JP2018052854A - Intestinal flora improving composition which comprises eggshell membrane-containing powder as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide intestinal flora improving compositions which comprise an eggshell membrane-containing powder or impalpable powder and which promote producing type III collagen in combination with hydrolyzed eggshell membrane component.SOLUTION: The present invention provides an intestinal flora improving composition which comprises a shell membrane-containing powder, particularly a shell membrane-containing impalpable powder having a volume mean particle diameter of 6 μm or less, or having a volume maximal particle diameter of 20 μm or less. Furthermore, a composition comprising also hydrolyzed eggshell membrane has improved III type collagen production promotion function and functions synergistically with intestinal flora improving effect to serve as a composition excellent in skin-beautifying and aging prevention.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a composition for improving intestinal flora containing eggshell membrane-containing powder or fine powder as an active ingredient. Furthermore, the present invention relates to a composition for improving intestinal flora and promoting the production of type III collagen, comprising an eggshell membrane-containing powder or fine powder and a hydrolyzed eggshell membrane component.

  The eggshell membrane is a membrane inside the eggshell of avian eggs such as chicken eggs. It consists of two inside and outside. The outer eggshell membrane adheres to the inside of the eggshell, and the inner eggshell membrane wraps the egg white and has antibacterial properties. Protects developing embryos from infection. The eggshell membrane has a net-like structure composed of tough fibrous proteins such as type I, type V and type X collagen, glucosamine, desmosine and hyaluronic acid, and these proteins contain cysteine Is relatively stable to acids, alkalis and proteases and is insoluble in water. It was not used as a waste product in the food industry before, but it was discovered as a cosmetic and nutritional supplement since it was discovered to promote skin regeneration and in particular to promote the production of type III collagen called fetal collagen. Is being developed and put to practical use.

  For example, in order to increase the digestion and absorption efficiency of the eggshell membrane component, the eggshell membrane powder is finely pulverized using a jet mill so that the volume average particle size is 6 μm or less or the maximum volume particle size is 20 μm (800 mesh) or less. It has been proposed to powder (Patent Document 1). In order to make insoluble eggshell membranes soluble in water, hydrolyzed eggshell membranes obtained by hydrolyzing proteins are dried into powder to improve handling and storage stability (Patent Document 2). It has been proposed that the hydrolyzed eggshell membrane powder is pH 3-6 to improve storage stability and safety (Patent Document 3).

JP2013-216652A Japanese Patent No. 5179847 JP-A-2006-98206

INDUSTRIAL APPLICABILITY The present invention is an intestinal flora that is highly safe and can be used as a food additive, food and drink, supplement, and pharmaceutical in a daily and simple manner, and can improve or maintain a living body in a good state with a single material. It is an object to provide a composition for improvement.
Furthermore, in addition to the function of improving intestinal flora, it has a function of promoting the production of type III collagen, and provides a composition capable of improving and / or maintaining the living body in a better state by the synergistic effect of both functions. This is the issue.

  Eggshell membranes or hydrolyzed eggshell membranes are used for skin beautification, skin regeneration, hair growth promotion, hair loss prevention, etc., as well as to improve daily health and maintain health, such as elimination of stiff shoulders, appetite enhancement, and fatigue recovery. It is supposed to be effective. In addition, eggshell membrane-containing fine powder (Patent Literature 1) and hydrolyzed eggshell membrane powder (Patent Literatures 2 and 3) are known to have a function of promoting the production of type III collagen in the body when digested and absorbed in the body. It has been.

  The present inventors have digested almost 100% of the hydrolyzed eggshell membrane and absorbed it into the blood, whereas about 54% of the eggshell membrane-containing powder and fine powder remained in the intestine without being digested and absorbed. The present inventors have found for the first time that the remaining components prevent the growth of bad bacteria among the intestinal bacteria and improve the intestinal flora, thereby completing the present invention.

