Compositions for the prevention or treatment of leaky gut syndrome comprising Ecklonia cava extract as an active ingredient

Abstract

The present invention relates to a composition for the prevention or treatment of leaky gut syndrome comprising a sea tangle extract as an active ingredient. The sea tangle extract of the present invention exhibits an excellent effect of suppressing inflammatory responses in the intestines, an effect of improving intestinal barrier function, and an effect of improving intestinal microbial imbalance, and thus can be used for the treatment and prevention of leaky gut syndrome.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention or treatment of leaky gut syndrome comprising Ecklonia cava extract as an active ingredient, and more specifically, to a composition for the prevention or treatment of leaky gut syndrome comprising Ecklonia cava extract that exhibits an effect of improving intestinal barrier function and an effect of increasing beneficial bacteria in the intestines. Background Technology

[0003] In addition to the basic functions of digesting, absorbing, and excreting food, the human intestinal tract performs an immunological function through its intestinal mucosa, acting as a barrier to prevent intraluminal microbes, their byproducts, antigens, and toxins from entering the bloodstream. In other words, the intestinal mucosa possesses a dual function: it acts as a barrier to block external substances while simultaneously allowing them to pass through and be absorbed. Since problems with these intestinal functions not only disrupt the body's nutrient supply but also significantly interfere with daily life due to recurring symptoms such as constipation, diarrhea, and abdominal pain, intestinal health is of paramount importance.

[0004] Leaky gut syndrome is a condition in which tiny holes form in the intestinal wall due to damage to intestinal cells or proteins acting between cells. In addition to the functions of digesting, absorbing, and excreting food, the intestines also act as a barrier to block the entry of harmful external substances, such as microorganisms and toxins. However, when inflammation occurs in the intestines, the permeability of the intestinal wall increases, allowing various bacteria and toxins to easily penetrate. Furthermore, the attack by endotoxins produced by toxins and bacteria that have entered the body due to the leak can lead to various diseases, including cancer, hypertension, diabetes, acne, cystitis, memory loss, premature aging, and hair loss. If left untreated, leaky gut syndrome can become the root cause of many illnesses.

[0005] Intestinal bacteria are known to be the cause of leaky gut syndrome. As beneficial bacteria decrease and harmful bacteria proliferate, toxin production increases, leading to stool abnormalities or indigestion and directly increasing the permeability of the intestinal wall. Inflammation of the intestinal mucosa occurs due to an imbalance of gut microorganisms, and when the dense connective tissue connecting mucosal cells weakens, harmful substances penetrate the body, triggering an immune response.

[0006] Gut microorganisms possess the following beneficial functions. First, they strengthen the immune system by educating and training the human immune system. When food is consumed, external antigens enter through the intestinal mucosa; intestinal microorganisms, which are primarily distributed in the outer layer of the mucosa, act as the primary defense against microorganisms in food, triggering a rapid and powerful immune response. During this process, intestinal microorganisms constantly interact with the human immune system to reinforce it. Second, through metabolic processes, they break down starches and polysaccharides that are not digested by the body's digestive enzymes, helping to supply the energy needed by the body and providing essential nutrients such as vitamins, folic acid, and short-chain fatty acids. They also participate in the metabolism of cholesterol, bile, and drugs, producing various metabolic byproducts. Third, they regulate gene expression by acting as a switch. Even if one has inherited harmful genes from parents, possessing beneficial gut microorganisms and consuming healthy foods can prevent the gene switch from turning on, thereby suppressing the expression of oncogenes.

[0007] The composition of the gut microbiome can be altered by environmental factors such as dietary habits, antibiotic misuse, and stress, in addition to genetic factors; dietary habits, in particular, play a decisive role. The traditional Korean diet, centered on grains and vegetables, provides adequate calories and is rich in vitamins, minerals, and fiber, resulting in a relatively low risk of adult diseases. However, with the widespread adoption of Western diets, nutritional imbalances have arisen—such as the excessive consumption of animal fats, proteins, refined sugars, and simple sugars, as well as deficiencies in vitamins, minerals, and fiber—leading to a significant increase in the incidence of adult diseases compared to the past. These changes in dietary intake can promote the proliferation of harmful gut microbes, causing a deterioration in gut health; representative intestinal diseases associated with this include inflammatory bowel disease and colorectal cancer. The misuse of antibiotics can also affect the composition of the gut microbiome; diversity decreases and composition changes within just a few days of starting antibiotic administration, and in some cases, the microbes may even acquire resistance to the antibiotics. Stress can also destroy the intestinal mucosa and cause an imbalance in gut microbes, leading to a deficiency in neurotransmitters such as serotonin and dopamine, which can induce depression, autism, ADHD, and dementia. In addition to the aforementioned diseases, gut microbes can affect the entire body, affecting not only various intestinal disorders such as functional bowel disorders, irritable bowel syndrome, and antibiotic-induced enteritis, but also neuropsychiatric disorders, fatty liver, diabetes, obesity, arteriosclerosis, allergic diseases, and cancer.

[0008] Consequently, various studies on microorganisms for gut health are being conducted. However, most research related to gut health focuses on bacterial flora, and the majority of gut health improvement products are limited to microbiome-based probiotics such as Bifidobacteria and Lactobacillus. Furthermore, while various functional ingredients with verified efficacy in improving gut health are being reported, there is a shortage of seaweed-derived ingredients certified as individually recognized ingredients.

[0009] Ecklonia cava is a brown algae belonging to the family Alariaceae of the order Laminariales, and in Korea, it mainly inhabits the coastal waters of Jeju at depths of around 10 meters. Research utilizing Ecklonia cava is being conducted on its antioxidant, anticancer, and anti-inflammatory properties, and it is reported to have sufficient value as a new functional material. In particular, phlorotannin, a major component of Ecklonia cava, is a polyphenol compound with phloroglucinol as its basic structural unit, and it has been reported to exhibit antioxidant and anti-inflammatory effects.

