JP2005075766A - エンドセリン−1産生抑制剤 - Google Patents
エンドセリン−1産生抑制剤 Download PDFInfo
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- JP2005075766A JP2005075766A JP2003307350A JP2003307350A JP2005075766A JP 2005075766 A JP2005075766 A JP 2005075766A JP 2003307350 A JP2003307350 A JP 2003307350A JP 2003307350 A JP2003307350 A JP 2003307350A JP 2005075766 A JP2005075766 A JP 2005075766A
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- endothelin
- extract
- luteolin
- production inhibitor
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Abstract
【解決手段】 本発明は、ルテオリンおよび/またはその配糖体を含有するエンドセリン−1産生抑制剤、ルテオリンおよび/またはその配糖体を含有する植物抽出物を含有するエンドセリン−1産生抑制剤、ならびに、クダモノトケイソウの抽出物、特に果皮抽出物または果肉抽出物、を含有するエンドセリン−1産生抑制剤に関する。
【選択図】 なし
Description
ネイチャー (Nature), (英国),2001年, 414巻, p.863-864 アテロスクレロシス (Atherosclerosis), (アイルランド), 2002年, 163巻, p. 339-347
(1) ルテオリンおよび/またはその配糖体を含有するエンドセリン−1産生抑制剤。
(2) ルテオリンおよび/またはその配糖体を含有する植物抽出物を含有するエンドセリン−1産生抑制剤。
(3) クダモノトケイソウ(Passiflora edulis、別名パッションフルーツ)の抽出物を含有するエンドセリン−1産生抑制剤。
(4) クダモノトケイソウの抽出物が、果皮抽出物または果肉抽出物である上記(3)に記載のエンドセリン−1産生抑制剤。
ブタ大動脈内皮細胞(継代数1、25 cm2培養フラスコ入り、大日本製薬(Cell Systems Corporation)より購入)をCS-C培地(D-MEM培地とハムF12培地を1:1に等比混合した培地に、10%ウシ胎児血清、15mM HEPES、酸性FGF、およびヘパリンを添加した培地、Cell Systems Corporationより購入)に分散させ、コラーゲンをコーティングした24穴(1穴2cm2)のプレート1枚に添加して、5% CO2存在下、37℃で4日間の培養を行った。次にプレートの各穴の培地を廃棄し、1mlのクレブス-リンガー重炭酸(KRB)緩衝液(NaCl 129 mM、NaHCO3 5 mM、KCl 4.8 mM、KH2PO4 1.2 mM、CaCl2 1.0 mM、MgSO4 1.2 mM、グルコース 2.8 mM、 ウシ血清アルブミン 0.1%、HEPES 10 mM、PH 7.4)にて各穴を2回洗浄した。その後、360μl のKRB緩衝液に40μlの試料(10%DMSO (ジメチルスルホキシド)水溶液中の63〜500μMルテオリン、10%DMSO水溶液中の250〜1000μMルテオリン配糖体、または対照としての10%DMSO水溶液)を加えた溶液(反応液)400μlを各穴に添加し、5%CO2存在下、37℃で0〜8時間静置し、溶液中のエンドセリン−1濃度を測定した。その結果を表1及び図1に示す。表1は3時間の間に溶液中へ放出されたエンドセリン−1量を、10%DMSO水溶液添加時(反応液中でのDMSOの終濃度1%)のエンドセリン−1量(対照)を100%として比較した値(n = 4の平均値)で示す。図1は試料添加後の各時点における溶液中のエンドセリン−1濃度(n = 3の平均値)を示す。ルテオリン及びルテオリン配糖体(ルテオリン-6-C-グルコシド)にエンドセリン−1産生抑制効果が認められた。
クダモノトケイソウの果実3個(那覇市内で購入)から剥いた果皮(150g)をミキサーで細かく破砕した後、メタノール200mlを加え、十分に攪拌した(電磁スターラー、400rpm、60分間)。次いで、濾紙(アドバンテック東洋製濾紙、No.2、直径30cm)による濾過を行って不溶物を除去し濾液を回収した。濾液をロータリエバポレーターで濃縮し、さらに真空ポンプによる減圧下で乾燥することにより、紫色の粉末3.6gを得た。
クダモノトケイソウの果実3個からゼリー状の果肉(種子を含む)110mlを採取し、これに同量の蒸留水を加え、十分に攪拌した(電磁スターラー、400rpm、60分間)。次いで、15,000rpmで20分間の遠心分離を行い不溶物を沈殿させ、上清を回収した。この溶液は限外濾過膜YM-10(分画分子量1万、Millipore)を通過させ、濾液を遠心式エバポレーターで濃縮し、さらに真空ポンプによる減圧下で乾燥することにより、薄いオレンジ色の粉末16gを得た。
試験例1と同様に24穴プレートで培養したブタ大動脈内皮細胞をKRB緩衝液で2回洗浄した後、360μl のKRB緩衝液に40μlの試料(2.5〜40 mg/mlのクダモノトケイソウ果皮抽出物水溶液、0.75〜7.5 mg/mlのクダモノトケイソウ果肉抽出物水溶液、または対照としての蒸留水)を加えた溶液(反応液)400μlを各穴に添加し、5%CO2存在下、37℃で0〜8時間静置した。なお、試料添加時における細胞数は1穴あたり4.0 x 105個である。その後、上清200μlを回収して、遠心により混入した細胞を除去した。上清中のエンドセリン−1量は、エンドセリン−1 ELISAシステム(Amersham Biosciences)を用いた酵素免疫測定法により測定した。その結果を表2及び図2に示す。表2は3時間の間に溶液中へ放出されたエンドセリン−1量を、蒸留水添加時のエンドセリン−1量(対照)を100%として比較した値(n = 4の平均値)で示した。図2は試料添加後の各時点における溶液中のエンドセリン−1濃度(n = 3の平均値)を示した。クダモノトケイソウ果皮抽出物及びクダモノトケイソウ果肉抽出物にエンドセリン−1産生抑制効果が認められた。
Claims (4)
- ルテオリンおよび/またはその配糖体を含有するエンドセリン−1産生抑制剤。
- ルテオリンおよび/またはその配糖体を含有する植物抽出物を含有するエンドセリン−1産生抑制剤。
- クダモノトケイソウの抽出物を含有するエンドセリン−1産生抑制剤。
- クダモノトケイソウの抽出物が、果皮抽出物または果肉抽出物である請求項3に記載のエンドセリン−1産生抑制剤。
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EP1763360A4 (en) * | 2004-04-02 | 2009-08-19 | Ind Res Ltd L L C | FRUIT EXTRACTS FROM PASSION AND USES THEREOF |
JP2010207125A (ja) * | 2009-03-09 | 2010-09-24 | Kao Corp | エンドセリン発現抑制剤の評価又は選択方法 |
WO2010113315A1 (ja) * | 2009-04-03 | 2010-10-07 | 森永製菓株式会社 | ピセアタンノール含有組成物及びピセアタンノール含有組成物の製造方法 |
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JP2013060404A (ja) * | 2011-09-15 | 2013-04-04 | Ichimaru Pharcos Co Ltd | リンパ管腔形成促進剤 |
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JP2018048103A (ja) * | 2016-09-16 | 2018-03-29 | オリザ油化株式会社 | 肌質改善剤 |
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