KR20210058730A - Composition for preventing or treating immune diseases comprising lactobacillus sp. strain and scfa - Google Patents

Abstract

The present invention relates to a composition for preventing and treating immune diseases which comprises a Lactobacillus sp. strain and short chain fatty acid (SCFA), and is effective in preventing and treating related immune diseases by alleviating arthritis pain, protecting cartilage, and suppressing inflammatory markers. The present invention provides a pharmaceutical composition for preventing or treating immune diseases, comprising a Lactobacillus sp. strain and SCFA as active components.

Description

Composition for the prevention and treatment of immune diseases including strains of the genus Lactobacillus and SCFA (Short fatty chain acid) {COMPOSITION FOR PREVENTING OR TREATING IMMUNE DISEASES COMPRISING LACTOBACILLUS SP. STRAIN AND SCFA}

The present invention provides a composition for the prevention and treatment of immune diseases including a strain of the genus Lactobacillus and a short fatty chain acid (SCFA).

Immune disease is a disease in which components of the mammalian immune system cause, mediate, or otherwise contribute to pathological conditions in mammals. In particular, inflammatory disorders are one of the most important health problems in the world. Inflammation is generally a localized protective response of body tissues against host invasion by foreign substances or harmful stimuli. The causes of inflammation include infectious causes such as bacteria, viruses and parasites; Physical causes such as burns or irradiation; Chemicals such as toxins, drugs or industrial agents; It may be an immune response, such as an allergic and autoimmune reaction, or a condition associated with oxidative stress.

Inflammation is characterized by pain, redness, swelling, fever, and eventual loss of function in the affected area. These symptoms are the result of a series of complex interactions between cells in the immune system. Cellular responses result in a network of interactions of several groups of inflammatory mediators: proteins (e.g. cytokines, enzymes (e.g. proteases, peroxidase)), major basic proteins, adhesion molecules (e.g. ICAM, VCAM), lipid mediators (e.g. eicosanoids, prostaglandins, leukotrienes, platelet activating factor (PAF)), reactive oxygen species (e.g. hydroperoxide, superoxide anion O2-, nitric oxide (NO), etc.) However, most of these mediators of inflammation are also modulators of normal cellular activity, so a lack of inflammatory response results in uncontrolled damage (ie infection) of the host, and thus partial damage due to chronic inflammation. As an inflammatory disease mediated by excessive production of several of the above-mentioned mediators, autoimmune disease, which is one of the immune diseases, is characterized in that the immune system attacks its own organs and causes a spontaneous response. These responses are due to the recognition of auto-antigens by T lymphocytes, which induces humoral (autoantigen production) and cellular (lymphocyte and macrophage cytotoxic activity increase) immune responses.

Microbiome refers to microorganisms that live in the human body, and this includes intestinal microorganisms. The number of microbiomes is more than twice the number of cells in a pure human body, and the number of genes is more than 100 times. Therefore, it is sometimes referred to as the second genome because it is impossible to discuss human genes except for microorganisms. Microbiome is a field that can be widely used in the development of new drugs and research on treatments for incurable diseases as it can analyze the relationship between the principle of generating beneficial bacteria and harmful bacteria and the relationship between diseases. In addition, microbiome is used in the development of food, cosmetics, and treatments. However, most researches on drug development using these are in the early stages, so it will take time to develop products.

Korean Patent Registration No. 10-1723265

In order to solve the above problems, the present inventors used a composition of Lactobacillus genus strain and short-chain fatty acid as an active ingredient, as a result, alleviating arthritis pain, protecting cartilage, inhibiting inflammatory markers, and related immune diseases. The present invention was completed by confirming that it is effective in the prevention and treatment of.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing or treating immune diseases, and a method for treating immune diseases using the same, comprising a strain of the genus Lactobacillus and a non-short chain fatty acid as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating immune diseases, comprising a strain of the genus Lactobacillus and a short-chain fatty acid as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving immune diseases, comprising a strain of the genus Lactobacillus and a short-chain fatty acid as an active ingredient.

In addition, the present invention provides a method for preventing or treating an immune disease, comprising administering a pharmaceutical composition for preventing or treating an immune disease according to the present invention to an individual in need of treatment.

