアミラーゼが好ましい。一方、細胞壁の分解処理は、例えば、セルラーゼ、キシラナーゼ等の酵素を用いた処理が好ましい。 Examples of the starch decomposition treatment include enzyme treatment and inoculation of aspergillus, and enzyme treatment is particularly preferable. Examples of the starch-degrading enzyme include amylases such as α-amylase, β-amylase, glucoamylase, and isoamylase. From the viewpoint of increasing fermentation efficiency, α-amylase and glucoamylase are particularly preferable, and glucoamylase is further preferable. .. On the other hand, the cell wall decomposition treatment is preferably a treatment using, for example, an enzyme such as cellulase or xylanase.
得られた処理液は、冷却、加熱、ｐＨ調整、殺菌などの所望の手段に付すことで酵素や麹菌を不活性化させてもよいし、そのまま発酵を行う微生物を接種してもよい。 The obtained treatment liquid may be subjected to desired means such as cooling, heating, pH adjustment, and sterilization to inactivate enzymes and aspergillus, or may be inoculated with microorganisms to be fermented as they are.
発酵を行う微生物はサッカロミセス・ヴェローナ（Saccharomyces veronae）（別名：ラカンセア・サーモトレランス（Lachancea thermotolerans））である。 The microorganism that ferments is Saccharomyces veronae (also known as Lachancea thermotolerans).
上記微生物を用いた発酵処理は、例えば、発酵原料と上記微生物とを発酵槽に入れ、２０〜４０℃、好ましくは２５〜３５℃の温度範囲で、１２〜７２時間、好ましくは１８〜４８時間処理することにより、行うことができる。また、発酵処理の開始時において、反応液のｐＨが５．０〜７．０に、好ましくは６．０〜７．０に調整されていると、発酵処理が好適に進行する。かかる発酵処理は、発酵液を冷却、加熱殺菌、ろ過などの所望の手段に付し、発酵菌を不活性化させることで終了させる。 In the fermentation treatment using the above microorganisms, for example, the fermentation raw material and the above microorganisms are placed in a fermenter and placed in a temperature range of 20 to 40 ° C., preferably 25 to 35 ° C. for 12 to 72 hours, preferably 18 to 48 hours. It can be done by processing. Further, when the pH of the reaction solution is adjusted to 5.0 to 7.0, preferably 6.0 to 7.0 at the start of the fermentation treatment, the fermentation treatment proceeds favorably. Such fermentation treatment is terminated by subjecting the fermentation broth to desired means such as cooling, heat sterilization, filtration and the like to inactivate the fermenting bacteria.
このようにして得られた発酵液は、そのまま発酵物として用いてもよく、または適宜希釈しもしくは濃縮して、発酵物として用いてもよい。さらには、濃縮物をさらに乾燥してもよく、粗精製などを行ってもよい。 The fermented liquid thus obtained may be used as it is as a fermented product, or may be appropriately diluted or concentrated and used as a fermented product. Furthermore, the concentrate may be further dried, or may be crudely purified.
このようにして得られるコメのサッカロミセス・ヴェローナによる発酵物は、優れたＨＡＳ３ ｍＲＮＡ発現促進作用を有するため、ＨＡＳ３ ｍＲＮＡ発現促進剤の有効成分として使用することができる。本実施形態に係るＨＡＳ３ ｍＲＮＡ発現促進剤は、医薬品、医薬部外品、化粧品、飲食品等の幅広い用途に使用することができる。 The fermented product of rice produced by Saccharomyces verona thus obtained has an excellent HAS3 mRNA expression promoting action, and therefore can be used as an active ingredient of a HAS3 mRNA expression promoting agent. The HAS3 mRNA expression promoter according to the present embodiment can be used in a wide range of applications such as pharmaceuticals, quasi-drugs, cosmetics, and foods and drinks.
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、コメのサッカロミセス・ヴェローナによる発酵物のみからなるものであってもよいし、コメのサッカロミセス・ヴェローナによる発酵物を製剤化したものであってもよい。 The HAS3 mRNA expression promoter of the present embodiment may consist only of a fermented product of rice saccharomyces verona, or may be a formulation of a fermented product of rice saccharomyces verona.
