JP6917164B2 - Composition for reducing endoplasmic reticulum stress and foods, pharmaceuticals, feeds containing the composition - Google Patents
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Description
本発明は、小胞体ストレス軽減用組成物及び該組成物を含む食品、医薬品、飼料に関する。 The present invention relates to a composition for reducing endoplasmic reticulum stress and foods, pharmaceuticals, and feeds containing the composition.
小胞体とは、細胞内に存在する膜の約半分を占める小器官であり、タンパク質合成、膜脂質合成、カルシウム貯蔵など、多岐にわたる生理機能を有している。
分泌タンパク質や膜タンパク質は小胞体膜に結合したリボソームにおいて合成され、小胞体に挿入される。これらのタンパク質は、小胞体において分子シャペロンなどの作用により様々な化学修飾を受け、糖鎖の付与、ジスルフィド結合の形成などを経て適正な立体構造が与えられる。
このとき、正常な立体構造を持ったタンパク質のみがゴルジ装置以降の分泌経路に進み、細胞膜や細胞外へと輸送され、異常な立体構造を持ったタンパク質は小胞体関連分解により処分される。
The endoplasmic reticulum is an organelle that occupies about half of the membrane existing in a cell, and has a wide range of physiological functions such as protein synthesis, membrane lipid synthesis, and calcium storage.
Secretory and membrane proteins are synthesized in the ribosome bound to the endoplasmic reticulum membrane and inserted into the endoplasmic reticulum. These proteins undergo various chemical modifications in the endoplasmic reticulum by the action of molecular chaperones and the like, and are given an appropriate three-dimensional structure through the addition of sugar chains and the formation of disulfide bonds.
At this time, only proteins having a normal three-dimensional structure proceed to the secretory pathway after the Golgi apparatus and are transported to the cell membrane or extracellularly, and proteins having an abnormal three-dimensional structure are disposed of by endoplasmic reticulum-related degradation.
虚血、低酸素、熱ショック、遺伝子変異などの要因により、小胞体内に正常な高次構造を持たないタンパク質（ｕｎｆｏｌｄｅｄ ｐｒｏｔｅｉｎ）が蓄積し、小胞体の機能障害を引き起こすことが知られている（小胞体ストレス）。
細胞はこの小胞体ストレスに対して、小胞体の状態を正常に向かわせるための防御システムである小胞体ストレス応答（ｕｎｆｏｌｄｅｄ ｐｒｏｔｅｉｎ ｒｅｓｐｏｎｓｅ）を働かせる。
小胞体ストレス応答は、ストレスが軽度のときは、タンパク質翻訳の抑制による小胞体への流入量の抑制、分子シャペロンの誘導による変性タンパク質の修復、修復できないタンパク質の小胞体外への排出と分解を行うことにより細胞機能を維持する方向へと向かわせるが、過度なストレスを受けたときにはアポトーシスが誘導され個体の保護が優先される。
It is known that due to factors such as ischemia, hypoxia, heat shock, and gene mutation, proteins (unfolded proteins) that do not have a normal higher-order structure accumulate in the endoplasmic reticulum and cause dysfunction of the endoplasmic reticulum. (Endoplasmic reticulum stress).
In response to this endoplasmic reticulum stress, cells activate the endoplasmic reticulum stress response (unfolded protein response), which is a defense system for normalizing the state of the endoplasmic reticulum.
The endoplasmic reticulum stress response, when the stress is mild, suppresses the influx into the endoplasmic reticulum by suppressing protein translation, repairs denatured proteins by inducing molecular chaperones, and excretes and decomposes irreparable proteins out of the endoplasmic reticulum. By doing so, the cell function is maintained, but when excessive stress is applied, apoptosis is induced and the protection of the individual is prioritized.
