JP2014152131A - Oral anti-aging agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an anti-aging agent having the action of suppressing or repairing tissue atrophy with aging, thereby preventing or improving hypofunction caused by tissue atrophy.SOLUTION: An anti-aging agent contains whey or whey processed product as an active ingredient.

Description

  The present invention relates to an anti-aging agent for oral administration and ingestion. More specifically, the present invention relates to an oral anti-aging agent that has an action of preventing or improving functional deterioration due to tissue atrophy by suppressing or repairing tissue atrophy due to aging.

  It is known that with aging (aging, aging), the volume of individual parenchymal cells decreases or the number of parenchymal cells decreases, and the tissue gradually shrinks. This is called physiological atrophy or senile atrophy. Such physiological atrophy of the tissue also reduces the function of the tissue, thereby causing various symptoms and pathologies in the body.

  For example, when glandular cells such as salivary glands and lacrimal glands involved in exocrine atrophy, the function of these glandular tissues decreases, and “dry mouth” (dry mouth), “dry eye” (dry eye disease), “dry nose” (nasal nose) Dryness symptoms such as dryness), “dry skin” (dry skin), and “driver jainer” (vaginosis) and disorders such as chronic pancreatitis, chronic gastritis and chronic bronchitis due to reduced exocrine secretion occur.

  In particular, salivary secretion disorders caused by salivary gland atrophy cause the aforementioned “dry mice” (xerostomia). In dry mice, the decrease in saliva secretion directly dries the lips and oral cavity, causing problems such as dry mouth, dry mucous membranes, or burning sensation. It may cause diseases, caries, masticatory disorders, dysphagia, aspiration pneumonia, atrophy of gastrointestinal mucosa and reflux esophagitis. It is also pointed out that saliva deficiency increases the prevalence of adult diseases (arteriosclerosis, myocardial infarction, stroke, cerebral infarction, diabetes). Therefore, salivary secretion disorder is a pathological condition deeply related to health maintenance not only for elderly people who are prone to aspiration pneumonia due to dysphagia, but also for elderly people. For the treatment of salivary secretion disorders, especially dry mice, pharmaceuticals are prescribed for critically ill patients, but for those with relatively mild symptoms, a method using commercially available mouse wash or artificial saliva, chewing gum is used. Only coping therapy such as a method of chewing and a method of drinking a liquid little by little throughout the day has been performed (Non-Patent Document 1), and the establishment of a radical treatment is required.

  On the other hand, whey is a by-product discharged in large quantities during the manufacture of fermented dairy products such as cheese and yogurt. In recent years, industrialization of whey degreasing, concentrating and powdering production has progressed and it has become effective, but this is limited to major dairy companies with comprehensive dairy factories. It has been. Not only small and medium-sized enterprises have not yet been able to effectively use whey, but there are concerns about the cost of disposal, and this solution is desired.

Lancet 366, 321-331, 2005

  An object of this invention is to provide the oral anti-aging agent used in order to improve or prevent the pathological condition and symptom which arise with an aging. In particular, the present invention relates to an oral anti-aging agent that is used to suppress or prevent and improve tissue atrophy due to aging, that is, a tissue atrophy suppression / restoration agent, and to suppress and prevent functional deterioration associated with tissue atrophy due to aging. Or it aims at providing the oral anti-aging agent used in order to improve, ie, a tissue function improving agent.

  The inventors of the present invention have made extensive studies to solve the above-mentioned problems, and suppress wrinkles that occur in glandular tissues such as salivary glands and organ tissues such as lungs in whey, which has been a problem in the past. It was found that there is an action to repair, and by actually ingesting whey in elderly rats with advanced aging, the atrophy of salivary gland tissue was suppressed or repaired compared to rats not fed whey, and the amount of saliva It was confirmed that gene expression related to salivary gland function and salivary gland function increased significantly, and lung function such as oxygen uptake into blood increased significantly by suppressing or repairing atrophy of lung tissue.

The present invention has been completed under further research based on such knowledge, and has the following embodiments.
(1) An oral anti-aging agent containing whey or processed whey as an active ingredient.
(2) The oral anti-aging agent according to (1), which is a tissue atrophy inhibiting / restoring agent for suppressing or repairing tissue atrophy due to aging.

In addition, the anti-aging agent contains “whey or processed whey product as an active ingredient, an agent for suppressing or restoring tissue atrophy due to aging consisting of an oral dosage form” or “whey or processed whey product as an active ingredient, In other words, it can be rephrased as an agent for suppressing or restoring senile atrophy of a tissue comprising an oral dosage form.
(3) The oral anti-aging agent according to (1), which is a tissue function improving agent for improving functional deterioration associated with tissue atrophy due to aging.

