Composition comprising bentonite as active ingredient for preventing, alleviating, or treating intestinal fibrosis

Abstract

The present invention relates to a composition for preventing, alleviating or treating fibrosis, the composition comprising bentonite as an active ingredient. Bentonite is highly effective in reducing a disease activity index, inhibiting shortening of colon length, regulating the expression level of a gene related to intestinal fibrosis, reducing a collagen deposition site in intestinal tissue, reducing the expression level of fibronectin and collagen proteins in intestinal tissue in an intestinal fibrosis animal model induced by dextran sulfate sodium (DSS), and reducing the expression level of ɑ-smooth muscle actin (ɑ-SMA) and collagen proteins in an intestinal fibrosis cell model induced by TGF-β, and is thus expected to be useful as a composition for preventing, alleviating, or treating intestinal fibrosis.

Description

A composition for the prevention, improvement, or treatment of intestinal fibrosis containing bentonite as an active ingredient

[0001] The present invention relates to a composition for the prevention, improvement, or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

[0002]

[0003] This research was conducted with support from the National Science and Technology Research Council Research Operation Expense Support Program of the Ministry of Science and ICT (Project No.: CAP23023-000).

[0004] Intestinal fibrosis is a disease caused by chronic and recurrent enteritis, leading to intestinal stenosis and obstruction. Although the exact cause of intestinal fibrosis has not yet been clearly identified, it is known that when intestinal microorganisms invade due to the destruction of the intestinal mucosa caused by genetic, environmental, or abnormal immune factors, the activation of intestinal immune cells progresses; this excessive activation leads to abnormal abnormalities in the immune cell regulatory system, progressing to chronic inflammation, and if such intestinal inflammation occurs continuously and repeatedly, intestinal fibrosis develops.

[0005] Currently, there are no reports of drugs that can cure intestinal fibrosis, but commonly prescribed medications include 5-aminosalicylic acid, adrenocortical steroids, immunomodulators (thiopurines), and anti-TNFα biological agents, which are prescribed and used in stages depending on the severity of symptoms; however, various side effects have been reported for these drugs. Therefore, there is a need for research on the development of treatments for intestinal fibrosis, which causes a decrease in quality of life and increases the mortality rate of patients with severe intestinal disease.

[0006] Meanwhile, Korean Registered Patent No. 2417640 discloses a "composition for the prevention or treatment of fibrotic diseases comprising a thiourea derivative" and Korean Registered Patent No. 1871949 discloses a "method for manufacturing an intestinal fibrosis cell model," but there is no description of a composition for the prevention, improvement, or treatment of intestinal fibrosis comprising the bentonite of the present invention as an active ingredient.

[0007] The present invention was derived from the above-mentioned needs, and the inventors confirmed that bentonite reduces the disease activity index, inhibits colon length shortening, regulates the expression of genes related to intestinal fibrosis, reduces collagen deposition sites in intestinal tissue, and reduces the expression of fibronectin and collagen proteins in intestinal tissue in an animal model of intestinal fibrosis induced by DSS (Dextran sulfate sodium), and complete the present invention by confirming that it reduces the expression of α-SMA (α-smooth muscle actin) and collagen proteins in a cell model of intestinal fibrosis induced by TGF-β.

[0008] To solve the above problem, the present invention provides a health functional food composition for the prevention or improvement of intestinal fibrosis comprising bentonite as an active ingredient.

[0009] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

[0010] In addition, the present invention provides a veterinary composition for the prevention or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

[0011] In addition, the present invention provides a feed additive for the prevention or improvement of intestinal fibrosis comprising bentonite as an active ingredient.

[0012] The bentonite according to the present invention is expected to be useful as a composition for the prevention, improvement, or treatment of intestinal fibrosis because it has excellent effects in reducing the disease activity index in an animal model of intestinal fibrosis induced by DSS (Dextran sulfate sodium), inhibiting shortening of the colon length, regulating the expression of genes related to intestinal fibrosis, reducing collagen deposition sites in intestinal tissue, reducing the expression of fibronectin and collagen proteins in intestinal tissue, and reducing the expression of α-SMA (α-smooth muscle actin) and collagen proteins in a cell model of intestinal fibrosis induced by TGF-β.

