KR102602070B1 - A composition for improving intestinal health comprising Salicornia herbacea mixed fermentation product - Google Patents

Abstract

The present invention relates to a composition for improving intestinal health containing a mixed fermented product of seaweed as an active ingredient.
The composition for improving intestinal health of the present invention contains fermented seaweed mixture as an active ingredient, thereby reducing intestinal inflammatory response and damage to intestinal crypts where intestinal stem cells are located, and maintaining the intestinal mucous layer thickness at a constant thickness. In addition, it can effectively improve intestinal health or intestinal function by increasing the content of short-chain fatty acids in the intestines.

Description

A composition for improving intestinal health comprising Salicornia herbacea mixed fermentation product}

The present invention is a mixed fermentation of seaweed that reduces intestinal inflammatory response and damage to crypts containing intestinal stem cells, maintains the intestinal mucous layer thickness at a constant thickness, and increases the content of short-chain fatty acids in the intestine. It relates to a composition for improving intestinal health containing water as an active ingredient.

Recently, with the entry into an aging society, maintenance and promotion of health has formed a social trend that is more important than anything else, and foods that are beneficial to health are becoming a subject of greater interest. In particular, one of the various health problems that occur as aging progresses. Interest in health is rapidly increasing.

In addition, as aging causes a decline in the adaptability of the intestines, it is very important to consume good foods that can help the elderly's intestinal health and prevent intestinal function decline.

The intestines have a very close and important relationship with immunity, as approximately 70% of the body's immune cells are gathered there, and are responsible for mechanical stress, chemical stress such as carcinogens from the external environment, and biological stress from harmful intestinal bacteria. Due to continuous damage, the intestinal epithelial cells are damaged or the production of stem cells and mucous substances are negatively affected, causing the intestines to age and pose a threat to intestinal health.

In other words, intestinal cells have a short lifespan of about 3-5 days because they are exposed to various stresses, so it is very important to create environmental conditions for stem cells to continuously generate cells.

Since intestinal stem cells are protected at the bottom of the crypts (intestinal crypts), there is a need to develop foods that help protect the crypts where intestinal stem cells exist and maintain the thickness of the intestinal mucous layer.

In addition, the number of patients with intestinal diseases such as inflammatory bowel disease and irritable bowel syndrome due to westernized and inappropriate eating habits is increasing, and many metabolic diseases such as diabetes and Alzheimer's disease are also reported to be ultimately related to intestinal health. Relevance is becoming more important.

In particular, a paper (Cell, 167, 1469, 2016; Nature, 544, 304, 2017) was recently published suggesting that the cause of degenerative brain diseases (Alzheimer's disease, Parkinson's disease, etc.) may be due to intestinal health problems, and metabolic diseases, etc. It has been revealed that gut health is involved in all health of the body.

Meanwhile, Salicornia is an annual herbaceous plant of the Salicornia herbacea L. family, and is a halophyte that grows naturally on the beaches of the west and south coasts of Korea along with horseradish herbacea and sea red herb. In particular, seaweed grows in large colonies around the salt farm, which produces sea salt, a specialty of Shinan-gun.

Unlike other plants, seaweed grows in high salinity, and its own salinity is also high. As it grows, it absorbs minerals beneficial to life from seawater, producing calcium (Ca), potassium (K), magnesium (Mg), iron (Fe), and zinc (Zn). ) and other minerals, so it has excellent nutritional value.

Recently, it has been reported that seaweed has various physiological activities such as antioxidant, anti-obesity, anti-diabetic, lowering blood cholesterol, hepatoprotective function, increasing immune regulation function, and whitening. Therefore, it is believed that the results of functional verification of the various physiological activities useful for the health of salicornia as well as its effect on intestinal health can help promote salicornia and create added value for salicornia farmers.

Therefore, we would like to use Salicornia herb to verify its functionality in improving intestinal health and attempt to create high added value from Salicornia herb from various angles.

KR 10-2011-0012397 A

The purpose of the present invention is to reduce intestinal inflammatory response and damage to intestinal crypts containing intestinal stem cells, maintain the intestinal mucous layer thickness at a constant thickness, and increase the content of short-chain fatty acids in the intestine. The goal is to provide health functional foods for improving intestinal health that contain mixed fermentation products as active ingredients.

In addition, another object of the present invention is to provide a pharmaceutical composition that can prevent or treat inflammatory bowel disease by containing the fermented seaweed mixture as an active ingredient.

In addition, another object of the present invention is to provide a feed composition for improving intestinal health containing the fermented seaweed mixture as an active ingredient.

In addition, another object of the present invention is to provide a pharmaceutical composition for animals that can prevent or treat inflammatory bowel disease by containing the fermented seaweed mixture as an active ingredient.

In addition, another object of the present invention is to provide a method for producing the fermented seaweed mixture for improving intestinal health.

In addition, another object of the present invention is to provide a health functional food for preventing or improving intestinal aging.

The health functional food for improving intestinal health of the present invention to achieve the above object may contain a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary materials.

The natural auxiliary material may be one or more selected from mistletoe, Cordyceps sinensis, pork potato, malt, turmeric, stevia, wilyeongseon, paejang, aloe, chicory, and burdock.

The health functional food for improving intestinal health includes health functional food for preventing or improving intestinal aging; Health functional foods for preventing or improving intestinal aging-related diseases; It may be a health functional food for improving the intestinal environment.

The intestinal aging-related disease may be one or more types selected from irritable bowel syndrome, inflammatory bowel disease, constipation, and ischemic enteritis.

The improvement of the intestinal environment includes an increase in butyric acid, a beneficial intestinal short-chain fatty acid, inhibition of damage to intestinal crypts, an increase in intestinal goblet cells, an increase in the thickness of the intestinal mucous layer, and a decrease in factors related to intestinal oxidative stress. There may be more than one species.

The natural auxiliary material may be 200 to 300 parts by weight based on 100 parts by weight of glasswort.

The natural material auxiliary materials include 10-15 parts by weight of mistletoe, 10-15 parts by weight of Cordyceps sinensis, 10-15 parts by weight of pork potato, 10-15 parts by weight of malt, 10-15 parts by weight of shellfish, and aloe, based on the total weight of the natural material auxiliary materials. It may include 10-15 parts by weight, 10-15 parts by weight of chicory, and 10-15 parts by weight of burdock.

The fermented seaweed mixture may be obtained by fermenting seaweed and natural auxiliary materials with lactic acid bacteria.

The fermented seaweed mixture includes (A) inoculating a mixture of seaweed, sugars, and purified water with lactic acid bacteria and then performing primary fermentation for 5 to 15 months; (B) mixing the primary fermentation product and organic acid and performing secondary fermentation for 20 to 30 days; (C) filtering the secondary fermentation and aging at low temperature for 10 to 30 days; (D) mixing 200 to 300 parts by weight of natural additives with 100 parts by weight of the low-temperature aged secondary fermentation product; and (E) subjecting the mixture to a third fermentation for 10 to 20 months.

In step (A), 10 to 20 parts by weight of sugars and 500 to 1,500 parts by weight of purified water are mixed with 100 parts by weight of salicornia, and lactic acid bacteria may be inoculated at 0.5 to 1.5 parts by weight.

The lactic acid bacteria include Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus plantarum, Leuconostoc kimchii, Leuconostoc citreum, Leuconostoc mesenteroides, and Leuconostoc acanthuscomitatum. , Leuconostoc lactis, Lactobacillus sakei, and Lactobacillus brevis.

Fermentation in steps (A) and (B) may be carried out at 18 to 22°C.

In step (B), the organic acid may be mixed in an amount of 2 to 6 parts by weight based on 100 parts by weight of the primary fermentation product.

In step (C), low-temperature aging may be performed at 2 to 10 °C.

In step (E), fermentation may be carried out at 18 to 22 ° C.

In addition, the pharmaceutical composition for preventing or treating inflammatory bowel disease of the present invention for achieving the above-mentioned other purposes may contain a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary materials.

The inflammatory bowel disease may be ulcerative colitis or Crohn's disease.

In addition, the feed composition for improving intestinal health of the present invention to achieve the above-described other purposes may contain a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary materials.

In addition, the pharmaceutical composition for animals for the prevention or treatment of inflammatory bowel disease of the present invention to achieve the above-mentioned other object may contain a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary materials.

In addition, the method for producing a fermented glasswort mixture for improving intestinal health of the present invention to achieve the above-described other purpose is (A) inoculating lactic acid bacteria into a mixture of glasswort, sugars, and purified water, followed by a primary treatment for 5 to 15 months. fermenting; (B) mixing the primary fermentation product and organic acid and performing secondary fermentation for 20 to 30 days; (C) filtering the secondary fermentation and aging at low temperature for 10 to 30 days; (D) mixing 200 to 300 parts by weight of one or more natural additives selected from mistletoe, Cordyceps sinensis, pork potato, malt, bean paste, aloe, chicory, and burdock with 100 parts by weight of the low-temperature-aged secondary fermentation product; and (E) subjecting the mixture to a third fermentation for 10 to 20 months.

The health functional food for improving intestinal health of the present invention contains a mixed fermented product of seaweed as an active ingredient, thereby reducing intestinal inflammatory response and damage to intestinal crypts where intestinal stem cells are located, and maintaining the intestinal mucous layer thickness to a certain thickness. Not only does it maintain the body's health, but it can also effectively prevent or improve the decline in intestinal function due to aging by increasing the content of short-chain fatty acids in the intestines.