近年、小胞体ストレスおよび小胞体ストレス応答の破綻は、パーキンソン病やアルツハイマー病、ポリグルタミン病、プリオン病、筋萎縮生側索硬化症（ＡＬＳ）等に代表される神経変性疾患、糖尿病、脂肪肝など非常に多様な疾患の発症に関わっていることが報告されている。
従って、小胞体ストレスを軽減することは、上記疾患の予防または治療に有効であることが期待される。
In recent years, the disruption of endoplasmic reticulum stress and endoplasmic reticulum stress response has been caused by neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, polyglutamine disease, prion disease, amyotrophic lateral sclerosis (ALS), diabetes, and fatty liver. It has been reported that it is involved in the development of a wide variety of diseases.
Therefore, reducing endoplasmic reticulum stress is expected to be effective in the prevention or treatment of the above-mentioned diseases.
上記のことから、小胞体ストレスを軽減する食品、医薬品、飼料の開発が望まれている。
特許文献１は黒米を有効成分とする小胞体ストレス軽減剤を開示している。
特許文献２はポリヌクレオチドを有効成分とする小胞体ストレス調節剤を開示している。
また、非特許文献１は、乳由来リン脂質混合物が小胞体ストレスにより誘導される神経細胞死を抑制することを開示している。
しかし、小胞体ストレスを軽減する組成物として、β−ラクトグロブリン及び／又は乳由来アルブミンを含む組成物を開示した文献等はない。さらに該組成物を含む食品、医薬品、飼料を開示した文献等もない。
From the above, the development of foods, pharmaceuticals, and feeds that reduce endoplasmic reticulum stress is desired.
Patent Document 2 discloses a endoplasmic reticulum stress regulator containing a polynucleotide as an active ingredient.
In addition, Non-Patent
However, there is no literature that discloses a composition containing β-lactoglobulin and / or milk-derived albumin as a composition for reducing endoplasmic reticulum stress. Furthermore, there is no literature that discloses foods, pharmaceuticals, and feeds containing the composition.
本発明は、新たな小胞体ストレスを軽減する組成物として、β‐ラクトグロブリン及び／又は乳由来アルブミンを含む組成物と該組成物を含む食品、医薬品、及び飼料を提供することを課題とする。 An object of the present invention is to provide a composition containing β-lactoglobulin and / or milk-derived albumin and a food, pharmaceutical, and feed containing the composition as a new composition for reducing endoplasmic reticulum stress. ..
上記課題を解決するため、本発明者らは、小胞体ストレスを軽減する有用な方法について検討を進めた結果、β−ラクトグロブリン及び／又は乳由来アルブミンが生体内の小胞体ストレスを軽減させることを見出し、本発明を完成するに至った。すなわち、本発明には以下の構成が含まれる。
（１）β−ラクトグロブリン及び／又はβ−ラクトグロブリン加水分解物を有効成分とする小胞体ストレス軽減用組成物。
（２）乳由来アルブミン及び／又は乳由来アルブミン加水分解物を有効成分とする小胞体ストレス軽減用組成物。
（３）β−ラクトグロブリン及び／又はβ−ラクトグロブリン加水分解物と、乳由来アルブミン及び／又は乳由来アルブミン加水分解物とを有効成分とする小胞体ストレス軽減用組成物。
（４）（１）から（３）のいずれかに記載の小胞体ストレス軽減用組成物を含む小胞体ストレス軽減用食品。
（５）（１）から（３）のいずれかに記載の小胞体ストレス軽減用組成物を含む小胞体ストレス軽減用医薬品。
（６）（１）から（３）のいずれかに記載の小胞体ストレス軽減用組成物を含む小胞体ストレス軽減用飼料。
In order to solve the above problems, the present inventors have studied a useful method for reducing endoplasmic reticulum stress, and as a result, β-lactoglobulin and / or milk-derived albumin reduce endoplasmic reticulum stress in vivo. The present invention has been completed. That is, the present invention includes the following configurations.
(1) A composition for reducing endoplasmic reticulum stress containing β-lactoglobulin and / or β-lactoglobulin hydrolyzate as an active ingredient.