In addition, the anti-aging agent includes “a tissue function improving agent for improving functional deterioration associated with tissue atrophy due to aging, comprising whey or a processed whey product as an active ingredient, or an oral administration form” or “whey or In other words, it can be rephrased as a “tissue function improving agent for improving functional deterioration associated with aging atrophy of a tissue, comprising an orally administered form containing a processed whey product as an active ingredient”.
(4) The oral anti-aging agent according to (1), which is used for improving a disease state or a condition caused by a functional decrease due to tissue atrophy due to aging.
(5) The oral anti-aging agent according to any one of (2) to (4), wherein the tissue is exocrine gland tissue or lung tissue.
(6) The oral anti-aging agent according to (5), wherein the exocrine gland tissue is salivary gland tissue.
(7) The oral anti-aging agent according to (4), wherein the symptom or pathology caused by the functional decline associated with tissue atrophy due to aging is an exocrine disorder or respiratory failure. In addition, the said anti-aging agent can be paraphrased as "the preventive or improving agent of the exocrine disorder or respiratory failure by aging which consists of an oral administration form which contains whey or a whey processed product as an active ingredient."
(8) The oral anti-aging agent according to (7), wherein the exocrine disorder is xerostomia (dry mouth). In addition, the said anti-aging agent can be paraphrased with "the prevention or improvement agent of the dry mouth of the mouth by age which consists of an oral administration form which contains whey or a whey processed product as an active ingredient."
(9) The oral anti-aging agent according to any one of (1) to (8), which has a solid preparation form selected from tablets, pills, powders (powder), granules, and capsules.
(10) The oral anti-aging agent according to any one of (1) to (9), which is a supplement.

  “Supplement” refers to nutrients (eg vitamins and minerals) that are often deficient in daily diet and functional ingredients that cannot be adequately supplemented in daily diet, separately from the normal diet. Ingested, solid preparations such as tablets, pills, powders (powder), granules, capsules (including soft capsules and hard capsules), and liquids such as drinks and syrups It refers to a preparation having a different form of preparation from general foods such as a preparation. In Japan, it is also called “health functional food” (nutrient functional food, food for specified health use) and in the United States “dietary supplement”. The “supplement” is a food having a purpose of health, health maintenance / promotion, etc. in a more positive sense than general food.

  According to the anti-aging agent of the present invention, it is possible to suppress a decrease in tissue function by preventing and repairing tissue atrophy due to aging. For this reason, the oral anti-aging agent of this invention can be used effectively in order to suppress and improve said various pathological conditions and states accompanying the said tissue function fall.

  In particular, according to the oral anti-aging agent of the present invention, it is possible to prevent a decrease in function by preventing or repairing atrophy of salivary glands (including sublingual glands and submandibular glands). It is effective in improving symptom (dry mouse). Improvement of xerostomia not only stabilizes oral function (antibacterial action, chewing / swallowing assistance, protection of oral mucosa, etc.) but also constancy of digestive tract function (immune action, prevention of reflux esophagitis) ), And thus, may be useful for systemic homeostasis (adult disease prevention).

JH test samples prepared from mature rats (Fig. 1 (a)) and elderly rats (Fig. 1 (b)) (above, tap water administration group) and jersey whey (JH) that were orally ingested with tap water for 2 months Shows hematoxylene / eosin-stained images of salivary gland tissues of mature rats (Fig. 1 (c)) and elderly rats (Fig. 1 (d)) (JH administration group) ingested orally for 2 months (Experimental example) 1). JH test samples prepared from mature rats (Fig. 2 (a)) and elderly rats (Fig. 2 (c)) (tap water administration group) and jersey whey (JH) that were orally ingested for 2 months. Rats orally ingested for 2 months (FIG. 2 (b)) (JH administration group) and elderly rats orally administered HH test samples prepared from Holstein whey (HH) for 2 months (FIG. 2 (d)) ) (HH administration group) shows a hematoxylene / eosin stained image of lung tissue (Experimental Example 1). The result of having measured the amount of proline rich protein in salivary glands and saliva is shown. FIG. 3 (a) shows the results of subjecting the salivary glands to SDS-PAGE (lane 1: mature rat control group (tap water administration) lane 2: mature rat JH administration group, lane 3: elderly rat control group (tap water administration), Lane 4: elderly rat JH administration group), FIG. 3B shows the results of applying saliva to SDS-PAGE (lane 1: elderly rat control group (tap water administration), lane 2: elderly rat JH administration group, lane 3: elderly rat HH administration group). FIG. 3 (c) shows the result of subjecting SDS-PAGE of saliva to Wester blotting (lane 1: elderly rat control group (tap water administration), lane 2: elderly rat JH administration group, lane 3: elderly rat HH administration group). Indicates.

  The oral anti-aging agent of the present invention contains whey or processed whey as an active ingredient, and has an oral dosage form (oral intake form).

(1) Whey or whey processed product The whey used as an active ingredient of the oral anti-aging agent of the present invention is a water fraction discharged after removing curd produced by adding rennet or acid to milk or skim milk. It is obtained as a by-product of cheese manufacture and casein manufacture.

