JP6541061B2 - 時計遺伝子発現促進用組成物 - Google Patents
時計遺伝子発現促進用組成物 Download PDFInfo
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- JP6541061B2 JP6541061B2 JP2015093534A JP2015093534A JP6541061B2 JP 6541061 B2 JP6541061 B2 JP 6541061B2 JP 2015093534 A JP2015093534 A JP 2015093534A JP 2015093534 A JP2015093534 A JP 2015093534A JP 6541061 B2 JP6541061 B2 JP 6541061B2
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Description
本発明の組成物は、時計遺伝子発現促進作用を奏する有効成分として黒生姜(Kaempferia parviflora)を少なくとも含有する。
全体を100質量部として、黒生姜抽出物 0.01質量部、生姜抽出物 0.01質量部、ケツメイシ 0.01質量部、グリセリン 10質量部、ジグリセリン 3質量部、1,3−ブチレングリコール 12質量部、ペンチレングリコール 3質量部、ヒアルロン酸ナトリウム 0.1質量部、クエン酸 0.01質量部、クエン酸ナトリウム 0.02質量部、キサンタンガム 0.1質量部、メチルパラベン 0.15質量部、カルボマー 0.2質量部、水酸化ナトリウム 0.03質量部及び水 残部を混合して、化粧水の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜抽出物 0.01質量部、ホップ抽出物 0.02質量部、カフェイン 0.01質量部、ラウレス硫酸ナトリウム 7.5質量部、コカミドプロピルベタイン 4.2質量部、コカミドDEA 3質量部、1,3−ブチレングリコール 0.1質量部、ポリクオタニウム−10 0.225質量部、クエン酸 0.15質量部、クエン酸ナトリウム 0.05質量部、フェノキシエタノール 0.9質量部及び水 残部を混合して、シャンプーの態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜粉砕物 0.5質量部、メリロート抽出物 0.2質量部、エラスチン 0.1質量部、グリセリン 2質量部、オリーブ油 1質量部、EDTA−4ナトリウム 0.1質量部、エチドロン酸4ナトリウム 0.2質量部及び石ケン素地 残部を混合及び固化することにより、石鹸の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜抽出物 0.1質量部、アラニン 0.1質量部、アルギニン 0.1質量部、ショ糖脂肪酸エステル 3質量部、グリセリン 12質量部、スクワラン 6質量部、ジメチルシリコーンオイル 24質量部、ポリプロピレングリコール 1質量部、増粘剤 0.06質量部、フェノキシエタノール 0.2質量部、エタノール 5質量部、水酸化ナトリウム 0.01質量部及び精製水 残部を混合して、乳液の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜抽出物 0.1質量部、イソマルトオリゴ糖 0.1質量部、フラクトオリゴ糖 0.1質量部、スクワラン 15.0質量部、ミリスチン酸オクチルドデシル 4.0質量部、水素添加大豆リン脂質 0.2質量部、ブチルアルコール 2.4質量部、硬化油 1.5質量部、ステアリン酸 1.5質量部、親油型モノステアリン酸グリセリン 1.5質量部、モノステアリン酸ポリグリセリル 0.5質量部、ベヘニルアルコール 0.8質量部、モノミリスチン酸ポリグリセリル 0.7質量部、サラシミツロウ 0.3質量部、d−δ−トコフェロール 0.1質量部、メチルパラベン 0.3質量部、C10〜30アルキル変性カルボキシビニルポリマー 0.2質量部、カルボキシビニルポリマー 0.1質量部、1,3−ブタンジオール 18.0質量部、水酸化ナトリウム 0.1質量部及び精製水 残部を混合して、化粧クリームの態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜抽出物 0.1質量部、没食子酸 0.01質量部、ポリビニルアルコール 20.0質量部、グリセリン 5.0質量部、エタノール 20.0質量部、カオリン 6.0質量部、防腐剤 0.2質量部、香料 0.1質量部及び精製水 残部を混合して、パック剤の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜粉末 10質量部、ショウガ抽出物 8質量部、カフェイン5質量部、メリロート 5質量部、結晶性セルロース 20質量部、乳糖 50質量部、ステアリン酸マグネシウム 4質量部及びコーンスターチ 残部を混合及び打錠することにより、錠剤の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜粉末 