JP4044975B2 - Diabetes therapeutic agent consisting of fermented lactic acid bacteria - Google Patents

Description

【００２０】
また、米糠や、玄米粉中には、フィチン酸が多く含まれていることが知られている（J.W.JR.Erdeman, J.Am.Oil.Chem.誌、56巻、736 頁、1979年参照）。
【００２１】
本発明の糖尿病治療剤及び免疫増強剤の原料としては、上記米糠、玄米粉から選ばれた少なくとも一種の他に、更に、小麦ふすま、トウモロコシ外皮、オカラ等を副原料として添加することもできる。これらの副原料の配合割合は、原料全体に対して１〜50重量％が好ましい。
【００２２】
本発明の糖尿病治療剤及び免疫増強剤の好ましい製造方法においては、まず、最初の工程において、米糠、玄米粉から選ばれた少なくとも一種に、必要に応じて副原料を配合した後、水を加え、次いで、液化型アミラーゼを加えてよく混合し、加熱処理することにより、予め澱粉をある程度まで分解して液化する。水の添加量は、原料全体に対して0.5 〜100 重量倍が好ましい。加熱処理は、始め10〜90℃までは、３〜60分間程度かけてゆっくり加熱し、その後、２〜30分間沸騰を続けるのが好ましい。このようにして加熱処理することにより、原料中の澱粉がα化されると共にある程度まで分解して液化され、後の工程で糊化するのが防止される。なお、液化型アミラーゼとしては、α−アミラーゼが好ましく用いられる。
【００２３】
このようにして最初の工程を終えた後、次の工程において、セルラーゼ、プロテアーゼ及び糖化型アミラーゼを加えて更に酵素処理する。セルラーゼにより糠の組織の細胞壁が分解され、プロテアーゼにより蛋白質が分解される。更に、糖化型アミラーゼにより、上記工程で液化された澱粉が、グルコースなどに糖化される。糖化型アミラーゼとしては、グルコアミラーゼが好ましく用いられる。なお、この工程においても、液化型アミラーゼと糖化型アミラーゼとを併用して分解してもよい。なお、これらの酵素は、いずれも市販されているので、その中から適宜選択して用いることができる。
【００２４】
酵素処理は、それぞれの酵素に応じた添加量、温度、ｐＨ、時間を選択して行えばよく、処理条件が同じであれば同時に添加して処理することもでき、また、一種又は二種以上を添加して、二回以上に分けて処理することもできる。なお、後に実施例で示すように、この酵素処理により、各種ビタミン量が増加する。
【００２５】
酵素処理を終えた処理液は、次の工程において、遠心分離又は濾過により、油脂部分を除去する。例えば遠心分離すると、水不溶性の沈殿物からなる下層と、水溶液からなる中・上層と、油脂部分からなる最上層とに分れるので、油脂部分からなる最上層を除くことができる。このようにして、油脂成分を除去しておくことにより、次の工程で行う乳酸発酵の際に、油脂成分が分解、酸化されて糠臭、腐敗臭となったり、有毒化するのが防止される。なお、下層の沈殿物と、中・上層の水溶液とは、混合した状態で次の乳酸発酵を行い、その後、沈殿物を除去することもできるが、この段階で、沈殿物をも除去し、茶褐色透明の水溶液のみを乳酸発酵させることがより好ましい。なお、上記沈殿物は、食物繊維質からなり、本発明の主旨とは異なるが、この部分も機能性食品として利用可能である。
【００２６】
こうして得られた酵素処理液を、次の乳酸発酵の工程における本培養液として用いるが、酵素処理を終了した段階、又は油脂部分を除去した段階において、必要に応じて、糖類を添加することもできる。糖類としては、例えば、６炭糖類、５炭糖類、２糖類等を用いることができる。
【００２７】
続いて、次の工程で、この処理液に乳酸菌を接種して発酵させる。なお、処理液は、本培養液として乳酸発酵に用いる前に、公知の手段により滅菌処理をしておくのが好ましい。通常は、オートクレーブ等に入れ、120 〜130 ℃で、５〜20分間程度処理することにより充分な滅菌がなされる。
【００２８】
乳酸菌としては、乳酸の生成量においてはラクトバシルス・アシドフィルス(Lactobacillus acidophilus) が、味の面ではラクトバシルス・ビフィズス(Lactobacillus bifidus) が好ましいが、その他、ストレプトコッカス・フェカリス(Streptococcus faecalis)、ラクトバシルス・ブルガリカス (Lactobacillus bulgaricus) 、ラクトバシルス・サンフランシスコ (Lactobacillus sanfrancisco) 、ラクトバシルス・カゼイ (Lactobacillus casei)、ストレプトマイセス・ラクチス(Streptomyces lactis) 等を用いることもできる。これらは、単独で、又は二種以上を併用して用いることができる。これらの乳酸菌の選択によって、最終的な発酵液の味、香り、栄養素等を変化させることができる。なお、これらの乳酸菌は、いずれも公知の菌で、容易に入手することができる。
【００２９】
これらの乳酸菌は、予め前培養した後に、上記本培養液に添加することが好ましい。例えば、牛乳培地等を用いて、35〜40℃で７日間程度培養した後、これを更にグルコース0.8 重量％、酵母エキス0.8 重量％、ラクトース0.7 重量％からなる培地に殖菌して、35〜40℃で２日間培養して前培養液を得る。そして、この前培養液を、本培養液に１〜20重量％添加することが好ましい。
【００３０】
米糠、玄米粉から選ばれた少なくとも一種を主原料として調製された本培養液に、乳酸菌を添加した後、常法にしたがって発酵を行う。培地は静置培養が好ましいが、その他、撹拌振とう培養、通気培養なども可能である。培養条件は、30〜40℃で、３〜10日間程度とするのが好ましい。
【００３１】
なお、油脂成分を除去した際に、沈殿物を除去していない場合は、乳酸発酵終了後、必要に応じて水不溶性部分を除去する。
【００３２】
乳酸発酵させることにより、後の実施例で示すように、各種水溶性ビタミンの量が増加し、特に、ニコチン酸又はニコチンアミド、パントテン酸、ピリドキシン（ビタミンＢ₆ ）、チアミン（ビタミンＢ₁ ）、リボフラビン（ビタミンＢ₂ ）は、顕著に増加する。
【００３３】
こうして得られた乳酸発酵液は、水溶性ビタミンを高含量で含有すると共に、セルロース、澱粉、蛋白質が低分子化され、水溶性となったもの、すなわち、水溶性の植物繊維、糖質、各種ペプチド、アミノ酸類も含有している。更に、各種のミネラルも豊富に含有している。
【００３４】
上記のようにして得られた発酵液は、そのままでも米糠又は玄米粉の有する特有の悪臭であるいわゆる糠臭はかなり低減されるが、更に以下に示す工程を行うと、残存する糠臭が更に著しく低減されるので好ましい。
【００３５】
すなわち、上記のようにして得られた発酵液に、ガス体を接触させるか、超音波を当てるか、又は放射線を照射する。
【００３６】
発酵液にガス体を接触させる方法としては、曝気、脱気、攪拌振盪等の方法が採用される。
【００３７】
発酵液を曝気する方法としては、人体に安全なガス体、例えば空気、窒素、酸素、アルゴン等のガス体を、例えばガス噴出器、スプレードライ機等を用いて接触させる方法が好ましく用いられる。ガス体としては、特に空気が好ましい。曝気をガス噴出器を用いて行なう場合は、発酵液100ml 当たり５〜3000ml/ 分のガス流量で、0.01〜25時間行うのが好ましい。
【００３８】
また、発酵液を脱気する方法としては、例えば、凍結乾燥器、真空乾燥器、加熱減圧乾燥器等を用いて減圧する方法が好ましい。脱気は、0.1 〜700mmHg 柱程度の減圧度で、0.01〜25時間程度行うのが好ましい。脱気すると、発酵液中に溶存しているガス体が放出され、そのときにガス体が発酵液と接触する。
【００３９】