  The whey targeted by the present invention includes acidic whey (fermented whey, casein whey), cheese whey, and heated (heat-denatured) whey described below.

  Examples of acidic whey include fermented whey obtained by fermentation using lactic acid bacteria, and casein whey obtained by removing all or most of the curd containing casein protein produced by adding acid to milk according to a conventional method.

  Fermented whey can be usually obtained by a method of fermenting milk with lactic acid bacteria or symbiotic fermentation with lactic acid bacteria and yeast. Examples of lactic acid bacteria include lactic acid bacteria belonging to the genus Streptococcus, Lactococcus, Lactobacillus, Bifidobacterium, and the like. Lactic acid bacteria belonging to the genus Lactobacillus are preferred. Specifically, for example, Lactobacillus bulgaricus (Lactobacillus hebuleticus), Lactobacillus helveticus, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus acidophilus, Lactobacillus fermentum ( Lactobacillus fermentum) and the like, and Lactobacillus helveticus can be particularly preferably used.

  The species of yeast used for symbiotic fermentation with lactic acid bacteria is not particularly limited, but preferred examples include yeasts of the genus Saccharomyces such as Saccharomyces cerevisiae.

Fermentation is usually carried out by standing or stirring culture, for example, at a fermentation temperature of 20 to 50 ° C. and a fermentation initial pH of about 6 to ⁷ , for example, when the number of bacteria is 10 ⁷ cells / ml or more and pH 5 or less. To stop. The card | curd is isolate | separated by normal isolation | separation operation from the fermented milk obtained in this way, and fermented whey is acquired.

  The casein whey has an acidity capable of mainly removing proteins such as casein by adding an acid such as lactic acid, citric acid, acetic acid, tartaric acid, fumaric acid, malic acid, gluconic acid, or adipic acid to milk. After adjustment, it is obtained by separating the whey component (water fraction) according to a conventional method such as membrane treatment. Moreover, the mixing ratio of an acid can be normally mixed in the ratio from which acidity will be 1-4% according to the kind of acid.

  The cheese whey can be obtained by separating the curd according to a conventional method after solidifying the curd with a rennet according to normal cheese production.

  The heated (heat-denatured) whey means a whey component (water fraction) obtained by heating to remove mainly proteins such as casein by heat denaturation. For example, milk is 80 ° C. or higher, preferably 150 ° C. It can be prepared by heating at the above temperature.

  Milk used as a raw material for whey used in the present invention is not limited to milk extracted from cows, and may be animal milk extracted from humans, goats, horses, camels, sheep, etc., and plant milk such as soy milk. It may be. Preferably it is milk extracted from cows. Here, the type of cattle is not particularly limited, Jersey, Holstein, Angler, Welsh Black, Ayrshire, Groningen, Kelly, South Devon, Simmental, Swedish Red and White, Dexter, Dairy Shorthorn, Norwegian Red, Norman, Pyrrouge Flanders, Finnish, Brown Swiss, Museline Ysell, Montbelière, Lincoln Red, Red Danish, Red Pole, and Red Grass The cows are preferably Jersey and Holstein cows, and more preferably Jersey cows.

  In addition to the raw milk described above, processed milk such as skim milk, reduced milk, powdered milk, condensed milk, and the like, which are processed products, can also be used as the raw milk, and can also be used as a mixture thereof. In addition, although the solid content concentration of the raw material milk used for manufacture of whey is not specifically limited, For example, when using skim milk, the nonfat milk solid content concentration is used about 9 mass% most often. In the case of solid milk such as powdered milk, the one dissolved in distilled water is used when preparing whey.

  For the preparation of the oral anti-aging agent of the present invention, the whey obtained by the above method is used as it is, and the whey is separated / fractionated, purified, concentrated, sterilized or / and dried. Whey processed products prepared for treatment can be used. The whey processed products include “casein-reduced whey processed products” in which casein, which accounts for the majority of the protein, is removed from whey, and the amount thereof is reduced, and “lipid-reduced whey processed products in which lipid is removed from whey and the amount thereof is reduced. "" Lactose-reduced whey processed product "in which lactose is removed from whey and the amount thereof is reduced.

  Although not limited, casein removal from whey can be reduced by immunoprecipitation of whey using an anti-casein antibody or by lowering the pH below the isoelectric point (about 4.6). In addition, as a method of removing lipid from whey and reducing its weight, the whey is subjected to centrifugation (for example, centrifugation at 3,500 g for 15 minutes, etc.) to remove the uppermost layer containing lipid, and lactose is removed from whey. Examples of the method for reducing the amount of weight to be removed include, but are not limited to, a method of gel filtration of whey and a method of subjecting to whey treatment (such as lactase).

