JP2018016624A - Oph active enhancer - Google Patents
Oph active enhancer Download PDFInfo
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- JP2018016624A JP2018016624A JP2017137758A JP2017137758A JP2018016624A JP 2018016624 A JP2018016624 A JP 2018016624A JP 2017137758 A JP2017137758 A JP 2017137758A JP 2017137758 A JP2017137758 A JP 2017137758A JP 2018016624 A JP2018016624 A JP 2018016624A
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- oph
- tea
- activity
- activity enhancer
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Abstract
Description
本発明は、酸化蛋白質を分解する生体内酵素である酸化蛋白質分解酵素（ｏｘｉｄｉｚｅｄ ｐｒｏｔｅｉｎ ｈｙｄｒｏｌａｓｅ：ＯＰＨ）の活性を増強させるＯＰＨ活性増強剤、及び、当該ＯＰＨ活性増強剤を含有する飲食品や医薬品などに関する。 The present invention relates to an OPH activity enhancer that enhances the activity of oxidized protein hydrolase (OPH), which is an in vivo enzyme that degrades oxidized protein, and foods and beverages and pharmaceuticals containing the OPH activity enhancer. About.
ＯＰＨは、蛋白のＮ末端アシル化アミノ酸を遊離するセリンプロテアーゼの一種で、アシルアミノ酸遊離酵素（ａｃｙｌａｍｉｎｏ−Ａｃｉｄ Ｒｅｌｅａｓｉｎｇ Ｅｎｚｙｍｅ：ＡＡＲＥ）、アシル化ペプチド分解酵素（ａｃｙｌｐｅｐｔｉｄｅ ｈｙｄｒｏｌａｓｅ：ＡＰＨ）などとも言われている。また、ＯＰＨは、アシル化だけでなく、ホルミル化（Ｆｏｒｍｙｌ）、アセチル化（Ａｃｅｔｙｌ）、ブチル化（Ｂｕｔｙｌ）、プロピル化（Ｐｒｏｐｙｌ）された蛋白質のＮ末端アミノ酸に対しても作用する。 OPH is a kind of serine protease that liberates N-terminal acylated amino acids of proteins, and is also referred to as acyl amino acid releasing enzyme (AARE), acylated peptide hydrolase (APH), and the like. . Further, OPH acts not only on acylation but also on the N-terminal amino acid of a protein that has been formylated (Formyl), acetylated (Acetyl), butylated (Butyl), or propylated (Propyl).
ＯＰＨはブタ肝臓、ラット脳、ヒト血液、角層などの生体組織に広く存在している。ＯＰＨは酸化蛋白質や糖化蛋白質を優先的に分解するとともにプロテアソームと協働して老化蛋白質を分解すること、アルツハイマー病の原因であるアミロイドβ蛋白質を減少させることが報告されている。またＯＰＨがＡＧＥｓ（終末糖化産物）を分解することも確認されている。これらのことから生体中のＯＰＨの活性を増強させることは、老化蛋白質の分解を促進し、老化や疾患を予防・治療できる可能性がある。 OPH is widely present in living tissues such as pig liver, rat brain, human blood, and stratum corneum. It has been reported that OPH preferentially degrades oxidized proteins and glycated proteins, degrades aging proteins in cooperation with the proteasome, and reduces amyloid β protein that causes Alzheimer's disease. It has also been confirmed that OPH degrades AGEs (terminal glycation products). From these facts, enhancing the activity of OPH in the living body promotes the degradation of aging proteins and may prevent or treat aging and diseases.
上記の事情を鑑み、本発明は、ＯＰＨの活性を増強させるＯＰＨ活性増強剤を提供することを課題とする。 In view of the above circumstances, an object of the present invention is to provide an OPH activity enhancer that enhances the activity of OPH.
上記課題を解決するための手段として、以下の発明などを提供する。すなわち、カキノキ、チコリ、シソ、ルイボス、サラシア、テンヨウケンコウシ、月見草、玄米、タンポポ、バラ、キャンドルブッシュ、モリンガ、アマチャ、オリーブ、ナタマメ、バナバのいずれか一種類以上を含有するＯＰＨ活性増強剤を提供する。 As means for solving the above problems, the following inventions and the like are provided. That is, an OPH activity enhancer containing any one or more of oyster, chicory, perilla, rooibos, salacia, velvet primrose, evening primrose, brown rice, dandelion, rose, candle bush, moringa, amacha, olive, jujube, banaba provide.
