JP2005021087A - Egg-derived bone-strengthening composition - Google Patents
Egg-derived bone-strengthening composition Download PDFInfo
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- JP2005021087A JP2005021087A JP2003190906A JP2003190906A JP2005021087A JP 2005021087 A JP2005021087 A JP 2005021087A JP 2003190906 A JP2003190906 A JP 2003190906A JP 2003190906 A JP2003190906 A JP 2003190906A JP 2005021087 A JP2005021087 A JP 2005021087A
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Abstract
Description
【０００１】
【発明の属する技術分野】本発明は、骨成長及び骨強化作用を有する卵由来水溶性画分を含有する組成物に関する。また、本発明は、この卵由来水溶性組成物を配合した飲食品、医薬品、又は飼料に関する。本発明の卵由来水溶性画分は骨芽細胞を増殖させ、骨の成長端における軟骨細胞の増殖も促進させる作用を有するので、骨代謝のバランスが崩れて起こる骨疾患の予防や治療や、学童など若年層において成長期の骨強化に有用である。
【０００２】
【従来の技術】近年の骨粗鬆症の増加や若年層での骨折率の増加など各種骨疾患の増加が社会問題となっている。骨粗鬆症の患者は全国に約１０００万人いると推定されており、高齢化の進行により２０１０年には患者数が１６００万人に達するとの予測もある。骨粗鬆症は、主に閉経後の女性において、女性ホルモンの減少の影響から骨形成と骨破壊のバランスが崩れ起こる疾患として注目されているが、男性でも老齢化に伴い骨塩量の減少が見られ、現在でも５０歳以上で約２００万人の骨粗鬆症患者がいる。また、最近２０年で小中学生の骨折率は２倍以上に増加しており、骨粗鬆症を予防すると言う見地からも若年期にしっかりと骨塩量を高めて、より高い最大骨量を得ることは非常に重要である。このように、いまや骨の健康を保つことは性別や年齢の関係なく全社会的な関心事である。
【０００３】
従来より骨の疾患を予防あるいは治療する方法として、食餌療法によるカルシウム補給、軽い運動、日光浴、薬物治療等が行われている。食餌療法によるカルシウム補給には、炭酸カルシウム、リン酸カルシウム等のカルシウム塩や、牛骨粉、卵殻、魚骨粉等の天然カルシウム剤が使用されているが、溶解性や吸収性、呈味性の点で必ずしも経口摂取に適している素材であるとはいえない。適度な運動は、骨量を増やし、骨を強化するので、散歩やウオーキングは骨の健康によいとされるが、体が弱っている場合は軽い運動も厄介なものである、まして寝たきりの老人ではほとんど運動できない。日光浴は活性化ビタミンＤ３の補給と言う点ではよいとされているが、これだけでは不十分である。薬物投与には、ビスホスホネートやカルシトニン製剤等が使用されており、骨粗鬆症の治療には有効であると言うことが知られている。しかし、これらの物質は医薬そのものであり、その服用には特別な注意が必要で、耳鳴り、頭痛、食欲不振など副作用の危険性もあり、継続的な食品素材として使用可能なものではない。
【０００４】
【発明が解決しようとする課題】
そこで、本発明では日常的に継続して摂取可能な骨強化剤として、骨強化作用を有し、安全、安価であり、飲食品に広く用いることが可能な食品素材を提供することを目的とする。
【０００５】
【課題を解決するための手段】
卵は、３７℃で３週間温められるとヒヨコに変わると言う現象が示すように、生体を形作るすべての材料を含み、次世代を生み出すバイオカプセルであり、まだ見出されていない生理活性も多いと考えられる。また、卵は古くからの食経験があり、そのものの栄養価も高く安全であって食品素材としての利用価値は非常に高い。
【０００６】
そこで、本発明者らは、卵の様々な画分について骨成長、骨強化を促進する成分の探索を行った結果、卵黄水溶性画分に骨芽細胞増殖及び石灰化促進活性があることを初めて見出し、また、この成分を経口摂取することによって骨成長を促進し、骨代謝を改善できることを見出し、本発明を完成するに至った。