  Although not limited, whey removes low molecular weight fractions by removing membranes such as ultrafiltration membranes or microfiltration membranes after removing lipids and other solids by centrifugation, and preferably has a molecular weight of about 14,000 or more. By concentrating the fraction, the anti-aging action (tissue atrophy suppression action, tissue atrophy repair action, tissue function improvement action) can be further enhanced.

  Sterilization treatment can be performed using sterilization treatment usually applied to whey and dairy products, but in order to avoid a decrease in anti-aging action (tissue atrophy suppression / restoration action, tissue function improvement action) due to heat treatment, Preferably low-temperature hold sterilization method (method of heat sterilization at about 63-65 ° C for about 30 minutes), ultra-high temperature short time sterilization method (high temperature sterilization (HTST method) of about 15 seconds at about 75 ° C, at 120-130 ° C Ultra-high temperature instantaneous sterilization (UHT method or the like) that is heated for 2 to 3 seconds, more preferably low pressure or slightly high pressure carbon dioxide gas sterilization.

  As the drying treatment, a drying treatment usually applied to whey and dairy products such as freeze-drying method and spray-drying method (spray drying method) can be used. In order to avoid a decrease in anti-aging action (tissue atrophy suppression action, tissue atrophy repair action, tissue function improvement action) due to heat, a vacuum spray drying method is preferred. The reduced pressure spray drying method is an improved method for drying at a lower temperature than the conventional spray drying method by reducing the pressure in the drying tower.

  Although not limited, whey processed products targeted by the present invention include whey (acid whey (fermented whey, casein whey), cheese whey, or heated (heat-denatured whey)), preferably acidic whey. It is subjected to centrifugation to obtain a liquid fraction, which is then subjected to microfiltration using a microfiltration membrane having a pore size of 0.1 to 10 μm, and the obtained filtrate is sterilized by sterilization using a high-pressure carbon dioxide gas, and then spray-dried powder. Whey processed products prepared by dry powdering by a drying method are included.

(2) Oral anti-aging agent The above-mentioned whey and processed whey products are orally administered (ingested) to mammals including humans, so that tissue atrophy due to aging (physiological atrophy, senile atrophy) ) Can be used as an “oral anti-aging agent”. In addition, whey and processed products of whey are used as “tissue atrophy suppression / restoration agents due to aging” (hereinafter simply referred to as “tissue atrophy suppression / restoration agents”), further suppressing and improving functional deterioration associated with tissue atrophy due to aging. Therefore, it can be used as a tissue function improving agent (hereinafter simply referred to as “tissue function improving agent”). In the present invention, the term “oral anti-aging agent” is used to encompass the above-mentioned “tissue atrophy inhibiting / restoring agent” and “tissue function improving agent”. Therefore, the “oral anti-aging agent” described below is also a description of “tissue atrophy prevention / restoration agent” and “tissue function improving agent”.

  Although content in particular of the said whey and whey processed product contained in the oral anti-aging agent of this invention is not restrict | limited, It can select suitably from the range of 1-100 mass% in conversion of solid substance. Preferably it is 10-100 mass%, More preferably, it is 30-100 mass%.

  The oral anti-aging agent of the present invention can be prepared in various dosage forms for oral administration or ingestion. Examples of such preparation forms include solid preparation forms such as tablets, pills, powders (powder), granules (fine granules), and capsules. Tablets, powders (powder), and granules (fine granules) are preferable. The solid preparation may be administered or ingested after being dissolved in water at the time of use.

  In preparing the oral anti-aging agent of the present invention in such a preparation form, various additives according to each preparation form can be blended. For example, excipients, binders, disintegrants, stabilizers, surfactants, plasticizers, lubricants, buffers, sweeteners, flavoring agents, antioxidants, coating agents (including sugar coatings), wetting Agent, wetting agent, filler, antifoaming agent, cooling agent, binder, coloring agent, flavoring agent, tonicity agent, softening agent, emulsifier, thickening agent, foaming agent, pH adjusting agent, dispersion An agent, a disintegrating aid, a fragrance, a moisture-proofing agent, a preservative, a solubilizing aid, a solvent, a fluidizing agent, a bulking agent, and the like can be blended. Examples of such additives are described in the Pharmaceutical Additives Dictionary, food additives, quasi-drug raw material standards, Japanese general-purpose cosmetic raw material collection, permission standards for cosmetic varieties, cosmetic raw material non-standard ingredient standards, CTFA, and the like. In addition, additives such as sugars, proteins, lipids, vitamins, minerals, flavors, or mixtures thereof can be added. Moreover, you may further contain milk components other than whey before isolate | separating whey.