  Further, by combining the hydrolyzed eggshell membrane component having a function of further promoting the production of type III collagen with the eggshell membrane-containing powder or fine powder, which is digested and absorbed only by about 46%, the eggshell membrane-containing powder or fine powder is used in combination. The function of improving internal flora and promoting the production of type III collagen act synergistically to improve immune function by improving intestinal flora, prevent adult diseases such as cancer, and promote the production of type III collagen Strengthening blood vessels, lungs, liver, and skin tissues has led to the completion of a composition that has an enhanced anti-aging effect and can further improve and / or maintain good health.

The present invention comprises the following technical matters.
(1) A composition for improving intestinal flora, comprising an eggshell membrane-containing powder as an active ingredient.
(2) The composition for improving intestinal flora according to (1), wherein the eggshell membrane-containing powder is an eggshell membrane-containing fine powder having a volume average particle size of 6 μm or less or a volume maximum particle size of 20 μm or less.
(3) The composition for improving intestinal flora according to (1) or (2), further comprising a hydrolyzed eggshell membrane component.
(4) The composition for improving intestinal flora according to (3), wherein the hydrolyzed eggshell membrane component is hydrolyzed eggshell membrane powder.
(5) The composition for improving intestinal flora according to (4), comprising 10 to 90% by weight of fine powder containing eggshell membranes and 90 to 10% by weight of hydrolyzed eggshell membrane powders.
(6) The composition for improving intestinal flora according to any one of (3) to (5), which is also a composition for promoting the production of type III collagen.
(7) The composition for improving intestinal flora according to any one of (1) to (6), which is a preparation to be administered orally.
(8) The composition for improving intestinal flora according to any one of (1) to (7), which is a food additive, food or drink, supplement, edible beauty agent, or pharmaceutical.
(9) The composition for improving intestinal flora according to (7) or (8), wherein the preparation is a tablet.
(10) The composition for improving intestinal flora according to any one of (3) to (9), wherein the total daily intake of the eggshell membrane-containing powder and the hydrolyzed eggshell membrane powder is 10 to 48,000 mg.
(11) The improvement of the intestinal flora is due to the fact that components remaining in the intestine without being absorbed in the intestine increase the diversity of intestinal bacteria and reduce the number of pathogenic bacteria (1) to (10) The composition for intestinal flora improvement in any one of.

  In the human intestine, more than 100 trillion species of intestinal bacteria live in a well-balanced manner, and a kind of ecosystem is maintained while good bacteria, bad bacteria, and opportunistic bacteria keep balance with each other in the intestine. (Intestinal flora) is formed. When the balance of this intestinal flora is lost, harmful substances accumulate in the intestine due to the increase of bad bacteria, etc., not only constipation, diarrhea symptoms, decline in immune function, but also by harmful substances absorbed in the blood, It is known to put a burden on the heart, liver and kidney, promote aging, and cause cancer.

  In an experiment using rats, about 54% of the eggshell membrane-containing fine powder remains in the intestine without being digested and absorbed, and the remaining components prevent the growth of bad bacteria among the intestinal bacteria, It has been found to improve. ADVANTAGE OF THE INVENTION According to this invention, the composition for intestinal flora improvement which can be easily ingested daily can be provided by various uses, such as a food additive, food-drinks, a supplement, an edible beauty agent, and a pharmaceutical. The composition for improving intestinal flora of the present invention has no irritation and can be used in combination with various components in various forms, and can also be used in combination with nutritional and cosmetic components in order to obtain further systemic effects.

In addition, about 46% of the eggshell membrane-containing fine powder is digested and absorbed, and almost 100% of the hydrolyzed eggshell membrane component is digested and absorbed, and the eggshell membrane component taken into the blood produces type III collagen in the skin. Not only can it be promoted, but it can also increase the elasticity of the entire blood vessel by promoting the production of type III collagen in the blood vessel. Therefore,
If the eggshell membrane-containing powder or fine powder and the hydrolyzed eggshell membrane component are used in combination, the intestinal flora improving function and the type III collagen production promoting function of the eggshell membrane-containing powder or fine powder, and the hydrolyzed eggshell membrane component III The synergistic action of the type I collagen production promotion function promotes the production of type III collagen in blood vessels, lungs, liver, skin, etc., and the improvement of intestinal flora enhances the skin beautifying effect and anti-aging effect, and health The state can be improved and / or maintained well. This composition can be used in various forms as food additives, foods and drinks, supplements, edible beauty agents, pharmaceuticals, and tableting is easy if hydrolyzed eggshell membrane powder is used as the hydrolyzed eggshell membrane component Can be done.

