JP2019517472A - 認知増強のためのセンシンレンの抽出物 - Google Patents
認知増強のためのセンシンレンの抽出物 Download PDFInfo
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
Description
本発明は、主成分として1〜4重量%の量のアンドログラホリド(AGP)、1〜3重量%の量のネオアンドログラホリド(NAG)、および0.05〜0.2重量%の量の14−デオキシ−11,12−10ジデヒドロアンドログラホリド(DDAG)を含有しているセンシンレン葉の水性抽出物を提供する。該抽出物は、0.2〜0.7重量%のタンパク質、0.01〜0.05重量%の多糖類、および15〜25重量%のグリコサポニンを含有する。
例1:抽出物の調製
センシンレンの葉を収穫し、流れる水道水で完全に洗浄し、そして、その後、3回連続して超高純度の蒸留水との交換に付し、配合機MX−800S(パナソニック)を使用して粉末にする前に、オーブン乾燥器中で、40℃で3日間にわたって乾燥した。粉末にした葉の試料の抽出は、水で1:20(w/v)の比率で行った。
モリス水迷路(MWM)試験は、動物モデルの空間学習および記憶を研究するための、十分に確立された行動タスクである(Morris 1984)。わずかな改変を加えた、Gulinelloら(2009)によって記載された2日間プロトコルを使用して、ラットの認知機能を試験した。簡潔には、AP、EGb、またはビヒクルによる7日間の前処置に続いて、動物を、暗い背景を有する水槽であって、水面より約2cm上の槽の内側の黒い表面とは異なるコントラストを有する見えるプラットホームを含む水槽のプールにおいて、訓練した。訓練は、最初の日の4件の試行から成り、そして、V1〜V4(見えるプラットホームの試行1〜4)またはD1V1〜4(1日目、見えるプラットホームの試行1〜4)に指定される。
製造業者(Cusabio Biotech Co. Ltd, Wuhan, China)によって提供されるプロトコルに従って、市販のELISAキットを使用して、免疫反応性IL−1β、TNF−α、およびIL−6レベルの存在について、脳サイトゾル上清を分析し、結果をpg/mLで表した。
脳切片溶解物中の細胞内ROSの生成を、それ以前にShinomol & Muralidhara (2011)によって記載されたとおりに2,7−ジクロロフルオレシンジアセタート(DCF−DA)を使用して測定した。簡潔には、反応混合物(1mlのLockes緩衝液pH7.4、0.2mlのサイトゾル分画、および10μlの5M DCFH−DA)を、室温で15分間インキュベートした。30分の更なるインキュベーションの後、分光蛍光光度計(484nm励起および530nm発光)を使用して、蛍光生成物DCFの形成を測定した。ROSの生成は、コントロールに対する倍率変化として報告した。
アセチルコリンエステラーゼ(AChE)阻害活性を、Ellmanら(1961)によって開発された原法を改変することによって決定した。96ウェルプレートを使用して、反応混合物(285μlのリン酸緩衝液(0.1M、pH 8.0)および5μl(10mM)のDTNB)を含有している各実験用ウェルに、5μlの試料(脳切片ホモジェネート)のまたはブランク(コントロール)を加えた。続いて、2μlのアセチルチオコリンヨーダイド(基質として使用される)(150mM)を加え、分光光度計を使用して、吸光度の変化を412nmで5分間モニターした。
ブチルコリンエステラーゼ酵素をアッセイするためにブチリルチオコリンヨーダイドを基質として使用した一方で、すべての他の試薬および条件が、上記のアセチルコリンエステラーゼ酵素アッセイについてのものと同じであった。
スーパーオキシドジスムターゼ(SOD)およびカタラーゼ(CAT)アッセイを、製造業者の指示に従ってELISAキット(Cayman Chemical Company, Ann Arbor, MI, USA)を使用して遂行した。VersaMaxマイクロプレートリーダー(Molecular devices, US)を使用して、SODおよびCATについてそれぞれ440および540nmで、光学濃度を読み取った。
水迷路試験は、プラットホームの位置を学習し、続いて記憶する、動物の認知能力の相対的な基準である(Gulinello et al. 2009)。すべての試験されたラットは、一般的に減少した、見えるプラットホームの試行(1日目)においてプラットホームを見つけるまでの潜時を示した。初期の見えるプラットホームの試験は、長期空間記憶を評価するための隠れたプラットホームの試験(24時間遅らせた後)の前に、逃避潜時に関する多数の利点の一部としての泳ぐ速度における、ベースラインの運動機能、馴化速度、ならびに個々のおよび処置群の差の評価を報告した、以前の研究に従う(chulz et al. 2007; Clinton et al. 2007; Galea et al. 2002)。LPSに対するラットの曝露は、MWM試験によって評価されたように空間学習および記憶における変性を誘導する(図1)。