Figure 1 is a graph showing the total phenolic compound content of the fermented seaweed mixture (GWⅡ) according to the Example and the fermented seaweed mixed product (GWⅠ) according to the comparative example.
Figure 2 is a graph showing the content of total flavonoid compounds in the fermented seaweed mixture (GWⅡ) according to the example and the fermented seaweed mixed product (GWⅠ) according to the comparative example.
Figure 3a is a graph showing the DPPH radical scavenging ability of the fermented seaweed mixture (GWII) according to the example and the fermented seaweed mixture (GWI) according to the comparative example, and Figure 3b is a graph showing the SC50 values of each experimental group.
Figure 4a is a graph showing the ABTS+ radical scavenging ability of the fermented seaweed mixture (GWII) according to the example and the fermented seaweed mixture (GWI) according to the comparative example, and Figure 4b is a graph showing the SC50 values of each experimental group.
Figure 5a shows body weight over the experimental period of the normal group (CON), aging-induced control (CD), positive control (PC), seaweed mixed fermentation low-concentration diet group (LGW), and seaweed mixed fermentation high-concentration diet group (HGW). This shows the increase, Figure 5b shows the dietary amount of each experimental group, and Figure 5c shows the water intake of each experimental group.
Figure 6a shows the colon of the normal group (CON), aging-induced control (CD), positive control (PC), Salicornia mixed fermentation low-concentration diet group (LGW), and Salicornia mixed fermentation high-concentration diet group (HGW) stained with H&E staining method. The crypts and goblet cells of the large intestine were morphologically analyzed, and Figure 6b shows a morphological analysis of the morphology of mucin-secreting goblet cells and intestinal tissue changes by staining the large intestine of each experimental group with Alcian blue staining. Figure 6c is a graph showing the number of goblet cells in colon tissue of each experimental group.
Figure 7a shows the mucous layer thickness of colonic tissue in the normal group (CON), aging-induced control (CD), positive control (PC), salicornia mixed fermentation low-concentration diet group (LGW), and salicornia mixed fermentation high-concentration diet group (HGW). This is a photograph showing, and Figure 7b is a graph showing the thickness of the mucous layer of the colon tissue of each experimental group.
Figure 8 shows the intestinal flora in the feces of the normal group (CON), aging-induced control (CD), positive control (PC), salicornia mixed fermentation low-concentration diet group (LGW), and salicornia mixed fermentation high-concentration diet group (HGW). This is a graph measuring the amount of short-chain fatty acids (SCFA) produced.
Figure 9a shows the lipid peroxide content in colon tissue of the normal group (CON), aging-induced control (CD), positive control (PC), salicornia mixed fermentation low-concentration diet group (LGW), and salicornia mixed fermentation high-concentration diet group (HGW). is a graph showing, and Figure 9b is a graph showing the content of protein oxides in the colon tissue of each experimental group.
Figure 10 shows TNF-, an inflammation-related factor, in colonic tissues of the normal group, the aging-induced control (CD), the positive control (PC), the salicornia mixed fermentation low-concentration diet group (LGW), and the salicornia mixed fermentation high-concentration diet group (HGW). Western blot results showing expression of α (a), NF-kB p65 (b), IL-1β (c), and IL-6 (d), and a graph showing the expression amount.

The present invention is a mixture of seaweed that reduces intestinal inflammatory response and damage to crypts containing intestinal stem cells, maintains the intestinal mucous layer thickness at a constant thickness, and increases the content of short-chain fatty acids in the intestine. It relates to a composition for improving intestinal health containing fermented product as an active ingredient.

In this specification, the health functional food for improving intestinal health includes health functional food for preventing or improving intestinal aging; Health functional foods for preventing or improving intestinal aging-related diseases; Alternatively, it may be a health functional food for improving the intestinal environment.

Hereinafter, the present invention will be described in detail.

Throughout the specification, when a part “includes” a certain component, this does not mean excluding other components, but rather means that it can further include other components.

In this specification, “fermented product” refers to a culture obtained by adding sugar sources and lactic acid bacteria to salicornia, which is the subject of fermentation, and fermenting and maturing it. This culture can be used as is, concentrated and/or freeze-dried and processed into liquid or powder form, or diluted with water and used as a beverage.

In this specification, “intestinal aging” refers to a decrease in intestinal health or intestinal function due to aging, such as increased intestinal inflammatory response, damage to the intestinal crypt, decreased intestinal mucous layer thickness, and decreased content of butyric acid, a beneficial short-chain fatty acid in the intestine.

The health functional food for improving intestinal health of the present invention contains the fermented seaweed mixture of the present invention as an active ingredient, and reduces intestinal inflammatory response increased due to aging and damage to the crypt where intestinal stem cells are located, and intestinal mucous decreased due to aging. It increases the layer thickness and the content of butyric acid, a beneficial short-chain fatty acid in the intestines, thereby effectively preventing or improving intestinal health or intestinal function that has deteriorated due to aging.

The composition of the present invention contains a mixed fermented product of salicornia as an active ingredient, and the mixed fermented glasswort is obtained by fermenting salicornia and natural auxiliary materials. The natural auxiliary materials include mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, and aloe. It may be one or more types selected from , chicory, and burdock. The natural product auxiliary materials may be one or more selected from mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, aloe, chicory, and burdock, and are not particularly limited. However, the natural product auxiliary material including all of the above natural product auxiliary materials may be used as a natural product-derived sugar source for fermentation. It is excellent in terms of source supply and is desirable in terms of efficacy in improving intestinal health, especially in preventing or improving intestinal aging.

In particular, the natural material auxiliary materials are 10-15 parts by weight of mistletoe, 10-15 parts by weight of Cordyceps sinensis, 10-15 parts by weight of pork potato, 10-15 parts by weight of shellfish, and 10-15 parts by weight of aloe, based on the total weight of the natural material auxiliary materials. , 10-15 parts by weight of chicory, and 10-15 parts by weight of burdock are excellent in terms of supplying natural product-derived sugar sources for fermentation, and have the best efficacy in preventing or improving intestinal aging, that is, improving intestinal health.

In the fermented seaweed mixture, the natural auxiliary materials may be included in an amount of 100 to 300 parts by weight, preferably 200 to 300 parts by weight, based on 100 parts by weight of seaweed.

If the content of the natural additives is less than the lower limit, the synergistic effect on the intestinal health improvement efficacy when fermented with seaweed may be minimal, and if it exceeds the upper limit, the intestinal health improvement efficacy may actually decrease.

The fermented seaweed mixture may be obtained by fermenting seaweed and natural auxiliary materials with lactic acid bacteria.

According to a preferred embodiment of the present invention, the fermented seaweed mixture includes the steps of (A) inoculating a mixture of seaweed, sugars, and purified water with lactic acid bacteria and then performing primary fermentation for 5 to 15 months; (B) mixing the primary fermentation product and organic acid and performing secondary fermentation for 20 to 30 days; (C) filtering the secondary fermentation and aging at low temperature for 10 to 30 days; (D) mixing 200 to 300 parts by weight of natural additives with 100 parts by weight of the low-temperature aged secondary fermentation product; and (E) subjecting the mixture to a third fermentation for 10 to 20 months.

First, in step (A), lactic acid bacteria are inoculated into a mixture of salicornia, sugars, and purified water, and then primary fermentation is performed for 5 to 15 months, preferably 9 to 11 months.

The seaweed may be a seaweed raw material, a seaweed extract, a seaweed extract, or a seaweed powder. However, the intestinal health improvement activity of the finally prepared seaweed mixed fermentation product is more excellent when used as a seaweed raw material.

The fermentation uses sugar as a substrate and ferments the seaweed using lactic acid bacteria and various enzymes contained in seaweed. When the seaweed is fermented, the nutrients of seaweed can be converted into a form that is easy to digest and absorb through various biochemical reactions, and enzyme action It is performed to express the function of improving intestinal health or intestinal function by the ingredients produced.

The sugars are mixed in an amount of 5 to 25 parts by weight, preferably 10 to 20 parts by weight, and purified water is mixed in an amount of 500 to 1,500 parts by weight, preferably 700 to 1,200 parts by weight, based on 100 parts by weight of Salicornia. If the sugar is added below the lower limit or exceeds the upper limit, unintended fermentation products may be produced.

The saccharide may be one or more selected from sugar, maltose, glucose, xylitol, sorbitol, mannitol, lactitol, polyglycitol, erythritol, maltitol, isomaltooligosaccharide, xylooligosaccharide, fructooligosaccharide, and galactooligosaccharide.

The lactic acid bacteria are preferably inoculated at 0.5 to 2 parts by weight, preferably 0.5 to 1.5 parts by weight, based on 100 parts by weight of Salicornia. If the lactic acid bacteria are added below the lower limit or exceed the upper limit, unintended fermentation products may be produced.

The lactic acid bacteria include Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus plantarum, Leuconostoc kimchii, Leuconostoc citreum, Leuconostoc mesenteroides, and Leuconostoc acanthuscomitatum. , Leuconostoc lactis, Lactobacillus sakei, and Lactobacillus brevis. Lactic acid bacteria of the genus Lactobacillus are preferable in terms of efficacy in improving intestinal aging, and more preferably Lactobacillus plantarum.

The lactic acid bacteria can be added as the lactic acid bacteria themselves or inoculated with a culture medium. The culture medium may include the culture medium itself obtained by culturing the strain in a liquid medium, a concentrate obtained by concentrating the culture medium, etc. In one embodiment, the amount of lactic acid bacteria is 1×10 ⁶ to 1×10 ⁹ cfu/mL, preferably 1×10 ⁶ to 1×10 ⁸ cfu/mL, more preferably 1×10 ⁷ to 1×10 ⁸ cfu. It may be inoculated with a culture medium contained at a concentration of /mL.

The primary fermentation may be carried out at 18 to 22 °C. If the temperature during fermentation is below the above lower limit, the fermentation period is prolonged, the growth of lactic acid bacteria is not promoted, and not only does it not have a significant effect on improving intestinal health, but it also cannot prevent or improve intestinal aging, and exceeds the above upper limit. In this case, abnormal growth of lactic acid bacteria may occur, rotting of the seaweed may occur, and the inflammatory reaction may become worse.

Next, in step (B), the primary fermentation product and organic acid are mixed and then subjected to secondary fermentation for 20 to 30 days. The addition of organic acids such as citric acid during secondary fermentation is intended to promote lactic acid bacteria fermentation and further increase the functionality of the fermented product.

The organic acid may be mixed in an amount of 2 to 6 parts by weight, preferably 3 to 5 parts by weight, based on 100 parts by weight of the primary fermentation product.

