JP5156298B2 - 筋肉細胞糖輸送促進作用組成物 - Google Patents
筋肉細胞糖輸送促進作用組成物 Download PDFInfo
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- JP5156298B2 JP5156298B2 JP2007218519A JP2007218519A JP5156298B2 JP 5156298 B2 JP5156298 B2 JP 5156298B2 JP 2007218519 A JP2007218519 A JP 2007218519A JP 2007218519 A JP2007218519 A JP 2007218519A JP 5156298 B2 JP5156298 B2 JP 5156298B2
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- cardamonin
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Description
食後、吸収された糖が門脈に入ると、膵臓から瞬時にインスリンの追加分泌が起こり、肝臓は糖の放出を抑え、逆に糖の取り込みを促進するようにその働きが切り替わる。
一方、食べ過ぎや運動不足でカロリー摂取過剰になるとエネルギーバランスがくずれ、肥満に伴い脂肪細胞の大きさもしくは数が増大し、また肝細胞や骨格筋で中性脂肪の過剰な蓄積が起こり、骨格筋の血糖取り込みを亢進させるインスリンの作用が低下するインスリン抵抗性の状態となる。
肝臓にインスリン抵抗性がある場合、食後、肝臓に糖が流入しても、肝臓への糖の取り込みはうまくいかず、また肝臓から糖の放出量も減らないため、血液中に流れ込む糖の量は増える一方となり、さらに、糖を利用する側の骨格筋にもインスリン抵抗性が存在すれば、糖がうまく骨格筋に取り込まれず、食後、急激に上昇した血糖値がなかなか下がらない状態に陥る。
この状態が長期間続くと、膵臓はインスリン分泌をさらに増やして血液中の糖の取り込みを促し、その結果、膵臓にかかる負担が限度を超えた場合、インスリン分泌機能が急激に衰え、糖尿病となる。
しかしながら、カルダモニンの骨格筋における作用は知られていない。
この糖尿病の予防または治療効果は、筋肉細胞にカルダモニンが作用する結果、筋肉内に直接働きかけて糖輸送を促進する作用をベースとしており、食事により上昇した血糖値を低下させることができ、また空腹時の血糖値を低下させることもできる。従って、本発明は、カルダモニンを含有することを特徴とする筋肉細胞糖輸送促進作用による血糖降下剤または、糖尿病の予防もしくは治療剤でもある。
また、インスリン依存的にグルコース取り込みを促進させる組織としては、脂肪と骨格筋が挙げられるが、脂肪細胞の糖取り込みを促進させると脂肪細胞は肥大し、インスリン抵抗性の原因となるアディポサイトカインの分泌不全および亢進につながるため、糖尿病の治療に関し本質的な解決にならない。しかし骨格筋は体内に吸収されたグルコースの85%を取り込むことが報告されていることから血糖値を低下させる上で最も重要な臓器であることが知られており、脂肪細胞のように悪影響は知られていない。よって、筋肉細胞の糖輸送を促進させる方法は食事摂取とは関係のない血糖の降下、糖尿病の予防または治療に関して最も有効な方法であるといえる。本発明者らは、これらの知見に基づいてさらに研究を進め、本発明を完成するに至った。
[1] カルダモニンを有効成分として含有することを特徴とする筋肉細胞糖輸送促進作用組成物、
[2] 医薬である前記[1]記載の組成物、
[3] 血糖降下剤である前記[2]記載の組成物、
[4] 糖尿病の予防または治療剤である前記[2]記載の組成物、
[5] 食品である前記[1]記載の組成物、および
[6] 食品素材である前記[1]記載の組成物
に関する。
本発明の組成物へのカルダモニンの配合量は、例えば、組成物を飲料等として用いる場合は、カルダモニンが約0.00001〜10重量%、好ましくは約0.0001〜1重量%含まれるように設定するのが望ましい。また、組成物が食品や食品素材または医薬品等として、固形状、粉末状、顆粒状、ペースト状等の形態で経口投与用として用いられる場合は、カルダモニンを約0.01重量%以上含有させればよく、上限は100重量%、すなわち、カルダモニンを単独で使用してもよい。また、組成物を非経口投与用として用いる場合は、カルダモニンを約0.00001〜10重量%、好ましくは約0.0001〜1重量%含まれるように設定するのが望ましい。なお、本明細書において摂取という用語には投与も含められるものとする。