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、デキストリン、シクロデキストリン等の薬学的に許容し得るキャリアーその他任意の助剤を用いて、常法に従い、粉末状、顆粒状、錠剤状、液状等の任意の剤形に製剤化することができる。この際、助剤としては、例えば、賦形剤、結合剤、崩壊剤、滑沢剤、安定剤、矯味・矯臭剤等を用いることができる。ＨＡＳ３ ｍＲＮＡ発現促進剤は、他の組成物（例えば、皮膚外用剤、経口組成物等）に配合して使用することができるほか、軟膏剤、外用液剤、貼付剤等として使用することができる。 The HAS3 mRNA expression promoter of the present embodiment uses a pharmaceutically acceptable carrier such as dextrin or cyclodextrin or any other auxiliary agent, and is optionally powdered, granular, tableted, liquid or the like according to a conventional method. It can be formulated into the dosage form of. At this time, as the auxiliary agent, for example, an excipient, a binder, a disintegrant, a lubricant, a stabilizer, a flavoring / odorant, and the like can be used. The HAS3 mRNA expression promoter can be used by blending with other compositions (for example, external preparations for skin, oral compositions, etc.), and can also be used as ointments, external liquids, patches, and the like.
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤を製剤化した場合、コメのサッカロミセス・ヴェローナによる発酵物の含有量は、特に限定されるものではなく、目的に応じて適宜設定することができる。 When the HAS3 mRNA expression promoter of the present embodiment is formulated, the content of the fermented product of rice by Saccharomyces verona is not particularly limited and can be appropriately set according to the purpose.
なお、本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、必要に応じて、ＨＡＳ３ ｍＲＮＡ発現促進作用を有する他の天然抽出物等を、コメのサッカロミセス・ヴェローナによる発酵物とともに配合して有効成分として用いることができる。 The HAS3 mRNA expression promoter of the present embodiment is used as an active ingredient by blending other natural extracts having a HAS3 mRNA expression promoting action together with a fermented product of rice saccharomyces verona, if necessary. Can be done.
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤の患者に対する投与方法としては、経皮投与、経口投与等が挙げられるが、疾患の種類に応じて、その予防・治療等に好適な方法を適宜選択すればよい。また、本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤の投与量も、疾患の種類、重症度、患者の個人差、投与方法、投与期間等によって適宜増減すればよい。 Examples of the method for administering the HAS3 mRNA expression promoter of the present embodiment to a patient include transdermal administration and oral administration. However, if a method suitable for prevention / treatment thereof is appropriately selected according to the type of disease. good. In addition, the dose of the HAS3 mRNA expression promoter of the present embodiment may be appropriately increased or decreased depending on the type and severity of the disease, individual differences of patients, administration method, administration period, and the like.
本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、有効成分であるコメのサッカロミセス・ヴェローナによる発酵物が有するＨＡＳ３ ｍＲＮＡ発現促進作用を通じて、ヒアルロン酸合成酵素３（ＨＡＳ３）の発現を促進することができるため、それによりヒアルロン酸の生成が促進され、皮膚の荒れ、しわ、くすみ、きめの変化、弾力性の低下及び保湿機能の低下等、皮膚の老化症状の予防、治療または改善等の用途に用いることができる。本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、これらの用途以外にもＨＡＳ３ ｍＲＮＡ発現促進作用を発揮することに意義のあるすべての用途に用いることができる。 Since the HAS3 mRNA expression-promoting agent of the present embodiment can promote the expression of hyaluronic acid synthase 3 (HAS3) through the HAS3 mRNA expression-promoting action of the fermented product of rice saccharomyces verona, which is the active ingredient. As a result, the production of hyaluronic acid is promoted, and it can be used for the prevention, treatment or improvement of skin aging symptoms such as rough skin, wrinkles, dullness, changes in texture, decreased elasticity and decreased moisturizing function. can. The HAS3 mRNA expression-promoting agent of the present embodiment can be used for all uses other than these uses, which are significant in exerting the HAS3 mRNA expression-promoting action.
また、本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、優れたＨＡＳ３ ｍＲＮＡ発現促進作用を有するため、例えば、皮膚外用剤、経口組成物等に配合するのに好適である。この場合に、コメのサッカロミセス・ヴェローナによる発酵物をそのまま配合してもよいし、コメのサッカロミセス・ヴェローナによる発酵物から製剤化したＨＡＳ３ ｍＲＮＡ発現促進剤を配合してもよい。 In addition, the HAS3 mRNA expression-promoting agent of the present embodiment has an excellent HAS3 mRNA expression-promoting effect, and is therefore suitable for blending with, for example, an external preparation for skin, an oral composition, or the like. In this case, the fermented product of Saccharomyces verona of rice may be blended as it is, or the HAS3 mRNA expression promoter formulated from the fermented product of Saccharomyces verona of rice may be blended.