[0013] Figure 1 is a schematic diagram showing the experimental process to confirm the effect of administering bentonite in an animal model of intestinal fibrosis induced by DSS (dextran sulfate sodium). Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT400 is the experimental group administered 400 mg / kg of bentonite to the animal model of intestinal fibrosis.

[0014] Figure 2 shows the results of confirming the disease activity index after administering bentonite to an animal model of intestinal fibrosis. Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis. ### indicates that the disease activity index of the animal model of intestinal fibrosis increased statistically significantly compared to the normal group, with p<0.001, and **, *** indicate that the disease activity index of the bentonite-administered group decreased statistically significantly compared to the animal model of intestinal fibrosis, with ** being p<0.01 and *** being p<0.001.

[0015] Figure 3 shows the results of determining the length of the large intestine (A), weight of the large intestine (B), the ratio of the large intestine (C), the weight of the liver (D), and the weight of the spleen (E) after administering bentonite to an animal model of intestinal fibrosis. Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT400 is the experimental group administered 400 mg / kg of bentonite to the animal model of intestinal fibrosis. ##, ### indicate that the length of the large intestine in the intestinal fibrosis animal model was statistically significantly reduced or the weight of the large intestine, the ratio of the large intestine, the weight of the liver, and the weight of the spleen were statistically significantly increased compared to the normal group, with ## being p<0.01 and ### being p<0.001, and *, **, *** indicate that the length of the large intestine in the bentonite-treated group was increased or the weight of the large intestine, the ratio of the large intestine, the weight of the liver, and the weight of the spleen were statistically significantly decreased compared to the intestinal fibrosis animal model, with * being p<0.05, ** being p<0.01, and *** being p<0.001.

[0016] Figure 4 shows the results of confirming the expression levels of intestinal fibrosis-related genes (Col3a1, Timp1, Mmp9, Mmp7, Klf4) after administering bentonite to an animal model of intestinal fibrosis. Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis. # and ### indicate that the expression levels of the Col3a1, Tim1, Mmp9, and Mmp7 genes or the Klf4 gene were statistically significantly increased or decreased compared to the normal group in the intestinal fibrosis animal model, with # being p<0.05 and ### being p<0.001. ** and *** indicate that the expression levels of the Col3a1, Tim1, Mmp9, and Mmp7 genes or the Klf4 gene were statistically significantly increased or decreased compared to the intestinal fibrosis animal model in the bentonite-treated group, with ** being p<0.01 and *** being p<0.001.

[0017] Figure 5 shows the results of evaluating the degree of intestinal fibrosis by staining collagen fibers deposited in the intestinal tissue using Sirius red staining after administering bentonite to an animal model of intestinal fibrosis. Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis. ### indicates that the area of ​​collagen deposition in the intestinal tissue of the intestinal fibrosis animal model increased statistically significantly compared to the normal group, p<0.001, and * indicates that the area of ​​collagen deposition in the intestinal tissue of the bentonite-treated group decreased statistically significantly compared to the intestinal fibrosis animal model, p<0.05.

[0018] Figure 6 shows the results of analyzing the expression levels of fibronectin and collagen I proteins in the intestinal tissue after administering bentonite to an animal model of intestinal fibrosis. Control is the normal group, DSS model is the animal model of intestinal fibrosis, DSS+BT50 is the experimental group administered 50 mg / kg of bentonite to the animal model of intestinal fibrosis, DSS+BT100 is the experimental group administered 100 mg / kg of bentonite to the animal model of intestinal fibrosis, and DSS+BT200 is the experimental group administered 200 mg / kg of bentonite to the animal model of intestinal fibrosis. ### indicates that the expression of fibronectin or collagen I protein in the intestinal tissue of the intestinal fibrosis animal model was statistically significantly increased compared to the normal group, p<0.001, and *** indicates that the expression of fibronectin or collagen I protein in the intestinal tissue of the bentonite-treated group was statistically significantly decreased compared to the intestinal fibrosis animal model, p<0.001.

[0019] Figure 7 shows the results of analyzing the expression levels of α-SMA (α-smooth muscle actin) and collagen proteins after administering bentonite to an intestinal fibrosis cell model. Con represents the normal group, and TGF-β is an intestinal fibrosis inducing substance. #### indicates that the expression level of α-SMA or collagen protein in the intestinal fibrosis cell model was statistically significantly increased compared to the normal group, with p<0.0001, and *, **** indicate that the expression level of α-SMA or collagen protein in the bentonite-administered group was statistically significantly decreased compared to the intestinal fibrosis cell model, with * being p<0.05 and **** being p<0.0001.

