JPWO2016032000A1 - 乳酸菌含有組成物 - Google Patents
乳酸菌含有組成物 Download PDFInfo
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- JPWO2016032000A1 JPWO2016032000A1 JP2016545660A JP2016545660A JPWO2016032000A1 JP WO2016032000 A1 JPWO2016032000 A1 JP WO2016032000A1 JP 2016545660 A JP2016545660 A JP 2016545660A JP 2016545660 A JP2016545660 A JP 2016545660A JP WO2016032000 A1 JPWO2016032000 A1 JP WO2016032000A1
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- Prior art keywords
- composition
- lactic acid
- acid bacteria
- bifidobacteria
- test
- Prior art date
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Abstract
Description
表1に記載の成分からなる乳酸菌含有組成物を製造した。なお、各成分は従来公知のものを用いることができる。
被験動物として、照明12時間、室温23±5℃、相対湿度60±10%の環境を維持した飼育室にて1週間馴化させた、7〜8週齢の雌性C57BL/6マウスを用いた。
被験動物を、1日1回、継続4時間、呼吸/排泄可能な処置を施したポリプロピレン製遠沈管(容量約60mL)内に拘束し、拘束中被験動物の顔面に送風(風速0.5〜1.0m/S)することによりストレス負荷処置を行った。ストレス負荷処置時間以外は、ケージ内で飼料(固型飼料、マウス・ラット・ハムスター用飼料 MF、製造元オリエンタル酵母工業株式会社)と飲水(水道水)は自由摂取とした。試験は1群5〜6匹で実施した。
ストレス負荷処置前に、被験動物の左右の外眼角に、それぞれ綿糸(ゾーンクイック(商標登録)、昭和薬品化工株式会社)を15秒挿入し、綿糸が涙液の浸透により褐色変色した長さを、0.5mmの精度で測定した。左右眼の平均値を個体の涙液分泌量とした。
データの統計解析には、統計ソフトであるSAS(SAS Institute Inc.製)及びStatLight(ユックムス株式会社製)を用いた。対照群を対照としたt−test又はDunnett test、及び処置前群を対照としたpaired t−testを実施した。いずれも有意水準は両側5%とし、P値が0.05未満を有意とみなした。また、平均値及び標準偏差を求めた。
実施例1の組成物を、ストレス負荷処置前日とストレス負荷処置期間中10mg/kg若しくは50mg/kgの量で、又はストレス負荷処置前5日間とストレス負荷処置期間中10mg/kgの量で1日1回経口投与した被験動物、及び対照群として実施例1の組成物を投与しなかった被験動物に、ストレス負荷処置を3日間実施した。各日の被験動物の涙液分泌量を測定し、統計解析を行った。
被験動物に実施例1の組成物を、ストレス負荷処置5日前からストレス負荷処置開始5日後まで、毎日10mg/kgを1日1回経口投与した。被験動物にストレス負荷処置を7日間実施した。各日の被験動物涙液分泌量を測定し、統計解析を行った。
被験動物に、実施例1の組成物を0.06%の濃度となるよう混合した飼料を、ストレス負荷処置5日又は14日前からストレス負荷処置終了まで自由摂取させた。被験動物にストレス負荷処置を5日間実施した。各日の涙液分泌量を測定し、統計解析を行った。
実施例1の組成物を混合した飼料を摂取した被験動物では、摂取期間が長い程、涙液分泌量の低下を抑制できることが判明した。この結果から、実施例1の組成物を食事として長期間継続して摂取することにより、特に高いドライアイ予防効果を達成できることが判明した。
ストレス負荷処置後涙液分泌量の低下が確認された被験動物に、実施例1の組成物を5mg/kg、10mg/kg又は50mg/kgの量で、1日1回、9日間経口投与した。対照群の被験動物には、実施例1の組成物を投与しなかった。実施例1の組成物の投与期間中ストレス負荷処置を実施した。各日の被験動物の涙液分泌量を測定し、統計解析を行った。
ストレス負荷処置後、涙液分泌量の低下が確認された被験動物に、実施例1の組成物を50mg/kgの量で、1日1回、9日間経口投与した。投与期間中及び投与終了後3日間ストレス負荷処置を実施した。各日の被験動物の涙液分泌量を測定し、統計解析を行った。
乳酸菌のドライアイ予防効果を明らかにするために、比較例1として、表1に記載された成分のうち乳酸菌のみを含まない組成物を製造した。
実施例1の組成物又は比較例1の組成物を、被験動物に、ストレス負荷処置前日に50mg/kgの量で単回経口投与した。対照群の被験動物にはストレス負荷処置前日に本発明の組成物を投与しなかった。ストレス負荷処置前日、ストレス負荷処置直前及びストレス負荷処置翌日に、被験動物の涙液分泌量を測定した。涙液分泌量について、統計解析を行った。