ここで、皮膚外用剤としては、その区分に制限はなく、経皮的に使用される皮膚化粧料、医薬部外品、医薬品等を幅広く含むものであり、具体的には、例えば、軟膏、クリーム、乳液、化粧水、美容液、ローション、ジェル、美容オイル、パック、ファンデーション、リップクリーム、入浴剤、ヘアートニック、ヘアーローション、シャンプー、リンス、石鹸、ボディシャンプー等が挙げられる。 Here, the external preparation for skin is not limited in its classification, and includes a wide range of skin lotions, non-pharmaceutical products, pharmaceuticals, etc. that are used transdermally. Examples include creams, milky lotions, lotions, beauty liquids, lotions, gels, beauty oils, packs, foundations, lip creams, bathing agents, hair nicks, hair lotions, shampoos, rinses, soaps, body shampoos and the like.
経口組成物とは、人の健康に危害を加えるおそれが少なく、通常の社会生活において、経口又は消化管投与により摂取されるものをいい、行政区分上の飲食品、医薬品、医薬部外品等の区分に制限されるものではない。したがって、本実施形態における「経口組成物」は、経口的に摂取される一般食品、飼料、健康食品、保健機能食品（特定保健用食品，栄養機能食品，機能性表示食品）、医薬部外品、医薬品等を幅広く含むものである。 Oral compositions are those that are less likely to harm human health and are ingested by oral or gastrointestinal administration in normal social life, such as foods and drinks, pharmaceuticals, quasi-drugs, etc. under administrative divisions. It is not limited to the classification of. Therefore, the "oral composition" in the present embodiment includes general foods, feeds, health foods, foods with health claims (foods for specified health use, foods with nutritional claims, foods with functional claims), and non-pharmaceutical products that are orally ingested. , Pharmaceuticals, etc. are widely included.
また、本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、優れたＨＡＳ３ ｍＲＮＡ発現促進作用を有するので、これらの作用機構に関する研究のための試薬としても好適に利用することができる。 Further, since the HAS3 mRNA expression-promoting agent of the present embodiment has an excellent HAS3 mRNA expression-promoting action, it can be suitably used as a reagent for research on these action mechanisms.
なお、本実施形態のＨＡＳ３ ｍＲＮＡ発現促進剤は、ヒトに対して好適に適用されるものであるが、それぞれの作用効果が奏される限り、ヒト以外の動物（例えば，マウス，ラット，ハムスター，イヌ，ネコ，ウシ，ブタ，サル等）に対して適用することもできる。 Although the HAS3 mRNA expression promoter of the present embodiment is preferably applied to humans, animals other than humans (for example, mice, rats, hamsters, etc.) as long as their respective actions and effects are exhibited. It can also be applied to dogs, cats, cows, pigs, monkeys, etc.).
以下、試験例を示し、本発明を具体的に説明するが、本発明は下記の各例に何ら制限されるものではない。 Hereinafter, the present invention will be specifically described with reference to test examples, but the present invention is not limited to the following examples.
〔製造例〕コメのサッカロミセス・ヴェローナによる発酵物の製造 [Production example] Production of fermented product by Saccharomyces verona of rice
白精米の米粉０．１ｋｇに水１ｋｇを加えて分散させ、１１０℃にて５分滅菌処理を行った後、グルコアミラーゼを加えて、６０℃にて１時間反応させた。 1 kg of water was added to 0.1 kg of rice flour of white polished rice to disperse it, and after sterilizing at 110 ° C. for 5 minutes, glucoamylase was added and the reaction was carried out at 60 ° C. for 1 hour.
その後、得られた酵素処理液にサッカロミセス・ヴェローナ（秋田今野商店社製）を植菌し、３０℃で２４時間発酵させた。得られた発酵液を１１０℃にて５分滅菌処理を行った。得られた培養液を珪藻土で濾過し、濾液を濃縮乾固することにより、コメのサッカロミセス・ヴェローナによる発酵物（約６４ｇ，試料１）を得た。 Then, Saccharomyces verona (manufactured by Akita Konno Shoten Co., Ltd.) was inoculated into the obtained enzyme-treated solution and fermented at 30 ° C. for 24 hours. The obtained fermentation broth was sterilized at 110 ° C. for 5 minutes. The obtained culture solution was filtered through diatomaceous earth, and the filtrate was concentrated to dryness to obtain a fermented product (about 64 g, sample 1) of rice by Saccharomyces verona.