Bacterial metagenomic analysis after ingesting eggshell membrane-containing fine powder supplements in mice with colitis induced by DSS: Enterobacterial profile Bacterial metagenomic analysis after ingesting eggshell membrane-containing fine powder supplements in mice with colitis induced by DSS: abundance of enterobacterial species Bacterial metagenomic analysis after ingesting eggshell membrane-containing fine powder supplements in mice with colitis induced by DSS: abundance ratio of enterobacteria Results after 17 days from oral ingestion experiment of hydrolyzed eggshell membrane powder supplement using mice. The expression level in the skin of the type III collagen gene by real-time PCR is shown.

  In the human intestine, there are about 100 trillion species of intestinal bacteria in a well-balanced inhabitant, and useful bacteria such as lactic acid bacteria and bifidobacteria (good bacteria) such as Escherichia coli and Clostridium bacteria. Harmful bacteria (bad bacteria) and opportunistic bacteria form a kind of ecosystem (intestinal flora) while maintaining a balance with each other in the intestine. If the balance of the intestinal flora is lost due to aging, stress, disease, environmental changes, dietary changes, harmful substances accumulate in the intestine due to the increase of bad bacteria, etc., causing constipation, diarrhea symptoms, immune function It is known that harmful substances absorbed in the blood not only lower but also burden the heart, liver and kidney, promote aging, and cause cancer.

  The composition for improving intestinal flora of the present invention comprises eggshell membrane-containing powder as an active ingredient, and in experiments using rats, 46% of the ingested eggshell membrane-containing fine powder is digested and absorbed, and the rest is absorbed. Remained in the intestine. Example 1 shows that this remaining component can increase the diversity of enteric bacteria and reduce the number of pathogenic bacteria such as E. coli and improve intestinal flora.

  The eggshell membrane used in the present invention can be any membrane as long as it is inside the eggshell of all terrestrial ovarian animals, particularly the eggs of birds. In particular, eggshell membranes of chicken eggs are preferred from the standpoint of availability and cost.

  The eggshell membrane-containing powder used in the present invention is not particularly limited as long as it contains at least an eggshell membrane, and the powder refers to any powder regardless of the particle size. The peeled eggshell membrane or the raw material with the eggshell membrane attached to the eggshell may be used for pulverization by any known method. A commercially available eggshell membrane-containing powder may also be used.

  The eggshell membrane-containing fine powder used in the present invention refers to a powder having a volume maximum particle size and / or a volume average particle size smaller than about 100 μm. In the present specification, “volume average particle diameter” and “volume maximum particle diameter” of a powder or fine powder mean values measured using a laser diffraction particle size distribution analyzer (manufactured by Seishin Co., Ltd., LMS-30). To do. In measuring the particle size, a measurement sample in which eggshell membrane-containing fine powder is dispersed in water using a surfactant is used. The “volume average particle size” means a particle size when the cumulative value from the small particle size side in the particle size distribution is 50%.

  As the eggshell membrane-containing fine powder used in the present invention, those having a volume average particle size of 6 μm or less or a volume maximum particle size of 20 μm or less are particularly preferred from the viewpoint of digestion and absorption efficiency. It is preferable to produce eggshell membrane-containing raw materials such as commercially available eggshell membrane-containing powders by a fine pulverization process using a jet mill that collides with each other in gas.

