JP5556945B1 - カテキン結合用組成物 - Google Patents
カテキン結合用組成物 Download PDFInfo
- Publication number
- JP5556945B1 JP5556945B1 JP2013192912A JP2013192912A JP5556945B1 JP 5556945 B1 JP5556945 B1 JP 5556945B1 JP 2013192912 A JP2013192912 A JP 2013192912A JP 2013192912 A JP2013192912 A JP 2013192912A JP 5556945 B1 JP5556945 B1 JP 5556945B1
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- Prior art keywords
- catechin
- binding
- composition
- present
- promoting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 50
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- 235000013325 dietary fiber Nutrition 0.000 claims abstract description 21
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Abstract
【解決手段】本発明のカテキン結合用組成物は、麦の葉、オリゴ糖、水溶性食物繊維及び乳酸菌を含有する。本発明のカテキン結合用組成物は、カテキン受容体蛋白質である67LRをコードする遺伝子の発現量を増加させることによって、カテキンの細胞への結合を有効に促進しうる。特に、本発明のカテキン結合用組成物は小腸上皮細胞において、67LRをコードする遺伝子の発現量を増加させることにより、カテキンの細胞へのシグナル伝達の促進に有効でありうる。
【選択図】図1
Description
しかしながら、これらの文献には、麦の葉を、オリゴ糖、水溶性食物繊維及び乳酸菌と組み合わせることによって、カテキンの細胞への結合を促進させることについて、何らの記載も示唆もない。
小腸上皮細胞におけるエピガロカテキンガレート受容体へのエピガロカテキンガレートの結合の促進に用いられる、エピガロカテキンガレートと受容体との結合促進用組成物を提供するものである。
また、麦の葉のエキスを得る方法としては、麦の葉又はその細片化物に、エタノール、水、含水エタノールなどの当業者が通常用いる抽出溶媒を加え、必要に応じて加温して抽出する方法を挙げることができる。抽出物は、必要に応じて濃縮してもよい。
大麦茎葉粉末、オリゴ糖、水溶性食物繊維、乳酸菌をそれぞれ、下記の<遺伝子発現解析試験>で説明する通りに用いた。
ここで、大麦茎葉粉末は、以下の方法で製造した。オリゴ糖として、イソマルトオリゴ糖を用いた。水溶性食物繊維として、難消化性デキストリンを用いた。乳酸菌として、Streptococcus faecalisの乾燥粉末を用いた。
原料として、背丈が約30cmで刈り取った二条大麦の茎及び葉(以下、茎葉という)を用いた。これを水洗いし、付着した泥などを除去し、5〜10cm程度の大きさに切断する前処理を行った。前処理した大麦茎葉を、送帯型蒸機を用いて、ブランチング槽で90〜100℃にて90秒間〜120秒間、1回のみブランチング処理し、その後、冷水で冷却した。続いて、得られた大麦茎葉を、水分量が5質量%以下となるまで、乾燥機中で、20分間〜50分間、80℃〜130℃の温風にて乾燥させた。乾燥した大麦茎葉を約5mmの大きさに切断し、殺菌処理した。得られた大麦茎葉を、200メッシュ区分を90%以上が通過するように気流式粉砕機(ジェットミル)を用いて粉砕処理し、大麦茎葉粉末を得た。
ウシ胎児血清を10質量%含有するDulbecco's modified Eagle's 培地(以下10%FBS/DMEMと略す)を用意した。この10%FBS/DMEMに、大麦茎葉粉末、オリゴ糖、難消化性デキストリン、乳酸菌をそれぞれ、800μg/mlになるように溶解し、4成分それぞれの溶液(サンプル溶液)を調製した。