【０００７】
すなわち、請求項１記載の発明（以下、本発明１という）の骨強化組成物は、骨芽細胞増殖促進活性、骨成長および骨強化作用を有する卵由来の水溶性画分を有効成分として含有することを特徴とする。
【０００８】
本発明２においては、前記卵黄水溶性画分がエタノール、イソプロパノール、ヘキサン、またはその他食品加工に用いられる有機溶媒のうちから選ばれた少なくともひとつの有機溶媒により脱脂された卵黄から抽出された水溶性画分であることが好ましい。
【０００９】
また、本発明３においては、前記卵黄水溶性画分が、全卵又は卵黄液にカラギナン液を加えることによって得られる抽出液であることが好ましい。
【００１０】
さらに、本発明４の飲食品、医薬または飼料は、本発明１から３で得られた卵黄水溶性組成物を配合して用いられることが好ましい。
【００１１】
本発明によれば、安全、安価で日常的に摂取可能な骨強化用素材及び飲食品、医薬、又は飼料を提供する事ができる。
【００１２】
【発明実施の形態】
本発明で用いられる卵は、鶏、アヒル、うずら等の卵があげられるが、生産性の点から、鶏卵が好ましく用いられる。
【００１３】
また、鶏卵を用いる場合には、全卵液、卵黄液、卵白液、全卵粉末、卵黄粉末、卵白粉末、脱脂全卵粉末、又は脱脂卵黄粉末を用いることができ、中でも脱脂全卵粉末、又は脱脂卵黄粉末が好ましく用いられる。
【００１４】
本発明の骨強化組成物は、全卵粉末から食品加工において用いられる有機溶媒を用いて脱脂して得られた粉末を、温水、又は０．５％塩化ナトリウムで抽出したときの上清として得ることができる。脱脂に用いる有機溶媒は、エタノール、イソプロパノール、ヘキサンと、ほかにも食品加工に用いられる有機溶媒であればいずれでも使用できるが、簡便性と安全性の点からエタノールが好ましく用いられる。
【００１５】
また、この骨強化組成物は、全卵液または卵黄液にアラビアガム、カラギナン、キサンタンガム等の天然ゴム質水溶液を加えることによって得られる抽出液として得ることができる。ここで用いる天然ガム質水溶液は、アラビアガム、カラギナン、キサンタンガム等のいずれでも使用できるが、抽出効率の点から、ラムダカラギナンが特に好ましく用いられる。
【００１６】
本発明の骨強化組成物は骨粗鬆症や各種骨疾患の予防または改善に適用され、その有効摂取量は、成人一日当たり卵黄水溶性画分粉末換算で１００ｍｇから６ｇである。また、本発明の骨強化組成物には、ラットによる急性毒性は認められなかった。
【００１７】
本発明の骨強化組成物は、必要に応じてタンパク質分解酵素により処理してもよい。上記タンパク質分解酵素としては、食品に用いることのできるタンパク質分解酵素であれば特に制限されず、酸性プロテアーゼ、中性プロテアーゼ、アルカリプロテアーゼのいずれも用いることができる。本発明においては、中性プロテアーゼが好ましく用いられる。中世プロテアーゼとしては、オリエンターゼ（商品名、株式会社エイチビイアイ製）、フレーバーザイム（商品名、ノボノルディスク製）等が挙げられる。
【００１８】
このようにして回収された骨強化組成物は、通常粉末化して用いられる。粉末化は、凍結乾燥によって、あるいは噴霧乾燥によって行ってもよく、通常は飲食品として経口的に投与される。
【００１９】
本発明の骨強化組成物は、上述した成分のほかに、賦形剤、乳化剤、安定剤、甘味料、ｐＨ調整剤、増粘剤、タンパク質、ペプチド、アミノ酸、脂質、多糖類、オリゴ糖等の通常食品に用いられる成分を含んでいてもよい。
【００２０】
本発明の骨強化組成物は、飲食品に加えるだけでなく投与形態に応じて種々の形に調製され、粉末（または顆粒）、ドリンク剤、錠剤、カプセル剤等として用いることもできる。卵は古くから食経験のある安全な素材であり、継続的に用いても副作用の問題のない素材として、医薬または飼料に用いることもできる。
【００２１】
【実施例】
以下、実施例を挙げて本発明をより詳細に説明するが、本発明はこれらの実施例により何ら限定されるものではない。
【００２２】
実施例１（有機溶媒による抽出）
卵黄粉末５００ｇにエタノール２．５ｌを加え、ブレンダーで３０分間攪拌したのち固形物を回収した。この操作を３回繰り返して卵黄から脱脂を行った。この結果、風乾したのちに２８４ｇの脱脂卵黄を得た。この脱脂卵黄粉末に３７℃の温水３ｌを加えて、温度を保ちながらスターラーでよく攪拌した後、８，０００ｒｐｍ、３０分間の遠心分離を行い、上清を得た。そして、凍結乾燥で粉末化して、１３．４ｇの卵黄水溶性組成物を得た。