  The hydrolyzed eggshell membrane component used in the present invention is a solution having protein-degrading action, such as an eggshell membrane or the aforementioned eggshell membrane-containing powder, an acid aqueous solution, an alkaline aqueous solution, an alkaline organic solvent solution, and a protease-containing solution. Obtained by hydrolyzing in. Although it can be used as an aqueous solution or extract, it is preferably in the form of a powder obtained by a drying means such as spray drying or freeze-drying for handling and storage stability. As the hydrolyzed eggshell membrane powder, a powder having a pH of 3 to 6 is particularly preferred from the viewpoint of storage stability and safety.

  Human skin is composed of an epithelial layer on the surface and a connective tissue dermis layer below it, and the protein constituting the dermis layer is mainly collagen, and it is also composed of elastin, albumin, globulin, and mucin. Collagen constituting the dermis layer is mainly α1 (I) type collagen and α1 (III) type collagen (hereinafter referred to as “type III collagen”). Type I collagen mainly maintains the structure of the skin, type III collagen It functions mainly differently from the provision of skin flexibility.

  The proportion of type I collagen and type III collagen in the skin changes with age. Type III collagen is the most common type of fetal collagen, also called fetal collagen, and more type III collagen in the fetal skin. If the ratio can be increased, skin aging due to aging can be stopped and the skin can be made smooth and moist. In addition to skin, type III collagen is contained in connective tissues such as blood vessels, lungs and liver.

  About 46% of the eggshell membrane-containing fine powder is digested and absorbed, almost 100% of the hydrolyzed eggshell membrane component is digested and absorbed, and the eggshell membrane component taken into the blood only promotes the production of type III collagen in the skin. In addition, the formation of type III collagen can be promoted in blood vessels to increase the elasticity of the entire blood vessels, and fibrosis can also be suppressed in the liver. When both the eggshell membrane component-containing powder and the hydrolyzed eggshell membrane component used in the present invention are digested and absorbed, the hydrolyzed eggshell membrane component has a function of promoting the production of type III collagen, but is almost 100% digested and absorbed. It turned out that the promotion effect is higher.

  Therefore, the combined use of eggshell membrane component-containing powder and hydrolyzed eggshell membrane component not only further enhances the effect of promoting the production of type III collagen, but also synergizes with the improvement of intestinal flora, synergistically enhances the skin and prevents aging. And a composition capable of improving and / or maintaining good health has been provided. The composition preferably contains 10 to 90% by weight of eggshell membrane-containing fine powder and 90 to 10% by weight of hydrolyzed eggshell membrane powder.

  The composition of the present invention can be an oral composition such as a tablet, powder, granule, capsule, or liquid. Since there is no irritation, there are no particular limitations on the dosage form, and various components and production methods for producing oral compositions of various dosage forms are appropriately selected from ingredients known in the production field of supplements, pharmaceuticals, etc. be able to. In particular, it is preferably used for tablets from the viewpoint that it is suitable for storage and distribution, is not deformed or disintegrated when taken, and can be easily taken orally.

  The total content of the eggshell membrane-containing powder and the hydrolyzed eggshell membrane powder (hereinafter also referred to as “eggshell membrane component”) contained in the tablet of the present invention is not particularly limited, but granulation and tableting into particles are possible. From the standpoint that the effect of improving the intestinal flora and the effect of promoting the production of type III collagen can be enhanced when the tablet is ingested smoothly, the proportion of 5-45% by weight relative to the total weight of the tablet is more preferable. Is preferably contained in a proportion of 10 to 40% by weight. When the content is 5% by weight or more, it is not necessary to take a large amount of tablets. On the other hand, when the content is 45% by weight or less, granulation and tableting are facilitated, and tablets can be easily produced.

As the various additives for forming the tablet, excipients, binders, disintegrants, lubricants, other nutrients, and the like can be added to the tablet of this embodiment.
As the excipient, known excipients can be used as appropriate, but at least one of modified starch and lactose should be used 0.3 to 3 times the weight of the eggshell membrane component from the viewpoint of formability. Preferably, it is 1 to 2.5 times. As the modified starch, one type or two or more types of dextrin such as roasted dextrin, oxidized starch, and low-viscosity modified starch can be used. When chemical starch and lactose are used in combination as an excipient, the use ratio (weight ratio) is preferably 1: 5 to 5: 1.
As the binder, known binders can be used as appropriate, and examples thereof include starch paste, gum arabic paste, and hydroxypropyl cellulose.
As the disintegrant, known disintegrants can be used as appropriate. For example, celluloses and the like can be used.
As the lubricant, known lubricants can be used as appropriate, and examples thereof include magnesium stearate, sucrose fatty acid ester, waxes, talc, vitamin C and the like.

