JP2020186215A - 更年期症状改善用組成物 - Google Patents
更年期症状改善用組成物 Download PDFInfo
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Abstract
Description
[1]カシス及びブルーベリー並びにそれらの抽出物からなる群から選ばれる少なくとも1種の有効成分を含有し、かつ、コラーゲン、エラスチン及びヒアルロン酸の減少による皮膚劣化以外の更年期症状を改善する作用を有する、更年期症状改善用組成物。
[2]前記更年期症状は、薄毛、血管硬化、体重増加及び脂肪増加からなる群から選ばれる少なくとも1種の更年期症状である、[1]に記載の組成物。
[3]前記有効成分は、カシス及びカシス抽出物からなる群から選ばれる少なくとも1種の有効成分である、[1]〜[2]のいずれか1項に記載の組成物。
[4]前記組成物は、経口用の組成物である、[1]〜[3]のいずれか1項に記載の組成物。
[5]カシス及びブルーベリー並びにそれらの抽出物からなる群から選ばれる少なくとも1種の有効成分を含有する、増毛用組成物、血管拡張用組成物、血管硬化改善用組成物、体重増加抑制用組成物、脂肪増加抑制用組成物又は抗肝炎用組成物。
[6]前記組成物は、成人女性に摂取されるための組成物である、[5]に記載の組成物。
[7][1]〜[6]のいずれか1項に記載の組成物を、更年期症状の雌性個体に投与することにより、該雌性個体の更年期症状を改善する工程を含む、更年期症状の改善方法。
[8][1]〜[6]のいずれか1項に記載の組成物を、成人女性に投与することにより、該成人女性に対して増毛する方法、血管を拡張する方法、血管を柔軟化する方法、体重を減少する方法、脂肪を減少する方法又は肝炎を減少する方法。
[9][1]〜[6]のいずれか1項に記載の組成物を製造するための、カシス及びブルーベリー並びにそれらの抽出物からなる群から選ばれる少なくとも1種の有効成分の使用。
(1−1)カシス抽出物及びブルーベリー果実粉末
カシス抽出物(BCE)として、市販のBCE粉末である「CaNZac−35」(光洋商会社)を用いた。BCEには、ポリフェノールが37.6g/100g−BCE、アントシアニンが38.0g/100g−BCEで含まれていた。また、BCEには、アントシアニンC3G(5.6質量%)、C3R(32.0質量%)、D3G(16.8質量%)及びD3R(45.3質量%)が含まれていた。
ヒト毛乳頭細胞(Hair follicle dermal papilla cells;HFDPC)はプロモセル社から購入した。HFDPCは、毛乳頭細胞増殖用培地(プロモセル社)を用いて維持した。細胞培養は、5%CO2を含む加湿インキュベーター内で37℃にて実施した。
培地には毛乳頭細胞基本培地(フェノールレッド不含;PromoCell社)を使用した。1μg/ml BCEを含有する被験培地及び毛乳頭細胞基本培地のみであるコントロール培地を用いて、HFDPCを24時間培養した後に細胞を回収し、次いで回収した細胞をPBSで2度洗浄した。洗浄後の細胞から「RNeasy mini kit」(キアゲン社)を用いて全RNAを抽出した。
マイクロアレイで変動が大きかった毛乳頭幹細胞マーカーであるkeratin 19(KRT19)の発現量を調べるために、ヒト毛乳頭細胞から抽出したRNAを用いて、「TB Green Premix Ex Taq II(登録商標) Tli RNaseH Plus」(TaKaRa社)によりリアルタイムPCRを行った。
更年期モデル動物として12週齢の卵巣除去ラット(「OVX female Sprague−Dawley」;クレアジャパン社)を用い、偽手術動物として12週齢のShamラット(「Sprague−Dawley」;クレアジャパン社)を用いた。卵巣除去ラット6匹を2群に分け、一方の3匹には「AIN−93M」(クレアジャパン社)のみからなる普通餌を摂取させ(「OVX control」群)、他方の3匹には3質量% BCEを含有する「AIN−93M」である被験餌を摂取させた(「OVX BCE」群)。また、手術行為による影響を排除するために、麻酔及び皮膚切開のみを行い、卵巣を除去していない偽手術ラットの3匹に、普通餌を摂取させた(「Sham」群)。各群のラットは餌及び水を自由に摂取できるようにした。
マイクロアレイ及びIPAによって解析して予測した、BCEを通じて発現が変動した遺伝子の上流因子を表1に示す。