更に、発酵液を攪拌振盪する方法としては、各種の攪拌混合機や、振盪機を用いて、発酵液を攪拌又は振盪する方法が採用される。振盪は、往復振盪、回転振盪のいずれでもよいが、往復振盪が好ましい。振盪条件は、10〜300 回／分で、10分間〜５時間行うのが好ましい。攪拌振盪によって、周囲の空気が発酵液中に混合して、発酵液と接触する。
【００４０】
一方、発酵液に超音波を当てる方法としては、公知の超音波発生機を用い、発酵液の容器や、発酵液中に浸漬した発振器等を介して、発酵液に超音波を付与すればよい。超音波の振動数は２万サイクル／秒以上が好ましい。超音波発生機として、例えば「Ultrasonic generator US300」（商品名、株式会社日本精機製作所製）を用いた場合、処理条件は、１〜500 μＡで、0.1 〜300 分間とするのが好ましい。
【００４１】
また、発酵液に放射線を照射する方法としては、人体に危険が少ないα線、β線を照射するのが好ましく、公知の放射線照射装置を用いて照射すればよい。
【００４２】
このようにして、発酵液に、ガス体を接触させるか、超音波を当てるか、又は放射線を照射することにより、無色透明に近く、米糠、玄米粉に起因する臭いが非常に少ない乳酸発酵液を得ることができる。
【００４３】
本発明の糖尿病治療剤は、上記発酵液そのものであってもよいが、必要に応じて上記発酵液を更に乾燥粉末化し、粉剤、錠剤、カプセル剤としたものであってもよい。乾燥粉末化する方法としては、噴霧乾燥、凍結乾燥、熱風乾燥、スプレードライなど、各種の方法が採用できる。
【００４４】
なお、飲料の形態にする場合には、上記発酵液をそのまま摂取することもできるが、そのままでは、味、香り、栄養価とも濃厚であるので、２〜２０倍程度に水で希釈して飲むのが好ましい。また、上記発酵液に、各種香料、着色料、呈味成分、ゲル化剤等を添加することもできる。
【００４５】
更に、本発明の糖尿病治療剤は、各種果汁、栄養ドリンク等の飲料に添加したり、ごはん、パン等の他の食品や、食品素材等に添加して摂取することもできる。
【００４６】
更にまた、本発明の糖尿病治療剤は、各種の飼料に添加することもできる。例えば、天然配合飼料中に、飼料重量の１／２０〜１０倍量の乳酸菌発酵液を加え、乾燥して飼料とすることもできる。
【００４７】
【実施例】
以下、本発明の実施例を挙げて説明する。以下の実施例において、液化型アミラーゼとしては、α−アミラーゼである「ユニアーゼＢＭ80」（商品名、株式会社ヤクルト本社製）を用い、糖化型アミラーゼとしては、グルコアミラーゼである「ユニアーゼ30」（商品名、株式会社ヤクルト本社製）を用い、セルラーゼとしては、「セルラーゼオノズカ３Ｓ」（商品名、株式会社ヤクルト本社製）を用い、プロテアーゼとしては、「パンチターゼＮＰ−２」（商品名、株式会社ヤクルト本社製）を用いた。
【００４８】
実施例１
新鮮米糠に、その重量の５倍量の水を加えた後、米糠に対して0.1 重量％のα−アミラーゼ（ユニアーゼＢＭ80）を加えて撹拌し、20分かけて90℃まで加熱し、その後沸騰させて、10分間保持した。
【００４９】
加熱終了後、50℃まで冷却し、米糠に対してそれぞれ0.5 重量％の、α−アミラーゼ（ユニアーゼＢＭ80）、グルコアミラーゼ（ユニアーゼ30）及びセルラーゼ（セルラーゼオノズカ３Ｓ）と、米糠に対して0.4 重量％のプロテアーゼ（パンチターゼＮＰ−２）とを添加し、50℃の恒温器中で、撹拌下に、24時間酵素処理を行った。
【００５０】
次いで、得られた酵素処理液を、５℃以下に冷却し、４℃、5000rpm の条件下に遠心分離し、最上層の油脂部分と、下層の沈殿部分とを分別除去して、茶褐色透明溶液を得た。
【００５１】
次に、得られた溶液を、１Ｌ（リットル）容三角フラスコに200 mlの割合で分注し、綿栓をした後、オートクレーブで、120 ℃の条件下に、15分間高圧滅菌処理した。これを乳酸発酵本培養液とする。
【００５２】
一方、ラクトバシルス・アシドフィルス、ラクトバシルス・ビフィズス、ストレプトコッカス・フェカリスを、それぞれ牛乳培地等を用いて、35〜40℃で７日間程度培養した後、この一白金耳を更にグルコース0.8 重量％、酵母エキス0.8 重量％、ラクトース0.7 重量％からなる培地に殖菌して、35〜40℃で２日間培養して３種類の前培養液を得た。
【００５３】
この３種類の前培養液を、それぞれ、上記乳酸発酵本培養液に、５重量％の割合で添加し、34〜40℃で、８日間発酵させて、３種類の乳酸発酵液を得た。
【００５４】
発酵の進行に伴ない、培養０時間、３２時間、８０時間、１４４時間、１９２時間で培養液の一部を採取して、ニコチン酸、パントテン酸、ピリドキシン（ビタミンＢ₆ ）、チアミン（ビタミンＢ₁ ）、リボフラビン（ビタミンＢ₂ ）の含量を分析した。
【００５５】
分析定量は、液体クロマトグラフ装置（株式会社島津製作所製、検知器：紫外分光光度計ＳＰＤ−６Ａ（210 nm）、カラム：シンパックＣＬＣ−ＯＤＳ［Ｍ］（4.6 ×250 mm）（40℃））を用い、溶媒として1.2 mMオクタスルフォン酸ナトリウムの0.1 M リン酸バッファー（ｐＨ2.1 ）溶液とアセトニトリルとの９：１混液（流速1.5 ml/ 分）を用いて行った。
【００５６】
この結果を表２に示す。なお、表２において、乳酸菌の欄のＡはラクトバシルス・アシドフィルス、Ｂはラクトバシルス・ビフィズス、Ｆはストレプトコッカス・フェカリスを表す。
【００５７】
【表２】

【００５８】
表２の結果から、乳酸発酵の進行に伴ない、各種の水溶性ビタミンが顕著に増大することがわかる。
【００５９】
次に、乳酸菌としてラクトバチルス・ビフィズスを用い、発酵時間を240 時間として、上記の方法で得られた発酵液について、ガス噴出器を用いて、発酵液100ml 当たり600ml/分の流量で空気を挿入し、５時間曝気することにより、糠臭を除去した。
【００６０】
こうして得られた乳酸菌発酵液について、食品としての安全性をみるための各種分析を行った結果を表３に示す。
【００６１】
【表３】

【００６２】
また、乳酸菌発酵液に含まれる蛋白質成分について、アミノ酸分析をした結果を表４に示す。なお、アミノ酸分析は、過ギ酸酸化処理後、塩酸加水分解して、アミノ酸自動分析法によって行い、トリプトファインだけは、高速液体クロマトグラフ法によって行った。
【００６３】
【表４】

【００６４】
表２〜４の結果から、この乳酸菌発酵液は、蛋白質又はその分解物、糖質、水溶性ビタミン、ミネラルを含む栄養豊富な飲料であることがわかる。
【００６５】
実施例２
実施例１の米糠を、ボールミルで微粉砕した玄米粉に替え、加える水の量を玄米粉の２重量倍とし、加熱処理の際のα−アミラーゼの量を玄米粉に対して0.6 重量％とした以外は実施例１と同様にして、乳酸菌発酵液を得た。
【００６６】
この乳酸菌発酵液の成分を分析したところ、実施例１の乳酸菌発酵液と同様に蛋白質又はその分解物、糖質、水溶性ビタミン、ミネラルを含み、糖質の含量がやや多いことがわかった。
【００６７】
試験例１
３週齢のＳＤ系雄性ラットを、各群６匹ずつＡ〜Ｄの合計４群用意し、通常の固形飼料（オリエンタル酵母製）と、飲料水として、Ａ群、Ｃ群については蒸留水のみ、Ｂ群、Ｄ群については実施例１で得られた乳酸菌発酵液を蒸留水で１／２に希釈した液（以下単に「１／２発酵希釈液」とする）とをそれぞれ自由に与えて、全群１７日間予備飼育した。全群とも、予備飼育の１７日目に１日絶食させ、Ａ〜Ｄ群について更に次のように飼育した。なお、１日絶食させ後、飼料については、全群とも上記と同じ固形飼料を自由摂取させた。
【００６８】
Ａ群：再び蒸留水のみを飲料水として飼育した。
Ｂ群：再び１／２発酵希釈液を飲料水として飼育した。
Ｃ群：１日絶食させた後、アロキサンを0.25ml(10mg)／100g体重の量で腹腔内投与し、その後、再び蒸留水のみを飲料水として飼育した。