The composition of the present invention can reduce pain in arthritis and reduce joint damage by using a strain of the genus Lactobacillus together with a short-chain fatty acid. In addition, the composition of the present invention can be usefully used in the prevention and treatment of immune diseases by inhibiting inflammatory markers and regulating immune cells.

1 is a result of confirming the pain evaluation (Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL)) according to the treatment of the mixed composition of the present invention in an arthritis animal model.
Figure 2 is a result of confirming the osteoarthritis pain control effect according to the treatment of the mixture composition of the present invention in an arthritis animal model.
3 is a CT analysis result confirming the ability to inhibit cartilage damage according to the treatment of the mixed composition of the present invention.
Figure 4 shows the CTX-II measurement results in plasma according to the treatment of the mixed composition of the present invention.
5 is a result of confirming pain evaluation (Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL)) according to sodium propionate treatment in an arthritis animal model.
6 is a result of confirming the osteoarthritis pain control effect according to sodium propionate treatment in an arthritis animal model.
7 is a result of confirming the Th1 inhibitory effect according to sodium propionate treatment in splenocytes of an arthritis animal model.
8 is a result of confirming the effect of Th17/Treg regulation according to sodium propionate treatment in splenocytes of an arthritis animal model.
9 is a result of confirming the Th1 inhibitory effect according to sodium propionate treatment in the blood of an arthritis animal model.
10 is a result of confirming the Th17 inhibitory effect according to sodium propionate treatment in the blood of an arthritis animal model.
11 is a result of confirming the pain evaluation (Paw withdrawal threshold (PWT) and paw withdrawal latency (PWL)) according to the treatment of the mixed composition of the present invention in an arthritis animal model.
12 is a result of confirming the Th1 inhibitory effect according to the treatment of the mixed composition of the present invention in splenocytes of an arthritis animal model.
13 is a result of confirming the Th17 inhibitory effect according to the treatment of the mixed composition of the present invention in splenocytes of an arthritis animal model.
14 is a result of confirming the Th1 inhibitory effect according to the treatment of the mixed composition of the present invention in the blood of an arthritis animal model.
15 is a result of confirming the Th17 inhibitory effect according to the treatment of the mixed composition of the present invention in the blood of an arthritis animal model.

Hereinafter, the present invention will be described in detail as an embodiment of the present invention. However, the following embodiments are presented as examples of the present invention, whereby the present invention is not limited, and the present invention is capable of various modifications and applications within the scope of equality interpreted from the description of the claims to be described later. .

The present invention provides a pharmaceutical composition for preventing or treating immune diseases, comprising a strain of the genus Lactobacillus and a short-chain fatty acid as an active ingredient.

The Lactobacillus strains are Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus Acidophilus, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei plantarum), Lactobacillus Helveticus, Lactobacillus fermentum, Lactobacillus paracasei, and Lactobacillus salivarius. , For example, the strain of the genus Lactobacillus may be Lactobacillus rhamnosus or Lactobacillus acidophilus, but is not limited thereto.

The short-chain fatty acid may be one or more selected from the group consisting of butyrate, propionate, and acetate, but is not limited thereto.

The strain of the Lactobacillus genus and the short-chain fatty acid may be mixed in a weight ratio of 5 to 10: 1 to 5, but is not limited thereto.

The composition may inhibit Th1 and Th17 cells and increase the activity of Treg cells, but is not limited thereto.

The strain may be used by culturing, and may be used in the form of a fermentation broth obtained by culturing in a culture medium, a concentrated fermentation broth, a dried product of the fermentation broth, a culture filtrate, a concentrated culture filtrate, or a dried product of the culture filtrate. It may be a culture filtrate containing or removing the strain after culturing. In addition, the formulation of the culture is not limited, and for example, may be a liquid or a solid.

In the present invention, the term "culture" means to grow microorganisms under appropriately artificially controlled environmental conditions.

The strain is capable of growing in a conventional medium, and contains nutrients required by a culture target, that is, a microorganism serving as a culture medium in order to cultivate a specific microorganism, and may be mixed by adding a substance for a special purpose. The medium is also referred to as a culture medium or a culture medium, and is a concept including all of a natural medium, a synthetic medium, or a selective medium.