In addition, the tablet of this embodiment preferably contains eggshell calcium as a hardness improver in order to increase the hardness of the tablet, prevent deformation and scratches, and improve handleability. Eggshell calcium is a fine powder obtained by pulverizing and drying the eggshells of birds such as chicken eggs, and the content of eggshell calcium contained in the tablet is preferably 0.1 to 20% by weight, more preferably 0.5 to 0.5%. 15% by weight.
Furthermore, the tablet of this embodiment is preferably covered with a coating film in order to prevent deterioration and decomposition of components contained in the tablet and to improve the scratch resistance of the tablet surface. As the coating film, a known coating film can be used as appropriate. For example, a trade name “Shellak” (manufactured by Gifu Shellac Co., Ltd.) can be used. The tablet of this embodiment may be covered with sugar coating, may be colored, and may be subjected to a glazing treatment after coloring.

  The size of the tablet of the present embodiment is not particularly limited, but in general, a circle or an ellipse having a diameter of about 7 to 10 mm is preferable from the viewpoint of handleability and ease of taking. The weight of one tablet is about 350 to 600 mg, and the eggshell membrane component is preferably contained in an amount of about 10 to 240 mg, more preferably about 20 to 150 mg. If the eggshell membrane component is contained in an amount of about 10 to 240 mg, adults can take 1 to 200 tablets per day (10 to 48,000 mg of eggshell membrane component per day).

  The tablet of the present embodiment can be produced by a known tablet production method using a raw material for tableting containing at least an eggshell membrane-containing fine powder and a hydrolyzed eggshell membrane powder. For example, after the tableting process of tableting raw materials to form a plain tablet, if necessary, the granulation process, protective coating process, sugar coating process, etc., and further coloring and polishing Also good.

  In addition, the effective dose of the pharmaceutical preparation as the composition for improving intestinal flora of the present invention varies depending on the degree of symptoms to be treated or prevented, the state of the administration subject (including age and sex), the dosage form, and the like. The oral dose of such pharmaceuticals to humans (adults weighing 60 kg) is preferably 1 to 100,000 mg per day in terms of eggshell membrane components. For example, the effective dose can be 10 to 48,000 mg in total of eggshell membrane components per day, and preferably 20 to 3,500 mg. Since the eggshell membrane-containing component of the present invention is highly safe and has no side effects, there is no problem even if the intake or application amount exceeds the above range as long as other components are appropriately selected.

  Furthermore, the composition of the present invention can be used as food additives such as confectionery, beverages, health foods, preserved foods and processed foods. Here, the subject of “medicine” and “food and beverage” is not limited to humans, but also includes pharmaceuticals and feeds for mammals such as pets and livestock. The concept of “food and drink” includes enteral nutritional foods, nutritional functional foods, functional display foods, foods for specified health use, and the like in addition to ordinary foods and drinks.

  Hereinafter, the present invention will be described in detail based on examples, but the present invention is not limited to these examples.

[Intestinal flora improving function and enteritis improving function of fine powder containing eggshell membranes]
(1) Production of Eggshell Membrane-Containing Fine Powder As an eggshell membrane-containing powder sample, a product name “EM Powder 300” (manufactured by QP Corporation) was finely pulverized with a jet mill. As the jet mill, a single track jet mill (manufactured by Seishin Co., Ltd., FS-4) was used, and the volume maximum particle size was 800 mesh (mesh size approximately about 80 mesh) at an air volume of 1.2 m ³ / min and a power of 11 kw. Grinding was carried out until the thickness became approximately 20 μm). When the particle size after pulverization was measured using a laser diffraction particle size distribution analyzer (manufactured by Seishin Co., Ltd., LMS-30), the volume maximum particle size was 19.6 μm, and the volume average particle size was 5.8 μm (hereinafter, "800 mesh fine powder").