長期ストレス負荷試験からの涙液分泌量回復(治療)効果試験を行った。被験動物にストレス負荷処置を連続35日間実施した。ストレス負荷処置開始後21日目から28日目まで、被験動物に比較例1の組成物を10mg/kg/単回経口投与した。その後、ストレス負荷処置開始後29日目から36日目までの1週間、実施例1の組成物を被験動物に10mg/kg/単回経口投与した。各日の被験動物の涙液分泌量を測定し、統計解析を行った。
長期ストレス負荷処置試験からの涙液分泌量回復(治療)効果試験を行った。被験動物にストレス負荷処置を連続40日間実施した。ストレス負荷処置開始後13日目から21日目まで、被験動物に実施例1の組成物を50mg/kg/単回経口投与した。ストレス負荷処置開始後22日目から28日目まで休薬期間とし、その後、29日目から40日目まで、被験動物に実施例1の組成物を10mg/kg/単回経口投与した。各日の被験動物の涙液分泌量を測定し、統計解析を行った。
(乳酸菌含有組成物およびその投与方法)
ドライアイ自覚症状のある22歳から59歳までの男女20名に、表2に記載の成分を含有するソフトカプセルを、1回2粒、1日1回夕食後に、8週間摂取させた。
ソフトカプセルの摂取前、摂取後の計2回、ドライアイの検査を行った。検査は、すべての眼で、眼症状3項目(シルマー試験第1法、BUT検査、フルオレセイン染色による角結膜上皮障害スコア)を実施し、さらに自覚症状アンケート2種(ドライアイQOL問診票(DEQS)、11項目の眼に関する自覚症状についてのVAS評価)を行った。シルマー試験第1法、BUT検査、フルオレセイン染色による角結膜上皮障害スコアは、2006年ドライアイ診断基準に従い検査を実施した。DEQSは、ドライアイ研究会にて開発された問診票を用いた。
疼痛等により検査が受けられない項目があった2名を除く、18名の結果を図9〜12、および表3に示した。ソフトカプセル摂取後は、眼症状の検査結果が全項目で改善した。また、自覚症状アンケートにおいても、ソフトカプセル摂取後はスコアが改善された。これより、本発明の組成物が、ドライアイ症状の改善に効果を示すことが示唆された。
(被験動物)
被験動物として、照明12時間、室温23±5℃、相対湿度60±10%の環境を維持した飼育室にて1週間馴化させた、7〜8週齢の雌性C57BL/6マウスを用いた。
被験動物を、1日1回、継続4時間、呼吸/排泄可能な処置を施したポリプロピレン製遠沈管(容量約60mL)内に拘束し、拘束中被験動物の顔面に送風(風速0.5〜1.0m/S)することによりストレス負荷処置を行った。ストレス負荷処置時間以外は、ケージ内で飼料(固型飼料、マウス・ラット・ハムスター用飼料 MF、製造元オリエンタル酵母工業株式会社)と飲水(水道水)は自由摂取とした。試験は1群5〜6匹で実施した。
ストレス負荷処置前に、被験動物の左右の外眼角に、それぞれ綿糸(ゾーンクイック(商標登録)、昭和薬品化工株式会社製)を15秒挿入し、綿糸が涙液の浸透により褐色変色した長さを、0.5mmの精度で測定した。左右眼の平均値を個体の涙液分泌量とした。
Streptococcus faecalis WB2000株(細菌学上はEnterococcus faecium WB2000株)、Enterococcus faecium JCM5804株、Lactobacillus salivarius WB21株、Lactobacillus acidophilus WB2001株、Lactobacillus pentosus TJ515株、又はBifidobacterium longum WB1001株の凍結乾燥末を、個別に、蒸留水0.5mLに0.34mg含むように懸濁した。ストレス負荷処置前日とストレス負荷処置期間中、前記懸濁液を凍結乾燥末の菌17mg/kgの量で1日1回経口投与した被験動物、及び対照群として上記乳酸菌及びビフィズス菌を投与しなかった被験動物に、ストレス負荷処置を4日間実施した。ストレス負荷処置前日、ストレス負荷処置2日目及びストレス負荷処置4日目における被験動物の涙液分泌量を測定した。
結果を図13に示した。対照群の被験動物は、ストレス負荷処置により涙液分泌量が大幅に低下した。上記乳酸菌又はビフィズス菌を前投与することにより、涙液分泌量の低下を抑えることができ、ドライアイ予防効果を示した。特にStreptococcus faecalis WB2000株は、他の菌と比較して特に涙液分泌量の低下を抑えることができるため、高いドライアイ予防効果が得られることが判明した。
Claims (7)
- ルテイン、魚油、ラクトフェリン、ビタミン、γアミノ酪酸及び亜鉛からなる群から選択される1以上の成分、並びに乳酸菌又はビフィズス菌を含有する組成物。
- ルテイン、魚油、並びに乳酸菌又はビフィズス菌を含有する請求項1に記載の組成物。
- さらにラクトフェリンを含有する請求項2に記載の組成物。
- 医薬組成物である請求項1〜3のいずれかに記載の組成物。
- 食品組成物である請求項1〜3のいずれかに記載の組成物。
- ドライアイ治療用又はドライアイ予防用の、請求項1〜5のいずれかに記載の組成物。
- Streptococcus属、Enterococcus属、Lactobacillus属、又はBifidobacterium属の微生物を含有する、ドライアイ治療用又はドライアイ予防用の組成物。
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