The medium used for cultivation must meet the requirements of a specific strain in an appropriate manner while controlling temperature, pH, etc. in a conventional medium containing an appropriate carbon source, nitrogen source, amino acid, vitamin, and the like. As a carbon source that can be used, a mixed sugar of glucose and xylose is used as the main carbon source. In addition, sugars and carbohydrates such as sucrose, lactose, fructose, maltose, starch, cellulose, soybean oil, sunflower oil, castor oil, coconut Oils and fats such as oil, fatty acids such as palmitic acid, stearic acid, and linoleic acid, alcohols such as glycerol and ethanol, and organic acids such as acetic acid are included. These materials can be used individually or as a mixture. Examples of nitrogen sources that can be used include inorganic nitrogen sources such as ammonia, ammonium sulfate, ammonium chloride, ammonium acetate, ammonium phosphate, ammonium carbonate, and ammonium nitrate; Amino acids such as glutamic acid, methionine, glutamine, and organic nitrogen sources such as peptone, NZ-amine, meat extract, yeast extract, malt extract, corn steep liquor, casein hydrolyzate, fish or its degradation products, skim soybean cake or its degradation products I can. These nitrogen sources may be used alone or in combination. The medium may contain first potassium phosphate, second potassium phosphate, and a corresponding sodium-containing salt as personnel.

Personnel that may be used include potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding sodium-containing salt. In addition, sodium chloride, calcium chloride, iron chloride, magnesium sulfate, iron sulfate, manganese sulfate and calcium carbonate may be used as the inorganic compound. Finally, in addition to the above substances, essential growth substances such as amino acids and vitamins can be used. In addition, precursors suitable for the culture medium may be used. The above-described raw materials may be added in a batch, fed-batch, or continuous manner to the culture during the cultivation process, but are not particularly limited thereto. Basic compounds such as sodium hydroxide, potassium hydroxide, ammonia, or acid compounds such as phosphoric acid or sulfuric acid can be used in an appropriate manner to adjust the pH of the culture.

The immune disease, the immune disease, the amount of CTX-II expression is increased; Treg cells are reduced; Th17 cells and Th1 cells may be diseases having increased conditions, and may include autoimmune diseases or inflammatory diseases.

In the present invention, the term "autoimmune disease" refers to an unresponsiveness of a living body to an autoantigen as immunologic unresponsiveness or tolerance, and if a problem arises in inducing or maintaining such self-tolerance An immune response occurs against the self-antigen, and as a result, a phenomenon of attacking the own tissue occurs, which means a disease caused by this process.

In the present invention, the term "inflammatory disease" is TNF-α (tumor necrosis factor-α), IL secreted by immune cells such as macrophages by excessively promoting the human immune system due to harmful stimulation such as an inflammation inducing factor or irradiation. It refers to diseases caused by inflammatory substances (inflammatory cytokines) such as -1 (interleukin-1), IL-6, prostagladin, luecotriene or nitric oxide (NO). .

The immune diseases include osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, spondyloarthritis, inflammatory bowel disease, psoriatic arthritis, gout, bacterial arthritis, childhood rheumatoid arthritis, lupus, scleroderma, multiple sclerosis, fibromyalgia, multiple myositis, dermatitis, Behcet's disease, Reiter's Syndrome, Lyme Arthritis, Adhesive Arthritis, Frozen Shoulder, Tendon Synovitis, Elbow Head Positis, DeQuavein Tendon Synovialitis, Recurrent Rheumatoid, Multiple Rheumatoid Pain, Adult Still's Disease, Autoimmune Hemocytopenia, Autoimmune Myocarditis, Atopic Dermatitis , Asthma, primary cirrhosis, Goodfighter syndrome, autoimmune meningitis, Sjogren syndrome, Addison's disease, alopecia areata, autoimmune hepatitis, autoimmune mumps, Crohn's disease, insulin dependent diabetes, dystrophic vesicular epidermolysis, epididymitis, glomeruli Nephritis, Graves' disease, Guillain-Barré syndrome, Hashimoto's disease, hemolytic anemia, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, psoriasis, rheumatic fever, sarcoidosis, skin sclerosis, spondyloarthrosis, thyroiditis, vasculitis, vitiligo, myxedema, pernicious anemia, mitochondria It may be one or more selected from the group consisting of related syndromes and ulcerative colitis, and may be, for example, osteoarthritis, but is not limited thereto.

