JP2019202969A - 骨格筋強化剤 - Google Patents
骨格筋強化剤 Download PDFInfo
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Abstract
Description
(植物抽出物の調整)
乾燥植物を市販のミキサーによって均一に細かくしたもの1gを焙煎後、10mlのヘキサンを加え、2日間静置して抽出後、濾紙(ADVANTEC No.2)を用いて濾過した。ヘキサン抽出残渣に1gにつき10mlのメタノールを加え、3日間静置して抽出後、濾紙(ADVANTEC No.2)を用いて濾過し、メタノール抽出画分を取得した。得られたメタノール抽出画分をロータリーエバポレーターにより蒸発乾固させたものを300mlの酢酸エチル、及び300mlの超純水にて分配濾過し、それぞれ酢酸エチル抽出画分(上層)、水抽出画分(下層)とした。酢酸エチル画分をロータリーエバポレーターにより蒸発乾固後、10mgあたり1mlのメタノールに溶解し、植物抽出物とした。
DMEM培地(10%FBS,(+P/S))を用いて、飽和状態の9〜10割に達したC2C12細胞を上記の植物抽出物(終濃度10μg/ml)を含むDMEM培地(2%HS,(P/S))(1g/L DMEM,2%HS,100units/ml penicillin G sodium,100μg/ml streptomycin,1g/l NaHCO3)で培養し、48時間毎に新鮮な培地に培地交換することにより分化誘導を行った。分化誘導4日目に細胞を回収し、ウエスタンブロット法によってMyHC発現レベルを評価した。この結果を図1に示した。図1から分かるように、当該植物抽出物は骨格筋分化誘導作用を有することが確認された。
次に、実施例1で得られた植物抽出物の酢酸エチル画分についてGC-MS分析を行ったところ、ステアラミド、パルミトアミド、オレアミド、ミリスチラミドが同定された(図2参照)。そして、同定されたステアラミド及びオレアラミドについて、市販品を用いて骨格筋の強化作用について調べた。
ステアラミド及びオレアラミドの分化誘導能を前記方法に従って調べた。その結果を図3に示した。ステアラミド及びオレアラミドのいずれも終濃度0.1μMで対照に比べて有意な差で分化誘導能を示した。
飽和状態の90〜100%に達したC2C12細胞を前記DMEM培地で分化誘導し、分化6日目の筋管細胞にオレアミドを2日間曝露した。曝露は、DMEM培地中に終濃度が0.1μMとなるようにオレアミドを添加し、37℃、5%CO2濃度下で培養した。その後、筋管細胞を蛍光免疫染色によりMyHCと核を染色した。核が2個以上存在する筋管細胞の短径を測定し、対照(vehicle)との相対比を求めた。その結果を図4に示した。
行動範囲を制限したマウスに(不活動マウス)対するオレアミドの影響を調べた。飼育ゲージの飼育面積を1/6に区画することでマウスの行動範囲を制限した。この環境下で飼育したマウスでは、種々の筋量及び筋力が低下することが事前に分かっている。そこで、オレアミドが不活動マウスの筋量や筋力に与える影響を調べた。
Claims (12)
- 化学式1に示される脂肪酸アミドからなる骨格筋強化剤。
- 前記骨格筋強化剤は、筋分化促進、骨格筋の筋量増加、骨格筋の筋量低下抑制、骨格筋の筋肥大の少なくとも何れか一つの作用を有する請求項1に記載の骨格筋強化剤。
- 化学式1に示される脂肪酸アミドを含む骨格筋強化のための組成物。
- 化学式1に示される脂肪酸アミドを含む天然物及び/又は化学式1に示される脂肪酸アミドを含む天然物の抽出物を含む骨格筋強化のための組成物。
- 前記天然物は、セリ科植物、ミル科植物、クロウメモドキ科植物、スベリヒユ科植物、キク科植物の少なくとも何れか1種以上の植物である請求項4に記載の組成物。
- 前記天然物は、セリ科オランダミツバ属の植物、セリ科ミツバ属の植物、ミル科ミル属の植物、クロウメモドキ科ナツメ属、スベリヒユ科スベリヒユ属の植物、キク科ゴボウ属の少なくとも何れか1種以上の植物である請求項4に記載の組成物。
- 前記組成物は、医薬組成物、食品組成物、化粧用組成物又は動物用飼料の何れかである請求項3〜6の何れか1項に記載の組成物。
- 前記組成物は、筋分化促進、骨格筋の筋量増加、骨格筋の筋量低下抑制、骨格筋の筋肥大の少なくとも何れか一つの作用を有する請求項3〜7の何れか1項に記載の骨格筋強化剤。
- 骨格筋強化剤として化学式1に示される脂肪酸アミドを含む医薬組成物、食品組成物、飲料、化粧用組成物又は動物用飼料。
- 化学式1に示される脂肪酸アミド、化学式1に示される脂肪酸アミドを含む天然物、当該天然物の抽出物の、骨格筋強化用組成物を製造するための使用。
- 前記骨格筋強化剤は、筋分化促進、骨格筋の筋量増加、骨格筋の筋量低下抑制、骨格筋の筋肥大の少なくとも何れか一つの作用を有する請求項10記載の使用。
- セリ科オランダミツバ属の植物、セリ科ミツバ属の植物、ミル科ミル属の植物、クロウメモドキ科ナツメ属の植物、スベリヒユ科スベリヒユ属の植物、キク科ゴボウ属の植物の何れか1種以上の植物体及び/又はそれら植物体の抽出物を有効成分とする骨格筋強化剤。
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