小腸上皮細胞のモデルとして、ヒト結腸腺癌由来細胞Caco2(DSファーマバイオメディカル社)を用いた。10%FBS/DMEM培地が入ったT75フラスコに、このCaco2を播種し、37℃、5%CO2インキュベーター内で4日間培養した。培養後のCaco2を、Phosphate Buffered Saline(以下PBSと略す)で洗浄後、トリプシン処理により浮遊させた。浮遊細胞を、5X104 cells/mlになるように、10%FBS/DMEM中に懸濁させて、細胞懸濁液を得た。この細胞懸濁液を6ウェル培養プレートに1.5ml/ウェル入れ、37℃、5%CO2インキュベーター内で一晩培養した。
ウェル中の培地を除去し、その代わりに、上記で調製した各サンプル溶液と、10%FBS/DMEMとを、大麦茎葉粉末、オリゴ糖、難消化性デキストリン、乳酸菌それぞれの最終濃度が下記の表1の記載の通りになるように、合計で1.6ml/ウェル入れた。サンプル入り培地を添加後、Caco2を、37℃、5%CO2インキュベーターで24hr培養した。培養後、各ウェル中の細胞Caco2から、それぞれ下記の方法でRNAを得た。得られたRNAを下記のDNAマイクロアレイに供することにより、Caco2における67LR遺伝子の発現量を測定した。サンプル入り培地を添加した実施例1、比較例1及び2のそれぞれにおける遺伝子の発現量を、未処理培地を添加した比較例3における遺伝子の発現量に対する比として、図1に示す。
ウェル中の培地を除去し、各ウェルに0.8mlのIsogen(ニッポンジーン社製)を加え、細胞を溶解したのち、1.5mlチューブへ移した。このチューブをボルテックスした後、0.16mlのクロロホルムを加え、再度ボルテックスした後、15000rpmで15分間遠心した。その後、上清を新しい1.5mlチューブへ移した。これに上清の0.8倍量のイソプロパノールを加え、転倒混和した後、15000rpmで15分間遠心した。次いで、上清を捨て、1mlの70%エタノールを加えた。更に15000rpmで15分間遠心した後、上清を捨て、チューブのフタを開けたまましばらく放置して乾燥させた。得られた乾燥物を30μlの水に溶解しOD260を測定してRNA濃度を算定した。算定した濃度に基づき、得られたRNAの濃度を水で100ng/mlに調整した。
DNAマイクロアレイはAgilent社製のキット「Whole Human Genome DNAマイクロアレイ4×44キットv2」を用いて行った。プローブとしては、このキットに含まれる同社製のもの(遺伝子の種類:19596種、プローブ長さ:60mer)を用いた。このプローブのうち、RPSAに対応するプローブについての発現量の結果を、67LR遺伝子の発現量とした。実験方法は、同社の取扱説明書に従った。
したがって、本発明のカテキン結合用組成物が、麦の葉、オリゴ糖、水溶性食物繊維及び乳酸菌による相乗効果により、小腸上皮細胞において67LRの量を増加しうること、それにより小腸上皮細胞におけるカテキンの結合を促進しうることが示された。
Claims (3)
- 大麦の葉、オリゴ糖、水溶性食物繊維及び乳酸菌を含有し、乳酸菌がストレプトコッカス・フェカリス(Streptococcus faecalis)である組成物であって、
小腸上皮細胞におけるエピガロカテキンガレート受容体へのエピガロカテキンガレートの結合の促進に用いられる、エピガロカテキンガレートと受容体との結合促進用組成物。 - 水溶性食物繊維が難消化性デキストリン、アルギン酸、グアガム加水分解物、グアガム、グルコマンナン、ガラクトマンナン、ペクチン、ポリデキストロース、及びカラギーナンから選ばれる一種又は二種以上である請求項1に記載のエピガロカテキンガレートと受容体との結合促進用組成物。
- オリゴ糖がイソマルトオリゴ糖、ラクチュロース、パラチノース、フラクトオリゴ糖、ラフィノース、スタキオース、キシロオリゴ糖、マルトオリゴ糖、トレハロース、ガラクトオリゴ糖、乳果オリゴ糖、ビートオリゴ糖、ゲンチオオリゴ糖、ニゲロオリゴ糖、スクロース、ラクトース、マルトース及びシクロデキストリンから選ばれる一種又は二種以上である請求項1又は2に記載のエピガロカテキンガレートと受容体との結合促進用組成物。
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JP2022173294A (ja) * | 2016-09-29 | 2022-11-18 | 株式会社東洋新薬 | 抗アレルギー剤、腸管免疫増強剤、乳酸菌の腸管接着性向上剤 |
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