(2) A composition for reducing endoplasmic reticulum stress containing milk-derived albumin and / or milk-derived albumin hydrolyzate as an active ingredient.
(3) A composition for reducing endoplasmic reticulum stress containing β-lactoglobulin and / or β-lactoglobulin hydrolyzate and milk-derived albumin and / or milk-derived albumin hydrolyzate as active ingredients.
(4) A food for reducing endoplasmic reticulum stress containing the composition for reducing endoplasmic reticulum stress according to any one of (1) to (3).
(5) A drug for reducing endoplasmic reticulum stress, which comprises the composition for reducing endoplasmic reticulum stress according to any one of (1) to (3).
(6) A feed for reducing endoplasmic reticulum stress containing the composition for reducing endoplasmic reticulum stress according to any one of (1) to (3).
本発明は、新たな小胞体ストレスを軽減する組成物として、β−ラクトグロブリン及び／又は乳由来アルブミンと該組成物を含む食品、医薬品、及び飼料を提供するものである。本発明の組成物の摂取により小胞体ストレスを安全かつ簡易に軽減することが可能となった。 The present invention provides foods, pharmaceuticals, and feeds containing β-lactoglobulin and / or milk-derived albumin and the composition as a novel composition for reducing endoplasmic reticulum stress. Ingestion of the composition of the present invention has made it possible to safely and easily reduce endoplasmic reticulum stress.
本発明の小胞体ストレス軽減用組成物について以下に詳細に説明する。 The composition for reducing endoplasmic reticulum stress of the present invention will be described in detail below.
（β‐ラクトグロブリン）
本発明の小胞体ストレス軽減用組成物に用いるβ‐ラクトグロブリンについて説明する。
本発明の小胞体ストレス軽減用組成物に用いるβ‐ラクトグロブリンは、ウシ、馬、羊、山羊等の獣乳中に存在するβ‐ラクトグロブリンを用いることができ、前記獣乳から得られたもの、合成したもののいずれも用いることができる。
また、β‐ラクトグロブリンをタンパク質分解酵素で処理したβ‐ラクトグロブリン分解物も用いることができる。β‐ラクトグロブリン分解物の分子量は２００Ｄａ程度以上１０ＫＤａ程度以下であればよく、２００Ｄａ程度以１ＫＤａ上程度以下がより好ましい。
(Β-lactoglobulin)
The β-lactoglobulin used in the composition for reducing endoplasmic reticulum stress of the present invention will be described.
As the β-lactoglobulin used in the composition for reducing endoplasmic reticulum stress of the present invention, β-lactoglobulin existing in animal milk of cows, horses, sheep, goats and the like can be used, and it was obtained from the animal milk. Both the one and the synthesized one can be used.
In addition, a β-lactoglobulin degradation product obtained by treating β-lactoglobulin with a proteolytic enzyme can also be used. The molecular weight of the β-lactoglobulin decomposition product may be about 200 Da or more and about 10 kDa or less, and more preferably about 200 Da or more and about 1 kDa or less.
（乳由来アルブミン）
本発明の小胞体ストレス軽減用組成物に用いる乳由来アルブミンについて説明する。
本発明の小胞体ストレス軽減用組成物に用いる乳由来アルブミンは、ウシ、馬、羊、山羊等の獣乳中に存在する乳由来アルブミンを用いることができ、前記獣乳から得られたもの、合成したもののいずれも用いることができる。
また、乳由来アルブミンをタンパク質分解酵素で処理した乳由来アルブミン加水分解物も用いることができる。乳由来アルブミン加水分解物の分子量は２００Ｄａ程度以上２０ＫＤａ程度以下であればよく、２００Ｄａ程度以上５ＫＤａ程度以下がより好ましい。
(Milk-derived albumin)
The milk-derived albumin used in the composition for reducing endoplasmic reticulum stress of the present invention will be described.