In the present invention, the term "treatment", unless otherwise stated, reverses, alleviates, inhibits or prevents the disease or disease to which the term applies, or one or more symptoms of the disease or disease. As used herein, the term “treatment” refers to the act of treating when “treating” is defined as above. Thus, the "treatment" or "therapeutic regimen" of an immune disease in a mammal may include one or more of the following:

(1) inhibiting the growth of immune diseases, that is, inhibiting their development,

(2) preventing the spread of immune diseases, i.e., preventing metastasis,

(3) Alleviate immune diseases.

(4) preventing recurrence of immune diseases, and

(5) Palliating the symptoms of immune diseases.

The pharmaceutical composition of the present invention may further include an adjuvant in addition to the active ingredient. Any of the adjuvants known in the art may be used without limitation, but, for example, Freund's complete adjuvant or incomplete adjuvant may be further included to increase its immunity.

The pharmaceutical composition according to the present invention may be prepared in a form in which an active ingredient is incorporated in a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients, and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention are not limited thereto, but lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

The pharmaceutical compositions of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions according to a conventional method. .

In the case of formulation, it may be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in the active ingredient, such as starch, calcium carbonate, sucrose, lactose, gelatin. It can be prepared by mixing and the like. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, and various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are commonly used diluents. I can. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like may be used. As a base for suppositories, witepsol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

The pharmaceutical composition according to the present invention can be administered to a subject by various routes. All modes of administration can be expected, for example oral, intravenous, intramuscular, subcutaneous, intraperitoneal injection.

The dosage of the pharmaceutical composition according to the present invention is selected in consideration of the individual's age, weight, sex, and physical condition. It is obvious that the concentration of the active ingredient contained in the pharmaceutical composition can be selected in various ways depending on the subject, and is preferably included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg/ml. When the concentration is less than 0.01 μg/ml, pharmaceutical activity may not appear, and when the concentration exceeds 5,000 μg/ml, toxicity to the human body may occur.

The pharmaceutical composition may be formulated in various oral or parenteral dosage forms.

Formulations for oral administration include, for example, tablets, pills, hard, soft capsules, solutions, suspensions, emulsifiers, syrups, granules, etc. These formulations include diluents (e.g., lactose, dextrose, water) in addition to the active ingredients. Krose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg, silica, talc, stearic acid and magnesium or calcium salts thereof and/or polyethylene glycol). In addition, the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, in some cases starch, agar, alginic acid Or a disintegrant or boiling mixture and/or absorbent, colorant, flavoring and sweetening agent such as its sodium salt. The formulation can be prepared by conventional mixing, granulating or coating methods.

In addition, a representative formulation for parenteral administration is a formulation for injection, and as a solvent for the formulation for injection, water, Ringer's solution, isotonic physiological saline, or a suspension may be mentioned. The sterile fixed oil of the injectable preparation can be used as a solvent or suspension medium, and any non-irritating fixed oil including mono-, di-glyceride can be used for this purpose.

In addition, the formulation for injection may use a fatty acid such as oleic acid.

In addition, the present invention provides a health functional food composition for preventing or improving immune diseases, comprising a strain of the genus Lactobacillus and a short-chain fatty acid as an active ingredient.

In addition to containing an active ingredient, the food composition of the present invention may contain various flavoring agents or natural carbohydrates as an additional ingredient, like a conventional food composition.

Examples of the above-described natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The above-described flavoring agent can be advantageously used as a natural flavoring agent (taumatin), stevia extract (eg, rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.). The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements, etc. There is this.

In addition, the food composition includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring and natural flavoring agents, coloring agents and thickeners (cheese, chocolate, etc.), pectic acid and salts thereof, in addition to the extract as an active ingredient. Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like may be contained. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juice and fruit juice beverages and vegetable beverages.