The organic acid may be one or more selected from citric acid, tartaric acid, malic acid, fumaric acid, succinic acid, and vitamin C.

The secondary fermentation may be carried out at 18 to 22 °C. If the temperature during fermentation is below the above lower limit, the fermentation period is prolonged, the growth of lactic acid bacteria cannot be promoted, and intestinal aging or decline in intestinal function due to intestinal aging cannot be improved. If the temperature exceeds the above upper limit, abnormal growth of lactic acid bacteria occurs. Otherwise, the seaweed may rot, and the inflammatory reaction may become worse.

Next, in step (C), the secondary fermentation product is aged at low temperature for 10 to 30 days.

The low-temperature ripening is a process for not only stabilizing the fermentation product by stopping or slowing down the fermentation, but also further increasing the effective bioactive components of the fermentation product and diluting the salty taste of the seaweed, preferably at 2 to 10 ° C. It can be done at low temperatures ranging from 3 to 6°C. Through the low-temperature aging process, it is possible to obtain a fermented product of seaweed that not only has the unique flavor and aroma of naturally fermented seaweed, but also has good nutritional value, including rich minerals and amino acids.

Next, in step (D), 200 to 300 parts by weight of natural additives are mixed with 100 parts by weight of the low-temperature-aged secondary fermentation product.

If the content of the natural additives is less than the lower limit, the synergistic effect on the intestinal health improvement efficacy when fermented with seaweed may be minimal, and if it exceeds the upper limit, the intestinal health improvement efficacy may actually decrease.

The natural auxiliary materials may be extracts, extracts or powders, but the intestinal health improvement activity of the finally prepared fermented seaweed mixture is more excellent when used in powder form.

Next, in step (E), the mixture is subjected to a third fermentation for 10 to 20 months.

The tertiary fermentation is to express the synergistic effect of the effective physiologically active ingredients in the salicornia and the natural product auxiliary materials on the efficacy of improving intestinal aging by fermenting the fermented product and the natural product auxiliary materials together, and can be carried out in the range of 18 to 22 ℃. there is. If the temperature and/or period of the tertiary fermentation is less than the above lower limit, the growth of lactic acid bacteria may be reduced due to the low temperature, and the synergistic effect on the intestinal health improvement effect may be minimal, and if it exceeds the above upper limit, unintended fermentation products may occur. can be created.

After step (E), the step of filtering and sterilizing the third fermentation product may be additionally performed.

The fermented seaweed mixture can be used as is, powdered, concentrated, or diluted.

The fermented seaweed mixture prepared as described above is a health functional food for improving intestinal health, a health functional food for preventing or improving intestinal aging, a health functional food for preventing or improving intestinal aging-related diseases, and suppressing the decline in intestinal function due to intestinal aging. Health functional food, health functional food for improving intestinal function, pharmaceutical composition for preventing or treating inflammatory bowel disease, feed composition for preventing or improving intestinal aging, feed composition for improving intestinal health, or preventing or treating inflammatory bowel disease. It can be used as a pharmaceutical composition for animals.

'Health functional food' of the present invention refers to food manufactured (including processing) in accordance with legal standards using raw materials or ingredients with functional properties useful to the human body (Article 3, Paragraph 1 of the Health Functional Food Act). The terminology or scope of the above 'health functional food' may vary depending on the country, but it is also called 'Dietary Supplement' in the United States, 'Food Supplement' in Europe, 'Health Functional Food' or 'Health Functional Food' in Japan. It may be classified as ‘Food for Special Health Use (FoSHU)’ or ‘Health Food’ in China.

The above health functional food may additionally contain food additives, and its suitability as a food additive shall be determined by the specifications and standards for the relevant item in accordance with the general provisions of the ‘Food Additive Code’ and general test methods, etc., unless otherwise specified.

In addition, the health functional food includes raw materials notified as 'functional raw materials' or individually recognized raw materials used in "health functional food for improving intestinal health" along with the fermented glasswort mixture, such as isomaltooligosaccharide, soybean oligosaccharide, raffinose, liquid A health functional food for improving intestinal health can be manufactured by combining health functional food ingredients such as fructooligosaccharide that help proliferate beneficial intestinal bacteria and/or increase short-chain fatty acids in the intestine.

When manufacturing a health functional food using the above seaweed mixed fermentation product as an active ingredient, there is no need to specifically limit the content of the seaweed mixed fermentation product as long as it shows intestinal health improvement activity, for example, 0.1 to 99% by weight, 0.5 to 95% by weight. , 1 to 90% by weight, 2 to 80% by weight, 3 to 70% by weight, 4 to 60% by weight, and 5 to 50% by weight.

The daily dosage of the fermented seaweed mixture, which is an active ingredient in the health functional food, varies depending on the condition, weight, presence, degree, and period of the disease of the administration subject, but can be appropriately selected by a person skilled in the art. However, for a desirable effect, the fermented seaweed mixture in the health functional food is 0.1 to 100 ㎍/kg, preferably 1 to 50 ㎍/kg, more preferably 5 to 35 ㎍/kg per day, based on solid content. More preferably, it is administered at 10 to 30 μg/kg, and even more preferably at 15 to 25 μg/kg. The number of administrations does not need to be particularly limited, but can be adjusted by a skilled artisan within the range of three times a day to once a week. In the case of long-term intake for health and hygiene purposes or health control, it may be below the above range.

The present invention relates to a feed composition for improving intestinal health containing a mixed fermented product of seaweed as an active ingredient.

The 'feed composition' can use food raw materials and food additives listed in the Food Additives Code as described in the Food Standards and Specifications ('Food Code'), in addition to the fermented glasswort mixture as an active ingredient, and can be used as food. Even if they are not food raw materials or food additives, raw materials that fall within the scope of single feed in Annex 1 of the ‘Standards and Specifications for Feed, etc.’ and raw materials that fall within the scope of supplementary feed in Annex Table 2 can be used.

The 'feed composition' may be an extractant among supplementary feeds according to 'Standards and specifications for feed, etc.', or may be a compound feed containing the supplementary feed.

When preparing a feed composition using the above-mentioned Salicornia mixed fermentation product as an active ingredient, the content of the Salicornia mixed fermentation product does not need to be specifically limited as long as it exhibits intestinal health improvement activity, but may be contained, for example, 0.1 to 99% by weight, 0.5 to 95% by weight, It may be included in 1 to 90% by weight, 2 to 80% by weight, 3 to 70% by weight, 4 to 60% by weight, and 5 to 50% by weight.

The fermented seaweed mixture, which is the active ingredient in the feed composition, varies depending on the condition, body weight, presence, degree, and period of disease of the ingesting animal, but may be appropriately selected by a person skilled in the art. For example, based on the daily dosage, 0.1 to 100 μg/kg, preferably 1 to 50 μg/kg, more preferably 5 to 35 μg/kg, more preferably 10 to 30 μg/kg, even more preferably. It is preferably administered at 15 to 25 μg/kg. The number of administrations does not need to be particularly limited, but can be adjusted by a skilled artisan within the range of three times a day to once a week. In the case of long-term intake for health and hygiene purposes or health control, it may be below the above range.

The present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory bowel disease containing a mixed fermented product of seaweed as an active ingredient. The inflammatory bowel disease may be ulcerative colitis or Crohn's disease.

In addition, the present invention relates to a pharmaceutical composition for animals for the prevention or treatment of inflammatory bowel disease, comprising fermented seaweed mixture as an active ingredient.

Additionally, the present invention provides a method of treating inflammatory bowel disease by administering the composition to humans or non-human animals.

In addition, the present invention provides a new use of the fermented seaweed mixture for the manufacture of medicine for the prevention or treatment of inflammatory bowel disease or medicine for animals.

The 'pharmaceutical composition', 'medicine', 'pharmaceutical composition for animals' or 'veterinary medicine' contains, in addition to the fermented seaweed mixture as an active ingredient, appropriate carriers, excipients and diluents commonly used in the production of pharmaceutical compositions, etc. It can be included.

The 'carrier' is a compound that facilitates the addition of the compound into cells or tissues. The 'diluent' is a compound diluted in water that not only stabilizes the biologically active form of the target compound, but also dissolves the compound.

There is no need to specifically limit the carrier, excipient, and diluent, but for example, lactose, glucose, sugar, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose. , methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

The amount of the pharmaceutical composition, medicine, pharmaceutical composition for animals, or medicine for animals may vary depending on the age, gender, and weight of the patient or animal to be treated, and, above all, the condition of the subject to be treated, the specific category of the disease to be treated, or It will depend on the type, route of administration, and properties of the therapeutic agent used.

The pharmaceutical composition, medicine, pharmaceutical composition for animals, or medicine for animals is appropriately selected depending on the absorption rate of the active ingredient in the body, the excretion rate, the age, weight, sex, and condition of the patient or animal to be treated, and the severity of the disease to be treated. , generally 0.1 to 100 μg/kg, preferably 1 to 50 μg/kg, more preferably 5 to 35 μg/kg, more preferably 10 to 30 μg/kg, more preferably 15 to 25 μg. It is preferable to administer /kg. The unit dosage form formulated in this way can be administered several times at regular time intervals as needed.

The pharmaceutical composition, medicine, pharmaceutical composition for animals, or medicine for animals may be administered individually as a preventive or therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

The pharmaceutical compositions, medicines, pharmaceutical compositions for animals, or medicines for animals can be used by formulating them into oral dosage forms such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc. according to conventional methods. there is. When formulated, it can be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, etc. These solid preparations include the compound with at least one excipient, such as starch, calcium carbonate, sugar or lactose, and gelatin. It can be prepared by mixing etc. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. .

The method of treating inflammatory bowel disease involves administering the composition to humans or non-human animals, especially mammals, for example, administering the composition to an individual to be treated with inflammatory bowel disease.

An individual subject to treatment with the inflammatory bowel disease may have inflammation in the intestine.

The dosage, administration method, and frequency of administration for the treatment may refer to the dosage, administration method, and frequency of administration of the pharmaceutical composition, medicine, pharmaceutical composition for animals, or medicine for animals.

