JP6161953B2 - 育毛剤、筋肉活性化剤及びエネルギー産生促進剤 - Google Patents
育毛剤、筋肉活性化剤及びエネルギー産生促進剤 Download PDFInfo
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- 108700004921 tetramethylrhodaminylphalloidine Proteins 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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- 239000001993 wax Substances 0.000 description 1
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Description
クロバナツルアズキの地上部5gに精製水50mLを加え、95〜100℃で1時間抽出した後、濾過し、その濾液を濃縮し、凍結乾燥してクロバナツルアズキ(地上部)の熱水抽出物を1.31g得た。
クロバナツルアズキの地上部5gに50(v/v)%エタノール50mLを加え、常温で5日間抽出した後、濾過し、その濾液を濃縮乾固して、クロバナツルアズキ(地上部)の50%エタノール抽出物を1.09g得た。
クロバナツルアズキの地上部5gにエタノール50mLを加え、常温で5日間抽出した後、濾過し、その濾液を濃縮乾固して、クロバナツルアズキ(地上部)のエタノール抽出物を0.99g得た。
処方 配合量
1.エタノール 60.0重量%
2.クロバナツルアズキ抽出液BG(丸善製薬) 2.0
3.グリセリン 2.0
4.精製水にて全量を100とする
[製造方法]成分2を成分1に溶解し、成分3及び成分4を加え、十分攪拌混合して製品とする。
処方例1において、クロバナツルアズキ抽出液BG(丸善製薬)を精製水に置き換えたものを、従来のヘアトニックとした。
処方 配合量
1.クロバナツルアズキ(地上部)の50%エタノール抽出物 0.2重量%
2.ステアリン酸 5.0
3.セチルアルコール 5.0
4.流動パラフィン 2.0
5.グリセリンモノステアレート 1.3
6.ソルビタンモノオレエート 1.5
7.ポリオキシエチレン(10)ソルビタンモノオレエート 0.8
8.グリセリン 6.0
9.防腐剤 適量
10.精製水にて全量を100とする
[製造方法]成分2〜7を加熱溶解して混合し、70℃に保ち油相とする。成分1及び8〜10を加熱溶解して混合し、75℃に保ち水相とする。油相に水相を加え、かき混ぜながら冷却して製品とする。
処方 配合量
1.クロバナツルアズキ(地上部)の熱水抽出物 0.1重量%
2.アルキル硫酸トリエタノールアミン 18.0
3.ラウリン酸ジエタノールアミド 3.0
4.メチルセルロース 0.5
5.香料 適量
6.精製水にて全量を100とする
[製造方法]成分6に成分4を均一に溶解した後、成分1及び成分2を加える。70〜75℃で加熱溶解した後、成分3を加える。冷却途中に成分5を加え、30℃まで冷却して製品とする。
処方 配合量
1.クロバナツルアズキ(地上部)の熱水抽出物 0.1重量%
2.ホホバ油 0.01
3.ベヘニルアルコール 3.0
4.1,3−ブチレングリコール 5.0
5.塩化ジステアリルジメチルアンモニウム(75%) 8.0
6.クエン酸 0.05
7.香料 適量
8.精製水にて全量を100とする
[製造方法]成分1〜8を60℃で溶解し、攪拌して30℃まで冷却して製品とする。
処方 配合量
1.クロバナツルアズキ(地上部)のエタノール抽出物 0.1重量%
2.カルボキシビニルポリマー 1.0
3.ヒドロキシエチルセルロース 0.5
4.1,3−ブチレングリコール 5.0
5.水酸化カリウム 0.5
6.95エタノール 5.0
7.エデト酸三ナトリウム 0.02
8.メチルパラベン 0.2
9.香料 適量
10.精製水にて全量を100とする
[製造方法]成分1〜4及び6〜10を均一混合した後、成分5を添加して製品とする。
処方 配合量
1.クロバナツルアズキ(地上部)のエタノール抽出物 0.5重量%
2.スクワラン 5.0
3.オリーブ油 5.0
4.ホホバ油 5.0
5.セタノール 1.5