〔試験例１〕ヒアルロン酸合成酵素３（ＨＡＳ３） ｍＲＮＡ発現促進作用試験
得られたコメのサッカロミセス・ヴェローナによる発酵物（試料１）について、以下のようにしてＨＡＳ３ ｍＲＮＡ発現促進作用を試験した。
[Test Example 1] Hyaluronic Acid Synthetic Enzyme 3 (HAS3) mRNA Expression Promoting Action Test The HAS3 mRNA expression promoting action of the obtained fermented rice produced by Saccharomyces verona (Sample 1) was tested as follows.
正常ヒト新生児表皮角化細胞（ＮＨＥＫ）を、正常ヒト表皮角化細胞用増殖培地（ＫＧＭ）を用いて前培養し、トリプシン処理により細胞を回収した。回収した細胞を２０×１０４cells／ｍＬの細胞密度になるようにＫＧＭ培地で希釈した後、３５ｍｍシャーレに２ｍＬずつ播種し（４０×１０４cells／シャーレ）、３７℃・５％ＣＯ２の条件下で一晩培養した。培養後、培地を正常ヒト表皮角化細胞基礎培地（ＫＢＭ，上記ＫＧＭ培地に増殖因子（ｈＥＧＦ，ＢＰＥ，インスリン）を添加していないもの）に交換し、さらに２４時間培養した。 Normal human neonatal epidermal keratinocytes (NHEK) were pre-cultured using a growth medium for normal human epidermal keratinocytes (KGM), and the cells were recovered by trypsin treatment. The collected cells were diluted with KGM medium to a cell density of 20 × 10 4 cells / mL, and then seeded in 2 mL each in a 35 mm petri dish (40 × 10 4 cells / petri dish) at 37 ° C. and 5% CO 2 . Incubated overnight under conditions. After culturing, the medium was replaced with a normal human epidermal keratinocyte basal medium (KBM, the above KGM medium to which growth factors (hEGF, BPE, insulin) were not added), and the cells were further cultured for 24 hours.
２４時間培養後、培地を除去し、ＫＢＭ培地に溶解した被験試料（試料１，試料濃度は下記表１を参照）を各シャーレに２ｍＬずつ添加し、３７℃・５％ＣＯ２の条件下にて２４時間培養した。なお、コントロールとして、試料無添加のＫＢＭ培地を用いて同様に培養した。培養後、培地を除去し、ＩＳＯＧＥＮ II（ニッポンジーン社製，Cat. No. 311-07361）にて総ＲＮＡを抽出し、それぞれのＲＮＡ量を分光光度計にて測定し、２００ｎｇ／μＬになるように総ＲＮＡを調製した。 After culturing for 24 hours, the medium was removed, and 2 mL of a test sample (sample 1, sample concentration, see Table 1 below) dissolved in KBM medium was added to each petri dish under the conditions of 37 ° C. and 5% CO 2. Was cultured for 24 hours. As a control, the cells were similarly cultured using KBM medium to which no sample was added. After culturing, the medium is removed, total RNA is extracted with ISOGEN II (manufactured by Nippon Gene, Cat. No. 311-07361), and the amount of each RNA is measured with a spectrophotometer so as to reach 200 ng / μL. Total RNA was prepared in.
この総ＲＮＡを鋳型とし、ＨＡＳ３および内部標準であるＧＡＰＤＨについて、ｍＲＮＡの発現量を測定した。検出はリアルタイムＰＣＲ装置Smart Cycler（Cepheid社製）を用いて、TaKaRa SYBR Prime Script RT-PCR kit（Perfect Real Time）（タカラバイオ社製，code No. RR063A）によるリアルタイム２Ｓｔｅｐ ＲＴ−ＰＣＲ反応により行った。ＨＡＳ３ ｍＲＮＡの発現量は、「被験試料添加」および「試料無添加」にてそれぞれ培養した細胞から調製した総ＲＮＡ標品を基にして、ＧＡＰＤＨの値で補正値を求めた。得られた値から、下記式によりＨＡＳ３ ｍＲＮＡ発現促進率（％）を算出した。 Using this total RNA as a template, the expression level of mRNA was measured for HAS3 and GAPDH, which is an internal standard. Detection was performed by real-time 2Step RT-PCR reaction using TaKaRa SYBR Prime Script RT-PCR kit (Perfect Real Time) (Takara Bio Inc., code No. RR063A) using a real-time PCR device Smart Cycler (manufactured by Cepheid). .. The expression level of HAS3 mRNA was corrected by the GAPDH value based on the total RNA preparation prepared from the cells cultured in "test sample addition" and "sample no addition", respectively. From the obtained values, the HAS3 mRNA expression promotion rate (%) was calculated by the following formula.