[0010] Prior art regarding the use of Ecklonia cava extract for improving gastrointestinal function includes Korean Published Patent No. 10-2022-0021237, "Composition for the prevention and treatment of intestinal diseases caused by fine dust containing Ecklonia cava extract," which discloses the effect of Ecklonia cava extract improving reduced body weight and food intake due to ultrafine dust and improving intestinal function; and Korean Published Patent No. 10-2022-0074353, "Composition for promoting gastrointestinal motility containing Ecklonia cava extract as an active ingredient," which discloses the effect of administering Ecklonia cava extract to animals with reduced gastrointestinal motility to promote gastrointestinal motility and stimulate the secretion of related hormones. However, there are no studies disclosing the effect of Ecklonia cava extract on improving intestinal barrier function and increasing beneficial intestinal bacteria.

[0011] The inventors verified the efficacy of Ecklonia cava extract in cell lines and animal-based inflammation models to discover a seaweed-derived raw material capable of preventing or treating leaky gut syndrome. Furthermore, by confirming that Ecklonia cava extract improves intestinal barrier function and increases beneficial gut bacteria, they completed the present invention by confirming that Ecklonia cava extract can be used for the treatment and prevention of leaky gut syndrome. The problem to be solved

[0013] The objective of the present invention is to provide a use of Ecklonia cava extract for the treatment and prevention of leaky gut syndrome. means of solving the problem

[0015] In order to achieve the above objective,

[0016] The present invention provides a pharmaceutical composition for the prevention or treatment of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient.

[0017] In addition, the present invention provides a health functional food composition for the prevention or improvement of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient.

[0018] In addition, the present invention provides a feed additive for the prevention or improvement of leaky gut syndrome containing Ecklonia cava extract as an active ingredient. Effects of the invention

[0020] The sea mustard extract of the present invention exhibits an excellent effect of suppressing inflammatory responses in the intestines, reduces the expression of proteins that increase intestinal barrier permeability, and improves intestinal barrier function by increasing the expression of tight junction proteins and intestinal mucin, and improves intestinal microbial imbalance by increasing beneficial bacteria and decreasing harmful bacteria, so it can be used for the treatment and prevention of leaky gut syndrome. Brief explanation of the drawing

[0022] Figure 1 shows the results of confirming the expression of inflammation-related proteins iNOS and COX2 after pre-treating macrophages with Ecklonia cava ethanol extract at different concentrations and stimulating them with LPS. Figure 2A is a figure confirming the expression of TNF-α, an inflammatory cytokine induced by LPS, in co-cultured Caco2 cells. Figure 2B shows the expression of IL-6, an inflammatory cytokine induced by LPS, in co-cultured Caco2 cells. Figure 2C shows the expression of IL-1β, an inflammatory cytokine induced by LPS, in co-cultured Caco2 cells. Figure 3A is a diagram showing the method for constructing a mouse animal model induced by Dextran sulfate Sodium (DSS). (Control: Normal group / DSS: Group with induced inflammatory bowel disease / ECE50: Group orally administered 50 mg / kg of Jeju Ecklonia cava extract starting 2 weeks before inducing colitis / ECE150: Group orally administered 150 mg / kg of Jeju Ecklonia cava extract starting 2 weeks before inducing colitis / ECE300: Group orally administered 300 mg / kg of Jeju Ecklonia cava extract starting 2 weeks before inducing colitis / DK: Group orally administered 2 mg / kg of Ecklonia cava diecol starting 1 week before inducing colitis) Figure 3B shows the colon length in a mouse animal model induced by Dextran sulfate Sodium (DSS). Figure 3C is a graph showing the results of measuring colon length in a mouse animal model induced by Dextran sulfate sodium (DSS). Figure 4 shows the results of measuring the concentration of Zonulin present in the blood using ELISA to determine the improvement effect of increased intestinal permeability, which is used as an indicator of intestinal immunity. Figure 5 shows the results of confirming the expression levels of inflammatory cytokines through ELISA to verify the anti-inflammatory efficacy of Ecklonia cava extract. Figure 6 shows the results of confirming the blood concentration of C-Reactive Protein (CRP), an inflammatory response factor induced by cytokines, through ELISA. Figure 7 shows the results of Hematoxylin-eosin (H&E) staining to determine the extent of lesions such as tissue necrosis, thickening, and inflammation of the mucous membrane. Figure 8A shows the results of staining with CD3 (T cell) antibodies to confirm the degree of infiltration of intestinal immune cells. Figure 8B is a graph showing the CD3 (T cell) IHC score to check the degree of infiltration of intestinal immune cells. Figure 8C shows the results of staining with CD4 (helper T cell) antibodies to confirm the degree of infiltration of intestinal immune cells. Figure 8D is a graph showing the CD4 (helper T cell) IHC score to check the degree of infiltration of intestinal immune cells. Figure 8E shows the results of staining with CD8 (cytotoxic T cell) antibody to confirm the degree of infiltration of intestinal immune cells. Figure 8F is a graph showing the CD8 (cytotoxic T cell) IHC score to check the degree of infiltration of intestinal immune cells. Figure 9A shows the staining results for Myeloperoxidase (MPO), a neutrophil enzyme activated by inflammation. Figure 9B is a graph showing the Myeloperoxidase (MPO) IHC score. Figure 10A is a figure showing changes in the expression of E-cadherin, a membrane-penetrating barrier protein connected to the intracellular cytoskeleton, to determine mucosal barrier integrity. Figure 10B is a figure showing changes in the expression of Occludin, a membrane-penetrating barrier protein connected to the intracellular cytoskeleton, to determine mucosal barrier integrity. Figure 11 shows the results of measuring the expression of Mucin2, a barrier mucus substance secreted from the intestinal mucosa that defends against substances entering from the outside, through Real-time PCR. Figure 12A shows the results of confirming the expression level of the inflammatory cytokine TNF-α by Real-time PCR to confirm the anti-inflammatory efficacy of Ecklonia cava extract. Figure 12B shows the results of confirming the expression level of the inflammatory cytokine IL-6 by Real-time PCR to confirm the anti-inflammatory efficacy of the Ecklonia cava extract. Figure 12C shows the results of confirming the expression level of the inflammatory cytokine IL-1b using Real-time PCR to confirm the anti-inflammatory efficacy of the Ecklonia cava extract. Figure 13 shows the results of confirming the expression of Inducible nitric oxide synthase (iNOS), which is increased by cytokines in inflammatory responses and regulates macrophage function, through Real-time PCR. Figure 14A is a figure showing the results of analyzing E. coli in feces to confirm changes in the intestinal microbial community. Figure 14B shows the results of analyzing bacteriodetes in feces to confirm changes in the intestinal microbial community. Figure 14C shows the results of analyzing Firmicutes in feces to confirm changes in the intestinal microbial community. Figure 14D is a figure showing the results of analyzing Lactobacillus in feces to confirm changes in the intestinal microbial community. Specific details for implementing the invention

[0023] The present invention will be described in detail below.