The functional food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, and the like. In the present invention, the term'health functional food composition' refers to a food manufactured and processed using raw materials or ingredients having useful functions for the human body pursuant to the Health Functional Food Act No.6727, and with respect to the structure and function of the human body. It means ingestion for the purpose of obtaining useful effects for health use such as controlling nutrients or physiological effects. The health functional food of the present invention may contain ordinary food additives, and whether it is suitable as a food additive is determined according to the general rules and general test methods of food additives approved by the Food and Drug Administration, unless otherwise specified. It is judged according to the standards and standards. Examples of the items listed in the'Food Additives Code' include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as reduced pigment, licorice extract, crystalline cellulose, high color pigment, and guar gum; Mixed preparations, such as a sodium L-glutamate preparation, an alkali additive for noodles, a preservative preparation, and a tar color preparation, etc. are mentioned. For example, in the health functional food in the form of a tablet, a mixture obtained by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant, and other additives is granulated by a conventional method, and then a lubricant is added thereto and compression molding, or the above The mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a mating agent or the like, if necessary. Among the health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture of the active ingredient of the present invention with additives such as excipients in a conventional hard capsule, and the soft capsules contain the active ingredient of the present invention with additives such as excipients. The mixture mixed with can be prepared by filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. Ring-shaped health functional foods can be prepared by molding a mixture of the active ingredient of the present invention, excipients, binders, disintegrants, etc. by conventionally known methods, and can be coated with white sugar or other coating agents as needed, Alternatively, the surface may be coated with a material such as starch or talc. The health functional food in the form of granules can be prepared in granular form by a mixture of the excipients, binders, disintegrants, etc. of the active ingredients of the present invention by a known method, and if necessary, may contain flavoring agents, flavoring agents, etc. I can.

In addition, the present invention provides a method for preventing or treating an immune disease comprising administering the pharmaceutical composition for preventing or treating the immune disease to an individual in need of treatment.

The treatment method of the present invention includes administering the pharmaceutical composition to a subject in a therapeutically effective amount. A specific therapeutically effective amount for a specific individual is the type and extent of the reaction to be achieved, the specific composition, including whether or not other agents are used in some cases, the individual's age, weight, general health status, sex and diet, administration time, It is preferable to apply differently according to various factors including the route of administration and the secretion rate of the composition, the treatment period, drugs used with or concurrently with the specific composition, and similar factors well known in the field of medicine. Therefore, it is preferable to determine an effective amount of a composition suitable for the purposes of the present invention in consideration of the foregoing.

The individual is applicable to any mammal, and the mammal includes humans and primates, as well as livestock such as cattle, pigs, sheep, horses, dogs and cats.

The present invention will be described in more detail through the following examples. However, the following examples are only for embodiing the contents of the present invention, and the present invention is not limited thereto.

[Example]

Example 1. Preparation of the mixed composition of the present invention

A microbiome mixture composition was prepared by mixing Lactobacillus rhamnosus and butyrate in a ratio of 5:1.

Example 2. The therapeutic effect of the composition of the present invention on osteoarthritis

Monosodium Iodoacetate (MIA, I2512, Sigma, Poole, UK) was dissolved in saline for injection at a concentration of 60 mg/ml and prepared on the day of the start of the experiment (day 0). At the start of the experiment, rats (Wistar rat, 6 weeks old, 180g~220g) were subjected to respiratory anesthesia using Isoflurane, and then MIA into the right knee joint using a gauge 0.5 inch needle (model 750; Hamiltion Companym, Reno, NV). Osteoarthritis (MIA) was induced by injecting 50ul (MIA 3mg/body) via infrapatellar ligament.

600 mg/Kg mixed composition treatment group of the present invention (L. rhamnosus 500 mg/kg + Butyrate 100 mg/kg); 30mg/Kg celecoxib treatment group; 500 mg/kg L. rhamnosus alone treatment group; And as an excipient (Vehicle) treatment group, each treatment was administered orally per rat every day from the day after MIA induction, and the progress of 0, 3, 8, 11, 15 and 18 days was confirmed.