すべての上記の知見より、標準化水性AP抽出物が、LPSによって誘発された神経炎症から脳を保護し、記憶を増強すると推論される。
Claims (9)
- 1〜4重量%の量のアンドログラホリド、1〜3重量%の量のネオアンドログラホリド、および0.05〜0.2重量%の量の14−デオキシ−11,12−5ジデヒドロアンドログラホリドを含有しているセンシンレン葉の水性抽出物であって、0.2〜0.7重量%のタンパク質、0.01〜0.05重量%の多糖類、および15〜25重量%のグリコサポニンを含有する、前記抽出物。
- 以下の工程:
−センシンレンの葉を乾燥させること、および
−少なくとも50℃の、好ましくは55〜70℃の温度で、水でそれらを抽出すること
を含むことを特徴とする、請求項1に記載の抽出物の生産方法。 - 抽出が、いかなる有機溶媒の添加もしないでなされる、請求項2に記載の方法。
- 葉を乾燥させる工程が、少なくとも30℃、好ましくは40〜70℃の温度で、オーブン乾燥器中で遂行され、その後に粉砕することを伴う、請求項2または3に記載の方法。
- 液体の水性抽出物が、固体の形の抽出物を得るために凍結乾燥に付される、請求項2〜4のいずれか一項に記載の方法。
- 抗神経炎症および抗酸化効果を有する認知増強剤としての使用のための、請求項1に記載のセンシンレン葉の水性抽出物。
- 使用が、記憶喪失、ストレス、痴呆またはアルツハイマー疾患などの認知障害の処置、改善、または予防のためである、請求項6に記載の使用のためのセンシンレン葉の水性抽出物。
- 請求項1に記載の抽出物、および、充填剤、結合剤または溶媒などの担体として作用する少なくとも1つの補助物質を含有することを特徴とする、認知障害の処置、改善または予防のための組成物、特に食品補助剤。
- 経口製剤、好ましくはタブレットまたはカプセルの形である、請求項8に記載の組成物。
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JP2007535542A (ja) * | 2004-04-28 | 2007-12-06 | ハッチソン メディファーマ エンタープライジズ リミテッド | センシンレン(Andrographispaniculata)由来の粗抽出物 |
JP2013517234A (ja) * | 2010-01-12 | 2013-05-16 | インデナ エッセ ピ ア | センシンレン(Andrographicspaniculata)抽出物と、リン脂質と複合体を形成した銀杏抽出物とを含む組成物 |
WO2014176149A1 (en) * | 2013-04-22 | 2014-10-30 | Innobioscience, Llc | Treatment of alzheimer's and cognitive impairment with andrographolides |
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JP2007535542A (ja) * | 2004-04-28 | 2007-12-06 | ハッチソン メディファーマ エンタープライジズ リミテッド | センシンレン(Andrographispaniculata)由来の粗抽出物 |
JP2013517234A (ja) * | 2010-01-12 | 2013-05-16 | インデナ エッセ ピ ア | センシンレン(Andrographicspaniculata)抽出物と、リン脂質と複合体を形成した銀杏抽出物とを含む組成物 |
WO2014176149A1 (en) * | 2013-04-22 | 2014-10-30 | Innobioscience, Llc | Treatment of alzheimer's and cognitive impairment with andrographolides |
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AM J CANCER RES., vol. 5(12), JPN6021032817, 2015, pages 3570 - 3587, ISSN: 0004577251 * |
JOURNAL OF COMMUNITY MEDICINE(社区医学雑誌), vol. 12(2), JPN6021032819, 2014, pages 1 - 3, ISSN: 0004577253 * |
JOURNAL OF FOOD PROTECTION, vol. 75(8), JPN6021032816, 2012, pages 1507 - 1511, ISSN: 0004577250 * |
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THE FASEB JOURNAL, EXPERIMENTAL BIOLOGY 2014 MEETING ABSTRACTS, vol. 28(S1), JPN6021032820, 2014, pages 1143 - 16, ISSN: 0004577254 * |
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