【００２３】
実施例２（カラギナンによる抽出）
卵黄粉末５００ｇに１％ガンマカラギナン液を３Ｌ加え、軽く攪拌した後、３０分間静置すると凝集が見られる。この後、８，０００ｒｐｍ、３０分間の遠心分離を行い上清を得た。これを凍結乾燥で粉末化して、１４．８ｇの卵黄水溶性組成物を得た。
【００２４】
実施例１、２で得られた水溶性組成物は、以下の条件のゲル濾過クロマトグラフィーで分子量の分析を行った。
【００２５】
カラム：ＹＭＣ−Ｐａｃｋ
Ｄｉｏｌ ２００ （６．０＆Ｉｕｍｌ；３００ ｍｍ） （商品名、ワイエムシイ社製）
溶出液：０．２Ｍリン酸カリウム緩衝液（ｐＨ６．９）
流速：０．７ｍｌ／分
検出波長：２８０ｎｍ
【００２６】
その結果、実施例１、２の卵黄水溶性組成物はいずれも分子量１０，０００〜２００，０００に同様の主要なピークの分布がみられた。
【００２７】
試験例１（骨芽細胞増殖活性の測定）
実施例１で得られた卵黄水溶性組成物の骨芽細胞増殖活性を以下のようにして測定した。ヒト骨芽細胞のＩｎＭ１．２細胞を、１０％ＦＢＳを含むα−ＭＥＭ培養液で３７℃、５％ＣＯ２−９５％ａｉｒの下で培養し、コンフルエントになるまで培養した後、トリプシン処理により細胞を集め、上記培養液を用いて１×１０４個／ｍＬに調製した。この細胞調製液を２４ｗｅｌｌ ｐｌａｔｅに０．５ｍＬずつ播種し、３７℃、５％ＣＯ２−９５％ａｉｒの下で培養した。翌日、試料が１００、２５、１２．５ ｐｐｍとなるように細胞に添加し、また石灰化促進剤であるα−グリセロリン酸塩（２ｍｍｏｌ／Ｌ）を添加した。試料は５０％ＤＭＳＯに溶解して、溶剤が培養液の１％になるように加えた。培養液は１日おきに交換し、２０日後にＭＴＴを用いて骨芽細胞の数を測定した。骨芽細胞の増殖促進活性は、対照群の増殖数を１００としたときの相対値であらわした。この結果を表１に示す。
【００２８】
【表１】
試験例２（骨成長促進活性）
実施例１で得られた卵黄水溶性組成物の骨成長促進活性を以下のようにして測定した。骨成長促進活性は、Ｓｐｒａｇｕｅ−Ｄａｗｌｅｙ系ラットを用いて評価した。試料を２０ｍｇ／ｍｌの濃度で精製水に溶かし、ラットに２０または１００ｍｇ／ｋｇ体重／１日摂取させた。試験期間中のラットの飲料水と飼料は自由摂取とし、基本飼料としてＭＦ（商品名、オリエンタル酵母社製）を用いた。試料摂取期間（１週間）中の３日目と５日目にそれぞれ蛍光物質としてテトラサイクリンを投与した。この試験例においてはテトラサイクリンがカルシウムをキレートし、骨に沈着される現象を利用して、骨に形成される蛍光ラインを観察することにより脛骨の卵タンパク質組成物による成長促進効果を測定した。脛骨を摘出し、テトラサイクリンを投与により脛骨成長板の下部、骨新生部に形成された各蛍光線間の長さ、すなわち成長した長さを測定した。卵黄由来水溶性組成物を与えた場合と基本試料（対照群）を与えた場合の骨の伸長を比較し促進効果を評価した。顕鏡観察の結果、卵黄由来水溶性組成物群でコントロール群に対して２本の染色バンドの間隔がコントロールと比較して３０％広く、骨伸長促進活性があると確認された。
【００３０】
試験例３（軟骨細胞増殖能を評価する動物試験）
実施例１、２で得られた卵黄水溶性組成物の生体における軟骨細胞増殖能を以下のようにして行った。軟骨細胞増殖能の評価は、細胞周期中のＤＮＡ合成能、細胞増殖能を表すブロモデオキシウリジン（ＢｒｄＵ）を用いた取り込み実験により行った。実施例１、２で得られた卵黄水溶性組成物をＳｐｒａｇｕｅ−Ｄａｗｌｅｙラットに対し１００ｍｇ／ｋｇ体重／一日の割合で摂取させた。試験期間中のラットの飲料水と飼料は自由摂取とし、飼料としてＭＦ（商品名、オリエンタル酵母社製）を用いた。摂取期間（１週間）の後、ＢｒｄＵを腹腔内投与（ＢｒｄＵ ５０ｍｇ／ｋｇ）し、投与２時間後に麻酔したラットから脛骨を摘出し成長板の軟骨細胞へのＢｒｄＵの取り込みを抗ＢｒｄＵ抗体を用いる免疫染色法により測定した。試料摂取群ではＢｒｄＵ取り込み活性がコントロールに比べて有意に高かった。このことは試料摂取により軟骨細胞の増殖活性が高くなることを示しており、骨の成長が促進されることが見出された。（図２）
【００３１】
実施例３（清涼飲料水）