ＨＡＳ３ ｍＲＮＡ発現促進率（％）＝Ａ／Ｂ×１００
式中の各項はそれぞれ以下を表す。
Ａ：被験試料添加での補正値
Ｂ：試料無添加での補正値
結果を表１に示す。
HAS3 mRNA expression promotion rate (%) = A / B × 100
Each term in the formula represents the following.
A: Correction value with the addition of the test sample B: Correction value without the addition of the sample The results are shown in Table 1.
表１に示すように、コメのサッカロミセス・ヴェローナによる発酵物（試料１）は、優れたＨＡＳ３ ｍＲＮＡ発現促進作用を有していた。 As shown in Table 1, the fermented product of rice by Saccharomyces verona (Sample 1) had an excellent HAS3 mRNA expression promoting effect.
〔配合例１〕
下記組成のクリームを常法により製造した。
コメのサッカロミセス・ヴェローナによる発酵物（製造例） ０．０５ｇ
クジンエキス ０．１ｇ
オウゴンエキス ０．１ｇ
流動パラフィン ５．０ｇ
サラシミツロウ ４．０ｇ
スクワラン １０．０ｇ
セタノール ３．０ｇ
ラノリン ２．０ｇ
ステアリン酸 １．０ｇ
オレイン酸ポリオキシエチレンソルビタン（20E.O.） １．５ｇ
モノステアリン酸グリセリル ３．０ｇ
油溶性カンゾウエキス ０．１ｇ
１，３−ブチレングリコール ６．０ｇ
パラオキシ安息香酸メチル １．５ｇ
香料 ０．１ｇ
精製水 残部（全量を１００ｇとする）
[Formulation Example 1]
A cream having the following composition was produced by a conventional method.
Fermented product of rice by Saccharomyces verona (Production example) 0.05g
Kujin extract 0.1g
Scutellaria extract 0.1g
Liquid paraffin 5.0g
Sarashi Beeswax 4.0g
Squalene 10.0g
Cetanol 3.0g
Lanolin 2.0g
Stearic acid 1.0g
Polyoxyethylene sorbitan oleate (20E.O.) 1.5g
Glyceryl monostearate 3.0 g
Oil-soluble licorice extract 0.1g
1,3-butylene glycol 6.0 g
Methyl paraoxybenzoate 1.5g
Fragrance 0.1g
Purified water balance (total amount is 100 g)
〔配合例２〕
下記組成に従い、乳液を常法により製造した。
コメのサッカロミセス・ヴェローナによる発酵物（製造例） ０．０１ｇ
ホホバオイル ４．００ｇ
１，３−ブチレングリコール ３．００ｇ
アルブチン ３．００ｇ
ポリオキシエチレンセチルエーテル（20E.O.） ２．５０ｇ
オリーブオイル ２．００ｇ
スクワラン ２．００ｇ
セタノール ２．００ｇ
モノステアリン酸グリセリル ２．００ｇ
オレイン酸ポリオキシエチレンソルビタン（20E.O.） ２．００ｇ
パラオキシ安息香酸メチル ０．１５ｇ
グリチルレチン酸ステアリル ０．１０ｇ
黄杞エキス ０．１０ｇ
グリチルリチン酸ジカリウム ０．１０ｇ
イチョウ葉エキス ０．１０ｇ
コンキオリン ０．１０ｇ
オウバクエキス ０．１０ｇ
カミツレエキス ０．１０ｇ
香料 ０．０５ｇ
精製水 残部（全量を１００ｇとする）
[Formulation Example 2]
A milky lotion was produced by a conventional method according to the following composition.