For the measurement of pain in rats, a Dynamic planter aesthesiometer (UgoBasile, Comerio, Itaily) was used. To measure the pain of the machine, a mesh plate was placed on the measuring machine, and a rat was placed in an animal holding frame made of acrylic on it, and then the right foot was pierced with the drug with a measuring machine. After stabbing, the time it took for the machine to automatically take off (s, seconds) and how much weight it took (g) was measured, and the time and weight were recorded, and a pain measurement graph was drawn.

As a result, as shown in Figs. 1 and 2, as a result of checking the behavioral experiment (paw withdrawal latency(s), paw withdrawal threshold(g) and weight on right hind rimb(%)), on the 3rd day after induction of osteoarthritis, each Although there was no significant difference in the pain response in the treatment group, it was confirmed that the degree of pain response was similar or remarkably recovered in the mixed composition treatment group of the present invention compared to the celecoxib treatment group after 8 days.

Example 3. The effect of protecting cartilage by the mixed composition of the present invention

In the animal model in which osteoarthritis was induced, it was confirmed through CT whether the joint damage was inhibited by the mixture composition of the present invention.

As a result, as can be seen in Figure 3, compared to the celecoxib treatment group and the L. rhamnosus alone treatment group in the knee joint of the rat treated with the composition of the present invention, the damage to the knee joint is less and the bone surface is wider, so it is excellent It was confirmed that the cartilage protection effect was shown.

Example 4. CTX-II inhibitory effect by the mixed composition of the present invention

Plasma was collected from the osteoarthritis-induced animal model as in Example 2, and CTX-II, an inflammatory marker related to cartilage damage, was measured.

As a result, as can be seen in Figure 4, compared to the celecoxib treatment group and L. rhamnosus alone treatment group, it was confirmed that the CTX-II expression level was significantly lower in the composition treatment group of the present invention.

Example 5. The therapeutic effect of sodium propionate on osteoarthritis

According to the same process as in Example 2, osteoarthritis (MIA) was induced in rats.

Then, 200 mg/Kg of sodium propionate (200 mpk); And as an excipient (Vehicle) treatment group, each treatment was administered orally per rat every day from the day after MIA induction, and then the progress was confirmed by day.

For the measurement of pain in rats, a Dynamic planter aesthesiometer (UgoBasile, Comerio, Itaily) was used. To measure the pain of the machine, a mesh plate was placed on the measuring machine, and a rat was placed in an animal holding frame made of acrylic on it, and then the right foot was pierced with the drug with a measuring machine. After stabbing, the time it took for the machine to automatically take off (s, seconds) and how much weight it took (g) was measured, and the time and weight were recorded, and a pain measurement graph was drawn.

As a result, as shown in FIGS. 5 and 6, paw withdrawal latency (s), paw withdrawal threshold (g), and weight on right hind rimb (%)) were confirmed. Although there was no significant difference in pain response, it was confirmed that the degree of pain response was recovered in the sodium propionate treatment group compared to the excipient treatment group after 7 days.

Example 6. Immunomodulatory effect by sodium propionate treatment

After the end of the experiment in Example 5, splenocytes and blood were obtained from rats and flow cytometric analysis was performed.

As a result, it was confirmed that Th1 and Th17 cells were suppressed and Treg cells increased in splenocytes by treatment with sodium propionate (FIGS. 7 and 8 ). In addition, it was confirmed that Th1 and Th17 cells were suppressed in blood (Figs. 9 and 10).

Example 7. Preparation of the mixed composition of the present invention

A microbiome mixture composition was prepared by mixing Lactobacillus acidophilus and sodium propionate at a ratio of 500mg/kg:200mg/kg (5:2).

Example 8. The therapeutic effect of sodium propionate on osteoarthritis

According to the same process as in Example 2, osteoarthritis (MIA) was induced in rats.

Then, 500 mg/kg of Lactobacillus acidophilus (L. acido); 500mg/kg+200mg/kg of the mixed composition of Example 7 (L. acido+Sodium propionate); And as an excipient (Vehicle) treatment group, each treatment was administered orally per rat every day from the day after MIA induction, and then the progress was confirmed by day.