実施例２で調製した卵黄水溶性組成物を含有する清涼飲料水を調製した。すなわち、組成が混合異性化糖１５．０％、果汁１０％、卵黄水溶性組成物２．０％、香料 ０．１％、カルシウム０．１％、水 ７２．８％である原料を混合し、プレート殺菌機を用いて９０℃、１５秒間殺菌し、骨強化作用を有する清涼飲料水を製造した。
【００３２】
実施例４（ヨーグルト）
実施例２で調整した卵黄水溶性組成物を含有するヨーグルトを調整した。すなわち、組成が卵黄水溶性組成物３．０％、蔗糖７％、香料０．１％、ヨーグルト８９．９％である原料を混合し容器に充填して、骨強化作用を有するヨーグルトを製造した。
【００３３】
実施例５（チーズ）
実施例２で調整した卵黄水溶性組成物を含有するプロセスチーズを調整した。すなわち、ゴーダチーズ３５％、チェダーチーズ３５％、パルメザンチーズ２０％、卵黄水溶性組成物２．０％、リン酸カルシウム１．０％、水７．０％を含むように各原料を混合後、乳化温度８５℃で乳化して、骨強化作用を有するプロセスチーズを製造した。
【００３４】
実施例６（カプセル剤）
実施例２で得られた卵黄水溶性組成物６０％、コーンスターチ３０％、乳糖１０％を含むように各原料を配合して、ゼラチンカプセルに充填（カプセル１個当たり２００ｍｇ）し、骨強化作用を有するカプセル剤を製造した。
【００３５】
実施例７（錠剤）
実施例２で得られた卵黄水溶性組成物６０％、還元麦芽糖１８％、結晶セルロース１８％、ショ糖エステル４％を含むように各原料を配合後打錠（１錠当たり３００ｍｇ）して、骨強化作用を有する錠剤を製造した。
【００３６】
【発明の効果】
以上のように、本発明の卵由来水溶性組成物は、骨芽細胞増殖促進、石灰化促進活性を有する。さらに、軟骨細胞の増殖を促進して骨の成長を促す活性を有している。また、本発明の骨強化剤は経口摂取が可能であり、チーズ、バター、発酵乳等の乳食品、飲料乳、ドリンクヨーグルト、コーヒー飲料、果汁等の飲料、ゼリー、プリン、クッキー、ビスケット、ウエハース等の菓子、冷凍食品等の各種の形態の飲食品に添加し、骨強化作用を有する飲食品とすることができる。また、カプセル、錠剤などの形態でサプリメントとして用いることもでき、骨粗鬆症の予防、症状改善を目的とした飲食品、医薬または飼料の素材として有用である。
【００３７】
【図面の簡単な説明】
【図１】通常飼料と卵黄水溶性画分を添加した飼料をラットに与えたときの、各飼料摂取群について頚骨端の伸長を比較したグラフである。
【図２】通常飼料、卵黄水溶性画分添加飼料の各飼育群においてラット頚骨成長板の軟骨細胞にＢｒｄＵが取り込まれる様子を表す図である。試料摂取群ではＢｒｄＵ取り込み活性がコントロールに比べて有意に高かった。図では通常飼料摂取群で３細胞の核が染色されているが、卵黄水溶性画分添加飼料摂取群では１１細胞で染色されている。[0001]
BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition containing an egg-derived water-soluble fraction having bone growth and bone strengthening actions. Moreover, this invention relates to the food-drinks, pharmaceuticals, or feed which mix | blended this egg origin water-soluble composition. The egg-derived water-soluble fraction of the present invention has the effect of proliferating osteoblasts and also promoting the proliferation of chondrocytes at the growth end of bone, so that the prevention and treatment of bone diseases caused by the loss of bone metabolism balance, It is useful for bone strengthening during the growth period in young people such as school children.
