JP2022173294A - 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 - Google Patents
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 Download PDFInfo
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
Description
また、茎葉の抽出物を得る方法としては、茎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。
Bifidobacterium属としては、Bifidobacterium bifidum、Bifidobacterium breve、Bifidobacterium infantis、Bifidobacterium lactis、Bifidobacterium longum、Bifidobacterium adolescentis、Bifidobacterium mongolienseが挙げられる。
Lactbacillus属としては、Lactbacillus brevis、Lactbacillus gasseri、Lactobacillus acidophilus、Lactobacillus buchneri、Lactobacillus bulgaricus、Lactobacillus delburvecki、Lactobacillus casei、Lactobacillus crispatus、Lactobacillus curvatus、Lactobacillus halivaticus、Lactobacillus pentosus、Lactobacillus plantarum、Lactobacilus paracasei、Lactobacillus rhamnosus、Lactobacillus salivarius、Lactobacillus sporogenes、Lactobacillus sakei、Lactobacillus fructivorans、Lactobacillus hilgardii、Lactobacillus reuteri、Lactobacillus fermentumが挙げられる。
Enterococcusとしては、Enterococcus faecalis(Streptococcus faecalis と称されることもある)、Enterococcus faesium(Streptococcus faesiumと称されることもある)、Streptococcus thermophilus、Lactococcus lactis(Streptococcus lactisと称されることもある) が挙げられる。
Leuconostoc属としては、Leuconostoc mesenteroides、Leuconostoc oenos が挙げられる。
Pediococcus属としては、Pediococcus acidilactici、Pediococcus pentosaceusが挙げられる。
Staphylococcus属としては、Staphylococcus carnosus、Staphylococcus xylosusが挙げられる。
Tetragenococcus属としては、Tetragenococcus halophilusが挙げられる。Bacillus属としては、Bacillus coagulans、及びBacillus mesentericusなどが挙げられる。
とりわけ、Bacillus coagulans、Enterococcus faecalis、Bifidobacterium bifidum、Enterococcus faesium、Lactobacillus acidophilusが好ましい。これらは、1種を単独で使用してもよいし、2種以上を併用してもよい。
このため、本発明の剤はこれを経口摂取することで、腸管における乳酸菌の作用を高めることができ、例えば腸管免疫増強剤として用いることができるほか、アトピー性皮膚炎、アレルギー性鼻炎、花粉症、湿疹、蕁麻疹等の発疹、下痢や嘔吐等のアレルギー症状の予防や改善を図ることができる。前記の腸管免疫増強剤の作用は、Th1細胞の働きを活発化したり、IgA抗体産生を促すことを含む。本発明の剤は、ヒトに対して好適に適用されるものであるが、それぞれの作用効果が奏される限り、ヒト以外の動物(例えば、マウス、ラット、ハムスター、イヌ、ネコ、ウシ、ブタ、サル等)に対して適用することもできる。本発明のアレルギー剤からは、インターロイキン4産生を抑制することにより抗アレルギー作用を発揮するものを除くものとする。
(1)96well plateを用いたmonolayer作製
37℃、5体積%CO2インキュベーター内で、75cm2フラスコを用いて、通常培地にて腸管上皮細胞株Caco-2細胞を培養した。この通常培地は、44.5 mL DMEM(1%ペニシリン-ストレプトマイシン含有)へ5mL FBS(Japan Bio Serum社)及び0.5 mL NEAA(Non-Essential Amino Acids、SIGMA社)を加えて調製した。トリプシン処理により浮遊させた細胞を、75 cm2フラスコから96well clear plateに4.0×104cells/wellの細胞密度で播種した。37℃、5体積%CO2インキュベーター内で72時間培養し、monolayerを作製した。
前培養したB.coagulansを採取し、室温・1000gで3分間遠心した。上清を除いて得たペレットを1mLのリン酸緩衝生理食塩液(PBS)へ懸濁し、室温・1000gで3分間遠心した。同様の懸濁及び遠心をもう1度繰り返し、上清を取り除いてペレットを1mLのPBSへ懸濁し、carboxyfluorescein diacetate(CFDA、同仁化学研究所社製)を15μL添加した。遮光し室温で30分間インキュベートした後、室温・1000gで3分間遠心した。上清を除き1mLのPBSへ懸濁し、室温・1000gで3分間遠心する操作を2回繰り返した。その後、上清を除き試験培地へ懸濁した。なおblankとしてCFDAを添加しない以外は同様の操作を行ったものを用意した。試験培地としては45 mL RPMI1640培地(1%ペニシリン-ストレプトマイシン含有)へ5 mL のFBS(Japan Bio Serum社)を加えたものを用いた。前記懸濁液をBlackplateへ100μL移し蛍光強度を測定した。励起波長は495nm、蛍光波長515nmとした。測定した蛍光強度からblankの値を差し引き、蛍光強度が200になるよう試験培地を用いて希釈しこれをB.coagulansサンプルとした。
(1)で作製したCaco-2細胞のmonolayerの培地を通常培地から前記試験培地へ置換した。培地を除き、被験物質を試験培地に分散させた分散液、及び(2)で調製したB.coagulansサンプルを100μLずつ96wellplateへ添加した(B.coagulans最終蛍光強度100)。37℃の5体積%CO2インキュベーター内で2時間培養後、各wellにホルマリン150μl添加し、遮光して4℃で30分間固定した。PBSで3回洗浄し、PBSを100μl添加したのちvarioskanで蛍光強度を測定した(励起波長495nm、蛍光波長515nm)。測定値に基づき、乳酸菌のみを添加したサンプル蛍光強度に対する相対値(%)を算出し、これを乳酸菌の腸管細胞monolayerへの接着率とした。結果を図1に示す。なお、被験物質のインキュベート時のCaco-2細胞培養液中の濃度は、下記表1に記載の通りである(表1の数値の単位はμg/mL)。
Claims (2)
- 大麦の茎及び/又は葉を含有するアレルギー症状の予防及び/又は改善用経口剤。
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食品であることを特徴とする、請求項1に記載の経口剤。
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