6.モノステアリン酸グリセリン 2.0
7.ポリオキシエチレンセチルエーテル(20E.O.) 3.0
8.ポリオキシエチレンソルビタンモノオレエート(20E.O.) 2.0
9.香料 適量
10.グリセリン 2.0
11.パラオキシ安息香酸メチル 0.2
12.プロピレングリコール 1.0
13.精製水にて全量を100とする
[製造方法]成分2〜8を加熱溶解して混合し、70℃に保ち油相とする。成分1及び10〜13を加熱溶解して混合し、75℃に保ち水相とする。油相に水相を加えて乳化し、かき混ぜながら冷却し、45℃で成分9を加え、30℃まで冷却して製品とする。
in vitro系における加齢モデルであるPDL(=population doubling level、細胞の分裂回数)の異なるヒト皮膚線維芽細胞(NB1RGB、PDL=29:若い、PDL=44:中程度及びPDL=65:老いた)から、RNAiso plus(TAKARA)を用いて総RNAの抽出を行った。総RNAを基に、リアルタイムRT−PCR法により、IGF−1mRNA発現量の測定を行った。同時に、内部標準として、β−アクチンmRNA発現量の測定を行った。尚、各遺伝子の発現量の測定に使用したプライマーは次の通りである。
AAGGAGGCTGGAGATGTATTGC(配列番号1)
CGGACAGAGCGAGCTGACTT(配列番号2)
β−アクチン用のプライマーセット
CACTCTTCCAGCCTTCCTTCC(配列番号3)
GTGTTGGCGTACAGGTCTTTG(配列番号4)
コンフルエントになったヒト皮膚線維芽細胞(NB1RGB)にクロバナツルアズキ抽出液BG(丸善製薬)を加え、FBSを含まないDMEM培地にて24時間培養した後、RNAiso plus(TAKARA)を用いて総RNAの抽出を行った。尚、添加濃度は30、100及び300μg/mLにて行った。総RNAを基に、リアルタイムRT−PCR法により、IGF−1mRNA発現量の測定を行った。同時に、内部標準として、β−アクチンmRNA発現量の測定を行った。尚、各遺伝子の発現量の測定に使用したプライマーは実験例1の通りである。
処方例1及び比較処方例1で得られる各ヘアトニックを用いて、使用試験を行った。男性型脱毛症患者である被験者20名の頭部に毎朝晩2回、連続6ヶ月間試料を塗布した後、毛髪の増加及び剛毛化を基準に育毛効果を評価した。
筋肉由来細胞株であるC2C12を96wellプレートに5×103個/well播種し、10%FBSを含むDMEM培地にて37℃、5%CO2条件下で培養した。72時間後、コンフルエントな状態になったところでクロバナツルアズキ抽出物BG(丸善製薬)1及び10μg/mLを添加したFBSを含まないDMEM培地にて更に24時間培養した。その後、MTT assayにより、ミトコンドリア内の脱水素酵素の活性を測定した。即ち、500μg/mLのMTT(3−[4,5−dimethylthiazol−2−yl]−2,5−diphenyl−tetrazolium bromide)(Sigma)を含むDMEM培地200μLにて2時間培養した後、培養上清を除去して生成されたフォルマザンの黒色結晶をイソプロパノール200μLに溶解し、1時間後に570nm及び630nmにおける吸光度を測定し、1wellあたりのフォルマザン生成量にて脱水素酵素の活性を評価した。
筋肉由来細胞株であるC2C12を、Cellmatrix I−C(新田ゼラチン)にてコラーゲンコートした12wellプレートに3×104個/well播種した。24時間培養した後、FBSを含まないDMEM培地に置換し、10μg/mLのクロバナツルアズキ抽出物BG(丸善製薬)を添加した。一日おきに培地交換を行い、10日程かけて筋管形成を誘導させた。細胞を固定した後、TRITC−phalloidinにてF−アクチンを染色し、蛍光顕微鏡にて観察した。
Claims (3)
- クロバナツルアズキの抽出物を有効成分として含有することを特徴とするインスリン様増殖因子1(IGF−1)産生促進剤。
- クロバナツルアズキの抽出物を有効成分として含有することを特徴とする筋肉活性化剤。
- クロバナツルアズキの抽出物を有効成分として含有することを特徴とする筋肉細胞におけるエネルギー産生促進剤。
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