  The oral anti-aging agent targeted by the present invention is an adult for the purpose of suppressing or repairing tissue atrophy due to aging, and for the purpose of suppressing and improving the functional deterioration of tissue associated with tissue atrophy due to aging. For males and females, preferably 30 years old or older, more preferably 35 years old or older, more preferably 40 years old or older, particularly preferably 45 years old or older, even more preferably 50 years old or older, more preferably 60 years old or older, As long as it has a pharmaceutical form that can be administered or ingested orally, it does not matter whether it is a pharmaceutical, a quasi-drug, or a food or drink. Preferably, it is a food product having a solid preparation form, and more preferably a supplement having a preparation form referred to as “health functional food”. Supplements targeted by the present invention include nutritional functional foods and foods for specified health use (including conditional foods for specified health use) that can specifically show the functions and effects of the foods on packaging containers, etc. However, the present invention is not limited to this, and includes foods that are labeled such that the functions and effects are imaged by consumers instead of the description of specific functions and effects.

  As a guideline of the administration (intake) amount of the oral anti-aging agent of the present invention, a range of 1 g to 50 g per day for an adult can be mentioned in terms of effective whey or processed whey (solid content). The amount can be administered (taken) once a day or divided into 2 to 10 times a day, for example, after a meal.

  The oral anti-aging agent of the present invention is used for the purpose of suppressing or repairing tissue atrophy due to aging as described above, and for the purpose of suppressing and improving the functional deterioration of the tissue associated with tissue atrophy due to aging. be able to. Here, the tissues include, but are not limited to, exocrine glands such as salivary glands (including sublingual gland, submandibular gland, parotid gland and minor salivary gland), gastric gland, sweat gland, lacrimal gland, mammary gland, vaginal gland and sebaceous gland; Mention may be made of organ tissues such as lung, brain, liver and kidney. Preferred are exocrine glands such as salivary glands and lung tissue.

  Examples of the pathological condition or condition caused by the functional decline associated with such tissue atrophy include, for example, “dry mouse” (xerostomia), “dry eye” (dry eye), “dry nose” due to glandular tissue functional decline related to exocrine secretion. (Nasal dryness), "dry skin" (dry skin), and "driver jainer" (vaginosis), and swallowing difficulty, vocalization, taste abnormalities, reflux esophagitis due to decreased exocrine secretion, Mention may be made of disorders such as chronic pancreatitis, chronic gastritis and chronic bronchitis. In addition, the ability of oxygen to be taken into the blood is reduced due to a decrease in lung function, resulting in problems such as shortness of breath, hypoxemia and respiratory failure. In addition, a decrease in the function of these organs also causes disorders such as cough and throat (burden).

  According to the oral anti-aging agent of the present invention, it is possible to suppress the functional deterioration of the tissue by preventing and repairing tissue atrophy due to aging. For this reason, the oral anti-aging agent of this invention can be used effectively in order to suppress and improve said various pathological conditions and states accompanying the said tissue function fall.

  Hereinafter, the configuration and effects of the present invention will be described using examples and experimental examples. However, the experimental examples and examples are examples of the present invention, and the present invention is not limited to these experimental examples and examples.

Experimental example 1
(1) Preparation of test sample As whey, whey prepared from Jersey cow milk (Jersey whey: JH) and whey prepared from Holstein cow milk (Holstein whey: HH) were used. Each whey was prepared by reacting with renin at 37 ° C for 1 hour.

  These whey (JH, HH) are centrifuged at 3,500g for 15 minutes at 4 ° C to remove the uppermost layer containing fat and precipitates containing solid components such as lactic acid bacteria. Minutes were collected. The following experiment was conducted using this as a JH test sample and an HH test sample.

(2) Experimental method To 3 month old Wistar male rats (mature rats) and 14 months old Wistar male rats (old rats), the JH test sample, HH test sample or tap water prepared above was supplied for 2 months. They were allowed to drink freely (JH administration group: 5 mature rats, 5 elderly rats; HH administration group: 3 mature rats, 3 elderly rats; tap water administration group: 5 mature rats, 5 elderly rats). During the study period, the same solid food (MF, Oriental) was used for the JH administration group (mature rats and elderly rats), the HH administration group (mature rats and elderly rats), and the tap water administration group (mature rats and elderly rats). Yeast Co., Ltd., Tokyo) was ingested freely.

  Two months later, rats in the test sample 1 administration group, the test sample 2 administration group, and the tap water administration group were killed by spinal cord dislocation, and salivary glands and lungs were removed. Each excised tissue was fixed with formalin according to a conventional method, and then a section was prepared from the tissue embedded in paraffin and stained with hematoxylin / eosin. This was observed with an optical microscope (Leica DMLS HC, Wetzlar Germany).

(3) Experimental results The results are shown in FIG. 1 for salivary gland tissue and in FIG. 2 for lung tissue.

  As for salivary gland tissue, JH test samples prepared from whey were administered orally, as is apparent from the comparison of the results of elderly rats in the tap water administration group (FIG. 1 (b)) and elderly rats in the JH administration group (FIG. 1 (d)). By ingestion, atrophy of salivary gland tissue, particularly atrophy of mucous cells in the sublingual gland (white portion in the figure) was significantly suppressed or repaired. This indicates that whey has an action of suppressing or repairing atrophy of salivary gland tissue due to aging.