[0002]
2. Description of the Related Art An increase in various bone diseases such as an increase in osteoporosis in recent years and an increase in the fracture rate in young people has become a social problem. It is estimated that there are about 10 million patients with osteoporosis nationwide, and it is predicted that the number of patients will reach 16 million in 2010 due to the progress of aging. Osteoporosis is attracting attention as a disease in which the balance between bone formation and bone destruction is disrupted mainly by postmenopausal women due to the effects of decreased female hormones. Currently, there are about 2 million osteoporosis patients over 50 years old. In addition, the fracture rate of elementary and junior high school students has more than doubled in the last 20 years, and from the standpoint of preventing osteoporosis, it is important to increase bone mineral content firmly in younger years and obtain a higher maximum bone mass. Very important. Thus, maintaining bone health is now a social concern regardless of gender or age.
[0003]
Conventional methods for preventing or treating bone diseases include dietary calcium supplementation, light exercise, sunbathing, and drug treatment. Calcium supplementation by dietary therapy uses calcium salts such as calcium carbonate and calcium phosphate, and natural calcium preparations such as beef bone meal, eggshell, and fish bone meal, but they are not necessarily soluble, absorbable, and tasty. It is not a material suitable for oral intake. Moderate exercise increases bone mass and strengthens bones, so walking and walking are said to be good for bone health, but light exercise is also troublesome if the body is weak, even bedridden elderly people I can hardly exercise. Sunbathing is considered good in terms of supplementing activated vitamin D3, but this alone is not sufficient. Bisphosphonates and calcitonin preparations are used for drug administration and are known to be effective in the treatment of osteoporosis. However, these substances are medicines themselves, and special care is required for taking them, and there is a risk of side effects such as tinnitus, headache, and loss of appetite, and they cannot be used as a continuous food material.
[0004]
[Problems to be solved by the invention]
Therefore, in the present invention, as a bone strengthening agent that can be ingested on a daily basis, an object is to provide a food material that has a bone strengthening action, is safe and inexpensive, and can be widely used for food and drink. To do.
[0005]
[Means for Solving the Problems]
The egg is a biocapsule that contains all the materials that form the living body and creates the next generation, as shown by the phenomenon that it turns into a chick when heated at 37 ° C for 3 weeks, and has many physiological activities that have not yet been discovered. it is conceivable that. Eggs have a long history of eating, their nutritional value is high and safe, and their value as a food ingredient is very high.
[0006]
Therefore, as a result of searching for components that promote bone growth and bone strengthening in various fractions of eggs, the present inventors have found that the yolk water-soluble fraction has osteoblast proliferation and calcification promoting activity. The inventors have found for the first time and found that oral intake of this ingredient can promote bone growth and improve bone metabolism, and have completed the present invention.
[0007]
That is, the bone strengthening composition of the invention according to claim 1 (hereinafter referred to as the present invention 1) contains an egg-derived water-soluble fraction having an activity of promoting osteoblast proliferation, bone growth and bone strengthening as an active ingredient. It is characterized by doing.
[0008]
In the present invention 2, the water soluble fraction extracted from egg yolk defatted with at least one organic solvent selected from ethanol, isopropanol, hexane, or other organic solvents used for food processing. A fraction is preferred.
[0009]
Moreover, in this invention 3, it is preferable that the said egg yolk water-soluble fraction is an extract obtained by adding a carrageenan solution to whole egg or egg yolk liquid.
[0010]
Furthermore, it is preferable that the food / beverage product, medicine or feed of the present invention 4 is used by blending the egg yolk water-soluble composition obtained in the present invention 1 to 3.
[0011]
ADVANTAGE OF THE INVENTION According to this invention, the bone strengthening raw material and food / beverage products, medicine, or feed which can be ingested safely and cheaply can be provided.
[0012]
DETAILED DESCRIPTION OF THE INVENTION
The eggs used in the present invention include eggs such as chickens, ducks and quails, and chicken eggs are preferably used from the viewpoint of productivity.