Ｄ群：１日絶食させた後、アロキサンを0.25ml(10mg)／100g体重の量で腹腔内投与し、その後、再び１／２発酵希釈液を飲料水として飼育した。
【００６９】
なお、アロキサンは、膵臓のβ細胞を破壊して、血糖低下作用を発揮するホルモン、インシュリンの分泌を抑制する物質として知られており、人工的に糖尿病を惹起する物質として各種実験に用いられているものである。
【００７０】
この飼育期間中に、各群の１匹当たりの平均体重、血糖値、尿糖値、１匹１日当たりの尿量、１匹１日当たりの平均飲水量、１匹１日当たりの平均摂餌量、尿中ウロビリノーゲン、尿蛋白、尿中ケトン体、尿ｐＨの各値について、経時的に測定した。
【００７１】
なお、血糖値は、試験紙として「アントセンスカートリッジ」（商品名、ダイキン工業株式会社製、小野薬品工業株式会社販売）を用い、小型電極血糖測定機器「アントセンス」（商品名、ダイキン工業株式会社製、三共株式会社販売）によって測定した。
【００７２】
また、尿糖値、尿中ウロビリノーゲン、尿蛋白値、尿中ケトン体、尿ｐＨは、テストペーパー「ウロラブスティクス」（商品名、マイルズ三共株式会社製）を用いて測定した。
【００７３】
図１は、各群の１匹当たりの平均体重を表すが、Ａ群とＤ群が同じ挙動を示し、また、Ｂ群とＣ群が同じ挙動を示した。注意すべき点は、１／２発酵希釈液を飲料水として飼育し、アロキサン投与したＤ群は、蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群に比べて、アロキサン投与後の体重増加が大きくなっており、アロキサン投与をしないＡ群と同様の挙動を示したことである。なお、Ｃ群の内の１匹は、アロキサン投与後の３日目にアロキサン毒性によって死亡した。
【００７４】
図２は、各群の血糖値の測定結果を示す。蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群は、顕著な血糖値の増加が見られたが、１／２発酵希釈液を飲料水として飼育し、アロキサン投与したＤ群は、血糖値の増加量が半減することがわかる。
【００７５】
図３は、各群の尿糖値の測定結果を示す。蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群は、顕著な尿糖値の増加が見られたが、１／２発酵希釈液を飲料水として飼育し、アロキサン投与したＤ群は、尿糖値の増加量がかなり減少することがわかる。
【００７６】
図４は、各群の尿量の測定結果を示す。蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群は、顕著な尿量の増加が見られたが、１／２発酵希釈液を飲料水として飼育し、アロキサン投与したＤ群は、その他の群と同様に正常な値を保ち続けた。
【００７７】
図５は、各群の飲水量の測定結果を示す。蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群のみが、顕著な飲水量の増加が見られた。
【００７８】
なお、飼料摂取量は、Ｂ群のみ実験後半にやや減少が見られたが、その他の群はほぼ同様の結果を示した。また、尿蛋白値は、各群全く錯綜した結果を示したが、全試験期間を通じて見た場合は、全群ほぼ同量の排泄量を示した。更に、尿ｐＨも各群錯綜していたが、全試験期間を通じて見た場合は、同様な結果であった。尿中ウロビリノーゲン、尿中ケトン体は、全群全く同じ結果を示し、何ら変化は見られなかった。
【００７９】
以上の結果を考慮すると、特に図２〜５の結果から、蒸留水のみを飲料水として飼育し、アロキサン投与したＣ群は、アロキサン投与後に顕著な糖尿病の症状が見られるが、１／２発酵希釈液を飲料水として飼育し、アロキサン投与したＤ群は、糖尿病の症状が顕著に抑制されることがわかる。
【００８０】
試験例２
５週齢の雄性ラットを各群１０匹ずつ、Ａ群、Ｂ群の２群用意し、Ａ群は蒸留水のみを飲料水とし、Ｂ群は試験例１と同じ１／２発酵希釈液を飲料水とし、飼料としてはいずれも試験例１と同じ通常の固形飼料を用い、飲料水及び飼料を自由摂取させて飼育した。
【００８１】
そして、飼育期間中の体重、飲水量、摂餌量を経時的に測定した。また、実験最終日に、0.05ml（２μｇ）のフィトヘムアグルチニン(Pytohaemagglutinin)を耳たぶの内皮に注入し、２４時間後の赤斑の程度を日本光電製測色色差計で測って免疫機能の程度を測定した。
【００８２】
なお、フィトヘムアグルチニンは、抗原として作用するもので、フィトヘムアグルチニンを注入した後の赤斑が濃くでるほど、抗原抗体反応による抗体産生力が強いと判断される。
【００８３】
図６は、体重の変化を示し、両群とも全く同じ挙動を示した。
【００８４】
図７は、１日当たりの飲水量の変化を示し、多少の変動はあるが、ほぼ試験全期間を通じてＢ群の摂取量が多いことがわかる。
【００８５】
図８は、Ａ、Ｂ両群の１日当たりの摂餌量を示している。また、Ｂ群については、１／２発酵希釈液からなる飲料水中に含まれる固液分の量を上記Ｂ群の摂餌量に加えた合計の摂餌量を計算し、図中Ｂ’（−△−のグラフ）で示した。
【００８６】
Ａ、Ｂ両群の摂餌量を比較すると、明らかにＡ群の方が多くなっているが、飲料水中に含まれる固液分の量を加えたＢ’で比較すると、両群ともほぼ同様な挙動を示すことがわかる。
【００８７】
すなわち、Ｂ群は、発酵希釈液中から固形分を摂取しているため、その分だけ摂餌量が減少することがわかる。逆に言えば、Ｂ群は、固形飼料の摂餌量を減らしてでも、発酵希釈液を好んで多飲することにより、摂餌量の不足分を補っていることがわかる。
【００８８】
図９は、フィトヘムアグルチニンを注入した翌日の免疫反応の結果を示し、図において、ａ値の増加は赤色が強いこと、ｂ値の増加は黄色が強いことを示している。免疫反応は、赤色が強いほど顕著であることを示すので、Ｂ群は、Ａ群に比べて、明らかに免疫反応が強く、免疫力が増強されていることがわかる。
【００８９】
試験例３
５週齢の雄性ラットを各群１０匹ずつ、Ａ群、Ｂ群の２群用意し、Ａ群は蒸留水のみを飲料水とし、Ｂ群は実施例１で得られた乳酸菌発酵液を蒸留水で１／３に希釈した液（以下単に「１／３発酵希釈液」とする）を飲料水とし、飼料としてはいずれも試験例１と同じ固形飼料を用い、飲料水及び飼料を自由摂取させて３５日間飼育した。
【００９０】
そして、この飼育期間の最終日（１０週齢）における各群の１匹当たりの平均体重と、最終日の１日における各群の１匹当たりの平均飲水量及び平均摂餌量とを測定した。
【００９１】
図１０は、平均体重を示し、Ａ、Ｂ両群ともほぼ同様であるが、Ｂ群の方が若干増加している。
【００９２】
図１１は、平均飲水量を示し、Ｂ群の方が明らかに多くなっており、成長ラットの場合も発酵希釈液を好んで飲用することがわかる。
【００９３】
図１２は、平均摂餌量を示している。なお、Ｂ群については、１／３発酵希釈液からなる飲料水中に含まれる固形分の量を上記Ｂ群の摂餌量に加えた合計の摂餌量を計算し、図中Ｂ’で示した。
【００９４】
Ａ、Ｂ両群の摂餌量を比較すると、明らかにＡ群の方が多くなっているが、飲料水中に含まれる固形分の量を加えたＢ’で比較すると、大きな差はないことがわかる。
【００９５】
したがって、１０週齢の成長ラットにおいても、発酵希釈液を好んで飲用し、それによって蒸留水を飲料水とした場合よりも体重が増加することがわかる。
【００９６】
試験例４
１３週齢の中老の雄性ラットを各群１０匹ずつ、Ａ群、Ｂ群の２群用意し、Ａ群は蒸留水のみを飲料水とし、Ｂ群は試験例３と同じ１／３発酵希釈液を飲料水とし、飼料としてはいずれも試験例１と同じ固形飼料を用い、飲料水及び飼料を自由摂取させて１日間飼育した。
【００９７】
そして、上記１日間における１匹当たりの平均飲水量を測定した。この結果を図１３に示す。
【００９８】
図１３に示されるように、中老ラットにおいても、発酵希釈液を好んで飲用することがわかる。
【００９９】