In addition, the present invention relates to a health functional food for suppressing decline in intestinal function due to aging containing a mixed fermented product of seaweed as an active ingredient. The “fermented seaweed mixed product” and “health functional food” are as described above.

In this specification, “deterioration of intestinal function due to aging” refers to a decrease in intestinal health or intestinal function due to aging, such as increased intestinal inflammatory response, damage to the intestinal crypt, decreased intestinal mucous layer thickness, and decreased content of butyric acid, a beneficial short-chain fatty acid in the intestine. means that

The health functional food for suppressing the decline in intestinal function due to aging of the present invention contains the mixed fermented product of seaweed of the present invention as an active ingredient, and reduces intestinal inflammatory response and intestinal crypt damage that increase due to aging, and reduces intestinal mucus decreased due to aging. It increases the layer thickness and the content of butyric acid, a beneficial short-chain fatty acid in the intestines, thereby effectively preventing or improving the decline in intestinal health or intestinal function due to aging.

In addition, the present invention relates to a health functional food for preventing or improving intestinal aging-related diseases containing fermented seaweed mixture as an active ingredient. The intestinal aging-related disease may be one or more types selected from irritable bowel syndrome, inflammatory bowel disease, constipation, and ischemic enteritis. The “fermented seaweed mixed product” and “health functional food” are as described above.

The health functional food for preventing or improving intestinal aging-related diseases of the present invention contains the fermented seaweed mixture of the present invention as an active ingredient, and is selected from irritable bowel syndrome, inflammatory bowel disease, constipation, and ischemic enteritis, which are intestinal diseases caused by aging. One or more types can be prevented or improved. The inflammatory bowel disease caused by aging may be ulcerative colitis or Crohn's disease.

In addition, the present invention relates to a pharmaceutical composition for preventing or treating intestinal aging-related diseases containing a mixed fermented product of seaweed as an active ingredient. The “salmonwort mixed fermentation product”, “intestinal aging-related disease”, and “pharmaceutical composition” are as described above.

Hereinafter, the present invention will be described in detail through examples, but the present invention is not limited to the following examples.

<Example>

Example: Preparation of glasswort mixed fermentation product (GWⅡ)

(1) After collecting seaweed with a moisture content of 88-92% from June to August in Shinan-gun, Jeollanam-do, the roots and hard stems were cut and washed 2-3 times with drinking water to remove dirt and foreign substances. The glasswort was cut into appropriate sizes with an average length of 5 cm.

And, Lactobacillus Plantarum strain (1×10 ⁹ cfu/g) was purchased from Wellbeing LS Co., Ltd.

(2) 100 parts by weight of salicornia were mixed with 15 parts by weight of sugar and 800 parts by weight of purified water. The mixture was inoculated with the lactic acid bacteria powder (1×10 ⁹ cfu/g) in an amount of 1 part by weight based on 100 parts by weight of Salicornia and then subjected to primary fermentation at 20° C. for 10 months.

(3) 4 parts by weight of citric acid were added to 100 parts by weight of the primary fermentation and then secondary fermentation was performed at 20° C. for 25 days.

(4) Afterwards, the secondary fermentation product was aged at low temperature for 25 days at 4°C.

(5) 12.5 parts by weight of mistletoe powder, 12.5 parts by weight of Cordyceps sinensis powder, 12.5 parts by weight of pork potato powder, 12.5 parts by weight of malt powder, 12.5 parts by weight of shell powder, 12.5 parts by weight of aloe powder, 12.5 parts by weight of chicory powder, 12.5 parts by weight of burdock powder Natural supplementary materials were prepared by mixing parts by weight.

250 parts by weight of the natural additives were mixed with 100 parts by weight of the low-temperature aged secondary fermentation product obtained in step (4) and homogenized.

(6) The mixture was subjected to a third fermentation at 20°C for 14 months.

The tertiary fermentation product was filtered using a filtration membrane of 300 mesh or more and then sterilized using a UV sterilization method to prepare a mixed fermentation product (GWII).

Comparative Example 1: Preparation of fermented glasswort (GWⅠ)

Fermented seaweed was prepared in the same manner as steps (1) to (4) of the example. The fermented Salicornia herbaceous product (GWI) was prepared by filtering the fermented salicornia product using a filtration membrane of 300 mesh or more and then sterilizing it using a UV sterilization method.

Comparative Example 2: Preparation of Salicornia extract

Distilled water, an extraction solvent, was added to Salicornia root powder at a ratio of 1:10 (w:v), then sufficiently immersed, and extraction was repeated three times for 3 hours at 80°C. The extract was Whatman No. 2 Filtered with filter paper. The filtrate was concentrated under reduced pressure at 60°C to prepare a powder sample and used in the experiment.

< Test example>

statistical processing

All statistical analyzes were performed using SPSS version 18.0 (IBM, Chicago, IL, USA). Differences in mean values between experimental groups (p <0.05) were determined using one-way ANOVA with Duncan's post-hoc test. The following experiments were each repeated at least three times, and each experimental data was expressed as mean ± standard deviation (SD).

Test Example 1: Evaluation of useful ingredient content and physiological activity

1-1: Measurement of phenolic compound content

The total phenolic compound content of the fermented glasswort (mixed) of Example and Comparative Example 1 was measured using Folin-Ciocalteu's reagent.

Specifically, 500 μL of 2 N Folin-Ciocalteu's reagent and 2.5 mL of 20% Na ₂ CO ₃ (Daejung Chemicals, Korea) were added to 100 μL of the fermented glasswort (mixed) of Example and Comparative Example 1, and reacted at room temperature for 20 minutes. Then, the absorbance of this reaction solution was measured at 735 nm using a UV/VIS spectrophotometer (HP-8452A, Hewlett Packard, Cheadle Health, Stockport, Cheshire, UK). The absorbance value was converted to the total phenolic compound content from the standard gallic acid curve, and the value was presented as gallic acid equivalent (GAE) (Figure 1).

As shown in Figure 1, the total phenolic compound content of the salicornia mixed fermentation product (GWⅡ) of the example was 144.6±96.7 mg GAE/g, and that of the salicornia fermentation product (GWⅠ) of Comparative Example 1 was 70.5±0.3 mg GAE/g. , it can be specifically confirmed that the total phenolic compound content of the glasswort mixed fermentation product (GWⅡ) according to an example of the present invention is more than two times higher than that of Comparative Example 1.

In the case of salicornia, it contains various phenolic compounds as a defense substance to respond to salt stress. In the case of salicornia mixed fermentation product (GWⅡ) according to the example, phenolic compounds are contained not only in salicornia but also in various natural auxiliary materials. It can be inferred that the compound content is higher than that of Comparative Example 1.

1-2: Measurement of total flavonoid compound content

Flavonoids are widely distributed in plants and have been used as anti-inflammatory and immunomodulatory agents. Recently, various physiological activities such as antibacterial, antiviral, vascular system regulation, anti-inflammatory, and anticancer have been revealed. In Salicornia, flavonoid glycoside compounds isorhamnetin 3-O-β-D-glucopyranoside, quercetin 3-O-β-D-glucopyranoside, and tungtungmadic acid (3-caffeoyl-4dihydrocaffeoly quinic acid) have been identified as antioxidant active substances.

The total flavonoid content was measured using a colorimetric method in which the flavonoids contained in the sample are chelated by aluminum and turn yellow when an alkaline solution is added.

Specifically, 1 mL of tertiary distilled water and 75 μL of 5% NaNO ₂ were added to 250 μL of the fermented glasswort (mixed) of Example and Comparative Example 1, and then reacted at room temperature for 5 minutes. Afterwards, 150 μL of 10 mM AlCl ₃ ㅇ6H ₂ O was mixed with the reaction solution and reacted again for 6 minutes, then 500 μL of 1 N NaOH was added, and the absorbance was measured at 510 nm after 11 minutes. The absorbance value was compared with a calibration curve using quercetin as a standard material, and the flavonoid content was expressed as quercetin equivalent (QE) (Figure 2).

As shown in Figure 2, the total flavonoid content of the fermented seaweed mixture (GWⅡ) of the example was 2.0 ± 0.1 mg QE/g, and the fermented seaweed mixture (GWⅠ) of Comparative Example 1 was 1.6 ± 0.1 mg QE/g. It was confirmed that the total flavonoid compound content of the glasswort mixed fermentation product (GWII) according to the example of the invention was significantly higher than that of Comparative Example 1. These results are similar to the total phenolic compound content, and in the case of the seaweed mixed fermentation product (GWⅡ) of the example, it is believed that the total flavonoid compound content was high due to the flavonoid compounds contained in various natural auxiliary materials as well as seaweed.

Most flavonoids contained in plants exist in the form of glycosides, but are known to be separated into aglycone form through a fermentation process, thereby increasing the absorption rate in the body and antioxidant activity. In the case of the fermented seaweed mixture of the present invention, it is believed that the content of aglycone-type flavonoid compounds produced during the fermentation process along with the useful components of seaweed itself during the fermentation process of seaweed and natural auxiliary materials was further increased.

1-3: DPPH radical scavenging ability

The DPPH radical scavenging ability of the fermented seaweed (mixed) product was measured using the principle that DPPH radicals are scavenged by bioactive substances with antioxidant activity such as tocopherol, ascorbate, flavonoid compounds, aromatic amines, and peptides, making them colorless.

Specifically, 900 μL of a 100 μM DPPH ethanol solution (final concentration 90 μM) was added to 100 μL of the Salicornia (mixed) fermented product of Example and Comparative Example 1, mixed lightly with a stirrer, and then reacted in the dark for 20 minutes and then stirred at 517 nm. The absorbance was measured. The DPPH radical scavenging ability was calculated using the formula below and shown in Figure 3a, and the SC50 value was calculated from the concentration at which the radical scavenging ability is 50% from the calibration curve for the radical scavenging ability at each concentration and shown in Figure 3b below.

