JP2017048150A - アレルギー抑制剤 - Google Patents
アレルギー抑制剤 Download PDFInfo
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- JP2017048150A JP2017048150A JP2015173451A JP2015173451A JP2017048150A JP 2017048150 A JP2017048150 A JP 2017048150A JP 2015173451 A JP2015173451 A JP 2015173451A JP 2015173451 A JP2015173451 A JP 2015173451A JP 2017048150 A JP2017048150 A JP 2017048150A
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Landscapes
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Abstract
Description
原料のプロテオグリカンは、市販の鮭由来プロテオグリカン((株)角弘プロテオグリカン研究所)を購入し、用いた。強酸性陽イオン交換樹脂(商品名:AG 50W−X8 resin、バイオラッド社)をガラス製カラムに充填し(内径2.5cm、高さ8.2cm)、樹脂を常法によりナトリウムイオン型に活性化した。原料の鮭由来プロテオグリカン0.40gを脱イオン水30mLに溶解した溶液を、室温でカラム上方から添加・流下した。その後、樹脂に脱イオン水を150mL流下し、得られた溶出液約180mLをエバポレーター(東京理科器械(株))にて濃縮した後、凍結乾燥(東京理科器械(株))し、0.37gの白色綿状固体であるプロテオグリカンのナトリウム塩を得た。
実施例1で得られたプロテオグリカンのナトリウム塩のタンパク質含量を、比色法であるローリー法にて、牛血清アルブミン(アクロス社)を標準物質とした検量線から求めたところ、5.7重量%であった。ウロン酸含量を、比色法であるカルバゾール硫酸法にて、グルクロン酸(シグマ社)を標準物質とした検量線から求めたところ、35.5重量%であった。原料の鮭由来プロテオグリカンについて同様に分析したところ、タンパク質含量は6.5重量%、ウロン酸含量は34.6重量%であった。これらの結果から、実施例1で得られたプロテオグリカンのナトリウム塩のタンパク質とウロン酸含量は、原料の鮭由来プロテオグリカンとほとんど差がないことが明らかとなった。
原料のプロテオグリカンは、市販の鮭由来プロテオグリカン((株)角弘プロテオグリカン研究所)を購入し、用いた。強酸性陽イオン交換樹脂(商品名:ダイヤイオンSK1B(三菱化学(株)))をガラス製カラムに充填し(内径2cm、高さ8cm)、樹脂を定法によりマグネシウムイオン型に活性化した。原料の鮭由来プロテオグリカン0.50gを脱イオン水40mLに溶解した溶液を、室温でカラム上方から添加・流下した。その後、樹脂に脱イオン水を160mL流下し、得られた溶出液約200mLをエバポレーター(東京理科器械(株))にて濃縮した後、凍結乾燥(東京理科器械(株))し、0.48gの白色綿状固体であるプロテオグリカンのマグネシウム塩を得た。
実施例3で得られたプロテオグリカンのマグネシウム塩のタンパク質含量を、比色法であるローリー法にて、牛血清アルブミン(アクロス社)を標準物質とした検量線から求めたところ、4.4重量%であった。ウロン酸含量を、比色法であるカルバゾール硫酸法にて、グルクロン酸(シグマ社)を標準物質とした検量線から求めたところ、31.3重量%であった。原料の鮭由来プロテオグリカンについて同様に分析したところ、タンパク質含量は6.5重量%、ウロン酸含量は34.6重量%であった。実施例3で得られたプロテオグリカンのマグネシウム塩のタンパク質とウロン酸含量は、原料の鮭由来プロテオグリカンとほとんど差がないことが明らかとなった。
発明品の安全性の確認に用いる試料は、実施例1で用いた市販の鮭由来プロテオグリカン((株)角弘プロテオグリカン研究所)、実施例1で得られたプロテオグリカンのナトリウム塩、および実施例3で得られたプロテオグリカンのマグネシウム塩を使用した。それぞれ蒸留水で0.2mg/mLとなるように希釈したものを使用した。対照として蒸留水を用いた。1群を8匹として、評価用のマウスには、6週齢のC57BL/6マウス(CLEA Japan製)を各群(対照群、鮭由来プロテオグリカン群、実施例1で得られたプロテオグリカンのナトリウム塩群および実施例3で得られたプロテオグリカンのマグネシウム塩群)に用いた。恒温、恒湿の一定環境の飼育室で、試料を自由飲水させ、固形飼料(CE―2、CLEA Japan製)を自由摂取させ飼育した。なお、実験動物の取り扱いは弘前大学動物実験委員会により承認され、弘前大学動物実験に関する規程に従った。
アレルギー抑制作用の測定に用いる試料は、市販の鮭由来プロテオグリカン、実施例1で得られたプロテオグリカンのナトリウム塩、および実施例3で得られたプロテオグリカンのマグネシウム塩で実施例5と同じものを使用した。それぞれ蒸留水で0.2mg/mLとなるように希釈したものを使用した。対照として蒸留水を用いた。1群を8匹として、作用評価用のマウスには、6週齢のC57BL/6マウス(CLEA Japan製)を各群(対照群、鮭由来プロテオグリカン群、実施例1で得られたプロテオグリカンのナトリウム塩群および実施例3で得られたプロテオグリカンのマグネシウム塩群)に用いた。恒温、恒湿の一定環境の飼育室で、試料を自由飲水させ、固形飼料(CE―2、CLEA Japan製)を自由摂取させ飼育した。なお、実験動物の取り扱いは弘前大学動物実験委員会により承認され、弘前大学動物実験に関する規程に従った。
好酸球率(%)=好酸球数/白血球総数×100
なお、統計処理には、Tukey法を用いた。対照-実施例1で得られたプロテオグリカンのナトリウム塩間および対照-実施例3で得られたプロテオグリカンのマグネシウム塩間でP値が0.05未満で有意差が認められた。なお、摂取飲水量は、4群とも一匹あたり一日約4mLで差はなかった。
Claims (2)
- プロテオグリカンのナトリウム塩、もしくはプロテオグリカンのマグネシウム塩の少なくとも一つを有効成分とするアレルギー抑制剤。
- 請求項1において、プロテオグリカンのナトリウム塩についてナトリウムを6.0重量%以上含み、かつカルシウムが1.0重量%未満であるプロテオグリカンのナトリウム塩、もしくはマグネシウムを5.0重量%以上含み、かつカルシウムが1.0重量%未満であるプロテオグリカンのマグネシウム塩のうち、少なくとも一つを有効成分とするアレルギー抑制剤。
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