以上の試験結果から、本発明の糖尿病治療剤及び免疫増強剤を含む発酵希釈液は、幼年ラットから成長ラット、中老ラットになるに従って好んで飲用する傾向があり、固形飼料の摂餌量を減らしても嗜飲することから、この発酵希釈液は、ラットにとって非常に美味な飲料となっていることがわかる。
【０１００】
また、発酵希釈液の摂取によって、年を取るに従って体重の増加を示すことから、この発酵希釈液は、美味であると共に、栄養豊富な飲料となっていることがわかる。
【０１０１】
そして、この発酵希釈液を飲ませることによって、免疫力が増強され、糖尿病誘因物質であるアロキサンを投与した後の糖尿病症状を顕著に軽減する効果がもたらされることがわかる。
【０１０２】
【発明の効果】
以上説明したように、本発明の糖尿病治療剤は、糖尿病症状を軽減すると共に、免疫力を増強させる効果を有し、美味であると共に栄養豊富であることから、糖尿病患者等の健康状態を回復させ、糖尿病の抜本的治療も期待できると考えられる。また、本発明の糖尿病治療剤は、前記表３に示したように、人体に危険な成分を含有しておらず、本来食品として自由に摂取できるものであるため、日常生活の中で手軽に、かつ好きなだけ摂取することができ、長期に亙って摂取することにより、治療効果を高めることができる。
【図面の簡単な説明】
【図１】試験例１における各群の平均体重の推移を示す図表である。
【図２】試験例１における各群の血糖値の推移を示す図表である。
【図３】試験例１における各群の尿糖値の推移を示す図表である。
【図４】試験例１における各群の尿量の推移を示す図表である。
【図５】試験例１における各群の飲水量の推移を示す図表である。
【図６】試験例２における各群の平均体重の推移を示す図表である。
【図７】試験例２における各群の飲水量の推移を示す図表である。
【図８】試験例２における各群の摂餌量の推移を示す図表である。
【図９】試験例２における各群の免疫反応の結果を示す図表である。
【図１０】試験例３における各群の平均体重を示す図表である。
【図１１】試験例３における各群の飲水量を示す図表である。
【図１２】試験例３における各群の摂餌量を示す図表である。
【図１３】試験例４における各群の飲水量を示す図表である。[0001]
BACKGROUND OF THE INVENTION
The present invention is a treatment for diabetes comprising a fermented lactic acid bacterium obtained using rice bran and / or brown rice flour as a raw material. To the agent Related.
[0002]
[Prior art]
As a therapeutic agent for diabetes, insulin is generally used. However, since insulin is decomposed in the intestine, it cannot be administered orally and must be administered by injection under the guidance of a doctor. For this reason, hospitalization or outpatient treatment is required for treatment with insulin. Moreover, when insulin administration is discontinued, the symptoms of diabetes reappear and are often not drastic therapeutic agents.
[0003]
In recent years, it has been found that food materials such as dietary fiber have an action to reduce symptoms of diabetes, and various therapeutic agents for diabetes have been proposed. For example, Japanese Patent Laid-Open No. 61-167622 proposes a diabetes control agent mainly composed of cell wall components of brewer's yeast.
[0004]
On the other hand, although lactic acid bacteria have long been used for the production of lactic acid bacteria beverages and have been known to be effective in improving intestinal bacterial flora, there are still few reports on the bioactive effects of lactic acid bacteria fermentation products on adult diseases, etc. Is the current situation.
[0005]
[Problems to be solved by the invention]
As described above, treatment with insulin has a problem that hospitalization or outpatient treatment is required and it is often not a drastic treatment.
[0006]
In addition, anti-diabetic agents such as dietary fiber can be easily ingested as food in daily life, and since it is a food material, there is no side effect, but dietary fiber is indigestible. Therefore, there are problems that it is difficult to take a large amount and other nutrient components are scarce.
[0007]
In addition, diabetes often causes various complications such as arteriosclerotic vascular disorders. In the treatment of diabetes, not only alleviating the symptoms of diabetes but also maintaining good nutrition and health, It is important to develop resistance to illness.
[0008]
Therefore, the object of the present invention is a diabetes treatment that can be easily ingested as a food and drink in daily life, contains a variety of nutritional components, and not only reduces diabetes symptoms but also enhances immunity. Agent It is to provide.