[0024] The present invention provides a pharmaceutical composition for the prevention or treatment of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient.

[0025] The *Ecklonia cava* of the present invention is an edible brown algae widely distributed along the southern coast of South Korea and in Japan. Although it is produced in abundance on Jeju Island, South Korea (30,000 tons / year), it is not utilized in Europe. This valuable brown algae is used in South Korea and Japan as a food ingredient, animal feed, fertilizer, and folk remedy for gynecological diseases. *Ecklonia cava* is produced in abundance on Jeju Island, Korea, and is widely used in the fields of food additives, livestock feed, fertilizer, and pharmaceuticals. It is known that *Ecklonia cava* contains components such as carotenoids, fucoidan, and phlorotannin, and many recent studies have reported that *Ecklonia cava* extracts possess antioxidant, anti-inflammatory, anti-allergic, tyrosinase inhibitory, and collagenase expression inhibitory activities.

[0026] The sea mustard extract of the present invention can be extracted with 40% to 70% ethanol, 40% to 60% ethanol, and preferably with 50% ethanol.

[0027] The above method for preparing the Ecklonia cava extract may utilize conventional extraction methods in the art. The extract can be obtained by immersing dried Ecklonia cava in water, alcohol, or a mixture thereof to obtain an extract, filtering the extract, and then freeze-drying or spray-drying it. In this case, methanol or ethanol may be used as the alcohol.

[0028] The term "leaky gut syndrome" in this invention refers to a phenomenon in which intestinal mucosal cells maintain a certain intercellular gap, but during the process of digestion and absorption, stimulation or damage occurs, causing increased permeability of the intestinal mucosa through which macromolecular substances can pass back and forth. It collectively refers to symptoms caused by a phenomenon (leaky gut condition) in which macromolecular substances in the blood leak into the lumen of the intestinal wall or macromolecular substances within the lumen enter the bloodstream directly due to the inability of the intestinal barrier to function properly. These symptoms manifest in various clinical conditions such as aging, allergies, multiple trauma, rheumatoid arthritis, inflammatory bowel disease, chronic fatigue syndrome, and irritable bowel syndrome. Furthermore, increased intestinal mucosal permeability or damage to the intestinal mucosa allows pathogens, antigens, and putrefactive substances to enter the intestinal mucosa, triggering an inflammatory response. Additionally, endotoxins enter the bloodstream, causing bacterial translocation and intestinal endotoxemia, which lead to various inflammatory and immune responses.

[0029] The leaky gut syndrome of the present invention can be caused by an inflammatory response in the intestine or an imbalance of the intestinal microbiota, and if the leaky gut syndrome worsens, it can become a cause of inflammatory bowel disease.

[0030] The term "inflammatory bowel disease" in this invention refers to chronic inflammation of unknown etiology occurring in the intestines. While it typically refers to idiopathic inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, Behcet's enteritis, which is relatively common in Korea, can also be considered to fall under this category. In a broader sense, infectious enteritis such as bacterial, viral, amoebic, and tuberculous enteritis, as well as ischemic bowel disease and radiation enteritis, are all inflammatory diseases occurring in the intestines and should be considered as differential diagnoses for inflammatory bowel disease. The symptoms of inflammatory bowel disease can be broadly classified into symptoms caused by inflammation of the digestive tract, general systemic symptoms, extraintestinal symptoms, and symptoms resulting from complications. Although inflammatory bowel disease is a chronic condition, symptoms are not always present; most patients experience a course of recurring periods of temporary worsening followed by periods of asymptomatic remission. In most cases, symptoms begin gradually, requiring a considerable amount of time to reach a diagnosis, whereas when symptoms appear acutely, it is difficult to distinguish from infectious enteritis.

[0031] The inflammatory bowel disease of the present invention may be selected from the group consisting of ulcerative colitis, Crohn's disease, collagenous colitis, lymphocytic colitis, ischemic colitis, transformative colitis, and Behcet's syndrome.

[0032] The term "Zonulin" in this invention refers to a protein that causes increased permeability of the intestinal barrier by destroying the intestinal barrier function through the breakdown of tight junction proteins that hold intestinal mucosal cells. Zonulin is produced by gluten and breaks down tight junction proteins in the intestine, creating a leaky gut that causes chronic inflammation and chronic diseases.

[0033] The tight junction mentioned above is a type of cell junction that restricts the movement of substances between connected cells and is wrapped around the cell in a band-like form. Tight junctions are formed when the cell membranes of adjacent cells are partially fused by proteins called claudin and occludin. They are primarily found in humans and vertebrates. Cell junction mechanisms play a crucial role in embryonic development, the determination of tissue and organ morphology, and the maintenance of normal tissue structure. They also form a blood-brain barrier, preventing harmful substances in the blood from reaching the cerebrospinal fluid. Other types of cell junctions include gap junctions and desmosomes.

[0034] The term "cadherin" in this invention refers to a type-1 transmembrane protein (TMP). It plays an important role in cell junctions, enabling cells to form tight junctions within tissues. Calcium ions (Ca 2+ It is dependent on ). E-cadherin (epithelial cadherin), one of the cadherins, is a 120 kD molecule and a membrane glycoprotein with extracellular and cytoplasmic regions; it is a protein that mediates cell junctions in epithelial cells. E-cadherin is an important component in maintaining the structure of the epithelium by attaching homologous cells; it not only acts as an adhesive between cells but also plays a crucial role in the formation and maintenance of tissue structure by mediating intercellular interactions. Therefore, the loss of E-cadherin expression induces tissue mutations in epithelial cells, leading to defects in intercellular binding function.