As the milk-derived albumin used in the composition for reducing endoplasmic reticulum stress of the present invention, milk-derived albumin existing in animal milk of cows, horses, sheep, goats and the like can be used, and those obtained from the above-mentioned animal milk, Any of the synthesized ones can be used.
In addition, a milk-derived albumin hydrolyzate obtained by treating milk-derived albumin with a proteolytic enzyme can also be used. The molecular weight of the milk-derived albumin hydrolyzate may be about 200 Da or more and about 20 KDa or less, and more preferably about 200 Da or more and about 5 KDa or less.
本発明の小胞体ストレス軽減用組成物には、β‐ラクトグロブリン及び／又は乳由来アルブミンを含む乳素材をそのまま用いることもできる。
前記乳素材はβ‐ラクトグロブリン及び／又は乳由来アルブミンを含むものであればどのようなものでも用いることができ、脱脂粉乳、甘性ホエイ粉、酸ホエイ粉、脱塩ホエイ粉などを例示できる。
このうち、β‐ラクトグロブリン、乳由来アルブミンの含量の高いＷＰＩ（Ｗｈｅｙ Ｐｒｏｔｅｉｎ Ｉｓｏｌａｔｅ、分離乳清タンパク質やＷＰＣ（Ｗｈｅｙ Ｐｒｏｔｅｉｎ Ｃｏｎｃｅｎｔｒａｔｅ、濃縮乳清タンパク質）を用いることが好ましい。
β‐ラクトグロブリン及び／又は乳由来アルブミンの分解物は、β‐ラクトグロブリン、乳由来アルブミン、あるいはβ‐ラクトグロブリン及び／又は乳由来アルブミンを含む乳素材をタンパク質分解酵素で処理したものを用いればよい。
タンパク質分解酵素はβ‐ラクトグロブリン及び／又は乳由来アルブミンのペプチド結合を加水分解できるものであればどのようなものでも用いることができるが、食品製造に用いられるペプシン、トリプシン、パパイン等が好ましい。
In the composition for reducing endoplasmic reticulum stress of the present invention, a milk material containing β-lactoglobulin and / or milk-derived albumin can be used as it is.
Any milk material can be used as long as it contains β-lactoglobulin and / or milk-derived albumin, and examples thereof include skim milk powder, sweet whey powder, acid whey powder, and desalted whey powder. ..
Of these, it is preferable to use WPI (Whey Protein Isolate, isolated whey protein, WPC (Whey Protein Concentrate, concentrated whey protein)) having a high content of β-lactoglobulin and milk-derived albumin.
As the degradation product of β-lactoglobulin and / or milk-derived albumin, β-lactoglobulin, milk-derived albumin, or a milk material containing β-lactoglobulin and / or milk-derived albumin treated with proteolytic enzyme can be used. good.
Any proteolytic enzyme can be used as long as it can hydrolyze the peptide bond of β-lactoglobulin and / or milk-derived albumin, but pepsin, trypsin, papain and the like used in food production are preferable.
（小胞体ストレス軽減用組成物）
本発明の小胞体ストレス軽減用組成物は、β−ラクトグロブリン及び／又は乳由来アルブミンを含むものである。小胞体ストレス軽減用組成物におけるβ−ラクトグロブリン及び／又は乳由来アルブミンの含量は、１重量％以上１００重量％以下であればよく、３０重量％以上１００重量％以下が好ましく、４０重量％以上１００重量％以下が最も好ましい。
(Composition for reducing endoplasmic reticulum stress)
The composition for reducing endoplasmic reticulum stress of the present invention contains β-lactoglobulin and / or milk-derived albumin. The content of β-lactoglobulin and / or milk-derived albumin in the composition for reducing endoplasmic reticulum stress may be 1% by weight or more and 100% by weight or less, preferably 30% by weight or more and 100% by weight or less, and 40% by weight or more. Most preferably 100% by weight or less.