また、クマザサ、カキノキ、テンヨウケンコウシ、バナバの各抽出物を含む混合物を含有するＯＰＨ活性増強剤を提供する。 Moreover, the OPH activity enhancer containing the mixture containing each extract of Kumazasa, Oyster mushroom, Tenno Kenkoushi, Banaba is provided.
また、マスリン酸、（＋）カテキン、イソクェルシトリンのいずれか１種類以上を含有するＯＰＨ活性増強剤を提供する。 Moreover, the OPH activity enhancer containing any one or more of maslinic acid, (+) catechin, and isoquercitrin is provided.
また、前記いずれか一のＯＰＨ活性増強剤を含有する食品、食品添加物、医薬品、医薬部外品及び化粧品を提供する。 The present invention also provides foods, food additives, pharmaceuticals, quasi drugs and cosmetics containing any one of the above OPH activity enhancers.
本発明により、酸化蛋白質を分解する生体内酵素であるＯＰＨの活性を増強させるＯＰＨ活性増強剤などを提供することができる。 The present invention can provide an OPH activity enhancer that enhances the activity of OPH, which is an in vivo enzyme that degrades oxidized protein.
以下、本発明の実施例について説明する。なお、本発明は、これらの実施例に何ら限定されるべきものではなく、その要旨を逸脱しない範囲において、種々なる態様で実施し得る。
＜実施形態＞
＜構成＞
Examples of the present invention will be described below. In addition, this invention should not be limited at all to these Examples, and can be implemented with various aspects in the range which does not deviate from the summary.
<Embodiment>
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本実施例に係るＯＰＨ活性増強剤は、カキノキ、チコリ、シソ、ルイボス、サラシア、テンヨウケンコウシ、月見草、玄米、タンポポ、バラ、キャンドルブッシュ、モリンガ、アマチャ、オリーブ、ナタマメ、バナバの抽出物のいずれか１種類以上を含有するＯＰＨ活性増強剤である。「ＯＰＨ活性増強剤」とは、酸化蛋白質の分解酵素であるＯＰＨの酸化蛋白質分解活性（以下、単にＯＰＨ活性）を増強させる剤を意味する。 The OPH activity enhancer according to the present example is any of the extracts of oyster, chicory, perilla, rooibos, Salacia, tempura, evening primrose, brown rice, dandelion, rose, candle bush, moringa, amacha, olive, jujube, banaba Or an OPH activity enhancer containing one or more kinds. The “OPH activity enhancer” means an agent that enhances the oxidized proteolytic activity (hereinafter simply referred to as OPH activity) of OPH, which is an enzyme that degrades oxidized protein.
本実施形態における植物の抽出物は、植物のどの部位から抽出したものであってもよく、例えば、全草、花、種子、果実、枝、茎、樹皮、根などから抽出したものであってよい。また、抽出物の性状を限定するものではない。以下に、本実施形態における各植物について説明する。 The plant extract in this embodiment may be extracted from any part of the plant, for example, extracted from whole grass, flowers, seeds, fruits, branches, stems, bark, roots, etc. Good. Moreover, the property of the extract is not limited. Below, each plant in this embodiment is demonstrated.
また、前記の各植物を水又は湯により抽出した物は、例えば、カキノキを湯や水で抽出した物は柿の葉茶といった称呼で茶外茶として喫飲されている。なお、茶外茶とは、チャノキ以外の植物の葉、芽、花、樹皮、根などを材料として水又は湯を注ぎ抽出した飲料のことをいう。 Moreover, the thing which extracted each said plant with water or hot water, for example, the thing which extracted the oyster tree with hot water or water is drunk as tea outside tea by the name of Kashiwanoha tea. In addition, tea outside tea means the drink which poured water or hot water using the leaves, buds, flowers, bark, roots, etc. of plants other than tea tree as a material.
「カキノキ（Ｄｉｏｓｐｙｒｏｓ ｋａｋｉ）」は、カキノキ科カキノキ属の植物である。サンプルとして、カキノキの葉を原料とする茶外茶である柿の葉茶を用いた。 “Diospyros kaki” is a plant of the genus Oysteraceae. As a sample, persimmon leaf tea, which is a tea outside tea made from oyster leaves, was used.