  As for the lung tissue, as shown in FIG. 2 (c), the elderly rats in the tap water administration group have a large alveolar space and atrophy and necrosis of tissue cells. As can be seen from the comparison of the results of the elderly rats (b) in the administration group, orally administering the JH test sample prepared from Jersey Whey (JH) reduced the alveolar space, resulting in lung tissue cell atrophy and necrosis. Suppressed or repaired. Blood vessel formation was also observed. In addition, as shown in FIG. 2 (d), the alveolar space is reduced in Holstein whey (HH) as in Jersey whey (JH) to suppress atrophy and necrosis of lung tissue cells. Or it was confirmed that there is an action to repair. From this, it was confirmed that whey has an action of suppressing or repairing atrophy of lung tissue due to aging.

  Based on the above results, whey has the effect of suppressing or repairing atrophy of tissues (eg, salivary gland, lung tissue) due to aging, and the action is related to the type of cattle and other animals such as Jersey and Holstein. Although it was observed, it was confirmed that it was particularly noticeable in whey derived from Jersey milk.

Experimental example 2
(1) Experimental method Three-month old Wistar male rats (mature rats) and 16-month old Wistar male rats (old rats) were divided into three groups (n = 5 in each group). The JH test sample (JH administration group), HH test sample (HH administration group), or tap water (control group) prepared in 1) was allowed to drink freely for 2 months.

(1-1) Measurement of saliva amount and cystatin amount, amylase amount, and mucin amount in saliva Two months after administration, the amount of saliva in each group, and the amount of cystatin, amylase, and mucin in saliva are as follows: It measured according to the method of (a)-(d).

  In addition, cystatin (cistatin S) in saliva is known to inhibit cysteine protease, exert antiviral action and antibacterial action, and suppress the growth of periodontal disease bacteria. In addition, amylase in saliva has a digestive action in the oral cavity and a self-cleaning action that decomposes components in the food residue. Furthermore, mucin in saliva contributes to the protection of the oral mucosa, lubrication, prevention of microbial adhesion, defense against infection, and adsorption and aggregation of bacteria. Usually, these ingredients contained in saliva combine to maintain the oral food intake function (digestive function, taste) and oral environment (self-cleaning, mucosal protection, antibacterial, anti-inflammatory, recalcification Chemistry and immunity) are well balanced.

(A) Measurement of saliva volume A muscarinic drug (cevimeline) was intraperitoneally administered at a rate of 10 mg / kg to each group of adult rats and elderly rats (JH administration group, HH administration group, control group). For ~ 20 minutes, saliva was collected from the mouth with a dropper and the total amount was measured.

(B) Measurement of cystatin amount Using the saliva collected above as a sample, the amount of cystatin in the saliva was measured according to the method of Abrahamson (Methods Enzymol, 244, 685-700, 1994).

(C) Measurement of amylase activity Using the saliva collected above as a sample, the amount of amylase in the saliva was measured according to Bernfeld's method (Methods Enzymol, 1, 149-154, 1955).

(D) Measurement of the amount of mucin Using the saliva collected above as a sample, the amount of mucin in the saliva was measured according to the Alcian blue method (Biochem Soc Trans 8, 72-73, 1980).

(1-2) Measurement of gene expression level Two months after administration of each sample, salivary glands were removed from each group of rats, and cystatin (cistatin S), lysozyme, amylase (amylase, α1A) in salivary gland (sublingual gland), And the expression level of mucin (similar to Mucin-4) and theobromine induced protein. The expression level of each gene was extracted from total RNA from the sublingual gland, biotin-labeled cRNA was prepared from this total RNA, and hybridized with Affymetrix Rat Gene 1.0ST Array set (Affymetrix) capable of analyzing 23,000 genes I let you. The biotin label was labeled with fluorescence, and microarray expression data was obtained from the fluorescence using an Agilent Technologies Microarray Scanner (manufactured by Agilent Technologies), and analyzed using GeneSpring GX10 Expression Analysis Software (Agilent Technologies, CA).

(2) Experimental results (2--1) Saliva volume, cystatin volume, amylase volume, and mucin volume in saliva For each group of mature and elderly rats (JH administration group, HH administration group, control group), saliva Table 1 shows the results of measuring the amount and the amount of cystatin, amount of amylase, and amount of mucin in saliva.

  As can be seen from this result, in the elderly rats, the whey intake (JH administration group, HH administration group) significantly increased in saliva volume compared to the rats (control group) ingestion of tap water. It was confirmed that the amount of cystatin, the amount of amylase, and the amount of mucin in saliva were also significantly increased. The effect of increasing these amounts was remarkably recognized by ingestion of jersey whey among whey. This indicates that whey has the effect of increasing the amount of saliva, the amount of cystatin, the amount of amylase, and the amount of mucin in saliva. From the results of Experimental Example 1, this action and effect is considered to be an action and effect obtained by suppressing or improving salivary gland atrophy due to aging and the resulting decrease in salivary gland function by oral intake of whey.