（小胞体ストレス軽減用組成物の摂取量）
本発明の小胞体ストレス軽減用組成物の有効摂取量は、β−ラクトグロブリン及び／又は、乳由来アルブミンの摂取量として２ｍｇ／日以上となるように摂取すればよい。
また、本発明の小胞体ストレス軽減用組成物は、以下に記載のとおり、食品、医薬品、飼料に添加することができるが、その際の小胞体ストレス軽減用組成物の添加量は前記した有効量が摂取できるよう適宜調整すればよい。
(Intake of composition for reducing endoplasmic reticulum stress)
The effective intake of the endoplasmic reticulum stress reducing composition of the present invention may be 2 mg / day or more as the intake of β-lactoglobulin and / or milk-derived albumin.
Further, the composition for reducing endoplasmic reticulum stress of the present invention can be added to foods, pharmaceuticals and feeds as described below, and the amount of the composition for reducing endoplasmic reticulum stress at that time is effective as described above. It may be adjusted appropriately so that the amount can be ingested.
（小胞体ストレス軽減用組成物を含む食品、医薬品、及び飼料）
本発明の小胞体ストレス軽減用組成物はそのまま使用してもよいが、食品、医薬品、及び飼料に通常含まれる他の原材料とともに使用できることから、常法に従い、粉末剤、顆粒剤、錠剤、カプセル剤、ドリンク剤等に用いることも出来る。また、ヨーグルト、乳飲料、ウエハース等の飲食品、及び飼料に配合することも可能である。
(Foods, medicines, and feeds containing compositions for reducing endoplasmic reticulum stress)
The composition for reducing endoplasmic reticulum stress of the present invention may be used as it is, but since it can be used together with other raw materials usually contained in foods, pharmaceuticals, and feeds, powders, granules, tablets, and capsules are used in accordance with a conventional method. It can also be used as an agent, a drink agent, and the like. It can also be added to foods and drinks such as yogurt, milk drinks and wafers, and feed.
（小胞体ストレス軽減用組成物中のβ‐ラクトグロブリンの定量方法）
本発明の小胞体ストレス軽減用組成物中のβ‐ラクトグロブリン含量は、Ｂｏｒｄｉｎらの方法（Ｊｏｕｒｎａｌ ｏｆ Ｃｈｒｏｍａｔｏｇｒａｐｈｙ Ａ.(２００１)９２８、１、６３‐７６）により測定することができる。
(Method for quantifying β-lactoglobulin in composition for reducing endoplasmic reticulum stress)
The β-lactoglobulin content in the composition for reducing endoplasmic reticulum stress of the present invention can be measured by the method of Bordin et al. (Journal of Chromatografy A. (2001) 928, 1, 63-76).
（小胞体ストレス軽減用組成物中の乳由来アルブミンの定量方法）
本発明の小胞体ストレス軽減用組成物中の乳由来アルブミン含量は、Ｓｔｕｒａｒｏらの方法（Ｊｏｕｒｎａｌ ｏｆ Ｄａｉｒｙ Ｓｃｉｅｎｃｅ (２０１６)９９、１、６８‐７６）により測定することができる。
(Method for quantifying milk-derived albumin in composition for reducing endoplasmic reticulum stress)
The milk-derived albumin content in the composition for reducing endoplasmic reticulum stress of the present invention can be measured by the method of Sturaro et al. (Journal of Dairy Science (2016) 99, 1, 68-76).