「チコリ（Ｃｉｃｈｏｒｉｕｍ ｉｎｔｙｂｕｓ）」は、キク科キクニガナ属の植物であり、菊苣とも称される。サンプルとして、乾燥させたチコリの根を原料とする茶外茶であるチコリ茶を用いた。なお、茶外茶とは、チャノキ以外の植物の葉、芽、花、樹皮、根などを材料として水又は湯を注ぎ抽出した飲料のことをいう。 “Chicorium intybus” is a plant belonging to the genus Asteraceae and is also referred to as chrysanthemum. As a sample, chicory tea which is a tea outer tea made from dried chicory roots was used. In addition, tea outside tea means the drink which poured water or hot water using the leaves, buds, flowers, bark, roots, etc. of plants other than tea tree as a material.
「シソ」は、シソ科シソ属の植物であり、この属には、アオジソ（Ｐｅｒｉｌｌａ ｆｒｕｔｅｓｃｅｎｓ ｖａｒ. ｃｒｉｓｐａ ｆ. ｖｉｒｉｄｉｓ）、アカジソ（Ｐｅｒｉｌｌａ ｆｒｕｔｅｓｃｅｎｓ ｖａｒ. ｃｒｉｓｐａ ｆ. ｐｕｒｐｕｒｅａ）などがある。サンプルとして、アカジソの葉を原料とする茶外茶であるシソ葉茶を用いた。 “Perilla” is a plant belonging to the genus Lamiaceae, which includes Perilla frescens var. Crispa f. Viridis, Akajiso (Perilla fluorescens var. Crispa f. As a sample, perilla leaf tea, which is a tea outside tea made from red radish leaves, was used.
「ルイボス（Ａｓｐａｌａｔｈｕｓ ｌｉｎｅａｒｉｓ）」は、マメ科アスパラトゥス属の植物である。サンプルとして、ルイボスの葉を原料とする茶外茶であるルイボス茶を用いた。 “Rosbos (Linearis)” is a plant of the genus Asparatus. As a sample, rooibos tea which is a tea outside tea made from rooibos leaves was used.
「サラシア」は、ニシキギ科サラシア属の植物であり、この属には、サラシア・オブロンガ（Ｓａｌａｃｉａ ｏｂｌｏｎｇａ）、サラシア・レティキュラータ（Ｓａｌａｃｉａ ｒｅｔｉｃｕｌａｔａ）などがある。サンプルとして、サラシア・レティキュラータの根や地下茎を原料とする茶外茶であるサラシア茶を用いた。なお、サラシア・オブロンガはインド産やブラジル産のものが知られており、サラシア・レティキュラータはスリランカ産のものが知られている。いずれのサラシアも基本的な性質は同様であるが、サラシア・レティキュラータの方が、糖尿病予防などについての効能がより高いと言われることがある。 “Salasia” is a plant belonging to the genus Salacia belonging to the asteraceae family. Examples of this genus include Salacia oblonga and Salacia reticulata. As a sample, Salacia tea, which is a tea outside tea made from Salacia reticulata roots and rhizomes, was used. Salacia oblonga is known from India and Brazil, and Salacia reticulata is known from Sri Lanka. Although all the basic properties of Salacia are the same, Salacia reticulata may be said to be more effective in preventing diabetes.
「テンヨウケンコウシ（Ｒｕｂｕｓ ｓｕａｖｉｓｓｉｍｕｓ）」は、バラ科キイチゴ属の植物である。サンプルとして、テンヨウケンコウシの葉を原料とする茶外茶である甜茶を用いた。 “Rubus suavissimus” is a plant belonging to the genus Rubiaceae in the family Rosaceae. As a sample, strawberry tea, which is a tea-outer tea made from the leaves of Tenno-komushi beef, was used.
「月見草（Ｏｅｎｏｔｈｅｒａ ｔｅｔｒａｐｔｅｒａ）」は、アカバナ科マツヨイグサ属の植物である。サンプルとして、月見草の葉と茎を原料とする茶外茶である月見草茶を用いた。 “Oenothera tetraptera” is a plant of the genus Oenothera. As a sample, evening primrose tea, a tea outer tea made from evening primrose leaves and stems, was used.