[0013]
When using chicken eggs, whole egg liquid, egg yolk liquid, egg white liquid, whole egg powder, egg yolk powder, egg white powder, defatted whole egg powder, or defatted egg yolk powder can be used. Alternatively, defatted egg yolk powder is preferably used.
[0014]
The bone strengthening composition of the present invention is obtained as a supernatant when a powder obtained by defatting whole egg powder using an organic solvent used in food processing is extracted with warm water or 0.5% sodium chloride. be able to. The organic solvent used for degreasing can be any ethanol, isopropanol, hexane, or any other organic solvent used for food processing, but ethanol is preferably used from the viewpoint of simplicity and safety.
[0015]
Moreover, this bone strengthening composition can be obtained as an extract obtained by adding a natural rubbery aqueous solution such as gum arabic, carrageenan, xanthan gum to whole egg liquid or egg yolk liquid. The natural gum aqueous solution used here can be any of gum arabic, carrageenan, xanthan gum and the like, but lambda carrageenan is particularly preferably used from the viewpoint of extraction efficiency.
[0016]
The bone strengthening composition of the present invention is applied to the prevention or improvement of osteoporosis and various bone diseases, and the effective intake is 100 mg to 6 g in terms of egg yolk water-soluble fraction powder per adult day. Moreover, the acute toxicity by a rat was not recognized by the bone strengthening composition of this invention.
[0017]
The bone reinforcing composition of the present invention may be treated with a proteolytic enzyme as necessary. The proteolytic enzyme is not particularly limited as long as it is a proteolytic enzyme that can be used in foods, and any of acidic protease, neutral protease, and alkaline protease can be used. In the present invention, neutral protease is preferably used. Examples of the medieval protease include orientase (trade name, manufactured by HIBI) and flavorzyme (trade name, manufactured by Novo Nordisk).
[0018]
The bone reinforcing composition collected in this way is usually used after being powdered. Powdering may be performed by freeze-drying or spray-drying, and is usually administered orally as a food or drink.
[0019]
In addition to the components described above, the bone reinforcing composition of the present invention includes excipients, emulsifiers, stabilizers, sweeteners, pH adjusters, thickeners, proteins, peptides, amino acids, lipids, polysaccharides, oligosaccharides, etc. Ingredients usually used in foods may be included.
[0020]
The bone reinforcing composition of the present invention is not only added to foods and drinks, but also prepared in various forms according to the dosage form, and can be used as a powder (or granule), a drink, a tablet, a capsule and the like. Egg is a safe material that has been eaten for a long time, and can be used in medicine or feed as a material that does not cause any side effects even when used continuously.
[0021]
【Example】
EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated in detail, this invention is not limited at all by these Examples.
[0022]
Example 1 (Extraction with organic solvent)
Ethanol (2.5 l) was added to egg yolk powder (500 g), and the mixture was stirred with a blender for 30 minutes, and then the solid was collected. This operation was repeated three times to degrease the egg yolk. As a result, 284 g of defatted egg yolk was obtained after air drying. To this defatted egg yolk powder was added 3 liters of warm water at 37 ° C. and well stirred with a stirrer while maintaining the temperature, followed by centrifugation at 8,000 rpm for 30 minutes to obtain a supernatant. And it pulverized by freeze-drying and obtained 13.4 g of egg yolk water-soluble composition.
[0023]
Example 2 (Extraction with carrageenan)
3 L of 1% gamma carrageenan solution is added to 500 g of egg yolk powder, and after gently stirring, agglomeration is observed when left for 30 minutes. Thereafter, centrifugation was performed at 8,000 rpm for 30 minutes to obtain a supernatant. This was pulverized by freeze-drying to obtain 14.8 g of an egg yolk water-soluble composition.
[0024]
The water-soluble compositions obtained in Examples 1 and 2 were analyzed for molecular weight by gel filtration chromatography under the following conditions.
[0025]
Column: YMC-Pack
Diol 200 (6.0 Ï 300 mm) (trade name, manufactured by YMC Corporation)
Eluent: 0.2M potassium phosphate buffer (pH 6.9)
Flow rate: 0.7 ml / min Detection wavelength: 280 nm
[0026]
As a result, the egg yolk water-soluble compositions of Examples 1 and 2 showed the same main peak distribution at a molecular weight of 10,000 to 200,000.