（小胞体ストレス軽減用組成物の小胞体ストレス軽減作用の評価）
本発明の小胞体ストレス軽減用組成物の小胞体ストレス軽減作用は、例えば、小胞体ストレス誘導体等により試験対象となる動物や細胞にストレスを与えた場合において、本発明組成物を付与した場合と付与しなかった場合におけるストレス応答を比較することにより評価できる。小胞体ストレス誘導剤には、例えばツニカマイシンが挙げられる。ツニカマイシンはタンパク質への糖鎖の付与を阻害することで、小胞体内に構造が異常なタンパク質を蓄積させ、小胞体ストレスを引き起こすことが知られており、小胞体ストレスを対象とした研究で広く用いられる。ストレス応答の評価指標は、細胞障害性、細胞生存率、小胞体ストレス応答関連遺伝子の発現などが挙げられる。より具体的には、実施例に示したｉｎ ｖｉｔｒｏにおける方法で評価することができる。
(Evaluation of endoplasmic reticulum stress reducing effect of composition for reducing endoplasmic reticulum stress)
The endoplasmic reticulum stress-reducing effect of the endoplasmic reticulum stress-reducing composition of the present invention is, for example, when the composition of the present invention is applied when stress is applied to an animal or cell to be tested by an endoplasmic reticulum stress derivative or the like. It can be evaluated by comparing the stress responses when not given. Examples of the endoplasmic reticulum stress inducer include tunicamycin. Tunicamycin is known to cause endoplasmic reticulum stress by accumulating proteins with abnormal structures in the endoplasmic reticulum by inhibiting the donation of sugar chains to proteins. Used. Evaluation indexes of stress response include cytotoxicity, cell viability, expression of endoplasmic reticulum stress response-related genes, and the like. More specifically, it can be evaluated by the in vitro method shown in the examples.
以下、本発明の実施例を詳細に説明するが、本発明はこれらに限定されるものではない。
〔実施例品１〕
チーズホエイ限外濾過に供することで得られる濃縮液を噴霧乾燥することで、タンパク質含量９２重量％、β−ラクトグロブリン含量６４．４重量％、乳由来アルブミン含量８重量％のＷＰＩを得た（実施例品１）。
Hereinafter, examples of the present invention will be described in detail, but the present invention is not limited thereto.
[Example product 1]
By spray-drying the concentrate obtained by subjecting to cheese whey ultrafiltration, WPI having a protein content of 92% by weight, a β-lactoglobulin content of 64.4% by weight, and a milk-derived albumin content of 8% by weight was obtained ( Example product 1).
〔実施例品２〕β‐ラクトグロブリン精製物の調製方法
実施例品１を用いて、Ａｌｏｍｉｒａｈらの方法（Ｉｎｔｅｒｎａｔｉｏｎａｌ Ｄａｉｒｙ Ｊｏｕｒｎａｌ、(２００４) １４、４１１-４１９）によりβ−ラクトグロブリン精製物（実施例品２）を得た。本β‐ラクトグロブリン精製物は固形あたり９９．９重量％以上のβ−ラクトグロブリンを含有していた。
[Example product 2] Method for preparing β-lactoglobulin purified product Using
〔実施例品３〕乳由来アルブミン精製物の調製方法
実施例品１を１０重量％で水に溶解し、井上らの方法（特開平７‐１７３１９０号）により乳由来アルブミン精製物（実施例品３）を得た。本乳由来アルブミン精製物は固形あたり９９．９重量％以上の乳由来アルブミンを含有していた。
[Example Product 3] Method for Preparing Milk-Derived Albumin Purified
〔試験例１〕
１．試験方法
細胞実験は、下記の手順により行なった。