「玄米」は、イネ科イネ属のイネ（Ｏ．ｓａｔｉｖａ．ｓｕｂｓｐ．ｊａｐｏｎｉｃａ）の果実である籾から籾殻を除去したものである。サンプルとして、焙煎した玄米を用いた。なお、番茶に玄米を混ぜたいわゆる玄米茶をサンプルとするものではない。 “Brown rice” is obtained by removing rice husks from cocoons which are the fruits of the rice of the genus Gramineae (O. sativa. Subsp. Japonica). As a sample, roasted brown rice was used. In addition, so-called brown rice tea mixed with brown rice and bancha is not used as a sample.
「タンポポ（Ｔａｒａｘａｃｕｍ）」は、キク科タンポポ属の植物である。サンプルとして、タンポポの花、葉、茎を原料とする茶外茶であるタンポポ茶を用いた。 "Taraxacum" is a plant belonging to the genus Dandelion. As a sample, dandelion tea which is a tea outside tea made from dandelion flowers, leaves and stems was used.
「ローズヒップ」は、バラ科バラ属のイヌバラ（Ｒｏｓａ ｃａｎｉｎａ）の果実である。サンプルとしては、ローズヒップを原料とする茶外茶であるローズヒップ茶を用いた。 “Rose hip” is the fruit of the Rosa rose of the genus Rosaceae. As a sample, rose hip tea which is tea outside tea made from rose hip was used.
「キャンドルブッシュ（Ｃａｓｓｉａ ａｌａｔａ）」は、マメ科カッシア属の植物であり、ゴールデンキャンドルとも呼ばれる。サンプルとしては、キャンドルブッシュの葉や茎を原料とする茶外茶であるキャンドルブッシュ茶を用いた。 “Candia bush” is a plant belonging to the genus Cassia, which is also referred to as a golden candle. As a sample, Candle Bush Tea, which is a tea outside tea made from candle bush leaves and stems, was used.
「モリンガ（Ｍｏｒｉｎｇａ ｏｌｅｉｆｅｒａ）」は、ワサビノキ科ワサビノキ属の植物である。サンプルとしては、モリンガの葉や茎を原料とする茶外茶であるモリンガ茶を用いた。 “Moringa oleifera” is a plant of the genus Wasabiaceae. As a sample, Moringa tea, which is a tea outside tea made from Moringa leaves and stems, was used.
「アマチャ（Ｈｙｄｒａｎｇｅａ ｍａｃｒｏｐｈｙｌｌａ Ｓｅｒ．ｖａｒ．ｔｈｕｎｂｅｒｇｉｉ）」は、アジサイ科アジサイ属の植物である。サンプルとしては、アマチャの葉や茎を原料とする茶外茶である甘茶を用いた。 “Amateur (Hydrangea macrophylla Ser. Var. Thunbergii)” is a plant belonging to the genus Hydrangea. As a sample, Amacha, which is a tea outside tea made from Amacha leaves and stems, was used.
「オリーブ（ｏｌｅａ ｅｕｒｏｐａｅａ）」は、モクセイ科オリーブ属の植物である。サンプルとしては、オリーブの葉を主原料とする茶外茶であるオリーブリーフ茶を用いた。 “Olivea europaea” is a plant belonging to the genus Oleaceae. As a sample, olive leaf tea, which is a tea outside tea mainly made of olive leaves, was used.
「ナタマメ（Ｃａｎａｖａｌｉａ ｇｌａｄｉａｔｅ）」は、マメ科ナタマメ属の植物である。サンプルとして、ナタマメの種子を主原料とする茶外茶であるナタマメ茶を用いた。 “Camellia (Canavalia gladiate)” is a plant belonging to the genus Leguminosae. As a sample, peanut tea, which is a tea outside tea mainly made of peanut seeds, was used.
「バナバ（Ｌａｇｅｒｓｔｒｏｅｍｉａ ｓｐｅｃｉｏｓａ）」は、ミソハギ科サルスベリ属の植物である。サンプルとして、バナバの葉や茎を原料とする茶外茶であるバナバ茶を用いた。 “Banaba (Lagerstromia speciosa)” is a plant of the genus Crape myrtle. As a sample, banaba tea, which is a tea outside tea made from banaba leaves and stems, was used.