[0009]
[Means for Solving the Problems]
The present inventors have focused on nutritional components contained in rice bran and / or brown rice flour, and have intensively studied to provide nutritious food and drink having a good flavor by fermenting them with lactic acid bacteria. In the process, the fermented lactic acid bacteria were found to have a diabetic symptom reducing effect and an immune enhancing effect, and the present invention was completed.
[0010]
That is, the present invention is to add water to rice bran and / or brown rice flour, add liquefied amylase and heat-treat, add cellulase, protease and saccharified amylase to the heat-treated solution, and perform enzyme treatment. The fat and oil part is removed from the treatment liquid by centrifugation or filtration, and the fermentation liquid itself obtained by inoculating and fermenting lactic acid bacteria in the treatment liquid or a dry powder of the fermentation liquid is used as an active ingredient. It is characterized by The present invention provides a therapeutic agent for diabetes.
[0012]
Main departure In the morning, It is preferable that the fermentation liquid is aerated after the completion of fermentation. Furthermore, the fermentation broth may have a water-insoluble portion removed after the end of fermentation.
[0013]
In addition, the protein or its degradation product as a component of the diabetes therapeutic agent and immunopotentiator of the present invention is meant to include proteins, peptides, amino acids and the like. In addition, the saccharide means to include polysaccharides such as starch and hemicellulose, partially decomposed products thereof, oligosaccharides, disaccharides, monosaccharides and the like.
[0014]
As shown in the animal test data described later, the therapeutic agent for diabetes and the immunopotentiator of the present invention have a remarkable effect of reducing the symptoms of diabetes, and a clear immune enhancing action is also observed. Therefore, it is considered that taking the diabetes therapeutic agent and immune enhancer of the present invention brings about not only a therapeutic effect on diabetes but also an effect of imparting resistance to various diseases.
[0015]
Moreover, the therapeutic agent for diabetes and the immunopotentiator of the present invention contain abundant nutritional components derived from rice bran and / or brown rice flour, and are given a good flavor by lactic acid bacteria fermentation. As much as you like, you can ingest as much as you like, thereby maintaining good nutrition and health.
[0016]
DETAILED DESCRIPTION OF THE INVENTION
The therapeutic agent for diabetes and the immunopotentiator of the present invention are fermented lactic acid bacteria obtained using rice bran and / or brown rice flour as a raw material, and contain a protein or a decomposition product thereof, a saccharide, a water-soluble vitamin, and a mineral. Any method can be used as the method for lactic acid bacteria fermentation. The preferable production method is described as follows.
[0017]
First, at least one selected from rice bran and brown rice flour is used as the main raw material. As rice bran, normal rice bran including rice peel, seed coat, glue layer, germs, etc. removed during the process of brown rice to white rice can be used as it is. What was made into a shape can be used. In addition, it is preferable to use fresh rice bran and brown rice flour.
[0018]
Rice bran and brown rice flour contain abundant nutrients as shown in Table 1 below. Table 1 shows the values of various components per 100 g of edible portion of brown rice, white rice and rice bran according to the “Japan Food Composition Table (4th revised edition), Resource Science and Technology Survey”.
[0019]
[Table 1]

[0020]
In addition, rice bran and brown rice flour are known to contain a large amount of phytic acid (see JWJR.Erdeman, J.Am.Oil.Chem., 56, 736, 1979). .
[0021]
As raw materials for the diabetes therapeutic agent and immune enhancer of the present invention, wheat bran, corn hulls, okara and the like can be added as auxiliary materials in addition to at least one selected from rice bran and brown rice flour. The blending ratio of these auxiliary raw materials is preferably 1 to 50% by weight with respect to the whole raw materials.
[0022]
In a preferred method for producing the diabetes therapeutic agent and immune enhancer of the present invention, first, in the first step, at least one selected from rice bran and brown rice flour is blended with auxiliary ingredients as necessary, and then water is added. Then, liquefied amylase is added and mixed well, followed by heat treatment, whereby starch is decomposed to some extent and liquefied in advance. The amount of water added is preferably 0.5 to 100 times the weight of the whole raw material. In the heat treatment, it is preferable that the heat treatment is first slowly heated to about 10 to 90 ° C. over about 3 to 60 minutes, and then the boiling is continued for 2 to 30 minutes. By performing the heat treatment in this manner, the starch in the raw material is gelatinized and decomposed to a certain extent to be liquefied, thereby preventing gelatinization in a later step. As the liquefied amylase, α-amylase is preferably used.
[0023]
After the first step is completed in this way, in the next step, cellulase, protease and saccharified amylase are added to perform further enzyme treatment. Cellulase degrades the cell wall of the cocoon tissue, and protease degrades the protein. Furthermore, the liquefied starch is saccharified into glucose or the like by the saccharification amylase. As the saccharified amylase, glucoamylase is preferably used. In this step as well, liquefaction amylase and saccharification amylase may be used in combination for decomposition. In addition, since these enzymes are all commercially available, they can be appropriately selected and used.
[0024]
Enzyme treatment may be performed by selecting the addition amount, temperature, pH, and time according to each enzyme. If the treatment conditions are the same, they can be added and treated at the same time. Can be added and processed in two or more steps. In addition, as shown in an Example later, the amount of various vitamins increases by this enzyme treatment.
[0025]
In the next step, the treatment liquid that has been subjected to the enzyme treatment removes oil and fat portions by centrifugation or filtration. For example, when it is centrifuged, it is divided into a lower layer composed of a water-insoluble precipitate, a middle / upper layer composed of an aqueous solution, and an uppermost layer composed of an oil / fat portion, so that the uppermost layer composed of an oil / fat portion can be removed. In this way, by removing the fat and oil component, the fat and oil component is prevented from being decomposed and oxidized to become a foul odor, a rotten odor or toxic during the lactic acid fermentation performed in the next step. The In addition, the lower layer precipitate and the middle and upper layer aqueous solution can be subjected to the next lactic acid fermentation in a mixed state, and then the precipitate can be removed, but at this stage, the precipitate is also removed, More preferably, only a brownish brown transparent aqueous solution is subjected to lactic acid fermentation. In addition, although the said deposit consists of dietary fiber and is different from the main point of this invention, this part can also be utilized as a functional food.
[0026]
The enzyme-treated solution thus obtained is used as the main culture solution in the next lactic acid fermentation process, but in the stage where the enzyme treatment is completed or the oil and fat part is removed, saccharides may be added as necessary. it can. As the saccharide, for example, hexose saccharide, pentose saccharide, disaccharide, and the like can be used.
[0027]
Subsequently, in the next step, the treatment solution is inoculated with lactic acid bacteria and fermented. The treatment solution is preferably sterilized by a known means before being used for lactic acid fermentation as the main culture solution. Usually, it is sufficiently sterilized by placing it in an autoclave or the like and treating it at 120 to 130 ° C. for about 5 to 20 minutes.
[0028]
As lactic acid bacteria, Lactobacillus acidophilus is preferred for the production of lactic acid, and Lactobacillus bifidus for taste, but Streptococcus faecalis, Lactobacillus bulgaricus), Lactobacillus sanfrancisco, Lactobacillus casei, Streptomyces lactis, and the like can also be used. These can be used alone or in combination of two or more. By selecting these lactic acid bacteria, the taste, aroma, nutrients and the like of the final fermentation broth can be changed. These lactic acid bacteria are all known bacteria and can be easily obtained.