Fermented product of rice by Saccharomyces verona (Production example) 0.01g
Jojoba oil 4.00g
1,3-butylene glycol 3.00 g
Arbutin 3.00g
Polyoxyethylene cetyl ether (20E.O.) 2.50 g
Olive oil 2.00g
Squalan 2.00g
Cetanol 2.00g
Glyceryl monostearate 2.00 g
Polyoxyethylene sorbitan oleate (20E.O.) 2.00 g
Methyl paraoxybenzoate 0.15 g
Stearyl glycyrrhetinate 0.10 g
Yellow 杞 extract 0.10g
Dipotassium glycyrrhizinate 0.10 g
Ginkgo leaf extract 0.10 g
Conchiolin 0.10g
Phellodendron extract 0.10 g
Chamomile extract 0.10g
Fragrance 0.05g
Purified water balance (total amount is 100 g)
〔配合例３〕
下記組成の美容液を常法により製造した。
コメのサッカロミセス・ヴェローナによる発酵物（製造例） ０．０１ｇ
カミツレエキス ０．１ｇ
ニンジンエキス ０．１ｇ
キサンタンガム ０．３ｇ
ヒドロキシエチルセルロース ０．１ｇ
カルボキシビニルポリマー ０．１ｇ
１，３−ブチレングリコール ４．０ｇ
グリチルリチン酸ジカリウム ０．１ｇ
グリセリン ２．０ｇ
水酸化カリウム ０．２５ｇ
香料 ０．０１ｇ
防腐剤（パラオキシ安息香酸メチル） ０．１５ｇ
エタノール ２．０ｇ
精製水 残部（全量を１００ｇとする）
[Formulation Example 3]
A beauty essence having the following composition was produced by a conventional method.
Fermented product of rice by Saccharomyces verona (Production example) 0.01g
Chamomile extract 0.1g
Carrot extract 0.1g
Xanthan gum 0.3g
Hydroxyethyl cellulose 0.1g
Carboxyvinyl polymer 0.1g
1,3-butylene glycol 4.0 g
Dipotassium glycyrrhizinate 0.1 g
Glycerin 2.0g
Potassium hydroxide 0.25g
Fragrance 0.01g
Preservative (methyl paraoxybenzoate) 0.15 g
Ethanol 2.0g
Purified water balance (total amount is 100 g)
〔配合例４〕
常法により、以下の組成を有する錠剤を製造した。
コメのサッカロミセス・ヴェローナによる発酵物（製造例） ５．０ｍｇ
ドロマイト（カルシウム20％、マグネシウム10％含有） ８３．４ｍｇ
カゼインホスホペプチド １６．７ｍｇ
ビタミンＣ ３３．４ｍｇ
マルチトール １３６．８ｍｇ
コラーゲン １２．７ｍｇ
ショ糖脂肪酸エステル １２．０ｍｇ
[Formulation Example 4]
Tablets having the following compositions were produced by a conventional method.
Fermented product of rice by Saccharomyces verona (Production example) 5.0 mg
Dolomite (containing 20% calcium and 10% magnesium) 83.4 mg
Casein phosphopeptide 16.7 mg
Vitamin C 33.4 mg
Maltitol 136.8 mg
Collagen 12.7 mg
Sucrose fatty acid ester 12.0 mg
〔配合例５〕
常法により、以下の組成を有する経口液状製剤を製造した。
＜１アンプル（１本１００ｍＬ）中の組成＞
コメのサッカロミセス・ヴェローナによる発酵物（製造例） ０．３質量％
ソルビット １２．０質量％
安息香酸ナトリウム ０．１質量％
香料 １．０質量％
硫酸カルシウム ０．５質量％
精製水 残部（１００質量％）
[Formulation Example 5]
An oral liquid preparation having the following composition was produced by a conventional method.
<Composition in 1 ampoule (100 mL per ampoule)>
Fermented product of rice by Saccharomyces verona (Production example) 0.3% by mass
Sorbitol 12.0% by mass
Sodium benzoate 0.1% by mass
Fragrance 1.0% by mass
Calcium sulfate 0.5% by mass
Purified water balance (100% by mass)
本発明に係るＨＡＳ３ ｍＲＮＡ発現促進剤は、皮膚の荒れ、しわ、くすみ、きめの変化、弾力性の低下及び保湿機能の低下等、皮膚の老化症状の予防、治療または改善等に大きく貢献できる。 The HAS3 mRNA expression promoter according to the present invention can greatly contribute to the prevention, treatment or improvement of skin aging symptoms such as rough skin, wrinkles, dullness, change in texture, decrease in elasticity and decrease in moisturizing function.
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