本実施例におけるＯＰＨ活性増強剤は、さらに既知の方法を用いることにより、当該ＯＰＨ活性増強剤を含有する食品、食品添加物、医薬品、医薬部外品、化粧品などとして提供することが可能である。 The OPH activity enhancer in this example can be provided as a food, food additive, pharmaceutical, quasi-drug, cosmetic or the like containing the OPH activity enhancer by using a known method. .
例えば、医薬品とする場合には、本実施例のＯＰＨ活性増強剤を粉体や粒体としカプセルに充填したり、あるいは、賦形剤、結合剤、崩壊剤などを添加して打錠機等を用いて製造することができる。また、食品とする場合には、各植物を適宜乾燥や破砕等を湯で煮出すことで提供できる。また、医薬品のようにカプセルや錠剤のような形態で提供してもよいし、他の飲料、調味料、菓子等の各種の食品にＯＰＨ活性増強剤を添加した態様で提供することもできる。 For example, in the case of a pharmaceutical product, the OPH activity enhancer of this example is filled with powder or granules, filled in a capsule, or added with an excipient, a binder, a disintegrant, etc. Can be used. Moreover, when setting it as a foodstuff, it can provide by simmering each plant suitably with a hot water such as drying or crushing. Moreover, you may provide in the form like a capsule and a tablet like a pharmaceutical, and can also provide in the aspect which added OPH activity enhancer to various foodstuffs, such as another drink, a seasoning, and confectionery.
また、美容液、クリーム、ローションなどの化粧品とすることもできる。例えば、美容液とする場合には、本実施例のＯＰＨ活性増強剤の他、水、コメヌカ油、ペンチレングリコール、グリセリン、スクワラン、パルミチン酸セチル、ダイマージリノール酸などを主成分とし、ヒアルロン酸Ｎａ、水添ナタネ油アルコール、カルボマー、キサンタンガム、水酸化カリウム、ジメチコン、ポリソルベート−６０、ステアリン酸グリセリル、水添ヒマシ油、フェノキシエタノール、尿素、アルギニン、アルブチン、クエン酸などを添加剤とする。そして、各成分を水溶性原料・油溶性原料に分けて溶解してから、それらを加熱して混合・乳化する。これを冷却しながらエキスなどの添加物を配合し、さらに低温になったところで精油や香料などの揮発性の高いものを添加する。その後、所定の安全性の検査（菌、ｐＨ、温度安定性、粘度等）を行い、瓶などに充填して製品として提供することができる。
≪試験１≫
Moreover, it can also be set as cosmetics, such as a cosmetic liquid, cream, and lotion. For example, when it is used as a cosmetic liquid, in addition to the OPH activity enhancer of this example, water, rice bran oil, pentylene glycol, glycerin, squalane, cetyl palmitate, dimer dilinoleic acid and the like as main components, hyaluronic acid Additives include Na, hydrogenated rapeseed oil alcohol, carbomer, xanthan gum, potassium hydroxide, dimethicone, polysorbate-60, glyceryl stearate, hydrogenated castor oil, phenoxyethanol, urea, arginine, arbutin, and citric acid. Then, after each component is dissolved in a water-soluble raw material and an oil-soluble raw material, they are heated and mixed and emulsified. While cooling this, an additive such as an extract is blended, and a highly volatile substance such as essential oil or fragrance is added when the temperature becomes lower. Thereafter, a predetermined safety test (bacteria, pH, temperature stability, viscosity, etc.) is performed, and the product can be provided as a product after being filled into a bottle.
