JP2017128520A - ルミカン産生促進剤およびそれを用いた皮膚外用剤、飲食品、並びに美容方法。 - Google Patents
ルミカン産生促進剤およびそれを用いた皮膚外用剤、飲食品、並びに美容方法。 Download PDFInfo
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Abstract
Description
検出器:紫外吸光光度計(214nm)
カラム:Superdex Peptide HR10/30(GEヘルスケア・ジャパン社製)
カラム温度:25℃
移動相:25mM トリス塩酸バッファー(pH7.5)、150mM NaCl
流量:0.5mL/min
流入量:50μL
分析周期:60min
スタンダード:Gel Filtration Calibration Kit LMW(低分子用、GEヘルスケア・ジャパン社製)、ヘキサグリシン(分子量360)、トリグリシン(分子量189)およびグリシン(分子量75)
ゲルろ過クロマトグラフィーによって測定される分子量特性が、後記の表1に示されるように異なる加水分解卵殻膜を準備し、それぞれを、精製水を用いて濃度10mg/mlの水溶液とした。これらのサンプルに対し、そのルミカン産生促進作用、MMP−1産生促進作用、I型コラーゲン産生促進作用、Endo180産生促進作用の有無を評価するために、以下の手順にしたがって各物質の産生量を測定した。そして、測定結果から、◎…顕著な作用を有する、〇…顕著とまではいかないが良好な作用を有する、△…作用がない、×…促進を阻害する、の4段階で評価を行い、それらの結果を、後記の表1に併せて示す。
ヒト線維芽細胞NHDF(NB)(クラボウ社製)をセミコンフルエントまで培養し、継代操作にて12ウェルプレートに1.0×105cells/wellとなるように播種し、72時間培養した。そして、血清を含まないダルベッコ改変イーグル培地(DMEM培地)にて調製した試験培地へ交換し、3日間培養した後、その培地を回収し、回収した培地中に含まれるルミカン量を、Lumican ELISA Kit,Human(UCSN LifeScience社製)あるいはLumican ELISA Kit,Human(Cusabio社製)を用いたELISA法にて測定した。試験はn=3で実施した。
ルミカン測定方法のELISA Kitを「Human Total MMP−1 DuoSet ELISA Kit(R&D Systems社製)」に代えて、同様の測定を行った。試験はn=3で実施した。
ルミカン測定方法のELISA Kitを「Pro−Collagen I alpha1 Duoset ELISA Kit(R&D Systems社製)」に代えて、同様の測定を行った。試験はn=3で実施した。
ヒト線維芽細胞NHDF(NB)(クラボウ社製)をセミコンフルエントまで培養し、継代操作にて12ウェルプレートに1.0×105cells/wellとなるように播種し、72時間培養した。そして、血清を含まないDMEM培地にて調製した試験培地へ交換し、3日間培養した後、培地を除き、リン酸緩衝食塩水(PBS)にて細胞を洗浄した。その後、M−PER(登録商標):Mammalian Protein Extraction Reagent(Thermo Fisher Scientific社製)に、終濃度1%(v/v)となるようにProtease inhibitor cocktail(Thermo Fisher Scientific社製)を配合し混合した液を200μl/well添加し、セルスクレーパー(ビーエム機器社製)等で細胞懸濁液を回収した。回収した細胞懸濁液に含まれるEndo180量を、C−type mannose receptor2 ELISA Kit(Cusabio社製)を用いたELISA法にて測定した。試験はn=3で実施した。
Claims (6)
- ゲルろ過クロマトグラフィーによって測定される分子量の最大ピーク値が150〜5500の範囲にある加水分解卵殻膜を含有することを特徴とするルミカン産生促進剤。
- 上記加水分解卵殻膜が、ゲルろ過クロマトグラフィーによって測定される分子量が10000以上の分画に1〜30%含まれるものである請求項1記載のルミカン産生促進剤。
- 上記加水分解卵殻膜の、ゲルろ過クロマトグラフィーによって測定される分子量100〜5500の分画に含まれるもののうち、その70〜100%が分子量3000以下の分画に含まれるものである請求項1または2記載のルミカン産生促進剤。
- 請求項1〜3のいずれか一項に記載のルミカン産生促進剤を含有することを特徴とする皮膚外用剤。
- 請求項1〜3のいずれか一項に記載のルミカン産生促進剤を含有することを特徴とする飲食品。
- 請求項5記載の飲食品を経口摂取させることを特徴とする美容方法。
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Citations (7)
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JPH0940564A (ja) * | 1995-07-26 | 1997-02-10 | Taiyo Kagaku Co Ltd | 悪酔予防改善用組成物 |
JP2001245634A (ja) * | 2000-03-03 | 2001-09-11 | Q P Corp | 卵製品 |
JP2004175684A (ja) * | 2002-11-25 | 2004-06-24 | Seiwa Kasei:Kk | 化粧料 |
JP2008118887A (ja) * | 2006-11-10 | 2008-05-29 | Q P Corp | 飲食品及び苦味マスキング剤 |
JP2012520340A (ja) * | 2009-03-16 | 2012-09-06 | ディーエスエム アイピー アセッツ ビー.ブイ. | トリペプチドの使用 |
JP2013135851A (ja) * | 2004-10-11 | 2013-07-11 | Sonovia Holdings Llc | 皮膚状態を処置するための装置 |
JP2014040402A (ja) * | 2012-08-24 | 2014-03-06 | Pharma Foods International Co Ltd | ヒアルロン酸産生促進剤 |
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Patent Citations (7)
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JPH0940564A (ja) * | 1995-07-26 | 1997-02-10 | Taiyo Kagaku Co Ltd | 悪酔予防改善用組成物 |
JP2001245634A (ja) * | 2000-03-03 | 2001-09-11 | Q P Corp | 卵製品 |
JP2004175684A (ja) * | 2002-11-25 | 2004-06-24 | Seiwa Kasei:Kk | 化粧料 |
JP2013135851A (ja) * | 2004-10-11 | 2013-07-11 | Sonovia Holdings Llc | 皮膚状態を処置するための装置 |
JP2008118887A (ja) * | 2006-11-10 | 2008-05-29 | Q P Corp | 飲食品及び苦味マスキング剤 |
JP2012520340A (ja) * | 2009-03-16 | 2012-09-06 | ディーエスエム アイピー アセッツ ビー.ブイ. | トリペプチドの使用 |
JP2014040402A (ja) * | 2012-08-24 | 2014-03-06 | Pharma Foods International Co Ltd | ヒアルロン酸産生促進剤 |
Non-Patent Citations (1)
Title |
---|
HUANG XI, ET AL.: "Chemiluminescence Evaluation of Antioxidant Activity and Prevention of DNA Damage Effect of Peptides", JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol. 58, no. 23, JPN6020012539, 3 November 2010 (2010-11-03), pages 12137 - 12142, ISSN: 0004247368 * |
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