ソウズク乾燥種子25gからメタノール400mlで抽出して抽出物4.5gを得た。抽出物を酢酸エチル/水で分配し酢酸エチル層を濃縮し乾固した(収量2.5g)。次に移動相を酢酸エチルとするシリカゲルカラムクロマトグラフィーで分画した後(収量1.5g)、さらにクロロホルムを移動相とするシリカゲルカラムクロマトグラフィーで分画し、カルダモニンを含む画分617mgを得た。これを逆相カラムクロマトグラフィー(ODS、50μm、120A、30×200mm、メタノール/水(70:30、v/v)、8ml/min)で分画し、カルダモニン170.5mgを得た。
(L6骨格筋細胞の作製)
L6筋芽細胞を2.5×104/mlの濃度になるようにα−MEM培地(10%v/v 牛胎児血清含有)で調整し、コラーゲンタイプIをコートした96穴組織培養プレートに1穴当り200μlを播種し、37℃の5%炭酸ガス培養器で2日間培養、コンフルエントになった後、培地をα−MEM培地(2%v/v 牛胎児血清含有)に変更し、37℃の5%炭酸ガス培養器で5日間培養することによりL6骨格筋細胞を作製した。
カルダモニンをジメチルスルホキシドに溶解させ、α−MEM培地(2%v/v 牛胎児血清含有)にて100倍希釈してカルダモニンをそれぞれ0,10,37μM含有するα−MEM培地を作製した。上記L6骨格筋細胞の培養プレートに、前記カルダモニンを含有するα−MEM培地をそれぞれ1穴当り170μl添加し、37℃の5%炭酸ガス培養器で4時間インキュベートした。ついで、0.1%BSAを含有したKrebs-Ringer-phosphate-Hepes(KRRH)バッファー(pH7.4、20mM Hepes、5mM KH2PO4、1mM MgSO4、1mM CaCl2、136mM NaCl、4.7mM KCl)にて1穴当り200μlで2回洗浄し、1mM 2−デオキシグルコース(2−DG)および0.1%BSAを含有したKRPHバッファー(pH7.4)を1穴当り100μl添加し、37℃の5%炭酸ガス培養器で20分間インキュベートした。さらに0.1%BSAを含有したKRPHバッファー(pH7.4)にて1穴当り200μlで2回洗浄した後、0.1N NaOHを1穴当り25μl添加し、冷凍庫にプレートを入れて凍結させた。凍結後、プレートを冷凍庫から出して室温に戻し、85℃、40分間湯浴中にて加熱処理し、室温になるまで放冷し、0.1N HClを1穴当り25μl添加し、続いて150mMトリエタノールアミンバッファー(pH8.1)を1穴当り25μl添加したものをサンプルとした。各サンプル中の2−DGを下記の通り定量し、コントロール(α−MEM培地(2%v/v牛胎児血清含有)のみ)の2−DG取込量と比較することにより、糖輸送促進作用を調べた。
前記サンプルを96穴組織培養プレートに1穴当り50μl添加し、アッセイカクテル 150μl(50mM トリエタノールアミンバッファー(pH8.1)、50mM KCl、0.5mM MgCl2、0.02% BSA、670μM ATP、120μM NADP+、25μM レサズリンナトリウム、5.5units/ml ヘキソキナーゼ、16units/ml グルコース−6−リン酸デヒドロゲナーゼ、1units/ml ジアフォラーゼ)を添加し、マイクロプレートミキサーにて攪拌、37℃の5%炭酸ガス培養器で90分間インキュベート後、マイクロプレートリーダーにて蛍光値(励起波長:530nm、測定波長:590nm)を測定することにより定量を行った。
アルピネチン(Alpinetin, 5-methoxy-7-hydroxy favanone)
カルダモニンおよび対照化合物の糖輸送促進作用(2−DG取込量)は、表1および図1に示す通りである。図1において、*は有意水準5%で有意差があったことを示し、**は有意水準1%で有意差があったことを示す。
下記表2に記載の配合量にて各原料をよく混合した後、打錠して1錠1g(カルダモニン50mg/錠含有)の錠剤を製造した。
下記表3に記載の配合量にて清涼飲料を製造した。
下記表4に記載の配合量にてソフトカプセルを製造した。
下記表5に記載の配合量にてヨーグルトを製造した。
下記表6に記載の配合量にてキャンディを製造した。
下記表7に記載の配合量にてグミを製造した。
下記表8に記載の配合量にてガムを製造した。
Claims (4)
- カルダモニンを有効成分として含有することを特徴とする筋肉細胞糖輸送促進作用組成物。
- 医薬である請求項1記載の組成物。
- 血糖降下剤である請求項2記載の組成物。
- 糖尿病の予防または治療剤である請求項2記載の組成物。
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