[0029]
These lactic acid bacteria are preferably added to the main culture solution after pre-culture. For example, after culturing for about 7 days at 35 to 40 ° C. using a milk medium or the like, this is further inoculated into a medium comprising 0.8% by weight of glucose, 0.8% by weight of yeast extract, and 0.7% by weight of lactose. Incubate at 40 ° C. for 2 days to obtain a preculture. And it is preferable to add 1 to 20 weight% of this preculture solution to the main culture solution.
[0030]
After adding lactic acid bacteria to the main culture liquid prepared using at least one selected from rice bran and brown rice flour as a main raw material, fermentation is performed according to a conventional method. The culture medium is preferably stationary culture, but stirring culture with shaking and aeration culture are also possible. The culture conditions are preferably 30 to 40 ° C. and about 3 to 10 days.
[0031]
In addition, when the deposit is not removed when removing the oil and fat component, the water-insoluble portion is removed as necessary after the end of lactic acid fermentation.
[0032]
By lactic acid fermentation, the amount of various water-soluble vitamins increases as shown in the following examples. In particular, nicotinic acid or nicotinamide, pantothenic acid, pyridoxine (vitamin B) ₆ ), Thiamine (vitamin B) ₁ ), Riboflavin (vitamin B ₂ ) Increases significantly.
[0033]
The lactic acid fermentation broth thus obtained contains a high content of water-soluble vitamins, and is made water-soluble by cellulose, starch and protein being reduced in molecular weight, that is, water-soluble plant fibers, carbohydrates, various It also contains peptides and amino acids. Furthermore, various minerals are also abundantly contained.
[0034]
The fermented liquor obtained as described above can reduce the so-called bad odor which is a peculiar bad odor of rice bran or brown rice flour as it is, but when the following steps are further performed, the remaining bad odor is further increased. This is preferable because it is significantly reduced.
[0035]
That is, the fermented liquid obtained as described above is brought into contact with a gas body, applied with ultrasonic waves, or irradiated with radiation.
[0036]
As a method of bringing the gas body into contact with the fermentation broth, methods such as aeration, deaeration, and stirring and shaking are employed.
[0037]
As a method for aeration of the fermentation broth, a method in which a gas body that is safe to the human body, for example, a gas body such as air, nitrogen, oxygen, and argon, is contacted using, for example, a gas ejector or a spray dryer is preferably used. As the gas body, air is particularly preferable. When aeration is performed using a gas ejector, it is preferably performed for 0.01 to 25 hours at a gas flow rate of 5 to 3000 ml / min per 100 ml of fermentation broth.
[0038]
Moreover, as a method of deaerating a fermentation broth, the method of pressure-reducing using a freeze dryer, a vacuum dryer, a heat | fever decompression dryer etc. is preferable, for example. Deaeration is preferably performed at a reduced pressure of about 0.1 to 700 mmHg column for about 0.01 to 25 hours. When degassing, the gas body dissolved in the fermentation broth is released, and the gas body then contacts the fermentation broth.
[0039]
Furthermore, as a method of stirring and shaking the fermentation broth, a method of stirring or shaking the fermentation broth using various stirring mixers and shakers is employed. The shaking may be either reciprocal shaking or rotary shaking, but reciprocal shaking is preferred. The shaking conditions are preferably 10 to 300 times / minute and 10 minutes to 5 hours. By stirring and shaking, ambient air is mixed into the fermentation broth and comes into contact with the fermentation broth.
[0040]
On the other hand, as a method of applying ultrasonic waves to the fermentation broth, a known ultrasonic generator may be used to apply ultrasonic waves to the fermentation broth via a fermentation broth container, an oscillator immersed in the fermentation broth, or the like. . The frequency of ultrasonic waves is preferably 20,000 cycles / second or more. When, for example, “Ultrasonic generator US300” (trade name, manufactured by Nippon Seiki Seisakusho Co., Ltd.) is used as the ultrasonic generator, the processing conditions are 1 to 500 μA, preferably 0.1 to 300 minutes.
[0041]
Moreover, as a method of irradiating the fermentation broth with radiation, it is preferable to irradiate α rays and β rays with less danger to the human body, and irradiation may be performed using a known radiation irradiation apparatus.
[0042]
In this way, by bringing the fermentation liquid into contact with a gas body, applying ultrasonic waves, or irradiating with radiation, the lactic acid fermentation liquid is almost colorless and transparent and has very little odor caused by rice bran and brown rice flour. Can be obtained.
[0043]
Diabetes treatment of the present invention Agent The fermented liquid itself may be used, but the fermented liquid may be further powdered into powder, tablets, and capsules as necessary. Various methods such as spray drying, freeze drying, hot air drying, spray drying and the like can be adopted as a method for dry powderization.
[0044]
In addition, when making into the form of a drink, although the said fermented liquor can also be ingested as it is, since taste, aroma, and nutritional value are rich as it is, it is diluted with water about 2 to 20 times and drinks. Is preferred. Moreover, various fragrance | flavors, a coloring agent, a taste component, a gelatinizer, etc. can also be added to the said fermented liquor.
[0045]
Further, the diabetes treatment of the present invention Agent It can also be added to beverages such as various fruit juices and nutritional drinks, or added to other foods such as rice and bread, food materials, and the like.
[0046]
Furthermore, the diabetes treatment of the present invention Agent It can also be added to various feeds. For example, a lactic acid bacterium fermentation solution having an amount of 1/20 to 10 times the weight of the feed can be added to the natural blended feed and dried to obtain a feed.
[0047]
【Example】
Examples of the present invention will be described below. In the following Examples, “Uniase BM80” (trade name, manufactured by Yakult Honsha Co., Ltd.) that is α-amylase is used as the liquefied amylase, and “Uniase 30” (product) that is glucoamylase is used as the saccharified amylase. "Cellulase Onozuka 3S" (trade name, manufactured by Yakult Honsha Co., Ltd.) is used as the cellulase, and "Pantase NP-2" (trade name, Co., Ltd.) is used as the protease. Yakult head office) was used.
[0048]
Example 1
After adding 5 times the weight of water to fresh rice bran, add 0.1 wt% α-amylase (Uniase BM80) to the rice bran, stir, heat to 90 ° C over 20 minutes, then boil And held for 10 minutes.
[0049]
After heating, the mixture is cooled to 50 ° C. and 0.5% by weight of rice bran, α-amylase (Uniase BM80), glucoamylase (Uniase 30) and cellulase (Cellulase Onozuka 3S), and 0.4% by weight of rice bran. % Protease (pantase NP-2) was added, and the enzyme treatment was performed in a thermostat at 50 ° C. for 24 hours with stirring.
[0050]
Next, the obtained enzyme-treated solution is cooled to 5 ° C. or lower, centrifuged at 4 ° C. and 5000 rpm, and the uppermost oil and fat portion and the lower layer precipitated portion are separated and removed to obtain a brown transparent solution. Got.
[0051]
Next, the obtained solution was dispensed into a 1 L (liter) Erlenmeyer flask at a rate of 200 ml, capped, and then autoclaved at 120 ° C. for 15 minutes in an autoclave. This is referred to as a lactic acid fermentation main culture solution.
[0052]
On the other hand, after culturing Lactobacillus acidophilus, Lactobacillus bifidos, Streptococcus faecalis for about 7 days at 35-40 ° C. using milk medium, etc., the platinum loop was further added with 0.8% glucose and 0.8% yeast extract. Inoculated in a medium consisting of 0.5% lactose and 0.7% by weight, and cultured at 35 to 40 ° C. for 2 days to obtain three types of precultured solutions.