[0020] To achieve the objective of the present invention, the present invention provides a health functional food composition for the prevention or improvement of intestinal fibrosis comprising bentonite as an active ingredient.

[0021] In the present invention, the bentonite is a general term for porous clay minerals composed of montmorillonite, and more precisely, means silicate minerals.

[0022] The above bentonite may include one or more selected from silicon dioxide (SiO2), titanium dioxide (TiO2), aluminum oxide (Al2O3), iron oxide (Fe2O3), manganese oxide (MnO), calcium oxide (CaO), magnesium oxide (MgO), potassium oxide (K2O), sodium oxide (Na2O), and phosphorus pentoxide (P2O5), and preferably may include 50 to 60 parts by weight of silicon dioxide, 0.1 to 1 part by weight of titanium dioxide, 10 to 15 parts by weight of aluminum oxide, 0.1 to 8 parts by weight of iron oxide, 0.1 to 1 part by weight of manganese oxide, 0.1 to 5 parts by weight of calcium oxide, 0.1 to 5 parts by weight of magnesium oxide, 0.1 to 1 part by weight of potassium oxide, 0.1 to 1 part by weight of sodium oxide, and 0.1 to 1 part by weight of phosphorus pentoxide, but is not particularly limited thereto. The above bentonite is characterized by a loss of ignition of 1.5% or less.

[0023] The above bentonite may be manufactured to a size of 0.2 to 20 μm through dry and wet purification, preferably to a size of 1 to 10 μm, but is not limited thereto. The above bentonite is characterized by not containing heavy metals.

[0024] In the health functional food composition of the present invention, the intestinal fibrosis may be caused by continuous and repetitive inflammation of the intestine, but is not limited thereto.

[0025] The health functional food composition of the present invention is preferably prepared in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0026] When the health functional food composition of the present invention is used as a food additive, the active ingredient may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient can be appropriately determined according to the purpose of use (improvement). Generally, when manufacturing food or beverages, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, relative to the raw material. However, in the case of long-term consumption for the purpose of health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount greater than the above range.

[0027] There are no specific restrictions on the types of the above-mentioned foods. Examples of foods to which the above-mentioned active ingredients may be added include meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense.

[0028] When the health functional food composition of the present invention is used as a health beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The aforementioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As sweeteners, natural sweeteners such as taumarin and stevia extract, or synthetic sweeteners such as saccharin or aspartame may be used. The proportion of the above natural carbohydrates is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. The composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination. Although the proportion of these additives is not critical, the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0029] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

[0030] In the pharmaceutical composition of the present invention, the bentonite and intestinal fibrosis are as described above.

[0031] The pharmaceutical composition of the present invention may further include, but is not limited to, pharmaceutically acceptable carriers, excipients, or diluents in addition to the active ingredient.

[0032] The pharmaceutical composition of the present invention may be prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols, but is not limited thereto.

[0033] Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in formulations and include, but are not limited to, saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol, lactose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, antioxidants, buffers, bacteriostatic agents, diluents, surfactants, binders, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, or preservatives. Suitable dosages of the pharmaceutical composition of the present invention may be prescribed in various ways depending on factors such as the formulation method, mode of administration, patient's age, body weight, sex, pathological condition, diet, time of administration, route of administration, excretion rate, and response sensitivity. The route of administration of the pharmaceutical composition for the prevention or treatment of intestinal fibrosis of the present invention may be any generally accepted route as long as it can reach the target tissue. The pharmaceutical composition of the present invention may be administered via routes such as intramuscular administration, ophthalmic administration, intraperitoneal administration, intravenous administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and rectal administration, although it is not particularly limited thereto, depending on the purpose, and specifically may be administered via the route of oral administration.

[0034] In addition, the present invention provides a veterinary composition for the prevention or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

[0035] The veterinary composition of the present invention may further include suitable excipients and diluents according to conventional methods. Examples of excipients and diluents that may be included in the veterinary composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben, and mineral oil. The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, spices, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using methods well known in the art to provide rapid, sustained, or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, etc.