(2−1)卵巣除去ラットを用いた体重増加抑制作用及び血中中性脂肪上昇抑制作用の評価
11週齢の卵巣除去ラット及びShamラットを用いた以外は、上記(1−5)と同様に各ラットを群分けし、それぞれの餌を用いて3ヵ月間飼育した。各群のラットについて、飼育前後の体重を測定した。
肝炎関連遺伝子の発現解析を行うために、肝臓から抽出したmRNAを用いて上記(1−4)と同様にしてリアルタイムPCRを行った。肝炎関連遺伝子としてはTNFα及びIL−6を対象とした。
3ヵ月間の飼育前後の体重の変化を図7に示す。餌の摂取前では体重に個体差は無かった。しかし、3ヵ月間の餌の摂取後では、OVX control群に対して、OVX BCE群及びSham群の体重は統計的に有意に減少した(p<0.05)。また、OVX BCE群の体重は、Sham群と大差がなかった。
(3−1)マイクロアレイ及びIPAによる遺伝子解析
細胞として、ヒト血管内皮細胞であるHUVEC(プロモセル社)を使用した以外は、上記(1−3)と同様にしてマイクロアレイ分析を実施した。また、1.5倍以上の発現上昇を示した遺伝子を対象とした以外は、上記(1−3)と同様にしてIPAソフトウェアを用いて分析した。
内皮一酸化窒素合成酵素(eNOS) mRNA発現を、RT−qPCR分析により評価した。HUVECと同じくヒト血管内皮細胞であるEA.hy926細胞(ATCC)を用いた。HUVEC及びEA.hy926細胞を12ウェル培養プレートに播種し、コンフルエントになるまで培養した。培地を交換し、次いで細胞をアントシアニン(10μM)、BCE(0.5又は1.0μg/mL)又は17β−エストラジオール(E2)(10nM)の存在下又は非存在下で24時間処理した。フルベストラントの実験系では、100nM フルベストラントの存在下又は非存在下で24時間インキュベートした後、処理した後の細胞をさらにアントシアニン及びBCEで24時間インキュベートした。PBSで2回洗浄した後「RNeasy mini kit」を用いて全RNAを抽出した。
EA.hy926細胞を12ウェル培養プレートに播種し、コンフルエントになるまで培養した。培地を交換し、次いで細胞を100nM フルベストラントの存在下又は非存在下で24時間インキュベートした後アントシアニン(10μM)、BCE(0.5又は1.0μg/mL)若しくはE2(10nM)の存在下又は非存在下で細胞をさらに5日間インキュベートした後、培地を回収し、SpeedVac濃縮装置(「SPD1010」;サーモフィッシャーサイエンティフィック社)で濃縮した。
上記(1−5)と同様に各ラットを群分けし、それぞれの餌を用いて3ヵ月間飼育し、次いで腹腔動脈組織を摘出し、組織切片を作製した。
測定結果は、少なくとも3回の独立した実験の平均±標準誤差(SEM)として表した。統計分析は、「ベルカーブ(BellCurve)エクセル バージョン2.13 ソフトウェア」(社会情報サービス社)を使用して実施し、スチールポストホックテスト;p<0.05を有するクラスカル−ウォリス分析は統計的有意性を示すとした。
マイクロアレイ及びIPAにより、BCEを通じて発現が変動した、血管系に関連する遺伝子の上流因子を表3に示す。
Claims (6)
- カシス及びブルーベリー並びにそれらの抽出物からなる群から選ばれる少なくとも1種の有効成分を含有し、かつ、コラーゲン、エラスチン及びヒアルロン酸の減少による皮膚劣化以外の更年期症状を改善する作用を有する、更年期症状改善用組成物。
- 前記更年期症状は、薄毛、血管硬化、体重増加及び脂肪増加からなる群から選ばれる少なくとも1種の更年期症状である、請求項1に記載の組成物。
- 前記有効成分は、カシス及びカシス抽出物からなる群から選ばれる少なくとも1種の有効成分である、請求項1〜2のいずれか1項に記載の組成物。
- 前記組成物は、経口用の組成物である、請求項1〜3のいずれか1項に記載の組成物。
- カシス及びブルーベリー並びにそれらの抽出物からなる群から選ばれる少なくとも1種の有効成分を含有する、増毛用組成物、血管拡張用組成物、血管硬化改善用組成物、体重増加抑制用組成物、脂肪増加抑制用組成物又は抗肝炎用組成物。
- 前記組成物は、成人女性に摂取されるための組成物である、請求項5に記載の組成物。
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