[Equation 1]

DPPH radical scavenging ability (%) = (1-A/B) x 100

(A: Absorbance of sample addition section, B: Absorbance of untreated section)

As shown in Figures 3a and 3b, the DPPH radical scavenging activity of the fermented seaweed mixture (GWII) of the example increased in a concentration-dependent manner. In particular, it was confirmed that the seaweed mixed fermentation product (GWⅡ), which was fermented by adding natural ingredients to seaweed, showed higher DPPH radical scavenging activity compared to the seaweed fermentation product (GWⅠ) of Comparative Example 1.

The antioxidant activity of the fermented seaweed mixture (GWII) was correlated with the total phenolic and total flavonoid compound contents. Salicornia contains various phenolic compounds as a defense mechanism to respond to salt stress, and these phenolic compounds are well known to have excellent antioxidant activity. Therefore, it is believed that the antioxidant activity of the fermented seaweed mixture is mainly due to the content of total phenolic compounds.

1-4: ABTS+ radical scavenging ability

The ABTS+ radical scavenging ability of each sample was measured by a colorimetric method using the principle that the ABTS+ radicals generated by the potassium persulfate reaction are scavenged by substances with antioxidant ability in the sample, causing the radical's unique blue-green color to become colorless.

Specifically, the ABTS+ radical solution was tested by mixing 1 mM AAPH solution with 2.5 mM ABTS solution dissolved in 100 mM potassium phosphate buffer (pH 7.4) containing 150 mM NaCl and reacting in a constant temperature bath at 70°C for 30 minutes. It was used in . At this time, the concentration of blue-green ABTS+ radical was adjusted to have an absorbance of 0.7 ± 0.02 at 734 nm using a UV/VIS spectrophotometer, and 20 μL of the fermented seaweed (mixed) of Example and Comparative Example 1 and the ABTS+ radical prepared above were adjusted. 980 μL of the solution was mixed and reacted at 37°C for 10 minutes, and then the absorbance was measured at 734 nm. The ABTS+ radical scavenging ability was calculated using the formula below and shown in Figure 4a, and the SC50 value was calculated from the concentration at which the radical scavenging ability is 50% from the calibration curve for the radical scavenging ability at each concentration and shown in Figure 4b below.

[Equation 2]

ABTS+ radical scavenging ability (%) = (1-A/B) x 100

(A: Absorbance of sample addition section, B: Absorbance of untreated section)

As shown in Figures 4a and 4b, the ABTS+ radical scavenging activity of the fermented seaweed mixture (GWII) of the example increased in a concentration-dependent manner. In particular, it was confirmed that the mixed fermented seaweed product (GWⅡ), which was fermented by adding natural ingredients to seaweed, showed higher ABTS+ radical scavenging activity compared to the fermented seaweed product (GWⅠ) in Comparative Example 1.

Therefore, it was confirmed again that the seaweed mixed fermentation product (GWⅡ) fermented by adding the natural auxiliary materials of the present invention had higher antioxidant activity than the seaweed fermentation product (GWI) of Comparative Example 1.

sintering

As a result of evaluating the useful components and physiological activity of the fermented seaweed product, the content of total phenolic compounds and total flavonoid compounds of the seaweed mixed fermented product (GWⅡ) fermented by mixing the natural auxiliary materials of the present invention was higher than that of the fermented seaweed product (GWⅠ). It was found to be significantly high. In addition, the mixed fermented product of seaweed (GWⅡ) was found to have higher antioxidant activity than the fermented seaweed (GWⅠ). Therefore, the efficacy of improving intestinal health was evaluated for the fermented seaweed mixture (GWII) of the example.

Animal testing: Evaluation of the effect of fermented seaweed mixture on intestinal health

In order to determine the effect of the fermented seaweed mixture (GWⅡ) prepared according to the above example on intestinal health, an intestinal health-related evaluation was performed on the large intestine of mice with induced aging.

(1) Selection of aging-induced animal model

As aging occurs, intestinal motility decreases, which has a negative impact on intestinal health, such as constipation. Recently, as the elderly population increases, the development of health functional foods targeting the elderly is increasing. Therefore, in this study, animal experiments were conducted under conditions that induce aging to develop products targeting the elderly.

The D-galactose-induced aging model, which is most commonly used in animal experiments on aging, uses the principle that excessive galactose generates excessive oxygen radicals in the body, causing aging.

Aging was induced using 10-week-old male Sparague-Dawley rats as experimental animals by intraperitoneally administering D-galactose (150 mg/kg) daily for 8 weeks. The SD rats used in this experiment were purchased from Gbio (Gwangju, Korea), approved by the Animal Experiment Ethics Committee of Mokpo University, and raised at the Animal Testing Center of Mokpo University (approval number, MNU-IACUC-2020-010).

The breeding environment was set at a temperature of 22 ± 1 ℃, humidity of 50 ± 5%, and light/dark cycle of 06:00 to 18:00. The experimental animals were raised with regular solid feed (purina) for one week after arrival, adapted to the environment, and aged for 7 weeks. At this point, each experimental group was divided according to body weight and the experiment was performed with two animals per cage.

(2) Experimental group classification

As shown below, the samples were divided into a total of 5 experimental groups (n=8) and each sample was orally administered while inducing aging for 8 weeks.

1) CON (normal group): untreated group

2) CD (Aging induced control group): D-galactose (150 mg/kg/day) intraperitoneal administration + saline

3) PC (aging induction + positive control group)

: D-galactose (150 mg/kg/day) intraperitoneal administration + fructooligosaccaride (0.1 mg/day)

4) LGW (Aging induction + Salicornia mixed fermentation product (GWⅡ) low concentration administration group)

: D-galactose (150 mg/kg/day) intraperitoneal administration + fermented glasswort mixture of the example (10 ㎍ solid content/kg/day)

5) HGW (Aging induction + Salicornia mixed fermentation product (GWⅡ) high concentration administration group)

: D-galactose (150 mg/kg/day) intraperitoneal administration + fermented glasswort mixture of the example (20 ㎍ solid content/kg/day)

6) GWⅠ (Aging induction + Salicornia fermented product (GWⅠ) administered group)

: D-galactose (150 mg/kg/day) intraperitoneal administration + fermented glasswort of Comparative Example 1 (20 ㎍ solid content/kg/day)

7) Salicornia extract (aging induction + Salicornia extract administration group)

: D-galactose (150 mg/kg/day) intraperitoneal administration + Salicornia extract of Comparative Example 2 (20 ㎍ solid content/kg/day)

Test Example 2: Changes in weight gain, diet, and water intake of experimental animals

The amount of feed and water consumed during the experiment period was calculated daily, excluding the remaining amount and spilled amount, and the amount of body weight gain was measured once a week for a total of 8 weeks, using the end of the stabilization period as the initial body weight and shown in Figure 5.

Figure 5a shows the weight gain of each experimental group, Figure 5b shows the dietary amount of each experimental group, and Figure 5c shows the water intake of each experimental group.

As shown in Figure 5, the body weight of all experimental groups continued to increase during the diet period, and in the case of the high concentration group of seaweed mixed fermentation (HGW), body weight tended to be low, but there was no statistically significant difference between the experimental groups.

Specifically, the body weight of the experimental group that consumed the fermented seaweed mixture was found to increase to a level similar to that of the normal group (CON), confirming that the diet proceeded normally. In addition, the experimental group that consumed fermented seaweed mixture showed a normal intake pattern overall, with a daily dietary intake of approximately 45 to 55 g. In addition, water intake showed a normal intake pattern, but decreased by about 10 mL in the 4th week of the experiment, but recovered again from the 5th week of the experiment, and the water intake of all experimental groups appeared to be at a similar level.

Test Example 3: Histological analysis of the large intestine

3-1: Shape of crypt and number of goblet cells in colon tissue

70% of immune cells are concentrated in the intestine, and the epithelial cells of the intestinal wall are continuously damaged by mechanical stress due to continuous intestinal movement, chemical stress due to environmental hormones from food, and biological stress from harmful bacteria in the intestine, shortening the lifespan. It is known to take about 3 to 5 days. Therefore, these intestinal cells must be continuously supplied from intestinal stem cells, and intestinal stem cells, which play such an important role, exist at the bottom of a narrow invagination called a crypt in the intestinal wall, thereby protecting them from harmful intestinal environments. there is. In addition, in the intestine, mucus-secreting cells called goblet cells cover the epithelium from the small intestine to the large intestine, forming a boundary between the intestinal lumen and the tissue. The intestinal mucosal epithelial tissue is responsible for the primary defense system, and in the case of goblet cells, the intestinal tract is protected. Secretes mucin, a major component of the mucus layer.

On the other hand, when the intestines are continuously stimulated by various stress factors such as mechanical, chemical, and biological, the crypt that protects the intestinal stem cells is damaged, and the amount of mucous substances produced and the number of goblet cells decrease, resulting in a decrease in the number of goblet cells. It has a negative impact on intestinal health.

In this study, we attempted to confirm histological changes in the crypt and goblet cells of the large intestine when consuming a mixed fermented product of Salicornia herbacea using H&E staining.

To observe histological changes in the experimental animals, after the experiment was completed, the experimental animals were sacrificed, the colon tissue was removed, and a part of the colon was fixed in a 10% formaldehyde solution, then dehydrated, transparent, and permeabilized, embedded in paraffin, and then placed in a tissue slicer. Serial sections were prepared by cutting into 5μm-thick sections using a microtome.

The serially sectioned sample was rehydrated after removing the paraffin embedded on the outside of the tissue with xylene, stained with haematoxylin solution for 10 minutes using H&E staining, rehydrated, and then stained with eosin solution for 20 seconds. Afterwards, after dehydration and clearing with alcohol, the tissue was encapsulated (mounted) with permount encapsulant, and morphological changes in the tissue were observed using an optical microscope (Olympus, Japan) (Figure 6).

Figure 6a shows the colon of the normal group (CON), aging-induced control (CD), positive control (PC), Salicornia mixed fermentation low-concentration diet group (LGW), and Salicornia mixed fermentation high-concentration diet group (HGW) stained with H&E staining method. The crypts and goblet cells of the large intestine were morphologically analyzed, and Figure 6b shows a morphological analysis of the morphology of mucin-secreting goblet cells and intestinal tissue changes by staining the large intestine of each experimental group with Alcian blue staining. Figure 6c is a graph showing the number of goblet cells in colon tissue.