[0036] The effective amount of the veterinary composition according to the present invention can be appropriately selected according to the individual animal. It may be determined based on the severity of the disease or condition, the sensitivity to the active ingredient of the present invention according to the individual's age, weight, health status or gender, the route of administration, the duration of administration, factors including other compositions combined with or used simultaneously with the composition, and other factors well known in the physiological or veterinary field.

[0037] In addition, the present invention provides a feed additive for the prevention or improvement of intestinal fibrosis comprising bentonite as an active ingredient.

[0038] The feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act. In the present invention, the term "feed" may refer to any natural or artificial prescribed food, single meal, etc., or the components of said single meal, intended for or suitable for animals to eat, consume, and digest. The types of said feed are not particularly limited, and feeds commonly used in the relevant technical field may be used. Non-limiting examples of said feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, single-cell proteins, zooplankton, or food waste. These may be used individually or in a mixture of two or more types.

[0039]

[0040] The present invention will be described in more detail below using manufacturing examples and embodiments. These manufacturing examples and embodiments are intended solely to explain the present invention more specifically, and it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

[0041]

[0042] Materials and Methods

[0043] 1. Establishment of an animal model of intestinal fibrosis

[0044] The intestinal fibrosis animal model used in the present invention was induced by continuous and repeated administration of DSS (dextran sulfate sodium). Specifically, an intestinal fibrosis animal model was established by sequentially providing 8-week-old C57BL / 6J mice with 1.75%, 2%, or 2.25% (w / v) DSS diluted with drinking water for 5 days, followed by providing regular drinking water for 10 days, repeating this cycle a total of 3 times (total 45 days) (Fig. 1).

[0045]

[0046] 2. Administration of Bentonite

[0047] During the 45 days of the experiment, bentonite diluted in saline was orally administered at doses of 50 mg / kg, 100 mg / kg, 200 mg / kg, or 400 mg / kg, respectively. The bentonite used in this invention was natural bentonite, which is calcium-based and was prepared to a size of 10 μm or less through dry / wet purification. Water was administered instead of DSS and saline instead of bentonite to serve as the control group (CON) (Fig. 1).

[0048]

[0049] Example 1. Measurement of disease activity index in an animal model of intestinal fibrosis

[0050] Bentonite was administered to an animal model of intestinal fibrosis, and the body weight of each animal was measured every 2 days for 44 days. The disease activity index (DAI) was calculated by observing the condition of the stool and the presence of rectal bleeding. The disease activity index was determined by calculating each score according to the criteria in Table 1 below as [(body weight loss rate + stool condition + rectal bleeding) / 3].

[0051] Disease Activity Index Measurement Criteria Score Weight Loss Rate (%) Stool Condition Rectal Bleeding 0-Normal Stool -11~5 Normal Stool Fragile 25~10 Loose Stool Hemoccult +3 10~20 Loose Stool Hemoccult ++4 >20 Diarrhea Gross Bleeding

[0052] As a result, as disclosed in Figure 2, it was confirmed that the disease activity index of the intestinal fibrosis-induced group increased compared to the normal group, whereas the disease activity index of the bentonite-administered group decreased compared to the intestinal fibrosis-induced group.

[0053]

[0054] Example 2. Analysis of organ changes in an animal model of intestinal fibrosis

[0055] After administering bentonite to an animal model of intestinal fibrosis, each animal was sacrificed on the 45th day, and the length of the large intestine, the weight of the large intestine, the weight of the liver, and the weight of the spleen were measured.

[0056] As a result, as shown in Figure 3, it was confirmed that the length of the large intestine in the intestinal fibrosis-induced group decreased compared to the normal group, while it increased in the bentonite-administered group, and the weight of the large intestine, liver, and spleen in the intestinal fibrosis-induced group increased compared to the normal group, while it decreased in the bentonite-administered group.

[0057]

[0058] Example 3. Analysis of Intestinal Fibrosis-Related Gene Expression Levels in an Animal Model of Intestinal Fibrosis

[0059] After administering bentonite to an animal model of intestinal fibrosis, each animal was sacrificed on the 45th day, RNA was isolated from samples of isolated intestinal tissue, and the relative mRNA expression levels of Col3a1 (collagen type III alpha 1), Tim1 (TIMP metallopeptidase inhibitor 1), Mmp9 (matrix metalloproteinase 9), Mmp7 (matrix metalloproteinase 7), and Klf4 (Krueppel-like factor 4) genes involved in intestinal fibrosis were determined through reverse transcription and Q-PCR.