For the measurement of pain in rats, a Dynamic planter aesthesiometer (UgoBasile, Comerio, Itaily) was used. To measure the pain of the machine, a mesh plate was placed on the measuring machine, and a rat was placed in an animal holding frame made of acrylic on it, and then the right foot was pierced with the drug with a measuring machine. After stabbing, the time it took for the machine to automatically take off (s, seconds) and how much weight it took (g) was measured, and the time and weight were recorded, and a pain measurement graph was drawn.

As a result, as shown in Figure 11, as a result of confirming the paw withdrawal latency (s) and paw withdrawal threshold (g), compared to the Lactobacillus acidophilus alone administration group Lactobacillus acidophilus and sodium propionate of Example 7 It was confirmed that the recovery of the degree of pain response was better in the combined administration group.

Example 9. Immunomodulatory effect by the mixture composition of the present invention

After the end of the experiment in Example 8, splenocytes and blood were obtained from rats and flow cytometric analysis was performed.

As a result, it was confirmed that Th1 and Th17 cells were inhibited in spleen cells (Figs. 12 and 13) and blood (Figs. 14 and 15) by the treatment of the mixed composition of Lactobacillus acidophilus and sodium propionate of Example 7.

So far, the present invention has been looked at around its preferred embodiments. Those of ordinary skill in the art to which the present invention pertains will be able to understand that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative point of view rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (10)

Lactobacillus (Lactobacillus) genus strain and short chain fatty acid (Short fatty chain acid; SCFA) containing as an active ingredient, a pharmaceutical composition for the prevention or treatment of immune diseases. The method of claim 1, wherein the Lactobacillus strain is Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus Acidophilus, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei In the group consisting of Lactobacillus plantarum, Lactobacillus Helveticus, Lactobacillus fermentum, Lactobacillus paracasei and Lactobacillus varicasei and Lactobacillus variobas One or more selected ones, the composition. The composition of claim 1, wherein the Lactobacillus genus strain is Lactobacillus rhamnosus or Lactobacillus Acidophilus. The composition of claim 1, wherein the short-chain fatty acid is at least one selected from the group consisting of butyrate, propionate, and acetate. The method of claim 1, wherein the immune disease is
The amount of CTX-II expression is increased;
Treg cells decrease;
The composition, wherein Th17 cells and Th1 cells have increased conditions. The method of claim 1, wherein the immune disease is osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, spondyloarthritis, inflammatory bowel disease, psoriatic arthritis, gout, bacterial arthritis, childhood rheumatoid arthritis, lupus, scleroderma, multiple sclerosis, fibromyalgia, multiple myositis, Dermatomyositis, Behcet's disease, Reiter's syndrome, Lyme arthritis, adhesive arthritis, frozen shoulder, tendon synovitis, elbow head pouchitis, dequeuvain tendon synovitis, recurrent rheumatism, rheumatoid multiple muscle pain, adult Still's disease, autoimmune hemocytopenia, Autoimmune myocarditis, atopic dermatitis, asthma, primary cirrhosis, Goodfighter syndrome, autoimmune meningitis, Sjogren syndrome, Addison's disease, alopecia areata, autoimmune hepatitis, autoimmune mumps, Crohn's disease, insulin-dependent diabetes, dystrophic blisters Epidermal detachment, epididymitis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Hashimoto's disease, hemolytic anemia, multiple sclerosis, myasthenia gravis, pemphigus vulgaris, psoriasis, rheumatic fever, sarcoidosis, skin sclerosis, spondyloarthritis, thyroiditis, vasculitis, vitiligo, mucus The composition is one or more selected from the group consisting of several species, pernicious anemia, mitochondrial-related syndrome and ulcerative colitis. The composition of claim 1, wherein the immune disease is osteoarthritis. The composition of claim 1, wherein the composition inhibits Th1 and Th17 cells and increases the activity of Treg cells. The composition of claim 1, wherein the Lactobacillus strain and the short-chain fatty acid are contained in a weight ratio of 5 to 10:1 to 5. Lactobacillus (Lactobacillus) genus strain and short chain fatty acid (Short fatty chain acid; SCFA) containing as an active ingredient, a health functional food composition for the prevention or improvement of immune diseases.

Images