[0027]
Test Example 1 (Measurement of osteoblast proliferation activity)
The osteoblast proliferation activity of the egg yolk water-soluble composition obtained in Example 1 was measured as follows. Human osteoblastic InM1.2 cells were cultured in α-MEM culture medium containing 10% FBS at 37 ° C. under 5% CO 2 -95% air, cultured until confluent, and then treated with trypsin. Were collected and adjusted to 1 × 10 4 cells / mL using the above culture solution. 0.5 mL each of this cell preparation was seeded on a 24 well plate and cultured at 37 ° C. under 5% CO 2 -95% air. On the next day, the sample was added to the cells at 100, 25, and 12.5 ppm, and α-glycerophosphate (2 mmol / L) as a mineralization accelerator was added. The sample was dissolved in 50% DMSO and added so that the solvent was 1% of the culture solution. The culture medium was changed every other day, and after 20 days, the number of osteoblasts was measured using MTT. The growth promoting activity of osteoblasts was expressed as a relative value when the growth number of the control group was 100. The results are shown in Table 1.
[0028]
[Table 1]
Test Example 2 (Bone growth promoting activity)
The bone growth promoting activity of the egg yolk water-soluble composition obtained in Example 1 was measured as follows. Bone growth promoting activity was evaluated using Sprague-Dawley rats. Samples were dissolved in purified water at a concentration of 20 mg / ml and rats were fed 20 or 100 mg / kg body weight / day. Rat drinking water and feed during the test period were freely consumed, and MF (trade name, manufactured by Oriental Yeast Co., Ltd.) was used as the basic feed. Tetracycline was administered as a fluorescent substance on the third and fifth days of the sample intake period (one week), respectively. In this test example, the growth promoting effect by the egg protein composition of the tibia was measured by observing the fluorescence line formed in the bone by utilizing the phenomenon that tetracycline chelates calcium and is deposited on the bone. The tibia was removed, and tetracycline was administered to measure the length between the fluorescent lines formed at the lower part of the tibia growth plate and at the new bone part, that is, the length of growth. The promotion effect was evaluated by comparing the bone elongation when the egg yolk-derived water-soluble composition was given and when the basic sample (control group) was given. As a result of microscopic observation, the interval between the two stained bands in the yolk-derived water-soluble composition group was 30% wider than that in the control group, and it was confirmed that there was activity to promote bone elongation.
[0030]
Test Example 3 (Animal test for evaluating chondrocyte proliferation ability)
The ability of the yolk water-soluble composition obtained in Examples 1 and 2 to proliferate chondrocytes in the living body was determined as follows. Evaluation of chondrocyte proliferative ability was performed by an uptake experiment using bromodeoxyuridine (BrdU) representing DNA synthesis ability and cell proliferative ability in the cell cycle. The egg yolk water-soluble composition obtained in Examples 1 and 2 was ingested at a rate of 100 mg / kg body weight / day to Sprague-Dawley rats. Rat drinking water and feed during the test period were freely ingested, and MF (trade name, manufactured by Oriental Yeast Co., Ltd.) was used as the feed. After the ingestion period (1 week), BrdU was intraperitoneally administered (BrdU 50 mg / kg), 2 hours after administration, the tibia was removed from the anesthetized rat, and BrdU incorporation into the chondrocytes of the growth plate was used using an anti-BrdU antibody. Measurement was performed by immunostaining. In the sample intake group, BrdU incorporation activity was significantly higher than that of the control. This indicates that the chondrocyte proliferation activity is increased by ingestion of the sample, and it has been found that bone growth is promoted. (Figure 2)
[0031]
Example 3 (soft drink)
A soft drink containing the egg yolk water-soluble composition prepared in Example 2 was prepared. That is, the raw materials whose composition is mixed isomerized sugar 15.0%, fruit juice 10%, egg yolk water-soluble composition 2.0%, fragrance 0.1%, calcium 0.1%, water 72.8% are mixed. Then, using a plate sterilizer, sterilized at 90 ° C. for 15 seconds to produce a soft drink having bone strengthening action.
[0032]
Example 4 (yogurt)
A yogurt containing the egg yolk water-soluble composition prepared in Example 2 was prepared. That is, yoghurt having a bone strengthening action was manufactured by mixing raw materials having a composition of 3.0% egg yolk water-soluble composition, 7% sucrose, 0.1% fragrance, and 89.9% yoghurt and filling the container. .
[0033]
Example 5 (cheese)
Processed cheese containing the egg yolk water-soluble composition prepared in Example 2 was prepared. That is, after mixing each raw material to include 35% Gouda cheese, 35% Cheddar cheese, 20% Parmesan cheese, 2.0% egg yolk water-soluble composition, 1.0% calcium phosphate, and 7.0% water, the emulsification temperature Emulsified at 85 ° C. to produce a processed cheese having a bone strengthening action.