[0060] As a result, as disclosed in Figure 4, it was confirmed that the expression levels of the Col3a1, Timp1, Mmp9, and Mmp7 genes in the intestinal fibrosis-induced group increased compared to the normal group, while the expression levels decreased in the bentonite-administered group, and it was confirmed that the expression levels of the Klf4 gene in the intestinal fibrosis-induced group decreased compared to the normal group, while the expression levels increased in the bentonite-administered group.

[0061]

[0062] Example 4. Analysis of collagen deposition sites in intestinal tissue in an animal model of intestinal fibrosis

[0063] After administering bentonite to an animal model of intestinal fibrosis, each animal was sacrificed on day 45, and intestinal tissue was isolated and stained with Sirius red. Sirius red staining is a method that specifically stains collagen fibers deposited in intestinal tissue and is used to evaluate the degree of intestinal fibrosis.

[0064] As a result, as shown in Figure 5, it was confirmed that the area of ​​collagen deposition in the intestinal tissue of the intestinal fibrosis-induced group increased compared to the normal group, while it decreased in the bentonite administration group.

[0065]

[0066] Example 5. Analysis of Fibronectin and Collagen Protein Expression in Intestinal Tissue in an Animal Model of Intestinal Fibrosis

[0067] After administering bentonite to an animal model of intestinal fibrosis, each animal was sacrificed on day 45, and intestinal tissue was isolated. The expression levels of fibronectin and collagen proteins were measured by Western blot and analyzed as relative expression levels to α-tubulin. Fibronectin and collagen proteins are major components of the extracellular matrix, and it is known that intestinal fibrosis progresses when they are excessively produced and accumulated within intestinal tissue.

[0068] As a result, as disclosed in Figure 6, it was confirmed that the expression levels of fibronectin and collagen proteins in the intestinal tissue of the intestinal fibrosis-induced group increased compared to the normal group, while they decreased significantly in the bentonite administration group.

[0069]

[0070] Example 6. Analysis of α-SMA (α-smooth muscle actin) and collagen protein expression in an intestinal fibrosis cell model

[0071] After subculturing human intestinal myofibroblasts in a culture medium (High glucose DMEM, FBS 10%, Penicillin-Streptomycin 1%, L-glutamine, HEPES), 2×10⁶ cells were placed in a 384-well plate. 3 Samples were inoculated at 50 µl per well. After 24 hours, the culture medium was changed to serum-free medium (High glucose DMEM, Penicillin-Streptomycin 1%, L-glutamine, HEPES) and cultured for an additional 24 hours. Subsequently, 5 ng / ml of TGF-β, a cytokine that induces intestinal fibrosis, was added along with bentonite at concentrations of 5 µg / ml, 10 µg / ml, 20 µg / ml, or 40 µg / ml, and the samples were cultured for 96 hours. Afterward, the samples were stained using antibodies against α-SMA and collagen proteins, and the staining intensity was analyzed using the Harmony 4.9 program. It is known that increased expression of α-SMA protein in intestinal tissue promotes the production of extracellular matrix, such as collagen, leading to the progression of intestinal fibrosis.

[0072] As a result, as disclosed in Figure 7, it was confirmed that the expression levels of α-SMA and collagen proteins increased in the intestinal fibrosis-induced group (TGF-β alone treatment group) compared to the normal group, while they decreased significantly in the bentonite administration group (TGF-β and bentonite simultaneous treatment group).

Claims

1. A health functional food composition for the prevention or improvement of intestinal fibrosis containing bentonite as an active ingredient.

2. A health functional food composition for the prevention or improvement of intestinal fibrosis according to claim 1, characterized in that the active ingredient is prepared in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages.

3. A pharmaceutical composition for the prevention or treatment of intestinal fibrosis comprising bentonite as an active ingredient.

4. A pharmaceutical composition for the prevention or treatment of intestinal fibrosis according to claim 3, further comprising, in addition to the active ingredient, a pharmaceutically acceptable carrier, excipient, or diluent.

5. A pharmaceutical composition for the prevention or treatment of intestinal fibrosis according to claim 3, characterized in that the composition is prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols.

6. A veterinary composition for the prevention or treatment of intestinal fibrosis containing bentonite as an active ingredient.

7. A feed additive for the prevention or improvement of intestinal fibrosis containing bentonite as an active ingredient.

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