As shown in Figures 6a and 6c, in the normal group, the crypt and goblet cells maintained their normal shape, but in the CD group, which is an aging-induced control group, severe damage to the colonic tissue was caused, leading to deformation of the crypt and goblet cells. It was confirmed that phenomena such as decrease occurred. Meanwhile, among the aging induction groups, the experimental group that consumed the fermented seaweed mixture showed less crypt damage and a higher number of goblet cells in the colon tissue compared to the CD group (the number of goblet cells was in the order of CON > HGW > PC ≒ LGW > CD). ). In particular, the number of goblet cells in the intestinal epithelial cells in the HGW group, which consumed a high concentration of seaweed mixed fermentation product, was significantly higher than that of the CD group, and the experimental group that consumed seaweed mixed fermentation product had a level similar to or higher than that of the positive control group.

In Table 1 below, the number of goblet cells in the intestinal epithelial cells of each experimental group, including the Example and Comparative Example diet groups, is compared and shown in specific numbers.

division Goblet cell count/10 crypts CON Jeongsang-gun 120.6±6.9 CD Aging-induced control group 83.6±3.4 PC Aging induction + positive control group 103.2±3.8 LGW Aging induction + glasswort mixed fermentation product of the example
Low dose group 100.6±4.2 HGW Aging induction + glasswort mixed fermentation product of the example
High dose group 113.1±9.2 GWⅠ Aging induction + Salicornia fermented product administration group of Comparative Example 1 90.2±1.6 Salicornia extract Aging induction + Salicornia extract administration group of Comparative Example 2 86.6±2.2

As shown in Table 1, it can be seen that the fermented seaweed mixed diet group of the present invention has a significantly higher number of goblet cells than the fermented seaweed diet group of Comparative Example 1 and the seaweed extract diet group of Comparative Example 2.

More specifically, the number of goblet cells in the aging-induced control group (CD) was reduced by about 30% compared to the normal group. On the other hand, among the seaweed mixed fermentation diet groups of the example, the low-concentration group (LGW) increased the number of goblet cells by about 20% or more compared to the aging-inducing control group (CD), and the high-concentration group (HGW) increased the number of goblet cells by more than 20% compared to the aging-inducing control group (CD). Compared to , the number of goblet cells increased by about 35%. In particular, in the high dose administration group (HGW), the number of goblet cells was found to be close to the level of the normal group, even though aging was induced.

Meanwhile, histological analysis was performed using alcian blue staining, which shows the acidic mucopolysaccharide mucin in goblet cells in blue to confirm mucin produced in goblet cells of the large intestine when consuming a mixed fermentation of seaweed (Figure 6b). As shown in Figure 6b, in the case of Alcian blue staining, since acidic mucopolysaccharide is stained, mucin in goblet cells is selectively stained, and changes in the morphology and intestinal tissue of goblet cells that secrete mucin can be confirmed. Looking at Figure 6b, mucin revealed by Alcian blue staining was observed to be similar to the shape and number of goblet cells observed by H&E staining.

From these results, it appears that the seaweed mixed fermentation product can effectively improve or prevent intestinal aging in aging-inducing conditions by reducing intestinal crypt damage and helping to increase the number of goblet cells and mucin secretion. It is expected that it can be usefully used as a health functional food material to improve intestinal health.

3-2: Analysis of mucus layer thickness in colon tissue

Mucus glycoprotein secreted from goblet cells is the main component of the mucous layer of the intestine, and paneth cells, which exist in the crypt of intestinal epithelial cells, secrete anti-microbial peptides, which are representative of defensive substances, to protect against external threats. It serves to defend the host by neutralizing invading pathogenic microorganisms. In addition, the intestinal mucosal epithelial tissue is covered with mucus, which is responsible for the primary defense function. When the inflammatory response in the intestine increases, mucin decreases quantitatively, and changes in the thickness of the mucin layer due to a decrease in the amount of mucin increase the risk of colon cancer. It is also known to be related.

Therefore, in this study, to observe changes in the thickness of the mucous layer of colonic tissue when consuming a mixed fermented product of seaweed, PAS oxidizes the mucous layer polysaccharide with periodic acid to release aldehyde, and stains the mucous layer red through the reaction between aldehyde and Schiff. Histological analysis was performed using (Periodic acid and Schiff) staining.

Specifically, after completing the experiment, the experimental animal was sacrificed, the colon tissue was removed, and a portion of the colon was fixed in Carnoy's solution (60% dry methanol, 30% chloroform, 10% glacial acetic acid) and then dehydrated in the order of methanol-alcohol-xylene. , transparent, permeabilized, embedded in paraffin, and then cut into thin sections of 5 μm using a microtome to produce serial sections.

The serially sectioned sample was treated with xylene-alcohol to remove the paraffin embedded on the outside of the tissue, and then hydrated and the mucous layer was stained using PAS staining. That is, it was stained with 0.5% periodic acid solution for 5 minutes, hydrated again, and then stained with Schiff's reagent solution for 15 minutes. Afterwards, after dehydration and clearing with alcohol, the mucous layer of colonic tissue was observed under an optical microscope (Olympus, Japan) by mounting with permount encapsulant (Figure 7). Figure 7a is a photograph showing the thickness of the mucous layer of colon tissue, and Figure 7b is a graph showing the thickness of the mucous layer of colon tissue.

Looking at Figure 7, the normal group maintained a constant mucous layer thickness of about 33 μm, and the CD group had the thinnest mucous layer thickness of about 20 μm, confirming that the colon tissue was damaged due to aging. In addition, it was confirmed that the thickness of the mucous layer of the experimental groups that consumed the fermented seaweed mixture was maintained at a constant level compared to the CD group, and that the thickness was similar to or thicker than that of the positive control group. In particular, in the case of the group with a high concentration of fermented seaweed mixture (HGW), the thickness of the mucous layer was about 33 μm, which was similar to that of the normal group.

In Table 2 below, the thickness of the mucous layer of the colon tissue of each experimental group, including the Example and Comparative Example diet groups, is compared and shown in specific values.

division Slime layer thickness (㎛) percentage(%) CON Jeongsang-gun 33.2±1.1 100 CD Aging-induced control group 20.5±1.6 61.75 PC Aging induction + positive control group 27.8±2.2 83.73 LGW Aging induction + glasswort mixed fermentation product of the example
Low dose group 30.7±0.5 92.47 HGW Aging induction + glasswort mixed fermentation product of the example
High dose group 33.1±0.7 99.70 GWⅠ Aging induction + Salicornia fermented product administration group of Comparative Example 1 26.9±1.3 81.02 Salicornia extract Aging induction + Salicornia extract administration group of Comparative Example 2 24.3±1.2 73.20

As shown in Table 2, it can be seen that the saltwort mixed fermentation diet group of the present invention had a significantly thicker mucous layer thickness than the saltwort fermentation diet group of Comparative Example 1 and the salicornia extract diet group of Comparative Example 2.

More specifically, the thickness of the mucous layer in the aging-induced control group (CD) was reduced by about 40% compared to the normal group. Meanwhile, among the seaweed mixed fermentation diet groups of the example, the low-concentration group (LGW) had an increase in the thickness of the mucous layer by about 50% compared to the aging-inducing control group (CD), and the high-concentration group (HGW) had an increase in the thickness of the mucous layer by about 50% compared to the aging-inducing control group (CD). Compared to , the thickness of the slime layer increased by about 60%. In particular, in the high concentration administration group (HGW), the thickness of the mucous layer was found to be close to the level of the normal group, even though aging was induced.

These results were similar to the above results of observing crypt and goblet cells, and it was confirmed once again that consumption of seaweed mixed fermentation had a positive effect on the intestinal environment under aging-inducing conditions, effectively improving intestinal aging and ultimately improving intestinal health. there was.

Test Example 4: Effect on the amount of short chain fatty acid (SCFA) production in the large intestine

The production of short-chain fatty acids (SCFA) by intestinal flora has a positive effect in terms of energy supply to the body. In particular, indigestible carbohydrates such as dietary fiber are decomposed into oligosaccharides and monosaccharides by intestinal microorganisms, and are ultimately fermented into short-chain fatty acids (e.g., acetic acid, butyric acid, etc.) and gas under anaerobic conditions. The produced short-chain fatty acids (SCFA) are absorbed into colon tissue and act as anions, or are used for various purposes such as an energy source for the host and immune regulation, and effects such as anti-inflammatory, increased intestinal motility, and anti-cancer effects have been reported.

Therefore, to investigate the effect of consuming seaweed mixed fermentation on the production of short-chain fatty acids (SCFA) by intestinal flora, acetic acid, propionic acid, and SCFAs such as butyric acid were quantitatively analyzed, and the results are shown in Figure 8.

Instrument Shimadzu GC-17A system (Kyoto, Japan) Column A J&W DB-FFAP capillary column
(30 m×0.53 mm×1-μm film thickness) Oven temp. Hold at 100 °C for 1 min, then increase at 8 °C/min to 180 °C, increase at 20 °C/min from 180 °C to 200 °C, then plateau at 200 °C for 5 min. Detector Flame ionization detector (FID) Injection port temp. 200℃ Detector temp. 240℃ Injection vol. 1.0 μL Carrier gas, flow H ₂ 30 mL/min, air 300 mL/min, He 25 mL/min

Figure 8 shows the intestinal flora in the feces of the normal group (CON), aging-induced control (CD), positive control (PC), salicornia mixed fermentation low-concentration diet group (LGW), and salicornia mixed fermentation high-concentration diet group (HGW). This is a graph measuring the amount of short-chain fatty acids (SCFA) produced.

Looking at Figure 8, among short-chain fatty acids, the content of butyric acid, which is a beneficial intestinal short-chain fatty acid, was significantly lower in the aging-inducing control group than in the normal group, and the glasswort mixed fermentation diet group showed that aging was induced. However, it was confirmed that the content was similar or significantly higher than that of the normal group.