ヒト肝癌由来細胞を５×１０４ｃｅｌｌｓ／ｗｅｌｌの密度で９６ウェルプレートに播種し、３７℃で２４時間培養後、無血清培地（Ｎｏｒｍａｌ）、小胞体ストレス誘導剤（ツニカマイシン（４０μｇ／ｍｌ））を混合した無血清培地（Ｃｏｎｔｒｏｌ）、ツニカマイシン（４０μｇ／ｍｌ）および実施例品１のＷＰＩ（０．２５ｍｇ／ｍｌ（β−ラクトグロブリン０．１６１ｍｇ／ｍｌ、乳由来アルブミン０．０２ｍｇ／ｍｌ），０．５ｍｇ／ｍｌ（β−ラクトグロブリン０．３２２ｍｇ／ｍｌ、乳由来アルブミン０．０４ｍｇ／ｍｌ），１．０ｍｇ／ｍｌ（β−ラクトグロブリン０．６４４ｍｇ／ｍｌ、乳由来アルブミン０．０８ｍｇ／ｍｌ））、実施例品２のβ−ラクトグロブリン（０．２５、０．５、１．０ｍｇ／ｍｌ）、実施例品３の乳由来アルブミン（０．２５、０．５、１．０ｍｇ/ｍｌ）、あるいは卵白アルブミン（以下ＯＶＡ、シグマ‐アルドリッチ；１ｍｇ／ｍｌ）を混合した無血清培地に培地交換した（各群ｎ＝３）。
その後、３７℃で２４時間培養し、その培養上清を回収し、ＬＤＨアッセイ（細胞障害性検出キットｐｌｕｓ（ＬＤＨ）、ロシュ・ライフサイエンス）に供し、細胞障害性を下記式により求め評価した。
〔式〕
Ａ：各サンプル上清の吸光度
Ｂ：高コントロール（細胞溶解物と培養上清の吸高度）
Ｃ：低コントロール（細胞を培養していない培地の吸光度）
細胞障害性（％）＝{（Ａ−Ｃ）／（Ｂ−Ｃ）}×１００
結果は、統計処理ソフトを用いて処理し、Ｄｕｎｎｅｔｔ型の多重比較によりコントロール群とそれ以外の群との差をそれぞれ検定した。
[Test Example 1]
1. 1. Test method The cell experiment was carried out according to the following procedure.
Human liver cancer-derived cells were seeded on a 96-well plate at a density of 5 × 10 4 cells / well, cultured at 37 ° C. for 24 hours, and then serum-free medium (Normal) and endoplasmic reticulum stress inducer (tunicamycin (40 μg / ml)). Serum-free medium (Control) mixed with Tunicamycin (40 μg / ml) and WPI of Example Product 1 (0.25 mg / ml (β-lactoglobulin 0.161 mg / ml, milk-derived albumin 0.02 mg / ml), 0.5 mg / ml (β-lactoglobulin 0.322 mg / ml, milk-derived albumin 0.04 mg / ml), 1.0 mg / ml (β-lactoglobulin 0.644 mg / ml, milk-derived albumin 0.08 mg / ml) )), β-lactoglobulin of Example product 2 (0.25, 0.5, 1.0 mg / ml), milk-derived albumin of Example product 3 (0.25, 0.5, 1.0 mg / ml) ) Or egg white albumin (hereinafter referred to as OVA, sigma-aldrich; 1 mg / ml) was replaced with a serum-free medium (n = 3 in each group).
Then, the cells were cultured at 37 ° C. for 24 hours, and the culture supernatant was collected and subjected to an LDH assay (cytotoxicity detection kit plus (LDH), Roche Life Science), and cytotoxicity was calculated and evaluated by the following formula.
〔formula〕
A: Absorbance of each sample supernatant B: High control (absorption altitude of cell lysate and culture supernatant)
C: Low control (absorbance of medium in which cells are not cultured)
Cytotoxicity (%) = {(AC) / (BC)} × 100
The results were processed using statistical processing software, and the differences between the control group and the other groups were tested by Dunnett's multiple comparison.
２．試験結果
図１に示すとおり、１ｍｇ／ｍｌのＯＶＡ添加によってツニカマイシン誘導性細胞障害は抑制されなかった。
これに対して、実施例品３の乳由来アルブミンは１ｍｇ／ｍｌの添加でツニカマイシン誘導性細胞障害を抑制した。実施例品２のβ−ラクトグロブリンは０．２５ｍｇ／ｍｌの添加で効果が確認され、用量依存的にツニカマイシン誘導性細胞障害を抑制した。
また、β‐ラクトグロブリンと乳由来アルブミンを含む実施例品１もツニカマイシン誘導性細胞障害を用量依存的に抑制した。
2. Test Results As shown in FIG. 1, addition of 1 mg / ml OVA did not suppress tunicamycin-induced cytotoxicity.
On the other hand, the milk-derived albumin of
In addition, Example 1 containing β-lactoglobulin and milk-derived albumin also suppressed tunicamycin-induced cytotoxicity in a dose-dependent manner.