本試験において、上記各植物の抽出物のＯＰＨ活性の増強作用について測定する。本測定ではＯＰＨとその反応基質であるＮ−ａｃｅｔｙｌ−Ｌ−ａｌａｎｉｎｅ ｐ−ｎｉｔｒｏ−ａｎｉｌｉｄｅ（ＡＡＰＡ）との反応系に試料溶液を添加し、ＯＰＨの酵素反応への影響を評価した。試料には、前述のカキノキ、チコリ、ルイボス、サラシア、テンヨウケンコウシ、月見草、植物のサンプルを用いた。併せて、イネ科ササ属のクマザサの葉を原料とするクマザサ茶、ミソハギ科サルスベリ属のバナバの葉を原料とするバナバ茶、フトモモ科バンジロウ属のガァバの葉を原料とするガァバ茶、バラ科バラ属のハマナスの花を原料とするメイグイ茶についても測定した。また、植物抽出物を組み合わせた場合のＯＰＨ活性増強作用を確認するために、「クマザサ茶、柿の葉茶、甜茶、バナバ茶」を等量で混合した混合茶についても測定した。 In this test, the effect of enhancing the OPH activity of each plant extract is measured. In this measurement, a sample solution was added to the reaction system of OPH and its reaction substrate, N-acetyl-L-alanine p-nitro-anilide (AAPA), and the influence of OPH on the enzyme reaction was evaluated. Samples of the aforementioned oyster mushrooms, chicory, rooibos, salacia, velvet primrose, evening primrose, and plants were used as samples. In addition, Kumazasa tea made from the grass of the genus Sasa genus, Banaba tea made from the banaba leaf of the genus Cranaceae, Gaba tea made from the leaves of the genus Vanilla genus, Rose family Measurements were also made on Meigui tea made from rose genus flowers. In addition, in order to confirm the effect of enhancing the OPH activity when combined with plant extracts, mixed tea in which equal amounts of “kumazasa tea, kashiwanoha tea, koji tea, banaba tea” were also measured.
また、バナバなどの含有成分であるマスリン酸（Ｍａｓｌｉｎｉｃ Ａｃｉｄ）（Ｃ３０Ｈ４８Ｏ４）、チャノキなどの含有成分である（＋）カテキン（Ｃ１５Ｈ１４Ｏ６）、柿の葉などの含有成分であるイソクェルシトリン（ｉｓｏｑｕｅｒｃｉｔｉｎ）（Ｃ２１Ｈ２０Ｏ１２）についても測定を行った。
＜測定方法＞
Further, maslinic acid (Maslinic Acid) (C 30 H 48 O 4) , which is a component such as banaba, (+) catechin (C 15 H 14 O 6 ), which is a component such as tea tree, and the like Measurements were also made for the component isoquercitin (C 21 H 20 O 12 ).
<Measurement method>
ＯＰＨとしてａｃｙｌａｍｉｎｏ−ａｃｉｄ ｒｅｌｅａｓｉｎｇ ｅｎｚｙｍｅ（ＡＡＲＥ）、ＯＰＨの反応基質としてＡＡＰＡ溶液を使用した。測定にはＯＰＨを０．０１Ｕ／ｍＬ、０．００５Ｕ／ｍＬ、０．００１Ｕ／ｍＬに調製して使用した。 Acylamino-acid releasing enzyme (AARE) was used as OPH, and AAPA solution was used as a reaction substrate for OPH. For the measurement, OPH was adjusted to 0.01 U / mL, 0.005 U / mL, and 0.001 U / mL.
９６ウェルマイクロプレートの各ｗｅｌｌにＯＰＨ、ＡＡＰＡ、試料溶液（ジメチルスルホキシドを溶媒）を混合添加し、３７℃に設定したインキュベーター内で４時間反応させた反応液の４０５ｎｍにおける吸光度をマイクロプレートリーダーで測定した。ＯＰＨの酵素活性は１時間当たりの吸光度変化量（反応速度）を求めた。同時にｒｅｆｅｒｅｎｃｅ（Ｒｅｆ）として試料無添加時の反応速度を求め、下式に従ってＲｅｆの反応速度を１００％とした時の活性増強作用を算出した。ＯＰＨ活性増強作用のネガティブコントロールにはエピガロカテキンガレート（ＥＧＣｇ）を使用した。
（式）ＯＰＨ活性増強作用（％）＝（試料のＯＰＨ反応速度／ＲｅｆのＯＰＨ反応速度）×１００
＜測定結果＞
Absorbance at 405 nm of the reaction solution obtained by mixing and adding OPH, AAPA, and sample solution (dimethyl sulfoxide as a solvent) to each well of a 96-well microplate for 4 hours in an incubator set at 37 ° C. was measured with a microplate reader. did. For the enzyme activity of OPH, the amount of change in absorbance per hour (reaction rate) was determined. At the same time, the reaction rate when no sample was added was determined as reference (Ref), and the activity enhancing action when the reaction rate of Ref was 100% was calculated according to the following formula. Epigallocatechin gallate (EGCg) was used as a negative control for enhancing the OPH activity.