In Table 4 below, the content (mM) of butyric acid contained in the feces of each experimental group, including the Example and Comparative Example diet groups, is compared and shown in specific values.

division Butyric acid content (mM) CON Jeongsang-gun 22.1±2.3 CD Aging-induced control group 19.4±1.9 PC Aging induction + positive control group 17.3±2.6 LGW Aging induction + glasswort mixed fermentation product of the example
Low dose group 22.1±1.8 HGW Aging induction + glasswort mixed fermentation product of the example
High dose group 25.8±1.1 GWⅠ Aging induction + Salicornia fermented product administration group of Comparative Example 1 20.4±1.5 Salicornia extract Aging induction + Salicornia extract administration group of Comparative Example 2 18.6±1.2

As shown in Table 4, the saltwort mixed fermentation diet group of the present invention had a significantly higher butyric acid content in feces than the saltwort fermentation diet group of Comparative Example 1 and the salicornia extract diet group of Comparative Example 2. You can check it.

More specifically, the aging induced control group (CD) had a decrease in fecal butyric acid content of about 10% or more compared to the normal group. On the other hand, among the saltwort mixed fermentation diet groups of the example, the low concentration administration group was found to have a butyric acid content in feces close to the level of the normal group, and the high concentration administration group (HGW) had a fecal butyric acid content that was lower than that of the normal group. It was found to have increased by more than 15%.

These results mean that consumption of seaweed mixed fermentation under aging conditions helps increase the production of short-chain fatty acids (especially butyric acid) in the intestine, ultimately helping to improve intestinal aging and/or intestinal health.

Test Example 5: Confirmation of inhibition of oxidative stress in the large intestine

Free radicals generated by various stress factors are very aggressive and are well known to cause various diseases in the body through cell damage and lipid oxidation. Therefore, when aging is induced with D-galactose, various reactive oxygen species are generated and intestinal inflammation is induced, resulting in high oxidative stress. Accordingly, the contents of lipid peroxides and protein oxides, which are indicators of oxidative stress, were measured in the intestines of rats that consumed fermented seaweed mixture under aging-inducing conditions.

Specifically, to measure lipid peroxide content, 1 mL of 0.01 M Tris-HCl buffer (pH 7.4) was added to colonic tissue (0.15 g) obtained from animal experiments, and then homogenized on ice with a Polytron (PT1200E, Kinematicca AG). Then, 1 mL of 0.15 M Tris-HCl buffer (pH 7.4) and 300 μL of 10 mM phosphate buffer (KH2PO4) were added to 100 μL of the supernatant obtained by centrifugation at 5,000 The reaction was performed at 37°C for 20 minutes using a shaking water bath. After the reaction was completed, 0.5 mL of 0.6 M HCl containing 20% trichloroacetic acid and 0.5 mL of 1 M NaOH solution containing 0.67% thiobarbituric acid were added, mixed, and reacted at 95°C for 15 minutes. This reaction solution was cooled, 0.8 mL of n-butanol was added, and the supernatant (n-butanol layer) obtained by centrifuging for 10 minutes at 1,500 , MDA) was converted to the lipid peroxide content in colon tissue using the standard curve.

In addition, to measure the content of protein oxide, a factor related to oxidative stress, in the colonic tissue of each experimental group, a protein solution (1.5 mg) obtained from the colonic tissue was added to 100 μL of a 2 M HCl solution containing 10 mM 2,4-dinitrophenylhydrazine (DNPH). After addition, it was reacted at room temperature and in the dark for 1 hour, with stirring at 15-minute intervals. To this reaction solution, 1 mL of ice-cooled 10% trichloroacetic acid (w/v) was added, mixed, and then centrifuged at 3000 xg for 10 minutes. The obtained protein pellet was washed three times with 2 mL of ethanol/ethyl acetate (1:1, v/v) solution, then dissolved in 1.5 mL of 6 M guanidine hydrochloride (pH 2.3) and incubated for 10 minutes at 37°C. After reacting for several minutes, absorbance was measured at 370 nm. This absorbance value was converted to the protein oxide (protein carbonyl) content in colonic tissue by dividing the molar extinction coefficient of DNPH (ε= 2.2

Figure 9a shows the lipid peroxide content in colon tissue of the normal group (CON), aging-induced control (CD), positive control (PC), salicornia mixed fermentation low-concentration diet group (LGW), and salicornia mixed fermentation high-concentration diet group (HGW). It is a graph showing, and Figure 9b is a graph showing the content of protein oxides in the colon tissue of each experimental group.

As shown in Figures 9a and 9b, the content of lipid peroxide (malondialdehyde, MDA), which is an indicator of lipid oxidation, was the highest in the aging-inducing control group, and the low-concentration diet group (LGW) of seaweed mixed fermentation and the high-concentration diet of seaweed mixed fermentation group. group (HGW) showed a low content. In addition, the protein oxide content was confirmed to be significantly lower in the low-concentration fermented seaweed mixed diet group (LGW) and the high-concentrated seaweed mixed fermented diet group (HGW) compared to the aging-inducing control group.

More specifically, the aging-induced control group (CD) increased the lipid peroxide content by about 10% or more and the protein oxide content by about 25% compared to the normal group. On the other hand, among the seaweed mixed fermentation diet groups of the example, the low concentration group (LGW) had a lipid peroxide content of about 10% or more and a protein oxide content of about 60%, compared to the aging induction control group (CD), and the high concentration group (HGW) The lipid peroxide content decreased by about 30% and the protein oxide content decreased by about 10% or more compared to the aging-inducing control group (CD). In particular, in the high concentration group (HGW), the lipid peroxide content decreased by about 20% compared to the normal group even though aging was induced, and in the low concentration group (LGW), the protein oxide content decreased by about 50% compared to the normal group even though aging was induced. It was found that

These results suggest that consumption of fermented seaweed mixture reduces oxidative stress caused by aging induced by D-galactose, thereby reducing lipid peroxide and protein oxide content.

Test Example 6: Confirmation of the effect of suppressing inflammation in the large intestine

Increased oxidative stress in the body induces an inflammatory response through the tumor necrosis factor-alpha (TNF-α) pathway. Along with an increase in TNF-α content, NF-kB expression increases and various cytokine secretion is promoted, which leads to an inflammatory response. It has been reported that in severe cases, it eventually causes cell death (apoptosis) or necrosis (necrosis), causing various diseases. The main function of Interleukin-1β (IL-1β) is as a mediator of inflammatory response similar to TNF, and it acts as a mediator of inflammatory response in various cells such as macrophages, NK cells, mast cells, neutrophils, and nerve cells through stimulation of various production-inducing substances. It is produced and the immune response through inflammatory response occupies a large area. Interleukin-6 (IL-6) is a multifunctional cytokine that functions in innate and adaptive immunity and is well known as an inflammatory response factor involved in immune responses and acute inflammatory responses.

Therefore, to confirm whether consumption of fermented seaweed mixture reduces intestinal inflammatory response, the expression levels of TNF-α, NF-kB p65, IL-1β, and IL-6, which are key factors in inflammation control, were measured in the colonic tissue of each experimental group. did.

Specifically, 0.01 M NaF, 0.01 M EDTA (pH 7.4), 2 mM phenylmethylsulfonyl fluoride, 0.01 M sodium pyrophosphate, 1 μg/mL pepstain, 1 μg/mL leupeptin, and 0.1 μg/mL aprotin were added to colon tissue (0.15 g). 1 mL of RIPA buffer prepared was added and then homogenized on ice with a polytron (PT1200E, Kinematicca AG, Switzerland). The homogenized solution was extracted by gently stirring at 4°C for 1 hour and centrifuged at 4°C for 40 minutes at 15,000 g to obtain the supernatant. The protein content of each treatment group was measured using the Bradford method. Next, 20 μL of 2x Trcine sample buffer (Biorad) was added to the same protein solution (100 μg, 20 μL) for each experimental group, denatured at 95°C for 5 minutes, and this solution was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS). Electrophoresis was performed using -PAGE). The developed proteins were transferred to polyvinylidene fluoride (PVDF) membrane I, then the membrane was washed with Tris buffered saline and blocked for 1 hour with 0.05% Tween-20 (TBST) buffer containing 3% bovine serum albumin (BSA). . Then, the membrane that was washed three times for 5 minutes each with TBST solution was placed in a lypophilized buffer containing primary antibodies and reacted overnight with gentle agitation at room temperature. After completing the primary antibody reaction, the membrane was washed three times for 5 minutes each with TBST solution, then horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobin G (Jackson Immuno Research, LABORATORIES, INC) was added and reacted at room temperature for 1 hour. . The washed membrane was reacted with an enhanced chemiluminescence (ECL) solution containing luminol, developed using Davinch Chemisystem (Core Bio System Co., Ltd), and analyzed using ImageJ software (NIH, Bethesda, Maryland, USA). The primary antibodies used were lypophilized buffer diluted with TNF-α (1:1,000), NF-kB p65 (1:1,000), IL-1β (1:1,000), and β-actin (1:1,000), respectively.

Figure 10 shows TNF-, an inflammation-related factor, in colonic tissues of the normal group, the aging-induced control (CD), the positive control (PC), the salicornia mixed fermentation low-concentration diet group (LGW), and the salicornia mixed fermentation high-concentration diet group (HGW). Western blot results showing expression of α (a), NF-kB p65 (b), IL-1β (c), and IL-6 (d), and a graph showing the expression amount.

As shown in Figure 10, the Salicornia mixed fermentation diet group reduced the expression level of inflammatory cytokines (TNF-α, NF-kB p65, IL-1β, IL-6) compared to the aging induction group (CD group). A trend was shown, and it was confirmed to be similar to or reduced to the positive control group.