[0035] In response to mucosal inflammation, functional and morphological changes, such as proliferation of crypt epithelial cells and atrophy of the villi, occur in intestinal epithelial cells. Inflammatory bowel diseases involve a variety of recurring cycles of chronic intestinal inflammation, damage to epithelial tissue, mucosal ulceration, and epithelial tissue recovery, leading to changes in defense mechanisms. During these processes, changes may occur in proteins associated with E-cadherin, which is responsible for cell-to-cell binding. Studies on E-cadherin expression in inflammatory bowel diseases have reported the loss of E-cadherin expression at the ulcer margin in ulcerative colitis and Crohn's disease.

[0036] The term "Occludin" in this invention refers to the earliest discovered representative protein component of tight junctions, an intrinsic membrane protein that penetrates the cell membrane four times, and is known to have a high correlation with the biological functionality of tight junctions. Ocludin plays an important role in the adhesion between epidermal cells and the function of tight junctions as a barrier, and the level of ocludin expression is correlated with the number or quantity of tight junctions and the level of biological barrier functionality of the tight junction structure.

[0037] The term "Mucin" in this invention refers to a type of glycoprotein secreted by all digestive organs of the human body. Among these, mucin secreted from the stomach wall aids in protein absorption and protects the stomach wall, while mucin secreted from the large intestine helps foreign substances in the intestines to be smoothly shed. The intestinal mucosa, respiratory mucosa, (capillary)vascular endothelial cells, blood-brain barrier, and renal glomerular endothelial barrier are covered by a single layer of epithelial cells / endothelial cells due to their clearance function. Therefore, they are vulnerable to attacks by external harmful substances compared to other organs. A mucin-mucus barrier exists over the mucosal cell layer, creating a biochemical barrier that protects the vulnerable mucosal cells, thereby preventing bacteria, viruses, and toxins from penetrating the epithelial cell layer.

[0038] The sea mustard extract of the present invention exhibits the effect of improving barrier function by reducing the expression of zonulin, which increases barrier permeability, increasing the expression of E-cadherin and ocludin, which are tight junction proteins of intestinal epithelial cells, and increasing the expression of mucin, which forms the immune barrier of the intestinal mucosa.

[0039] The term "inflammation" in this invention refers to a protective response involving immune cells, blood vessels, and inflammatory mediators, which is one of the biological responses to harmful stimuli. Inflammation is a non-specific response mediated by innate immunity; when caused by pathogens, it occurs regardless of the type of pathogen or prior infection. In addition to innate immunity, some local adaptive immunity is also involved in inflammation. While inflammation possesses protective functions such as healing infections or promoting tissue regeneration, tissue damage or disease can simultaneously occur as a result of inflammation.

[0040] The Ecklonia cava extract of the present invention exhibits the effect of suppressing intestinal inflammatory responses by reducing the expression of inflammation-related factors and reducing the infiltration of immune cells within intestinal tissues.

[0041] Intestinal microorganisms perform beneficial functions for the human body by synthesizing short-chain fatty acids and essential vitamins. When an imbalance in the composition of intestinal microorganisms occurs, pathogenic harmful bacteria become involved. This can lead to the activation of the intestinal mucosal immune system, increased intestinal epithelial permeability due to mucosal damage, resulting in exposure to microbial antigens and invasion by various intestinal microorganisms. Consequently, these processes induce and sustain an intestinal mucosal immune response, potentially resulting in inflammatory bowel disease (IBD). Recent studies on the association between IBD and intestinal microorganism composition have reported distinct changes in the composition and metabolic activity of intestinal microorganisms in IBD. Intestinal microorganisms continuously stimulate antigens, thereby inducing pathogenic immune responses in individuals with a genetic predisposition. Genetic defects in mucosal barrier function or intracellular bacterial clearance lead to a high volume of microbial antigen stimulation. Furthermore, genetic defects in the body's immune regulation result in abnormal and excessive T-cell activation against microbial antigens, accompanied by a loss of immune tolerance in the mucosal immune system.

[0042] The sea mustard extract of the present invention has the effect of improving the imbalance of the intestinal microbial composition by increasing beneficial intestinal bacteria such as Firmicutes and Lactobacillus and decreasing harmful bacteria such as Escherichia coli (E. coli) and Bacteroidetes.

[0043] Intestinal bacteria consist of beneficial bacteria such as Bifidobacterium and Lactobacillus, harmful bacteria such as E. coli, Proteobacteria, and Clostridium, and intermediate bacteria that make up the majority of the intestinal microbiome. Beneficial bacteria facilitate peristalsis, which moves food, and suppress harmful bacteria, thereby reducing intestinal toxins and supporting the immune system, which enables the normal functioning of the large intestine. In contrast, harmful bacteria generate waste products and toxins harmful to the body, causing diarrhea or enteritis and leading to a weakening of large intestine function. Therefore, to maintain a healthy gut, beneficial and harmful bacteria must coexist in appropriate proportions.

[0044] The composition of the present invention may contain 0.1 to 99.9 weight% of the active ingredient of the present invention as an active ingredient based on the total weight of the composition for administration. In addition, the composition of the present invention may additionally include one or more active ingredients exhibiting the same or similar functions in addition to the above active ingredient.

[0045] The pharmaceutical composition of the present invention may further include pharmaceutically acceptable additives.

[0046] The above additive may further include suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions.

[0047] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is one that is commonly used in formulations and includes, but is not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0048] The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, or sterile injectable solutions, each according to conventional methods. Specifically, when formulating, it may be prepared using diluents or excipients such as fillers, weighting agents, binders, wetting agents, disintegrants, and surfactants that are commonly used.

[0049] Solid dosage forms for oral administration include, but are not limited to, tablets, pills, powders, granules, and capsules. These solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc., in addition to the active ingredient. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. In addition to liquids and liquid paraffin for oral administration, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be added.

[0050] Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and the subject. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrosol, Tween 61, cocoa paste, laurin paste, glycerogelatin, etc. may be used.

[0051] The pharmaceutical composition of the present invention may additionally include a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, etc., in addition to the above components.