(2) Experimental animals After purchasing 7-week-old male C57BL6 / J mice and adapting them for 3 days, they were divided into three groups (n = 8) so that the average body weights were equal. For 7 days, either the control AIN-93G basic regular diet (DSS group) or the diet supplemented with 8% eggshell membrane 800 mesh fine powder (D-ESM group 8) was fed. Then, for 9 days, while giving both groups the same experimental feed as before, drinking water containing 1.5% (W / V) dextran sulfate sodium (DSS) (molecular weight 40 KDa; MP Biomedicals, Irvine, CA) Colitis was induced. Normal control mice (CON group) were fed AIN-93G basal diet and normal drinking water throughout the course of the experiment.
The 8% eggshell membrane 800 mesh fine powder reinforced diet was adjusted with corn starch and casein to maintain calorie and protein balance. The disease activity index (DAI) was monitored daily from the day when DSS water was given, using the three scores of mouse weight, stool hardness, and blood stool.

(3) Bacterial metagenomic analysis of the cecum Metagenomic DNA was extracted from the contents of the cecum of mice using the QIAamp Stool Min kit (Qiagen, Hilden, Germany). In order to amplify the variable regions 3 and 4 of the 16S rRNA gene, amplification was performed using primers of (5′-CCTACGGGGGCWGCAG-3 ′) and (5′-CAGGACTACTCHGGGGTATCTAATCC-3 ′). Sequencing was performed on DNA sequences with an amplified length of 250 bp or more and an average quality score of 30 or more.
Representative sequences were classified using the Ribosomal Database Project (RDP) classification according to the Greengenes OTU database.
Quantitative real-time PCR was performed to detect changes in segmented bacteria and several pathogenic bacteria.

(4) Results of Bacterial Metagenome Analysis According to the analysis at the level of bacteria, the relative amounts of the three most widely spread bacteria are significantly different among the three experimental groups (D-ESM8, DSS, CON). It was. As shown in FIG. 1, Bacteriodetes decreased from 67.0% in the CON group to 34.6% in the DSS group and recovered to 55.2% in the D-ESM8 group. Firmicutes decreased from 26.5% in the CON group to 13.7% in the DSS group and recovered to 18.6% in the D-ESM8 group. Proteobacteria increased from 0.9% in the CON group to 40.2% in the DSS group and recovered to 8.0% in the D-ESM8 group.
Next, two classes of α diversity and β diversity were evaluated. According to alpha diversity analysis, mice in the DSS group were the least abundant in bacterial species among the three groups (FIG. 2).
This result was as expected because the bacterial species in the intestine were reduced in relation to disease states including inflammatory bowel disease in humans and DSS-induced colitis in mice. The abundance of bacterial species recovered to the same level as the CON group.

As shown in the upper part of FIG. 3, the increased Enterobacteriaceae, which is a marker of enteritis and a marker of human inflammatory bowel disease and oxidative stress in mouse colitis, is an inflammatory DSS group compared to the CON group (Proteobacteria genus is a whole bacterium). 39%) was detected very frequently. In contrast, Enterobacteriaceae was very rarely detected in the D-ESM8 group (about 6%). Similarly, DSO administration increased the number of Enterococcaceae, but it was reduced to near normal levels by supplementation with eggshell membrane fine powder.
Metabolites of intestinal bacteria such as short chain fatty acids (SCFA) are the primary energy source of colon cells and are thought to directly affect host gene expression, so the presence of SCFA producing bacteria was evaluated ( (Lower part of FIG. 3). Although SCFA-producing bacteria such as Ruminococcaceae and Porphyromonadaceae, which are known to decrease in inflammatory bowel disease, decreased in the DSS group, these improved with the intake of eggshell membranes.