[0053]
Each of these three types of precultures was added to the lactic acid fermentation main culture solution at a rate of 5% by weight and fermented at 34 to 40 ° C. for 8 days to obtain three types of lactic acid fermentation broths.
[0054]
As the fermentation progresses, a portion of the culture solution is collected at 0, 32, 80, 144, and 192 hours of culture, and nicotinic acid, pantothenic acid, pyridoxine (vitamin B ₆ ), Thiamine (vitamin B) ₁ ), Riboflavin (vitamin B ₂ ) Content was analyzed.
[0055]
Analytical quantification is a liquid chromatograph apparatus (manufactured by Shimadzu Corporation, detector: UV spectrophotometer SPD-6A (210 nm), column: Sinpack CLC-ODS [M] (4.6 × 250 mm) (40 ° C.) ) And a 9: 1 mixture (flow rate of 1.5 ml / min) of 1.2 mM sodium octasulfonate 0.1 M phosphate buffer (pH 2.1) and acetonitrile as a solvent.
[0056]
The results are shown in Table 2. In Table 2, A in the column of lactic acid bacteria represents Lactobacillus acidophilus, B represents Lactobacillus bifidus, and F represents Streptococcus faecalis.
[0057]
[Table 2]

[0058]
From the results shown in Table 2, it can be seen that various water-soluble vitamins significantly increase with the progress of lactic acid fermentation.
[0059]
Next, using Lactobacillus bifido as lactic acid bacteria and fermentation time of 240 hours, air was inserted into the fermentation broth obtained by the above method at a flow rate of 600 ml / min per 100 ml of fermentation broth using a gas blower. Then, the odor was removed by aeration for 5 hours.
[0060]
Table 3 shows the results of various analyzes conducted on the lactic acid bacteria fermentation broth obtained in this way to check the safety of food.
[0061]
[Table 3]

[0062]
Table 4 shows the results of amino acid analysis of protein components contained in the lactic acid bacteria fermentation broth. In addition, the amino acid analysis was performed by hydrolyzing hydrochloric acid after performing the formic acid oxidation treatment, and the amino acid automatic analysis method was performed.
[0063]
[Table 4]

[0064]
From the results of Tables 2 to 4, it can be seen that the lactic acid bacteria fermentation broth is a nutrient-rich beverage containing protein or a decomposition product thereof, sugar, water-soluble vitamin, and mineral.
[0065]
Example 2
The rice bran of Example 1 is replaced with brown rice flour finely pulverized with a ball mill, the amount of water added is twice the weight of the brown rice flour, and the amount of α-amylase during the heat treatment is 0.6% by weight with respect to the brown rice flour. A lactic acid bacteria fermentation broth was obtained in the same manner as in Example 1 except that.
[0066]
Analysis of the components of the lactic acid bacteria fermentation broth revealed that it contained a protein or a degradation product thereof, a saccharide, a water-soluble vitamin, and a mineral as in the case of the lactic acid bacterium fermentation broth of Example 1, and had a slightly high sugar content.
[0067]
Test example 1
Three groups of SD male rats of 3 weeks of age are prepared in total, 4 groups of 6 to 6 for each group, and only distilled water is used for Group A and Group C as normal solid feed (made by Oriental yeast) and drinking water. For the B group and the D group, a liquid obtained by diluting the lactic acid bacteria fermentation liquid obtained in Example 1 to 1/2 with distilled water (hereinafter simply referred to as “1/2 fermentation diluted liquid”) was freely given. All groups were preliminarily raised for 17 days. All groups were fasted for 1 day on the 17th day of preliminary breeding, and further reared for groups A to D as follows. In addition, after fasting for one day, all the groups were allowed to freely consume the same solid feed as described above.
[0068]
Group A: Only distilled water was again raised as drinking water.
Group B: The 1/2 fermentation dilution was again raised as drinking water.
Group C: After fasting for 1 day, alloxan was intraperitoneally administered in an amount of 0.25 ml (10 mg) / 100 g body weight, and then again distilled water alone was raised as drinking water.
Group D: After fasting for 1 day, alloxan was intraperitoneally administered in an amount of 0.25 ml (10 mg) / 100 g body weight, and then the 1/2 fermentation dilution was again raised as drinking water.
[0069]
Alloxan is known as a hormone that destroys β-cells of the pancreas and exerts a hypoglycemic action, a substance that suppresses insulin secretion, and is used in various experiments as a substance that artificially induces diabetes. It is what.
[0070]
During this breeding period, the average body weight per animal, blood sugar level, urinary sugar level, urine volume per day, average water consumption per day, average food consumption per day, Each value of urinary urobilinogen, urine protein, urinary ketone body, and urine pH was measured over time.
[0071]
The blood glucose level was measured using “Antosense Cartridge” (trade name, manufactured by Daikin Industries, Ltd., sold by Ono Pharmaceutical Co., Ltd.) as a test paper. Measured by Sanyo Co., Ltd.).
[0072]
The urinary sugar level, urinary urobilinogen, urinary protein level, urinary ketone body, and urine pH were measured using a test paper “Urolabs Sticks” (trade name, manufactured by Miles Sankyo Co., Ltd.).
[0073]
FIG. 1 shows the average body weight per animal in each group. The A group and the D group showed the same behavior, and the B group and the C group showed the same behavior. It should be noted that group 1/2, which was bred 1/2 drinking dilution as drinking water and administered alloxan, was fed only distilled water as drinking water, and weight after administration of alloxan compared to group C administered alloxan. The increase is large, and the same behavior as that of the group A not administered with alloxan is shown. One animal in Group C died of alloxan toxicity on the third day after alloxan administration.
[0074]
FIG. 2 shows the blood glucose level measurement results for each group. Group C, which was bred only as distilled water and administered with alloxan, showed a marked increase in blood glucose level. Group D, which was bred as a half-fermented dilution as drinking water and administered with alloxan, It can be seen that the increase in value is halved.
[0075]
FIG. 3 shows the measurement results of the urine sugar value of each group. Group C, which was bred only as distilled water and administered with alloxan, showed a marked increase in urine sugar level, but group D which was bred with 1/2 fermentation dilution as drinking water and administered with alloxan, It can be seen that the increase in the urinary sugar value is considerably reduced.
[0076]
FIG. 4 shows the measurement results of urine volume in each group. Group C, which was bred only as distilled water and administered with alloxan, showed a marked increase in urine volume. However, Group D, which was bred with 1/2 fermented dilution as drinking water and administered with alloxan, The normal value continued to be maintained as in the group.
[0077]
FIG. 5 shows the measurement results of the amount of drinking water in each group. Only the distilled water was raised as drinking water and only the group C administered with alloxan showed a significant increase in the amount of drinking water.
[0078]
The feed intake was slightly decreased in the latter half of the experiment only in group B, but the other groups showed almost the same results. Moreover, although the urine protein value showed the result which was completely complicated for each group, when it looked through the whole test period, it showed the excretion amount of the whole group almost the same amount. Furthermore, although the urine pH was complicated for each group, the same results were obtained when viewed throughout the entire test period. Urinary urobilinogen and urinary ketone bodies showed the same results in all groups, and no change was observed.
[0079]
Considering the above results, especially from the results of FIGS. 2 to 5, group C, which is fed only distilled water as drinking water and administered with alloxan, shows remarkable diabetic symptoms after alloxan administration. It can be seen that Group D, which was bred as a drinking water and administered with alloxan, significantly suppressed the symptoms of diabetes.