[0052] Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed, 1995).

[0053] The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered via intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc.

[0054] A suitable dosage of the pharmaceutical composition of the present invention depends on factors such as the formulation method, method of administration, age, body weight, sex, pathological condition, food, time of administration, route of administration, excretion rate and response responsiveness, and the degree of bone loss in the patient, and can be appropriately selected by a person skilled in the art, specifically 0.001 mg / kg to 50 mg / kg, and can be administered once or several times a day as needed.

[0055] The pharmaceutical composition of the present invention may be prepared in a unit volume form or contained in a multi-volume container by formulation using pharmaceutically acceptable carriers and / or excipients according to a method that can be easily practiced by a person skilled in the art to which the invention belongs. In this case, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granule, tablet, or capsule, and may additionally include a dispersant or a stabilizer.

[0056] The "subjects of administration" of the present invention include, but are not particularly limited to, humans, monkeys, cattle, horses, pigs, sheep, chickens, cats, dogs, mice, rabbits, etc.

[0058] The present invention provides a health functional food composition for the prevention or improvement of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient.

[0059] The term "health functional food" in this invention refers to a food manufactured using raw materials or ingredients (functional raw materials) that have functions useful to the human body or nutrients that are easily deficient in daily meals, and which maintains and improves health by maintaining normal bodily functions or activating physiological functions, but is not limited thereto.

[0060] The health functional food of the present invention can be manufactured by a method commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry during the manufacturing process.

[0061] The health functional food of the present invention can be manufactured by adding the active ingredient of the present invention within an appropriate effective concentration range according to various manufacturing methods known in the art.

[0062] In addition, the formulation of the above-mentioned health functional food may also be manufactured without restriction as long as it is a formulation recognized as a health functional food.

[0063] The health functional food of the present invention can be manufactured in the form of powder, granules, tablets, capsules, or liquid.

[0064] In addition, the health functional food composition according to the present invention may also contain, in addition to the active ingredient, other ingredients that can preferably provide a synergistic effect to the main effect intended by the present invention, within a range that does not impair the main effect intended by the present invention.

[0065] Unlike general pharmaceuticals, the health functional food composition of the present invention uses food as a raw material, offering the advantage of not causing side effects that may occur with long-term use, and is highly portable, allowing it to be consumed as an adjuvant to alleviate symptoms of disease.

[0066] The health functional food composition of the present invention can prevent inflammatory colitis and improve inflammatory colitis that has already occurred by having an individual who intends to prevent or improve inflammatory colitis consume it consistently.

[0067] The above-mentioned health functional food is not specifically limited to any individual intended for the prevention or improvement of disease and can be applied to any individual. For example, it can be applied to any individual, such as non-human animals like monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, birds, or fish.

[0069] The present invention provides a feed additive for the prevention or improvement of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient.

[0070] The term "feed additive" in this invention refers to a substance added to feed for the purpose of various effects, such as supplementing nutrients and preventing weight loss, enhancing the digestibility and utilization of fiber in the feed, improving milk quality, preventing reproductive disorders and improving conception rates, and preventing high-temperature stress during the summer.

[0071] When supplying the above feed additive, it may be supplied to poultry and livestock, etc., alone or mixed with feed.

[0072] In addition, the feed additive of the present invention can be manufactured by adding the active ingredient of the present invention within an appropriate effective concentration range according to various feed manufacturing methods known in the art.

[0073] The above additive is not specifically limited to any individual intended for the prevention or improvement of disease and can be applied to any individual. For example, it can be applied to any individual, such as non-human animals like monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, birds, or fish.

[0075] In a specific example, the inventors pretreated macrophages with an ethanol extract of Ecklonia cava at various concentrations and confirmed the expression of inflammation-related proteins iNOS and COX2 after LPS stimulation. As a result, protein analysis confirmed that the expression of iNOS and COX2, which had been increased by LPS, was reduced in a concentration-dependent manner by the ethanol extract of Ecklonia cava, and that the phosphorylation of NF-κB, ERK, and p38 induced by LPS was inhibited by the ethanol extract of Ecklonia cava (see Fig. 1). In addition, Real-time PCR analysis confirmed that the expression of TNF-α, IL-6, and IL-1β, representative inflammatory cytokines induced by LPS in co-cultured Caco2 cells, was significantly reduced by the ethanol extract of Ecklonia cava (see Fig. 2).

[0076] In addition, when measuring the colon length in a mouse animal model induced by Dextran sulfate Sodium (DSS), it was confirmed that the colon length, which had been shortened by DSS, gradually lengthened with increasing doses of Ecklonia cava extract (see Fig. 3). Furthermore, to investigate the effect of improving increased intestinal permeability, which is used as an indicator of intestinal immunity, the concentration of Zonulin present in the blood was measured using ELISA, and it was confirmed that the concentration, which had been increased by DSS, was improved by the administration of Ecklonia cava extract (see Fig. 4). Moreover, to confirm the anti-inflammatory efficacy of Ecklonia cava extract, the expression levels of inflammatory cytokines were measured, and a decrease in blood concentration was verified via ELISA (see Fig. 5). Additionally, when checking the blood concentration of C-Reactive Protein (CRP), an inflammatory response factor induced by cytokines, using ELISA, it was confirmed that the increase caused by DSS was reduced by Ecklonia cava extract (see Fig. 6).

[0077] Hematoxylin-eosin (H&E) staining was performed to assess the extent of lesions such as mucosal tissue necrosis, thickening, and inflammation. The results showed that the mucosa, which had been necrotic due to DSS, recovered upon administration of Ecklonia cava extract (see Fig. 7). To determine the degree of infiltration of intestinal immune cells, staining was performed using CD3 (T cell), CD4 (helper T cell), and CD8 (cytotoxic T cell) antibodies. The results confirmed that the infiltration, which had been increased by DSS, decreased upon administration of Ecklonia cava extract (see Fig. 8). Staining for Myeloperoxidase (MPO), a neutrophil enzyme activated by inflammation, confirmed that the inflammation, which had been increased by DSS, was significantly reduced by Ecklonia cava extract (see Fig. 9).