(5) Summary From the results of (4) above, the intake of eggshell membrane component-containing fine powder not only increases the diversity of enterobacteria, but also reduces the absolute number of pathogenic bacteria such as E. coli. It was shown to suppress the abnormal growth of transformed filamentous fungi.
Thus, ingestion of eggshell membrane-containing fine powder has been shown to improve intestinal symbiosis imbalance and enterotoxemia, and improve intestinal flora.

[Promoting function of type III collagen production by fine powder containing eggshell membrane and hydrolyzed eggshell membrane powder]
Each of the eggshell membrane 800 mesh fine powder and the hydrolyzed eggshell membrane powder was orally ingested by mice once a day for 17 days, and a supplement oral intake experiment was conducted. Hydrolyzed eggshell membrane powder having a pH of 3 to 6 (Patent Document 3) was used as the hydrolyzed eggshell membrane powder.

  For 8-week-old hairless mice, hydrolyzed eggshell membrane powder was mixed with MediGel and voluntarily ingested as supplements once a day for 17 days during the active period of the animals. 1/2, 1 and 2 times the amount of human daily intake (n = 6 each), and eggshell membrane 800 mesh fine powder is similarly used as the amount equivalent to 1 time of human daily intake. Ingested (n = 6). As a control, hairless mice without supplements were used. After 17 days, the expression of type III collagen gene in the skin and liver of each mouse was examined by quantitative real-time PCR.

Quantitative real-time PCR analysis was performed as follows.
Back skin and liver samples were taken from each mouse and ground in liquid nitrogen. After homogenization of whole skin tissue or whole liver tissue, total RNA was isolated and cDNA synthesis was performed using a trade name “Takara PrimeScript RT reagent kit”. The real-time PCR method was performed using the trade name “SYBR Premix Ex TaqTM II (Takara) on Thermal Cycler Dice Real Time System” (Takara). As a primer, a primer designed for amplification of a gene encoding Col3a1 (type III collagen) was used. As an internal standard, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, which is a housekeeping gene, was similarly amplified. In the PCR cycle, initial denaturation was performed at 95 ° C. for 30 seconds and 40 cycles (denaturation at 95 ° C. for 5 seconds, annealing and extension reaction at 60 ° C. for 1 minute).

The expression level of type III collagen gene in the skin is shown in FIG.
It was shown that the expression of the gene encoding type III collagen increased in a dose-dependent manner in the dorsal skin of mice that had been orally ingested with hydrolyzed eggshell membrane powder (p = 0.08). The expression of the gene encoding type III collagen was increased more effectively than the mice that orally ingested the eggshell membrane 800 mesh fine powder.

  Regarding the expression level of type III collagen gene in mice ingested with hydrolyzed eggshell membrane powder in the liver, mice given 1-fold equivalents as supplements consumed by humans did not change from the control group. When the double conversion amount was given, the expression of type III collagen gene was significantly increased (p = 0.007). Increased expression of type III collagen gene in the liver is expected to suppress liver fibrosis and improve the vascular environment.

[Manufacture of tablets]
The eggshell membrane 800 mesh fine powder and hydrolyzed eggshell membrane powder used in Example 2 were each 20.0 parts by weight, trade name “Wakia” (manufactured by Solar Eclipse Co., Ltd.), 10.0 parts by weight, and trade name “Pine”. "Fiber" (Matsuya Chemical Co., Ltd.) 20.0 parts by weight, lactose (Megure Co., Ltd.) 25.9 parts by weight, eggshell calcium Product name "Cal Hope" (Kewpie Co., Ltd.) 10.0 parts by weight, β-carotene 5 A raw material mixture was prepared by mixing 0.0 part by weight, 0.05 part by weight of vitamin B2, 0.05 part by weight of vitamin E, and 2.0 parts by weight of niacin using a V-type mixer. 15 parts by weight of ethyl alcohol is mixed with 93 parts by weight of this raw material mixture, and the resulting mixture is granulated using a wet granulator, then dried at a temperature of 50 ° C. for about 16 hours, and tableted. Granules for use were produced.