(2-2) Gene expression level in salivary glands For elderly rats administered jersey whey (JH), Holstein whey (HH) and tap water, and mature rats administered tap water, Table 2 shows the results of comparing the expression levels of genes (cystatin S), lysozyme, amylase (amylase, α1A), mucin (similar to Mucin-4), and theobromine induced protein.

  From the results of Table 2, it was confirmed that oral intake of whey, particularly jersey whey (JH), caused cystatin S and lysozyme having antibacterial action, amylase having digestive action (amylase α1A), and mucin having mucosal protective action in salivary glands. It can be seen that the expression level of is significantly increased. Among them, Jersey whey (JH) was confirmed to recover and increase the gene expression levels of cystatin S and mucin (similar to Mucin-4) in the salivary gland (sublingual gland) decreased by aging.

  From this, it can be seen that whey suppresses or improves salivary gland functional decline also from a genetic aspect.

Experimental example 3
(1) Experimental method (1-1) Measurement of proline-rich protein mass in saliva and salivary glands Rats prepared in Experimental Example 2 (aged rat JH administration group, HH administration group, and control group (tap water administration); Saliva and salivary glands were collected from a control group of adult rats (tap water administration), and the amounts of proline-rich protein contained in these saliva and salivary glands were measured and compared.

  Proline-rich protein accounts for about 40% of saliva protein and binds to dietary components such as tannin, which not only protects the oral cavity, stomach, and intestines, but also binds calcium ions to form tartar and salivary stone. It prevents or adsorbs to the tooth enamel crystal surface (forms a pellicle) to protect the teeth from acids and prevent bacteria from adhering to the tooth surface. It is also a protein that plays a role of lubrication.

  The amount of proline-rich protein in the salivary gland is obtained by subjecting the salivary gland to SDS-PAGE, and using a Western blotting method using anti-proline-rich protein 1-3 antibody, the concentration of the band obtained from the known amount of proline-rich protein It was calculated by comparing with the intensity of the band. In addition, the amount of proline-rich protein in saliva is the same as that of salivary glands by subjecting the saliva to SDS-PAGE and the density of the band obtained by Western blotting. Calculated by comparison.

(1-2) Measurement of gene expression level of proline-rich protein in salivary gland (sublingual gland) Two months after administration of each sample, rats in each group (JH administration group, HH administration group, and control group of elderly rats) (Tap water administration); Remove the salivary gland (sublingual gland) from the control group of adult rats (tap water administration), and protein rich protein (proline-rich protein 2, proline-rich glycoprotein) , Proline-rich protein 15, proline-rich proteoglycan 1, and proline-rich protein BstN1) were measured. The gene expression level was measured by extracting total RNA from the sublingual gland of each group rat and subjecting it to microarray analysis according to a standard method.

(2) Experimental results (2-1) Measurement of proline-rich protein in saliva and salivary glands Fig. 3 (a) shows the results of SDS-PAGE of salivary glands, and Fig. 3 (b) shows the results of SDS-PAGE of saliva. Fig. 3 (c) shows the result of Western blotting of saliva. In FIG. 3A, lane 1 is a mature rat control group (tap water administration), lane 2 is an adult rat JH administration group, lane 3 is an elderly rat control group (tap water administration), and lane 4 is an elderly rat JH administration. Group results are shown. In addition, in FIG. 3 (b, c), lane 1 shows the results of the elderly rat control group (tap water administration), lane 2 shows the results of the elderly rat JH administration group, and lane 3 shows the results of the elderly rat HH administration group.

  As can be seen from this result, administration of jersey whey (JH) resulted in a thicker or larger protein band containing salin gland and proline-rich protein in saliva (Fig. 3 (a), lanes 2 and 2). Lane 4, lane 2) in FIG. 3 (b).

(2-2) Measurement of gene expression level of proline-rich protein in salivary glands For elderly rats administered with jersey whey (JH), Holstein whey (HH) and tap water, and mature rats administered with tap water, Table 3 shows the results of comparison of gene expression levels of various proline-rich proteins (proline-rich protein 2, proline-rich glycoprotein, proline-rich protein 15, proline-rich proteoglycan 1, and proline-rich protein BstN1). .

  From the results in Table 3, it can be seen that the ingestion of whey, particularly jersey whey, significantly increases the gene expression level in the salivary gland (sublingual gland) of proline-rich protein having a mucosal and dental protective action. From this, it can be seen that whey suppresses or improves salivary gland functional decline also from a genetic aspect.