[0078] In addition, to assess the integrity of the mucosal barrier, changes in the expression of transmembrane barrier proteins (E-cadherin, Occludin) connected to the intracellular cytoskeleton were confirmed via Real-time PCR, and it was confirmed that the decrease caused by DSS was increased by the administration of Ecklonia cava extract (see Fig. 10). Furthermore, the expression of Mucin2, a barrier mucus substance secreted from the intestinal mucosa that defends against substances entering from the outside, was measured via Real-time PCR, and it was confirmed that the decrease caused by DSS was restored by the administration of Ecklonia cava extract (see Fig. 11).

[0079] In addition, to confirm the anti-inflammatory efficacy of the Ecklonia cava extract, the expression levels of inflammatory cytokines were measured, and a decrease in gene expression in intestinal tissue was confirmed by Real-time PCR (see Fig. 12). Furthermore, the expression of Inducible nitric oxide synthase (iNOS), which is increased by cytokines in inflammatory responses and regulates macrophage function, was confirmed by Real-time PCR, and it was confirmed that the expression, which had been increased by DSS, was reduced by the Ecklonia cava extract (see Fig. 13).

[0080] In addition, to confirm changes in the gut microbial community, DNA was extracted from feces and analyzed using microbial community-specific primers, and the results showed that E. coli, which had increased due to DSS ( E. coli ) and the Bacteroidetes group were reduced, and the reduced Firmicutes and Lactobacillus groups were verified to be increased by Ecklonia cava extract (see Fig. 14).

[0082] The present invention will be explained in detail below through examples.

[0083] However, the following examples are illustrative of the present invention, and the scope of the present invention is not limited thereto; various modifications and improvements by those skilled in the art using the basic concept of the present invention as defined in the claims are also included within the scope of the present invention.

[0085] <Example 1> Preparation of Jeju Ecklonia cava ethanol extract

[0086] The Ecklonia cava used in the present invention was collected from the waters off Jeju Island and naturally dried in the shade. Then, 10 times the amount of 50% ethanol was added, and the mixture was extracted for 6 to 24 hours. After vacuum filtration through a decanter, the mixture was vacuum concentrated at 60°C or below and sterilized at 87°C or above for 60 minutes. 20% dextrin was mixed and dissolved to homogenize the mixture, which was then sterilized again at 65–85°C for 40 minutes, spray-dried at 180°C, and cooled for use in experiments.

[0088] <Experimental Example 1> Evaluation of Anti-inflammatory Efficacy of Ecklonia cava Using Cell Lines

[0089] To construct an ex vivo model mimicking the intestinal environment, a system was established to co-culture Caco-2 intestinal epithelial cell line and RAW264.7 macrophage cell line. Inflammatory signals were activated by treating the medium containing the macrophage cell line with LPS (1 μg / ml), and the anti-inflammatory efficacy was confirmed by treating with the Ecklonia cava extract of Example 1, measuring the expression levels of IL-1β, IL-6, and TNF-α by RT-PCR, and confirming the activity of iNOS, COX2, NK-κB, ERK, and p38 by Western blot.

[0091] <1-1> Cell Culture

[0092] Intestinal epithelial cell line Caco-2 cells and macrophage cell line RAW264.7 cells were cultured in DMEM media (10% FBS + 1% penicillin / streptomycin) and maintained at 37°C and 5% CO2 conditions.

[0094] <1-2> Co-culture

[0095] To construct an ex vivo model mimicking the intestinal environment, a system was used in which Caco-2 cells and RAW264.7 cells were co-cultured. Caco-2 cells were cultured in the insert region, while RAW264.7 cells were cultured below it. Inflammatory signals were activated by treating Caco-2 cells with Ecklonia cava extract and RAW264.7 cells with LPS (1 μg / ml). RNA and protein were extracted from the Caco-2 cells in the insert region.

[0097] <1-3> Confirmation of inflammatory factor expression

[0098] When macrophages were pretreated with Ecklonia cava ethanol extract at various concentrations and the expression of inflammation-related proteins iNOS and COX2 was checked after LPS stimulation, protein analysis confirmed that the expression of iNOS and COX2, which had been increased by LPS, was reduced in a concentration-dependent manner by the Ecklonia cava ethanol extract, and that the phosphorylation of NF-κB, ERK, and p38 induced by LPS was inhibited by the Ecklonia cava ethanol extract (Fig. 1). Additionally, Real-time PCR analysis confirmed that the expression of TNF-α, IL-6, and IL-1β, representative inflammatory cytokines induced by LPS in co-cultured Caco2 cells, was significantly reduced by the Ecklonia cava ethanol extract (Fig. 2).

[0100] <Experimental Example 2> Evaluation of the Efficacy of Ecklonia Cinnamomum Extract Using an Inflammatory Bowel Disease Animal Model

[0101] An inflammatory bowel disease model was established by feeding 2.5% DSS (dextran sodium sulfate, MW 36,000~5,000 kD, MP Biomedicals) to 5-week-old BALB / c male mice. To verify preventive efficacy, the Ecklonia cava extract of Example 1 was orally administered daily starting 2 weeks prior to the establishment of the model, or to evaluate therapeutic efficacy, the extract was orally administered from the time the inflammation model was established. The efficacy of the Jeju Ecklonia cava ethanol extract in restoring intestinal health was evaluated by analyzing selected biomarkers using feces, blood, and tissue collected on the 28th.

[0103] <2-1> Establishment of a Mouse Model of Inflammatory Bowel Disease Caused by DSS

[0104] 5-week-old male BALB / c mice were purchased and acclimatized for one week before the experiment was conducted. To verify the preventive efficacy, Ecklonia cava extract (50, 150, 300 mg / kg) and Diecol (2 mg / kg) were orally administered starting two weeks before DSS feeding (administered until sacrifice). Subsequently, inflammatory bowel disease was induced by consuming drinking water containing 2.5% DSS (dextran sodium sulfate) for five days, after which the water was switched to normal drinking water, and the mice were sacrificed 28 days later.