  9 parts by weight of vitamin C and 1 part by weight of sucrose fatty acid ester are mixed with 100 parts by weight of granules for tableting, and the resulting mixture is 200 mg using a tableting device. Of uncoated tablets. An aqueous solution of the trade name “Serak” (manufactured by Gifu Shellac Co., Ltd.) was applied to the surface of the uncoated tablet using a coating apparatus, and dried at a temperature of 40 ° C. for 2 hours to obtain a protective-coated tablet. Using a sugar coating device, a sugar coating paste A (70 parts by weight granulated sugar, 3 parts by weight gum arabic, 4 parts by weight gelatin, 3 parts by weight calcium eggshell and After coating a paste mixed with 65 parts by weight of water, it was dried at a temperature of 40 ° C. for about 4 hours. After that, the sugar-coating paste A obtained by diluting the sugar-coating paste A with water was coated using a sugar-coating apparatus and then dried at a temperature of 40 ° C. for 4 hours to obtain sugar-coated tablets.

  After applying a colorant containing the trade name “SR Red K3” (manufactured by Saneigen Co., Ltd.) on the surface of the sugar-coated tablet, it is dried at 40 to 50 ° C. to produce a red-colored tablet. Polishing was performed using carnauba wax. The tablet thus obtained weighed 400 mg and contained eggshell membrane components at a ratio of about 80 mg per tablet. The polished tablets were selected to remove defective products, weighed after product inspection, and packaged in a double bag containing a desiccant. The tablets had sufficient hardness and shape retention, and did not deform or disintegrate during sorting, inspection and packaging, and were excellent in handleability.

ADVANTAGE OF THE INVENTION According to this invention, the composition for intestinal flora improvement containing eggshell membrane containing powder or a fine powder which is non-irritating and can be used together with various components in various forms can be provided.
When used in combination with the hydrolyzed eggshell membrane component, the intestinal flora improving function and the type III collagen production promoting function of the eggshell membrane-containing fine powder, and the type III collagen production promoting function of the hydrolyzed eggshell membrane component Synergistically promotes the production of type III collagen in blood vessels, lungs, liver, skin, etc., improves intestinal flora, enhances skin beautification and anti-aging effects, improves health and / or Or it becomes the composition which can be maintained favorable. This composition can be used in various uses and forms such as food additives, foods and drinks, supplements, edible beauty agents, and pharmaceuticals, and can be easily taken on a daily basis.

Claims (11)

  A composition for improving intestinal flora, comprising a eggshell membrane-containing powder as an active ingredient.   The composition for improving intestinal flora according to claim 1, wherein the eggshell membrane-containing powder is an eggshell membrane-containing fine powder having a volume average particle size of 6 µm or less or a volume maximum particle size of 20 µm or less.   The composition for improving intestinal flora according to claim 1 or 2, further comprising a hydrolyzed eggshell membrane component.   The composition for improving intestinal flora according to claim 3, wherein the hydrolyzed eggshell membrane component is hydrolyzed eggshell membrane powder.   The composition for improving intestinal flora according to claim 4, comprising 10 to 90 wt% of eggshell membrane-containing fine powder and 90 to 10 wt% of hydrolyzed eggshell membrane powder.   The composition for improving intestinal flora according to any one of claims 3 to 5, which is also a composition for promoting the production of type III collagen.   The composition for improving intestinal flora according to any one of claims 1 to 6, which is a preparation to be administered orally.   The composition for improving intestinal flora according to any one of claims 1 to 7, which is a food additive, food or drink, supplement, edible beauty agent, or pharmaceutical product.   The composition for improving intestinal flora according to claim 7 or 8, wherein the preparation is a tablet.   The composition for improving intestinal flora according to any one of claims 3 to 9, wherein the total daily intake of the eggshell membrane-containing powder and the hydrolyzed eggshell membrane powder is 10 to 48,000 mg.   The improvement in intestinal flora is due to the fact that the components remaining in the intestine without being absorbed in the intestine increase the diversity of intestinal bacteria and reduce the number of pathogenic bacteria. A composition for improving intestinal flora according to any one of the above.