Experimental Example 4 Measurement of Gene Expression Level of Immunological Substance in Salivary Gland (1) Experimental Method Rats prepared in Experimental Example 2 (2 months after administration of each sample) ((JH administration group, HH administration group of old rats, and control) Group (tap water administration); adult rat control group (tap water administration), salivary glands were collected, and immune substances contained in salivary glands (immunoglobulin δ heavy chain, immunoglobulin γ heavy chain, α-2u globulin PGCL4) , Α-2u globulin PGCL3, α-2u globulin PGCL1, albumin promoter) were measured and compared.

(2) Experimental results Table 4 shows the results.

  From the results of Table 4, the gene expression level in the salivary gland of immune substances decreased with aging increased by oral intake of whey, particularly jersey whey, and the gene expression in salivary glands of immune substances increased with aging. It can be seen that the expression level tends to decrease. From this, it can be seen that whey suppresses or improves salivary gland functional decline also from a genetic aspect.

Experimental Example 5
(1) Measurement of oxygen concentration in blood
14-month-old Wistar male rats (old rats) were divided into 3 groups (n = 3 in each group), and test samples 1 (jersey whey-derived samples) prepared in (1) of Experimental Example 1 (JH samples) Administration group), test sample 2 (holstein whey-derived sample) (HH sample administration group), or tap water (control group) was allowed to drink freely for 2 months. Similarly, 3 months old Wistar male rats (mature rats) (n = 3) were allowed to freely drink tap water (comparison group) for 2 months.

  After drinking for 2 months, blood was collected from each group of rats, and the oxygen concentration in the blood was measured with a blood gas analyzer.

  The results are shown in Table 5.

  As can be seen from the results of the aged and mature rat tap water administration groups shown in Table 5, the blood oxygen concentration significantly decreases with aging, but this decrease is significant due to oral intake of whey, particularly jersey whey. To recover. From the results of Experimental Example 1, this effect is considered to be an effect brought about by whey suppressing or improving the atrophy of lung tissue with aging and the resulting decrease in lung tissue function.

(2) Measurement of collagen concentration in lung tissue For the above-mentioned JH administration group, HH administration group, control group, and comparison group, after drinking each sample for 2 months, lung tissue was collected from each group of rats, and Sircol Using a Soluble Collagen Assay Kit (Biocolor Ltd., Northern Ireland), the collagen concentration in lung tissue was measured according to the manual.

  The results are shown in Table 6.

  As can be seen from the results of the aged rats and adult rats treated with tap water shown in Table 6, the collagen concentration in the lung tissue decreases markedly with aging, but this decrease is due to oral intake of whey, especially jersey whey. , Recover significantly. From the results of Experimental Example 1, this effect is considered to be an effect brought about by whey suppressing or improving the atrophy of lung tissue with aging and the resulting decrease in lung tissue function.

Experimental Example 6 Measurement of Granzyme Gene Expression Level in Lung Tissue It is known that cell necrosis (necrosis) and apoptosis (cell death) are induced by an increase in granzyme (serine protease).

  In this experimental example, the JH-administered group, the HH-administered group, the control group (above, elderly rats), and the comparative group (mature rats) prepared in Experimental Example 5 were used for 2 months for each sample, and then the rats in each group. Lung tissue was collected from the cells, total RNA was extracted, and subjected to microarray analysis, thereby measuring the gene expression level of granzyme (granzyme B, C and K) in the lung tissue.

  The results are shown in Table 7.

  From the results of Table 7, it can be seen that oral intake of whey, particularly jersey whey, significantly reduces the gene expression level in lung tissues of granzymes (granzymes B, C and K), which increases with age. From this, it can be seen that whey suppresses atrophy and cell necrosis of lung tissue and suppresses or improves a decrease in function of lung tissue also from a genetic side.

Claims (10)

An oral anti-aging agent containing whey or processed whey as an active ingredient. The oral anti-aging agent according to claim 1, which is a tissue atrophy suppression / restoration agent for suppressing or repairing tissue atrophy due to aging. The oral anti-aging agent according to claim 1, which is a tissue function improving agent for improving functional deterioration associated with tissue atrophy due to aging. The oral anti-aging agent according to claim 1, which is used for improving a disease state or a condition caused by a functional decrease associated with tissue atrophy due to aging. The oral anti-aging agent according to any one of claims 2 to 4, wherein the tissue is exocrine gland tissue or lung tissue. The oral anti-aging agent according to claim 5, wherein the exocrine gland tissue is salivary gland tissue. The oral anti-aging agent according to claim 4, wherein the symptom or pathology caused by functional deterioration associated with tissue atrophy due to aging is exocrine disorder or respiratory failure. The oral anti-aging agent according to claim 7, wherein the exocrine disorder is xerostomia. The oral anti-aging agent according to any one of claims 1 to 8, which has a solid preparation form selected from tablets, pills, powders (powder), granules, and capsules. The oral anti-aging agent according to any one of claims 1 to 9, which is a supplement.

Images