[0105] Control : Normal group

[0106] DSS : Group that caused inflammatory bowel disease

[0107] ECE50 : Group orally administered 50 mg / kg of Jeju Ecklonia cava extract starting 2 weeks prior to inducing colitis

[0108] ECE150 : Group orally administered 150 mg / kg of Jeju Ecklonia cava extract starting 2 weeks prior to inducing colitis

[0109] ECE300 : Group orally administered 300 mg / kg of Jeju Ecklonia cava extract starting 2 weeks prior to inducing colitis

[0110] DK : Group orally administered 2 mg / kg of Ecklonia cava diecol starting one week before inducing colitis

[0112] <2-2> Measuring Length

[0113] As a result of measuring the colon length in a mouse animal model induced by Dextran sulfate Sodium (DSS), it was confirmed that the colon length shortened by DSS gradually increased with the dosage of Ecklonia cava extract administered (Fig. 3).

[0115] <2-3> ELISA Measurement

[0116] After obtaining blood from mice, the serum was separated using a centrifuge (4°C, 3000g, 10 min) and stored at -80°C until use. After adding 50 µl of serum to each well, the mixture was reacted at room temperature for 2 hours, washed, and subjected to a conjugate reaction, washing, substrate reaction, washing, and stop solution reaction. The absorbance was then measured at 450 nm using a microplate reader.

[0117] To investigate the effect of improving increased intestinal permeability, which is used as an indicator of intestinal immunity, the concentration of Zonulin present in the blood was measured by ELISA, and it was confirmed that the concentration increased by DSS was improved by the administration of Ecklonia cava extract (Fig. 4).

[0118] To confirm the anti-inflammatory efficacy of the Ecklonia cava extract, the expression levels of inflammatory cytokines in the blood of animals were checked using ELISA, and it was confirmed that the expression of IL-6 and IL-1b was reduced by the Ecklonia cava extract (Fig. 5). Additionally, the blood concentration of C-Reactive Protein (CRP), an inflammatory response factor induced by cytokines, was checked using ELISA, and it was confirmed that the increase caused by DSS was reduced by the Ecklonia cava extract (Fig. 6).

[0120] <2-4> Histological Examination (H&E)

[0121] To confirm the histopathological nature of the colon, the colon was fixed in 10% neutral formalin, and a paraffin block was prepared. Hematoxylin-eosin (H&E) staining was performed to assess the degree of inflammation and necrosis. For deparaffinization, the sample was immersed in xylene for 10 minutes (twice), and then immersed in ethanol (100% to 50%) for 5 minutes each to remove the xylene, followed by hematoxylin staining. Differential decolorization was performed using HCl alcohol, followed by blueing with 0.3% ammonia water. Cytoplasm and other areas not stained by hematoxylin were stained using an eosin solution, and after dehydration with ethanol (50%->100%), the sample was placed in xylene to remove the alcohol and the staining was completed.

[0122] Hematoxylin-eosin (H&E) staining was performed to determine the extent of lesions such as tissue necrosis, thickening, and inflammation of the mucosa, and it was observed that the mucosa necrotic due to DSS recovered with the administration of Ecklonia cava extract (Fig. 7).

[0124] <2-5> Immunohistochemistry (IHC)

[0125] After fixing the colon in 10% neutral formalin, a paraffin block was prepared to make slides. For deparaffinization, the slides were immersed in xylene for 10 minutes (twice), and to remove xylene, they were reacted in ethanol (100%->95%->90%->80%->70%) for 5 minutes each, followed by washing with PBS (+1% tween 20). The slides were placed in antigen retrieval buffer (Tris-EDTA, pH 9.0) and heated in a microwave for 5 minutes, after which they were cooled with cold distilled water for 10 minutes. After reacting in a 0.3% H2O2 solution for 30 minutes, they were washed with PBS (+1% tween 20) and blocked with 1% BSA for 1 hour. The primary antibody (1% BSA) was reacted overnight, washed with PBS (+1% tween20), and the secondary antibody was reacted for 1 hour. After washing with PBS (+1% tween20), the pre-prepared vector ABC reagent was reacted for 30 minutes, followed by washing with PBS (+1% tween20). The DAB solution was reacted until color development occurred, washed with PBS (+1% tween20), reacted with hematoxylin solution for 1 minute, washed with distilled water, and then immersed in ethanol (70%->80%->90%->95%->100%) for 5 minutes each. After adding the mounting solution, the surface was covered with a cover slide to complete the staining process.

[0126] To confirm the degree of infiltration of intestinal immune cells, staining was performed using CD3 (T cells), CD4 (helper T cells), and CD8 (cytotoxic T cells) antibodies, and it was confirmed that the infiltration increased by DSS was reduced by the administration of Ecklonia cava extract (Fig. 8).

[0127] In addition, staining results for Myeloperoxidase (MPO), a neutrophil enzyme activated by inflammation, confirmed that the inflammation increased by DSS was significantly reduced by the Ecklonia cava extract (Fig. 9).

[0129] <2-6> Real-time PCR

[0130] To confirm the expression patterns of the genes, RNA was isolated by adding Trizol reagent to colon tissue, and the quantified 1 µg RNA was converted into cDNA using a cDNA synthesis kit (Takara). Real-time PCR was performed using the synthesized cDNA, primers corresponding to each gene, and SYBR (Takara), and gene amplification was analyzed using the 2-△△Ct method. The base sequences of the primers used in the real-time PCR are listed in Table 1 below.