[0080]
Test example 2
10 male rats of 5 weeks of age are prepared for each group, 2 groups of Group A and Group B. Group A uses only distilled water as drinking water, Group B uses the same 1/2 fermentation dilution as in Test Example 1. As the drinking water, the same normal solid feed as in Test Example 1 was used as the feed, and the drinking water and the feed were fed freely.
[0081]
Then, body weight, water consumption, and food intake during the breeding period were measured over time. On the last day of the experiment, 0.05 ml (2 μg) of phytohaemagglutinin was injected into the earlobe endothelium, and the degree of red spots 24 hours later was measured with a color photometric colorimeter manufactured by Nihon Kohden. The degree was measured.
[0082]
Note that phytohemaglutinin acts as an antigen, and it is determined that the higher the red spot after injecting phytohemagglutinin, the stronger the antibody-producing ability by antigen-antibody reaction.
[0083]
FIG. 6 shows the change in body weight, and both groups showed exactly the same behavior.
[0084]
FIG. 7 shows changes in the amount of drinking water per day, and although there are some fluctuations, it can be seen that the intake of group B is large throughout the entire test period.
[0085]
FIG. 8 shows the amount of food consumed per day for both groups A and B. For group B, the total amount of food added by adding the amount of solid and liquid contained in the drinking water consisting of 1/2 fermentation dilution to the amount of food in group B was calculated, and B ′ ( -Δ- graph).
[0086]
When comparing the amount of food consumed by both groups A and B, it is clear that group A has more food, but when comparing B 'with the amount of solid liquid contained in the drinking water, both groups are almost the same. It can be seen that it shows a good behavior.
[0087]
That is, it can be seen that group B is ingesting the solid content from the diluted fermentation solution, so that the amount of food intake decreases accordingly. In other words, it can be seen that group B compensates for the shortage of food intake by favoring the fermented diluent even if the food intake of the solid feed is reduced.
[0088]
FIG. 9 shows the result of the immune reaction on the next day after injecting phytohemaglutinin. In the figure, an increase in a value indicates that red is strong and an increase in b value indicates that yellow is strong. Since the immune response is more pronounced as the red color is stronger, it can be seen that the group B clearly has a stronger immune response and enhanced immunity compared to the group A.
[0089]
Test example 3
Two 5-week-old male rats, each of group A and group B, are prepared. Group A uses only distilled water as drinking water. Group B uses the lactic acid bacteria fermentation solution obtained in Example 1 as a distilled water. A liquid diluted to 1/3 with water (hereinafter simply referred to as “1/3 fermentation diluted liquid”) is used as drinking water, and the same solid feed as in Test Example 1 is used as the feed, and drinking water and feed are ingested freely. The animals were reared for 35 days.
[0090]
Then, the average body weight per animal in each group on the last day (10 weeks of age) of this breeding period, and the average amount of water consumed and the average food intake per animal in each group on the first day of the last day were measured. .
[0091]
FIG. 10 shows the average body weight, which is almost the same in both groups A and B, but the group B is slightly increased.
[0092]
FIG. 11 shows the average amount of drinking water, and the group B is clearly increased, and it can be seen that in the case of growing rats, the fermented diluted solution is preferred.
[0093]
FIG. 12 shows the average food intake. In addition, about the B group, the total feed amount which added the amount of solid content contained in the drinking water which consists of 1/3 fermentation dilution liquid to the feed amount of the said B group was calculated, and it shows by B 'in a figure It was.
[0094]
When comparing the amount of food consumed by both groups A and B, it is clear that group A has more food, but there is no significant difference when comparing B 'with the amount of solids contained in the drinking water. Recognize.
[0095]
Therefore, it can be seen that even in a 10-week-old growing rat, the body weight is increased as compared with the case where the fermented diluted solution is favored and the distilled water is used as drinking water.
[0096]
Test example 4
Two groups of 13-week-old middle-aged male rats, 10 groups each, Group A and Group B are prepared. Group A uses only distilled water as drinking water, and Group B uses the same 1/3 fermentation as in Test Example 3. The diluted solution was used as drinking water, and as the feed, the same solid feed as in Test Example 1 was used, and the drinking water and feed were freely ingested and reared for 1 day.
[0097]
And the average amount of water consumed per animal in the said 1 day was measured. The result is shown in FIG.
[0098]
As shown in FIG. 13, it can be seen that middle-aged rats also prefer to drink the fermentation dilution.
[0099]
From the above test results, the fermented diluent containing the therapeutic agent for diabetes and the immunopotentiator of the present invention tends to be preferably consumed as it grows from a young rat to a grown rat and a middle aged rat. Even if it decreases, it will be drunk, and it turns out that this fermentation dilution liquid is a very delicious drink for rats.
[0100]
Moreover, since an increase in body weight is shown with an ingestion of fermentation dilution liquid, it turns out that this fermentation dilution liquid is not only delicious but also a nutrition-rich beverage.
[0101]
And it turns out that immunity is strengthened by drinking this fermentation dilution liquid, and the effect which remarkably reduces the diabetes symptom after administering alloxan which is a diabetes inducer is brought about.
[0102]
【The invention's effect】
As explained above, the diabetes treatment of the present invention Agent Because it is effective in reducing diabetes symptoms and enhancing immunity, it is delicious and rich in nutrients, it can be expected to restore the health of diabetics, etc. . The diabetes treatment of the present invention Agent As shown in Table 3, since it does not contain any dangerous ingredients in the human body and can be taken freely as a food, it can be taken easily and as much as you like in daily life. The effect of treatment can be enhanced by taking for a long time.
[Brief description of the drawings]
1 is a chart showing changes in average body weight of each group in Test Example 1. FIG.
FIG. 2 is a chart showing the transition of blood glucose level in each group in Test Example 1.
3 is a chart showing changes in urine sugar values of each group in Test Example 1. FIG.
4 is a chart showing changes in urine volume in each group in Test Example 1. FIG.
5 is a chart showing changes in the amount of drinking water in each group in Test Example 1. FIG.
6 is a chart showing changes in average body weight of each group in Test Example 2. FIG.
7 is a chart showing changes in the amount of drinking water in each group in Test Example 2. FIG.
8 is a chart showing changes in food intake of each group in Test Example 2. FIG.
9 is a chart showing the results of immune responses in each group in Test Example 2. FIG.
10 is a chart showing the average body weight of each group in Test Example 3. FIG.
11 is a chart showing the amount of water consumed in each group in Test Example 3. FIG.
12 is a chart showing food intake in each group in Test Example 3. FIG.
13 is a chart showing the amount of water consumed in each group in Test Example 4. FIG.

Claims (3)

After adding water to rice bran and / or brown rice flour, liquefied amylase is added and heat-treated, and cellulase, protease and saccharified amylase are added to the heat-treated solution and subjected to enzyme treatment, and the enzyme-treated solution is subjected to centrifugation or A therapeutic agent for diabetes characterized in that the fat or oil part is removed by filtration, the fermented liquid itself obtained by inoculating and fermenting lactic acid bacteria in this treated liquid, or a dried powdered form of the fermented liquid, as an active ingredient . It said fermentation liquid, antidiabetic agent according to claim 1, wherein those which are aerated after the end of fermentation. The therapeutic agent for diabetes according to claim 1 or 2 , wherein the fermented liquid is obtained by removing a water-insoluble part after completion of fermentation.

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