〔実施例品４〕小胞体ストレス軽減用カプセル剤
表１に示す配合で原材料を混合後、常法により造粒し、カプセルに充填して、小胞体ストレス軽減用カプセル剤を製造した。このカプセル剤にはβ−ラクトグロブリンが２０．０重量％含まれていた。
[Example product 4] Capsules for reducing endoplasmic reticulum stress After mixing the raw materials with the formulation shown in Table 1, granulated by a conventional method and filled into capsules to produce capsules for reducing endoplasmic reticulum stress. The capsule contained 20.0% by weight of β-lactoglobulin.
〔実施例品５〕小胞体ストレス軽減用カプセル剤
表２に示す配合で原材料を混合後、常法により造粒し、カプセルに充填して、小胞体ストレス軽減用カプセル剤を製造した。このカプセル剤には乳由来アルブミンが５．０重量％含まれていた。
[Example product 5] Capsules for reducing endoplasmic reticulum stress The raw materials were mixed according to the formulation shown in Table 2, granulated by a conventional method, and filled into capsules to produce capsules for reducing endoplasmic reticulum stress. The capsule contained 5.0% by weight of milk-derived albumin.
〔実施例品６〕小胞体ストレス軽減用飲料
実施例品１のホエイタンパク質濃縮物５０ｇを６６０ｇの脱イオン水に添加し、５０℃まで加熱後、９，５００ｒｐｍで３０分間撹拌混合した。
前記混合物に表３の原材料を添加した後、１００ｍｌのガラス瓶に充填し、９５℃、１５秒間殺菌後、密栓し、小胞体ストレス軽減用飲料１０本（１００ｍｌ入り）を調製した。この飲料にはβ−ラクトグロブリンが３．２２重量％含まれており、乳由来アルブミンが４重量％含まれていた。
[Example product 6] Beverage for reducing endoplasmic reticulum stress 50 g of the whey protein concentrate of
After adding the raw materials shown in Table 3 to the mixture, the mixture was filled in a 100 ml glass bottle, sterilized at 95 ° C. for 15 seconds, and then sealed to prepare 10 beverages for reducing endoplasmic reticulum stress (containing 100 ml). The beverage contained 3.22% by weight of β-lactoglobulin and 4% by weight of milk-derived albumin.
〔実施例品７〕イヌ用小胞体ストレス軽減用飼料
実施例品１のホエイタンパク質濃縮物２０ｋｇを８０ｋｇの脱イオン水に添加し、５０℃まで加熱後、３，６００ｒｐｍで４０分間撹拌混合して、実施例品１のホエイ粉を２０ｇ／１００ｇ含有する溶液を得た。この溶液１０ｋｇに、表４の原材料の全量を混合し、１２０℃、４分間殺菌して、本発明のイヌ用小胞体ストレス軽減用飼料１００ｋｇを製造した。この飼料にはβ−ラクトグロブリンが１．２９重量％含まれており、乳由来アルブミンが０．１６重量％含まれていた。
[Example product 7] Feed for reducing endoplasmic reticulum stress for dogs Add 20 kg of whey protein concentrate of
本発明は、新たな小胞体ストレスを軽減する組成物として、β−ラクトグロブリン及び／又は乳由来アルブミンを含む組成物と該組成物を含む食品、医薬品、及び飼料を提供する。本発明の組成物等を摂取することにより小胞体ストレスを安全かつ簡易に軽減することが期待できる。 The present invention provides a composition containing β-lactoglobulin and / or milk-derived albumin as a novel composition for reducing endoplasmic reticulum stress, and foods, pharmaceuticals, and feeds containing the composition. It can be expected that endoplasmic reticulum stress can be safely and easily reduced by ingesting the composition of the present invention.
Claims (6)
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