[0131] Gene names primer sequence human TNF-α sense 5'-GAG ATC AAT CGG CCC GAC TA-3' human TNF-α antisense 5'-ACA GGG CAA TGA TCC CAA AG-3' human IL-6 sense 5'-GTA CAT CCT CGA CGG CAT CTC-3' human IL-6 antisense 5'-GTG CCT CTT TGC TGC TTT CAC-3' human IL-1β sense 5'-GCT GCT CTG GGA TTC TCT TCA-3' human IL-1β antisense 5'-TGG CGA GCT CAG GTA CTT CTG-3' mouse TNF-α sense 5'-CAT CTT CTC AAA ATT CGA GTG ACA A-3' mouse TNF-α antisense 5'-TGG GAG TAG ACA AGG TAC AAC CC-3' mouse IL-6 sense 5'-AGT TGC CTT CTT GGG ACT GA-3' mouse IL-6 antisense 5'-TCC ACG ATT TCC CAG AGA AC-3' mouse IL-1β sense 5'-GGAAGGTCCACGGGAAAGAC-3' mouse IL-1β antisense 5'-AGGCAGGCAGTATCACTCATTGT-3' mouse iNOS sense 5'-CAG CAG GAG AGA GAT CCG ATT TA-3' mouse iNOS antisense 5'-GCA TTA GCA TGG AAG CAA AGA-3 mouse E-cadherin sense 5′-GGC TGG CTG AAA GTG ACA CA-3′ mouse E-cadherin antisense 5'-ACG GCA TGA GAA TAG AGG ATG TAC T-3 mouse Occludin sense 5'-TGA AAG TCC ACC TCC TTA CAG A-3 mouse Occludin antisense 5'-CCG GAT AAA AAG AGT ACG CTG G-3'

[0133] To investigate the integrity of the mucosal barrier, changes in the expression of transmembrane barrier proteins (E-cadherin, Occludin) connected to the intracellular cytoskeleton were confirmed via Real-time PCR, and it was confirmed that the decrease caused by DSS was increased by the administration of Ecklonia cava extract (Fig. 10).

[0134] Real-time PCR was used to measure the expression of Mucin2, a mucus substance that acts as a barrier to defend against substances entering from the outside and is secreted from the intestinal mucosa. The results confirmed that the decrease caused by DSS was restored by the administration of Ecklonia cava extract (Fig. 11).

[0135] To confirm the anti-inflammatory efficacy of the Ecklonia cava extract, the expression levels of inflammatory cytokines were measured, and a decrease in gene expression was confirmed in the intestinal tissue (Fig. 12).

[0136] Real-time PCR was used to confirm the expression of Inducible nitric oxide synthase (iNOS), which is increased by cytokines in inflammatory responses and regulates macrophage function. It was found that the expression, which was increased by DSS, was reduced by the Ecklonia cava extract (Fig. 13).

[0138] <2-7> 장내 미생물 조성 확인

[0139] To verify changes in the gut microbial community, DNA was extracted from feces and analyzed using community-specific primers. Feces were placed in lysis buffer and incubated at 70°C to effectively lyse the microorganisms present in the feces and to separate DNA degradation products and PCR inhibitors present in the feces from the DNA. The fecal homogenate was centrifuged, and the supernatant containing microorganisms was transferred to a silica-membrane column for purification. Residual impurities bound to the DNA were completely removed using wash buffer. The DNA attached to the silica membrane was purified and concentrated by adding low-salt buffer. Real-time PCR was performed on the purified DNA (5 ng) using 16S rRNA-based community-specific primers. The proportion of each microbial community was expressed as a relative value compared to the total gut microbial community. The primer sequences are listed in Table 2 below.

[0140] 미생물 명칭 first 염기 서열 Universal (Eubacteria) sense 5'-AAA CTC AAA GAA TTG ACG G-3' Universal (Eubacteria) antisense 5'-CTC ACC ACG AGC TGA C-3' E.coli subgroup sense 5'-CAT GCC GCG TGT ATG AAG AA-3' E.coli sub-group antisense 5'-CGG GTA ACG TCA ATG AGC AAA-3' Bacteroidetes sense 5'-GTT TAA TTC GAT GAT ACG CGA G-3' Antisense bacteroidetes 5'-TTA ASC CGA CAC CTC ACG G-3' Firmicutes sense 5'-GGA GAT GTG GTT TAA TTC GAA GCA-3 Firmicutes antisense 5'-AGC TGA CGA CAA CCA TGC AC-3 Lactobacillus sense 5'-CGA TGA GTG CTA GGT GTT GGA-3' Antisense lactobacillus 5'-CAA GAT GTC AAG ACC TGG TAA G-3'

[0141] Analysis results showed that E. coli increased by DSS ( E. coli ) and the Bacteroidetes group were reduced, and the reduced Firmicutes It was confirmed that the Lactobacillus group was increased by the Ecklonia cava extract (Fig. 14).

[0143] Through the above examples and experimental examples, it was confirmed that the sea tangle extract of the present invention reduces inflammatory responses, improves intestinal barrier function, and increases beneficial bacteria in the intestines, which means that the sea tangle extract can be used for the prevention, treatment, or improvement of inflammatory bowel disease and leaky gut syndrome.

Claims

Claim 1 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, characterized by the overexpression of zonulin protein, containing Ecklonia cava extract as an active ingredient. Claim 2 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the leaky gut syndrome is caused by an inflammatory response in the intestine or an imbalance of the intestinal microbiota. Claim 3 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the composition reduces the expression of a protein that increases barrier permeability. Claim 4 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, characterized in that, in paragraph 3, the protein that increases the barrier permeability is zonulin. Claim 5 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the composition increases the expression of tight junction proteins. Claim 6 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, characterized in that, in claim 5, the tight junction protein is either E-cadherin or ocludin. Claim 7 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the composition increases the expression of mucin in the intestinal mucosa. Claim 8 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the composition reduces an inflammatory response in the large intestine. Claim 9 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, wherein, in claim 1, the composition is characterized by increasing beneficial bacteria in the intestines. Claim 10 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, characterized in that, in claim 9, the beneficial intestinal bacteria are one or more of Firmicutes and Lactobacillus. Claim 11 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, wherein, in claim 1, the composition is characterized by reducing harmful bacteria in the intestines. Claim 12 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome, characterized in that, in claim 11, the harmful intestinal bacteria are one or more of Escherichia coli (E. coli) and Bacteroidetes. Claim 13 A pharmaceutical composition for the prevention or treatment of leaky gut syndrome according to claim 1, characterized in that the composition reduces the infiltration of immune cells within intestinal tissue. Claim 14 A health functional food composition for the prevention or improvement of leaky gut syndrome, containing Ecklonia cava extract as an active ingredient and characterized by the overexpression of zonulin protein. Claim 15 A feed additive for the prevention or improvement of leaky gut syndrome, characterized by the overexpression of zonulin protein, containing Ecklonia cava extract as an active ingredient.

Images