10質量部、ケツメイシ末 8質量部、メリロート 5質量部、イソマルトオリゴ糖 0.5質量部、フラクトオリゴ糖 0.5質量部、結晶性セルロース 20質量部、乳糖 50質量部、ステアリン酸マグネシウム 4質量部及びコーンスターチ 残部を混合及び打錠することにより、錠剤の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜粉末 10質量部、ホップ末 15質量部、ケツメイシ抽出物 5質量部、乳糖 10質量部、ステアリン酸カルシウム 1質量部及び結晶性セルロース 残部を混合及び顆粒化することにより、顆粒剤の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜抽出物 10質量部、生姜抽出物 20質量部、エラスチン5質量部、レシチン 8質量部及びオリーブ油 残部を混合して調製したものを内容液として、これをカプセル殻に内包することにより、カプセル剤の態様で本発明の組成物を調製した。
全体を100質量部として、黒生姜エキス粉末 0.84質量部、アルギニン 1質量部、アラニン 1質量部、果糖ブドウ糖液糖 10質量部、クエン酸 1質量部、安息香酸ナトリウム 0.02質量部、香料製剤 2質量部、スクラロース 0.05質量部、アセスルファムカリウム 0.03質量部、及び精製水 残部を混合して、液剤の態様で本発明の組成物を調製した。
(1)被験試料群1の調製
黒生姜の乾燥根茎 0.5gを50%エタノール 10mLに加えたものを、30秒のヴォルテックス撹拌処理及び10秒の超音波処理に供し、次いで10分間の振盪処理に供し、次いで遠心分離処理に供した。遠心分離処理により得られた上清を黒生姜50%エタノール抽出物とした。同様に、黒生姜の代わりに黒豆、マッシュルーム、シログワイ、ブロッコリー、サツマイモ、ゴボウ又はトマトを用いた50%エタノール抽出物を調製した。これらの50%エタノール抽出物を被験試料群1とし、後述する遺伝子発現解析に用いた。
黒生姜の乾燥根茎 100gを95%エタノール 1,000mLに加えたものを、6日間の非連続撹拌処理に供し、次いでろ過処理に供した。得られたろ液を蒸発乾燥することにより、黒生姜95%エタノール抽出物(6.8g)を得た。黒生姜95%エタノール抽出物は、後述する分画に用いた。
マウス繊維芽細胞であるNIH−3T3細胞を3日培養したものに、100nM デキサメタゾンを用いて、2時間同調化させた後、被験試料群1を0.5mg/mLとなるようにNIH−3T3細胞に添加した。添加後、所定の時間でNIH−3T3細胞から全RNAを抽出した。時計遺伝子(Per2、Cry1、Bmal1、及びClock)の発現を定量的RT−PCRにより調べた。なお、被験試料群1の代わりに0.5%となるようにエタノールを加えたものをコントロールとした。
黒生姜における時計遺伝子調節機能を有する成分を同定するために、黒生姜95%エタノール抽出物を用いて分取HPLCにより分画を行った。HPLC条件は以下のとおりとした。また、分画した画分のうち、時計遺伝子調節機能を有する成分を含有する画分の同定は、FT−MS及び1H−NMRを用いて行った。
Column:YMC−DispoPack AT ODS−25
Eluent:ACN/H2O(55/45)
Flow Rate:15mL/min
[HPLC条件2]
Column:Wakosil−II5C18 RS−Prep,5μm,20x250mm
Eluent:ACN/MeOH/H2O(20/30/35)
Flow Rate:13mL/min
[HPLC条件3]
Column:Wakosil−II 5C18 RS−Prep,5μm,20x250mm
Eluent:ACN/MeOH/H2O(20/30/30)
Flow Rate:13mL/min
(1)遺伝子発現解析
被験試料群1を添加した6時間後に、時計遺伝子の発現をコントロールに対する相対量として調べた結果を図1に示す。黒生姜を被験試料として用いた場合の4時間毎に経時的に相対的mRNA量を比較した結果を図2に示す。黒生姜の最終濃度を0.005、0.01、0.03及び0.05mg/mLに変化させて、黒生姜を添加した6時間後(Per2及びCry1)又は20時間後(Bmal1)に、時計遺伝子の発現をコントロールに対する相対量として調べた結果を図3に示す。
黒生姜95%エタノール抽出物について、HPLC条件1により分画を実施した。結果を図4に示す。図4に示すとおり、Fr−1〜Fr−5までの5つの画分が得られ、これらを乾燥させることにより、それぞれ0.154g、0.537g、0.012g、0.041g及び0.066gの乾燥物を得た。これらの合計回収率は81%であった。
Claims (3)
- 黒生姜(Kaempferia parviflora)を含有する、時計遺伝子発現促進用組成物。
- 前記時計遺伝子が、Per2、Cry1、Bmal1及びClockからなる群から選ばれる少なくとも1種の時計遺伝子である、請求項1に記載の組成物。
- 前記黒生姜が黒生姜抽出物である、請求項1又は2に記載の組成物。
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