[0034]
Example 6 (capsule)
Each raw material was blended so as to contain 60% egg yolk water-soluble composition obtained in Example 2, corn starch 30%, and lactose 10%, and filled into gelatin capsules (200 mg per capsule) to provide bone strengthening action. The capsule which has was manufactured.
[0035]
Example 7 (tablet)
Each raw material was mixed so as to contain 60% egg yolk water-soluble composition obtained in Example 2, reduced maltose 18%, crystalline cellulose 18%, sucrose ester 4%, and then tableted (300 mg per tablet). Tablets with bone strengthening action were produced.
[0036]
【The invention's effect】
As described above, the egg-derived water-soluble composition of the present invention has osteoblast proliferation promotion and calcification promoting activity. Furthermore, it has the activity of promoting the growth of chondrocytes and promoting bone growth. In addition, the bone strengthening agent of the present invention can be taken orally, and milk products such as cheese, butter, fermented milk, beverage milk, drink yogurt, coffee beverage, fruit juice beverage, jelly, pudding, cookies, biscuits, wafers It can be added to foods and drinks of various forms such as confectionery and frozen foods, etc. to make foods and drinks having a bone strengthening action. It can also be used as a supplement in the form of capsules, tablets, etc., and is useful as a material for foods, beverages, medicines or feeds for the purpose of preventing osteoporosis and improving symptoms.
[0037]
[Brief description of the drawings]
FIG. 1 is a graph comparing the extension of the tibia at each feed intake group when a normal feed and a feed supplemented with an egg yolk water-soluble fraction were given to rats.
FIG. 2 is a view showing a state in which BrdU is taken into chondrocytes of a rat tibia growth plate in each feeding group of a normal feed and a yolk water-soluble fraction-added feed. In the sample intake group, BrdU incorporation activity was significantly higher than that of the control. In the figure, the nucleus of 3 cells is stained in the normal feed intake group, but it is stained in 11 cells in the feed intake group supplemented with egg yolk water-soluble fraction.
Claims (4)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003190906A JP2005021087A (en) | 2003-07-03 | 2003-07-03 | Egg-derived bone-strengthening composition |
| KR1020040024785A KR20050003989A (en) | 2003-07-03 | 2004-04-10 | Egg derivatives having bone-strengthening effects |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003190906A JP2005021087A (en) | 2003-07-03 | 2003-07-03 | Egg-derived bone-strengthening composition |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2005021087A true JP2005021087A (en) | 2005-01-27 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2003190906A Pending JP2005021087A (en) | 2003-07-03 | 2003-07-03 | Egg-derived bone-strengthening composition |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP2005021087A (en) |
| KR (1) | KR20050003989A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006075558A1 (en) * | 2005-01-11 | 2006-07-20 | Pharma Foods International Co., Ltd. | Egg-derived bone-strengthening composition |
| WO2011122127A1 (en) * | 2010-03-31 | 2011-10-06 | 株式会社ファーマフーズ | Peptide having osteoblast proliferation promotion activity and use thereof |
| CN103584106A (en) * | 2013-11-29 | 2014-02-19 | 哈药集团中药二厂 | Composition capable of improving bone mineral density |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20230131437A (en) * | 2022-03-04 | 2023-09-13 | (주)메디언스 | Composition for increasing height growth and bone density derived from egg yolk protein |
-
2003
- 2003-07-03 JP JP2003190906A patent/JP2005021087A/en active Pending
-
2004
- 2004-04-10 KR KR1020040024785A patent/KR20050003989A/en not_active Application Discontinuation
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006075558A1 (en) * | 2005-01-11 | 2006-07-20 | Pharma Foods International Co., Ltd. | Egg-derived bone-strengthening composition |
| WO2011122127A1 (en) * | 2010-03-31 | 2011-10-06 | 株式会社ファーマフーズ | Peptide having osteoblast proliferation promotion activity and use thereof |
| JP2011211979A (en) * | 2010-03-31 | 2011-10-27 | Pharma Foods International Co Ltd | Peptide having osteoblast proliferation promotion activity and use thereof |
| CN103584106A (en) * | 2013-11-29 | 2014-02-19 | 哈药集团中药二厂 | Composition capable of improving bone mineral density |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20050003989A (en) | 2005-01-12 |
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