(Expression) OPH activity enhancing action (%) = (OPH reaction rate of sample / OPH reaction rate of Ref) × 100
<Measurement results>
図１は、測定結果を示す表である。表に示すように、本実施形態に係る、カキノキ、チコリ、シソ、ルイボス、サラシア、テンヨウケンコウシ、月見草の抽出物はいずれもＯＰＨ活性増強作用を有することが分かった。また、混合茶についてもＯＰＨ活性増強作用を有することが分かった。また、マスリン酸、（＋）カテキン、イソクェルシトリンについてもＯＰＨ活性増強作用を有することが分かった。
≪試験２≫
FIG. 1 is a table showing measurement results. As shown in the table, it was found that all the extracts of oyster mushroom, chicory, perilla, rooibos, Salacia, velvet primrose and evening primrose according to the present embodiment have an OPH activity enhancing action. It was also found that mixed tea has an effect of enhancing OPH activity. It was also found that maslinic acid, (+) catechin, and isoquercitrin have an effect of enhancing OPH activity.
上述の試験１と同様に本実施形態の植物抽出物のＯＰＨ活性の増強作用について測定した。前述の玄米、タンポポ、バラ、キャンドルブッシュ、モリンガ、アマチャ、オリーブ、ナタマメ、バナバを測定対象とし、併せてジャスミン茶、紅茶、メイグイ茶、緑茶についても測定対象とした。
In the same manner as in
試料として、以下の通り植物抽出液を調整した。まず、秤量した２ｇの各茶葉を４０ｍＬの熱水（８０℃）で１時間抽出し、常温まで冷ましてから市販のお茶パックで濾して上清を回収して抽出液とした。また、抽出液原液だけでなく１０倍希釈液も測定対象に加えた。さらに、希釈して活性化率がアップしそうな試料については１００倍希釈液でも測定した。
＜測定方法＞
A plant extract was prepared as a sample as follows. First, 2 g of each weighed tea leaf was extracted with 40 mL of hot water (80 ° C.) for 1 hour, cooled to room temperature, filtered through a commercially available tea pack, and the supernatant was recovered to obtain an extract. In addition to the extract stock solution, a 10-fold diluted solution was also added to the measurement target. Furthermore, the sample that was likely to increase the activation rate by dilution was also measured with a 100-fold diluted solution.
<Measurement method>
ＯＰＨとしてａｃｙｌａｍｉｎｏ−ａｃｉｄ ｒｅｌｅａｓｉｎｇ ｅｎｚｙｍｅ（ＡＡＲＥ）（タカラバイオ）を、５０ｍｍｏｌ／Ｌ リン酸緩衝液（ｐＨ７．２）にて０．０２５Ｕ／ｍＬに調製した。また、反応基質としてＡＡＰＡ溶液（ＢＡＣＨＥＭ）を、５０％エタノール液で２５ｍｍｏｌ／Ｌに調製した。また、反応緩衝液として、１２０ｍｍｏｌ／Ｌ Ｔｒｉｓ−ＨＣｌ（ｐＨ７．４）を用いた。また、ポジティブコントロールとして１ｍｇ／ｍＬのルブソシド、ネガティブコントロールとして１ｍｇ／ｍＬのＥＧＣｇ（エピガロカテキンガレート）を用いた。 As OPH, acylamino-acid releasing enzyme (AARE) (Takara Bio) was adjusted to 0.025 U / mL with 50 mmol / L phosphate buffer (pH 7.2). In addition, an AAPA solution (BACHEM) was prepared as a reaction substrate to 25 mmol / L with a 50% ethanol solution. Further, 120 mmol / L Tris-HCl (pH 7.4) was used as a reaction buffer. Further, 1 mg / mL rubusoside was used as a positive control, and 1 mg / mL EGCg (epigallocatechin gallate) was used as a negative control.
そして、マイクロプレート１ウェルに、試料１０ｕＬ、０．０２５Ｕ／ｍＬ ＯＰＨ １０ｕＬ、反応緩衝液２１０ｕＬを入れて、３７℃で６０分間予備加温（プレインキュベーション）した。一旦、室温に戻してから、２５ｍｍｏｌ／Ｌ ＡＡＰＡ溶液（基質液）２０ｕＬを加え、よく混合してＯＰＨ反応を開始した。反応は３７℃の恒温槽で行った。 Then, 10 uL of sample, 10 uL of 0.025 U / mL OPH, and 210 uL of reaction buffer were added to one well of the microplate, and pre-warmed (preincubation) for 60 minutes at 37 ° C. Once returned to room temperature, 20 uL of 25 mmol / L AAPA solution (substrate solution) was added and mixed well to initiate OPH reaction. The reaction was carried out in a constant temperature bath at 37 ° C.
３０分、あるいは６０分間隔でマイクロプレートを恒温槽から取り出し、マイクロプレートリーダーにて４０５ｎｍの吸光度を測定し、ＯＰＨ反応によって生じるｐＮＡ（パラニトロアニリン）を測定した。 The microplate was taken out from the thermostatic chamber at intervals of 30 minutes or 60 minutes, the absorbance at 405 nm was measured with a microplate reader, and pNA (paranitroaniline) generated by the OPH reaction was measured.
ここで、試料のうちキャンドルブッシュについてのＯＤ（光学密度）が０．５を上回ったので、吸光度が０．５以下に収まるように蒸留水で２倍希釈したものを原液として用いることにした。 Here, since the OD (optical density) of the candle bush among the samples exceeded 0.5, it was decided to use a sample diluted twice with distilled water so that the absorbance was 0.5 or less.
そして、ＯＰＨの活性の増強作用は、試験１と同様に１時間当たりの吸光度変化量（反応速度）を求めた。同時にｒｅｆｅｒｅｎｃｅ（Ｒｅｆ）として試料無添加時の反応速度を求め、下式に従ってＲｅｆの反応速度を１００％とした時の活性増強作用を算出した。
（式）ＯＰＨ活性増強作用（％）＝（試料のＯＰＨ反応速度／ＲｅｆのＯＰＨ反応速度）×１００
＜測定結果＞
Then, for the enhancement of the activity of OPH, the amount of change in absorbance (reaction rate) per hour was determined in the same manner as in
(Expression) OPH activity enhancing action (%) = (OPH reaction rate of sample / OPH reaction rate of Ref) × 100
<Measurement results>
図２は、測定結果を示す表である。図中の「濃度１」は抽出液原液を試料として用いた場合のＯＰＨ活性増強作用を示している。また、「濃度２」は１０倍希釈液を、「濃度３」は１００倍希釈液を用いた結果を示している。ただし、キャンドルブッシュについては上記の通り「濃度１」において２倍希釈液を、「濃度２」において２０倍希釈液を用いた。また、最大活性増強作用（％）を下線を付した斜字で示した。表に示すように、本実施形態に係る、玄米、タンポポ、バラ、キャンドルブッシュ、モリンガ、アマチャ、オリーブ、ナタマメ、バナバの抽出物はいずれもＯＰＨ活性増強作用を有することが分かった。
＜効果＞
FIG. 2 is a table showing the measurement results. “
<Effect>
試験１及び試験２の結果として示すように、本実施形態により、カキノキ、チコリ、シソ、ルイボス、サラシア、テンヨウケンコウシ、月見草、玄米、タンポポ、バラ、キャンドルブッシュ、モリンガ、アマチャ、オリーブ、ナタマメ、バナバのいずれか一種類以上を含有するＯＰＨ活性増強剤などを提供することができる。
As shown as the results of
また、クマザサ、カキノキ、テンヨウケンコウシ、バナバの各抽出物を含む混合物を含有するＯＰＨ活性増強剤などを提供することができる。 Moreover, the OPH activity enhancer etc. containing the mixture containing each extract of Kumazasa, Oyster mushroom, Tenno-komushi and Banaba can be provided.
また、マスリン酸、（＋）カテキン、イソクェルシトリンのいずれか１種類以上を含有するＯＰＨ活性増強剤などを提供することができる。 In addition, an OPH activity enhancer containing at least one of maslinic acid, (+) catechin, and isoquercitrin can be provided.
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