More specifically, the senescence-induced control group (CD) showed an increase in NF-kB p65 expression by about 25%, IL-1β expression by about 50%, and IL-6 expression by about 10% compared to the normal group. On the other hand, among the seaweed mixed fermentation diet groups of the example, the high concentration administration group (HGW) had an expression level of TNF-α of about 10%, an expression level of NF-kB p65 level of about 20%, and an expression level of IL-1β compared to the aging control group (CD). The expression level of IL-6 decreased by about 25% and about 5%. In particular, in the high dose administration group (HGW), despite inducing aging, the expression levels of NF-kB p65 and IL-1β were reduced to levels close to those of the normal group.

It is believed that consumption of fermented seaweed mixture reduces the expression of inflammatory response factors by reducing inflammatory response and oxidative stress in aging-induced conditions.

conclusion

In this study, the effect of improving intestinal aging and intestinal health of the fermented seaweed mixture of the present invention was verified using an animal model under conditions that induce aging.

Specifically, when observing the colon tissue of experimental animals by staining, it was confirmed that when the fermented seaweed mixture was consumed, there was less damage to the crypt that protects the intestinal stem cells and there was a large number of mucin-secreting goblet cells that form the mucin layer of the intestine. . In addition, when the fermented seaweed mixture was consumed even under aging-inducing conditions, the intestinal mucous layer that protects the intestines from harmful environments was maintained thick, and the production of butyric acid, a short-chain fatty acid used in the intestines for various purposes such as energy source and immune regulation of the host, was increased. appeared to be increasing. It has also been shown to reduce intestinal oxidative stress and inflammatory responses.

Therefore, consumption of the fermented seaweed mixture of the present invention has a positive effect on the intestinal environment under aging-inducing conditions and has been shown to be effective in improving intestinal health, especially in preventing, improving or overcoming intestinal aging.

Below, a preparation example of a composition containing the mixed fermented product of Salicornia of the present invention is described, but the present invention is not intended to be limited, but merely explained in detail.

Formulation Example 1: Preparation of powder

Glasswort mixed fermentation product (solid content) of the example: 20 ㎍/kg

100 mg lactose

Talc 10 mg

The above ingredients are mixed and filled into an airtight bubble to prepare a powder.

Formulation Example 2: Preparation of tablets

Glasswort mixed fermentation product (solid content) of the example: 20 ㎍/kg

Corn starch 100 mg

100 mg lactose

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are manufactured by compressing them according to a typical tablet manufacturing method.

Formulation Example 3: Preparation of capsules

Glasswort mixed fermentation product (solid content) of the example: 20 ㎍/kg

3 mg crystalline cellulose

Lactose 14.8 mg

Magnesium stearate 0.2 mg

Capsules are prepared by mixing the above ingredients and filling them into gelatin capsules according to a typical capsule manufacturing method.

Formulation Example 4: Preparation of granules

Glasswort mixed fermentation product (solid content) of the example: 20 ㎍/kg

Vitamin mixture dosage

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 μg

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture appropriate amount

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Monobasic Potassium Phosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above vitamin and mineral mixture is a mixture of ingredients suitable for health functional foods in a preferred embodiment, but the mixing ratio may be modified arbitrarily, and the above ingredients are mixed according to a normal health functional food manufacturing method. Next, granules can be prepared and used to manufacture a health functional food composition according to a conventional method.

Formulation Example 5: Preparation of beverage formulation

Glasswort mixed fermentation product (solid content) of the example: 20 ㎍/kg

1,000 mg citric acid

100 g oligosaccharides

2 g plum concentrate

1 g taurine

Add purified water to make a total of 900 mL.

After mixing the above ingredients according to a typical beverage production method, stirring and heating at 85° C. for about 1 hour, the resulting solution was filtered, placed in a sterilized 2 L container, sealed, sterilized, and stored in the refrigerator. Then, the solution according to the present invention was mixed. It is used in the production of functional beverage compositions.

Formulation Example 6: Preparation of feed composition

Example: Glasswort mixed fermentation product (solid content) 20 ㎍/kg, corn 25.5 kg, wheat 15.04 kg, wheat flour 8.15 kg, rice bran 7.4 kg, soybean meal 18 kg, jade gluten 1kg, chicken by-product 14 kg, animal fat 9 kg, processed salt An animal (dog, pet) feed composition was prepared by mixing 0.3 kg, 0.3 kg of tricalcium phosphate, 1 kg of limestone, 0.01 kg of choline chloride, 0.05 kg of vitamins, 0.05 kg of minerals, and 0.1 kg of digestive enzymes.

Although the present invention has been described in terms of the above-mentioned preferred embodiments, various modifications and variations can be made without departing from the gist and scope of the invention. Additionally, the appended claims cover such modifications or variations as fall within the subject matter of the present invention.

Claims (18)

It is a health functional food for improving intestinal health containing a mixed fermentation product of salicornia obtained by fermenting salicornia and natural auxiliary ingredients,
The natural ingredients include mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, aloe, chicory, and burdock,
The improvement of intestinal health is the prevention or improvement of intestinal aging or intestinal aging-related diseases,
The intestinal aging is caused by one or more types selected from the following: increased intestinal inflammatory response due to aging, increased damage to intestinal crypts, decreased number of intestinal goblet cells, decreased intestinal mucous layer thickness, and decreased content of butyric acid, a short-chain fatty acid in the intestine. Deterioration of intestinal health or intestinal function occurs,
A health functional food for improving intestinal health, wherein the intestinal aging-related disease is at least one selected from irritable bowel syndrome, inflammatory bowel disease, and ischemic enteritis. delete delete According to paragraph 1,
A health functional food for improving intestinal health, characterized in that the natural supplementary material is 200 to 300 parts by weight based on 100 parts by weight of the seaweed. According to paragraph 1,
The natural material auxiliary materials include 10-15 parts by weight of mistletoe, 10-15 parts by weight of Cordyceps sinensis, 10-15 parts by weight of pork potato, 10-15 parts by weight of malt, 10-15 parts by weight of shellfish, and aloe, based on the total weight of the natural material auxiliary materials. A health functional food for improving intestinal health, comprising 10-15 parts by weight, 10-15 parts by weight of chicory, and 10-15 parts by weight of burdock. According to paragraph 1,
The fermented seaweed mixture is a health functional food for improving intestinal health, characterized in that it is obtained by fermenting seaweed and natural auxiliary ingredients with lactic acid bacteria. According to paragraph 1,
The fermented seaweed mixture includes the steps of (A) inoculating a mixture of seaweed, sugars, and purified water with lactic acid bacteria and then performing primary fermentation for 5 to 15 months; (B) mixing the primary fermentation product and organic acid and performing secondary fermentation for 20 to 30 days; (C) aging the secondary fermentation product at low temperature for 10 to 30 days; (D) mixing 200 to 300 parts by weight of natural additives with 100 parts by weight of the low-temperature-aged secondary fermentation product; and (E) subjecting the mixture to a third fermentation for 10 to 20 months. A health functional food for improving intestinal health, characterized in that it is manufactured by a method comprising: In clause 7,
In the step (A), sugars are mixed at 10 to 20 parts by weight, purified water is mixed at 500 to 1500 parts by weight, and lactic acid bacteria are inoculated at 0.5 to 1.5 parts by weight based on 100 parts by weight of seaweed. Health functional food for improving intestinal health. . In clause 7,
The lactic acid bacteria include Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus plantarum, Leuconostoc kimchii, Leuconostoc citreum, Leuconostoc mesenteroides, and Leuconostoc acanthuscomitatum. A health functional food for improving intestinal health, characterized in that it is one or more selected from among , Leuconostoc lactis, Lactobacillus sakei and Lactobacillus brevis. In clause 7,
A health functional food for improving intestinal health, characterized in that fermentation in steps (A) and (B) is carried out at 18 to 22 ℃. In clause 7,
In the step (B), the organic acid is mixed in an amount of 2 to 6 parts by weight based on 100 parts by weight of the primary fermentation product. In clause 7,
A health functional food for improving intestinal health, characterized in that low-temperature ripening in step (C) is carried out at 2 to 10 ° C. In clause 7,
A health functional food for improving intestinal health, characterized in that fermentation in step (E) is carried out at 18 to 22 ℃. Contains a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary ingredients,
A pharmaceutical composition for the prevention or treatment of inflammatory bowel disease, characterized in that the natural additives are mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, aloe, chicory, and burdock. The pharmaceutical composition for preventing or treating inflammatory bowel disease according to claim 14, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease. A feed composition for improving intestinal health containing a mixed fermentation product of salicornia obtained by fermenting salicornia and natural auxiliary materials,
The natural ingredients include mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, aloe, chicory, and burdock,
The improvement of intestinal health is the prevention or improvement of intestinal aging or intestinal aging-related diseases,
The intestinal aging is caused by one or more types selected from the following: increased intestinal inflammatory response due to aging, increased damage to intestinal crypts, decreased number of intestinal goblet cells, decreased intestinal mucous layer thickness, and decreased content of butyric acid, a short-chain fatty acid in the intestine. Deterioration of intestinal health or intestinal function occurs,
A feed composition for improving intestinal health, wherein the intestinal aging-related disease is at least one selected from irritable bowel syndrome, inflammatory bowel disease, and ischemic enteritis. Contains a mixed fermented product of salicornia obtained by fermenting salicornia and natural auxiliary ingredients,
A pharmaceutical composition for animals for the prevention or treatment of inflammatory bowel disease, characterized in that the natural additives are mistletoe, Cordyceps sinensis, pork potato, malt, shellfish, aloe, chicory, and burdock. (A) inoculating a mixture of glasswort, sugars, and purified water with lactic acid bacteria and then performing primary fermentation for 5 to 15 months;
(B) mixing the primary fermentation product and organic acid and performing secondary fermentation for 20 to 30 days;
(C) aging the secondary fermentation product at low temperature for 10 to 30 days;
(D) mixing 200 to 300 parts by weight of natural auxiliary materials consisting of mistletoe, Cordyceps sinensis, pork potato, malt, bean paste, aloe, chicory and burdock with 100 parts by weight of the low-temperature-aged secondary fermentation product; and
(E) a third fermentation of the mixture for 10 to 20 months. A method of producing a fermented seaweed mixture for improving intestinal health, comprising: