KR102445903B1 - Lactobacillus plantarum LRCC5314 having an effect of blood glucose lowering-activity and controlling stress and its application - Google Patents

Abstract

The present invention relates to a killed Lactobacillus plantarum LRCC5314 strain having excellent blood glucose lowering and stress controlling effects, and use thereof. According to the present invention, the Lactobacillus plantarum LRCC5314 strain has alpha-amylase or alpha-glucosidase inhibitory activity, has acid resistance, and has bile resistance or intestinal adhesion, and is a novel lactic acid bacterium isolated from kimchi, which is a traditional fermented food with blood sugar glucose, antidiabetic, anti-inflammatory, or anti-stress effects, thereby being usefully used as probiotic compositions and a useful food and medicine material for preventing, mitigating, or treating diseases caused by diabetes or inflammation.

Description

Heat-treated Lactobacillus plantarum LRCC5314 strain and its use with excellent effect of inhibiting blood sugar rise and controlling stress {Lactobacillus plantarum LRCC5314 having an effect of blood glucose lowering-activity and controlling stress and its application}

The present invention relates to a heat-treated Lactobacillus plantarum strain LRCC5314 having an effect of inhibiting blood sugar rise and regulating stress, and uses thereof.

Recently, with the westernization of dietary habits in Korea, excessive fat intake is increasing, as well as adult diseases such as obesity and hyperlipidemia, as well as type 2 diabetes. In Korea, the incidence of diabetes among those in their 70s has increased significantly in recent years, at a rate of 10%, and the resulting diabetes mortality rate is the highest among OECD countries. It belongs to 6th place. In general, in the case of diabetic patients, since a sudden rise in blood sugar after a meal causes serious complications, suppression of the rise in blood sugar after a meal is essential for a therapeutic agent.

Currently, drugs for treating diabetes include biguanide drugs, but side effects include gastrointestinal symptoms such as abdominal bloating, nausea, and diarrhea.

Stress is caused by a pervasive factor in our daily life and is known to contribute to the occurrence of almost all diseases and medical conditions. Specifically, there are various causes of stress and physiological changes in the human body, but in common, stress hormones are secreted by stimulating the pituitary and adrenal system. It causes symptoms such as defects, which develop into secondary diseases such as gastroduodenal ulcer, high blood pressure, irritable colitis, and angina pectoris. Glucocorticoids, hormones secreted by the adrenal cortex, are derived from their role in regulating glucose metabolism. It is called cortisol in primates and corticosterone in rodents, and is known as a stress hormone secreted when stressed. Specifically, when we receive a stress stimulus, our body activates the sympathoadrenal system and the hypothalamus-pituitary-adrenal axis (HPA axis), and the hypothalamus increases corticotropin-releasing hormone (CRH) secretion, and the pituitary Stimulates the production of ACTH (adrenocorticotropin), which in turn secretes glucocorticoids in the adrenal cortex.

Currently, people rely on antidepressants and anti-anxiety drugs to treat stress, and people are often reluctant to take them because of their fear of these drugs. In addition, when used for a long period of time, there is a risk of tolerance and side effects to the drug, so continuous use is not possible.

Representative factors related to blood sugar and stress control include glucose transpoter 4 (GLUT4), Neuropeptide Y (NPY), and Neuropeptide Y receptors type 2 (Y2R).

First, GLUT4 (glucose transpoter 4) factor is a major transporter used for facilitated diffusion to increase intracellular glucose levels, and its expression is increased by insulin in muscle or fat and is known to be involved in glucose homeostasis by suppressing postprandial blood glucose rise. .

Next, NPY (Neuropeptide Y) factor is a kind of peptide related to appetite regulation and is known to be closely related to reactivity to stress. That is, it was found that when stress increases the secretion of NPY from nerves, the growth and differentiation of adipocytes is promoted to increase abdominal fat.

Neuropeptide Y receptors type 2 (Y2R) factor, like NPY factor, is also activated when under stress and causes metabolic syndrome in which blood sugar, blood pressure, and triglycerides increase by increasing body weight. It is known to reduce fat and improve metabolic syndrome.

Lactobacillus inhabits the human digestive tract, activates the immune system of our body, synthesizes vitamin K and vitamin B group, and is known to regulate the synthesis of harmful ingredients, absorption into the body, and energy metabolism. In particular, live lactic acid bacteria and probiotics have traditionally been known to be excellent for intestinal function through normalization of intestinal flora, but recently, various effects such as immune enhancement, liver cirrhosis improvement, allergy and atopy improvement, and cholesterol lowering are known. In addition, recently, research on postbiotics has been increasing, and postbiotics are metabolites or dead cells from lactic acid bacteria that have biological activity in the host and only act as probiotics without risk of live bacteria. It is a substance that is expected to be used. Butyrate and short-chain fatty acids as metabolites of probiotics and heat-killed Lactobacillus rhamnosus GG as dead cells have been studied to prevent necrotizing enteritis and sepsis.

Accordingly, the present inventors have focused on the urgent need to develop natural medicines or functional food materials that can minimize the side effects caused by drugs, improve insulin resistance, and control stress at the same time. 10-2091757 patent was registered for Lactobacillus plantarum LRCC5314 strain by confirming the functionality of lactic acid bacteria involved in blood sugar control.

However, the above patent only confirms the effect of blood sugar and inflammation-related factors on live cells.

Korean Patent No. 10-2091757

Following the above study, the present inventors tried to confirm the functionality through animal experiments using dead cells through Tindalhwa instead of live cells as a follow-up study. More excellent efficacy was confirmed for the stress control function.

Accordingly, the inventors of the present invention can be used as probiotics from fermented food, can relieve stress, and as a result of earnest efforts to derive a strain that can solve the problem of a product for preventing and / or improving conventional diabetes, traditional fermented food The present invention was completed through the dead cellization of the strain isolated from kimchi.

An object of the present invention is to provide a dead cell through Tyndalization of the Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P.

Another object of the present invention is to provide a probiotic composition comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide a composition for lowering blood sugar comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating diabetes, comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide a food composition for preventing or improving diabetes, comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide an anti-inflammatory pharmaceutical composition comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide an anti-inflammatory food composition comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

Another object of the present invention is to provide a food composition for relieving stress, comprising, as an active ingredient, a dead cell of the Lactobacillus plantarum LRCC5314 strain, a culture thereof, or a lysate thereof.

The object of the present invention is not limited to the object mentioned above. The objects of the present invention will become more apparent from the following description, and will be realized by means and combinations thereof described in the claims.

In order to achieve the above object, the following solutions are provided.

One aspect of the present invention is a Lactobacillus plantarum LRCC 5314 (Lactobacillus plantarum LRCC 5314) strain having a hypoglycemic, antidiabetic, anti-inflammatory or stress-relieving effect, wherein the strain is a heat-treated dead cell.

In one aspect of the present invention, the strain is deposited with accession number KCCM12540P, provides a strain.

In one aspect of the present invention, the strain is isolated from kimchi, and provides a strain comprising the 16S rDNA base sequence of SEQ ID NO: 1.

In one aspect of the present invention, the strain modulates inflammatory factors of TNF-a, IL-6 or CCL-2, or the strain modulates GLUT4, Adiponectin or Leptin's blood glucose-related genes, or the strain is NPY or It provides a strain that expresses Y2R stress improvement related gene, or the strain regulates excessive secretion of cortisol in the blood.

Another aspect of the present invention is the strain according to any one of the aspects of the present invention; lysate thereof; its culture; its ferment; and extracts of the strains, lysates, cultures or fermented products; It provides a microbial preparation comprising one or more of.

In another aspect of the present invention, the strain provides a dead cell, microbial preparation.

Another aspect of the present invention provides a probiotic composition comprising, as an active ingredient, the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a composition for lowering blood sugar comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating diabetes, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

Another aspect of the present invention provides a food composition for preventing or improving diabetes, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

Another aspect of the present invention provides an anti-inflammatory pharmaceutical composition comprising, as an active ingredient, the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof.

Another aspect of the present invention provides an anti-inflammatory food composition comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a food composition for improving cortisol hypersecretion, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

The heat-treated Lactobacillus plantarum LRCC5314 strain according to one aspect of the present invention is isolated from kimchi, a traditional fermented food, and has high safety, It has the effect of lowering blood sugar by controlling and regulating absorption.

Since the heat-treated Lactobacillus plantarum LRCC5314 strain according to one aspect of the present invention exhibits an effect of inhibiting cortisol hypersecretion, it can be used in foods for relieving stress.

The heat-treated Lactobacillus plantarum LRCC5314 strain according to one aspect of the present invention is superior to NPY and Y2R factors, which are stress-related factors, compared to the same live bacteria.

The heat-treated Lactobacillus plantarum LRCC5314 strain according to an aspect of the present invention has superior mRNA expression levels of GLUT4, Adiponectin, and Leptin factors compared to the same live cell, and thus has a more excellent effect on blood sugar control.

The heat-treated Lactobacillus plantarum LRCC5314 strain according to one aspect of the present invention has excellent anti-inflammatory effect because it regulates the expression of genes related to inflammation regulation of TNF-a, IL-6, and CCL-2 compared to the same live cells.

The heat-treated Lactobacillus plantarum LRCC5314 strain according to an aspect of the present invention is more effective in inhibiting cortisol hypersecretion than the same live bacteria.

The effects of the present invention are not limited to the above-mentioned effects. It should be understood that the effects of the present invention include all effects that can be inferred from the following description.

1 is a result of acute toxicity of Example 1 Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain dead cells.
2 is a result of measuring the change in body weight of mice administered with Example 1 Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain dead cells.
3 is a result of fasting blood glucose measurement in mice administered with Example 1 Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) dead cells.
4 is a glucose tolerance test (IPGTT) results after administering Example 1 Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain dead cells to diabetes-induced mice.
5 is an insulin tolerance test (ITT) result after administering Example 1 Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain dead cells to diabetes-induced mice.
6 is a result of (6) inflammation regulation-related gene expression analysis of Example 2, FIG. 6a TNF-a, FIG. 6b is IL-6, FIG. 6c is CCL-2.
7 is a result of analysis of the (7) blood glucose control-related gene expression level of Example 2, FIG. 7a GLUT4, FIG. 7b is Adiponectin, and FIG. 7c is the mRNA expression level of Leptin factor.
8 is a result of (8) stress improvement-related gene expression analysis of Example 2, FIG. 8a is NPY, FIG. 8b is Y2R factor, and FIG. 8c is the concentration of corticosterone in the blood.

The above objects, other objects, features and advantages of the present invention will be easily understood through the following preferred embodiments in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments introduced herein are provided so that the disclosed subject matter may be thorough and complete, and that the spirit of the present invention may be sufficiently conveyed to those skilled in the art.

In the present specification, terms such as “comprise” or “have” are intended to designate that a feature, number, step, operation, component, part, or combination thereof described in the specification exists, but one or more other features It is to be understood that it does not preclude the possibility of the presence or addition of numbers, steps, operations, components, parts, or combinations thereof.

In this specification, when a range is described for a variable, the variable will be understood to include all values within the stated range including the stated endpoints of the range. For example, a range of “5 to 10” includes the values of 5, 6, 7, 8, 9, and 10, as well as any subranges such as 6 to 10, 7 to 10, 6 to 9, 7 to 9, etc. It will be understood to include any value between integers that are appropriate for the scope of the recited range, such as 5.5, 6.5, 7.5, 5.5 to 8.5 and 6.5 to 9, and the like. Also for example, a range of “10% to 30%” includes values such as 10%, 11%, 12%, 13%, and all integers up to and including 30%, as well as 10% to 15%, 12% to It will be understood to include any subrange, such as 18%, 20% to 30%, etc., as well as any value between reasonable integers within the scope of the recited range, such as 10.5%, 15.5%, 25.5%, and the like.

The present invention relates to Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain, its dead cells and uses thereof, and more particularly, to lower blood sugar, antidiabetic, anti-inflammatory, or anti-stress effect, a traditional fermented food It relates to a novel Lactobacillus plantarum LRCC5314 strain isolated from kimchi and uses thereof.

One aspect of the present invention provides a Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P.

The "Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain" is a novel lactic acid bacteria strain isolated from kimchi, deposited on June 20, 2019 at the Korea Microorganism Conservation Center (overseas) with accession number KCCM12540P. The strain not only has alpha-amylase and alpha-glucosidase inhibitory activity, but also has acid resistance, bile resistance and intestinal adhesion characteristics related to the function of the probiotic strain. In addition, the strain lowers blood sugar by controlling and regulating the digestion and absorption of sugar through alpha-amylase and alpha-glucosidase inhibitory action, and thus ultimately has an antidiabetic effect through blood sugar control. In addition, the strain also has an anti-inflammatory effect as it inhibits the production of nitric oxide (NO), which is an important mediator in the inflammatory process. In addition, the strain has an anti-stress effect by inhibiting the secretion of cortisol, which secretion is promoted when stress is added.

On the other hand, the strain contains the base sequence shown in SEQ ID NO: 1 as 16S rDNA, and carbon sources are D-glucose (D-glucose), D-fructose (D-fructose), D-mannose (D-mannose), N -Acetyl glucosamine, amygdalin, arbutin, esculin, salicin, D-celiobiose, D-maltose, D-saccharose (sucrose), D-trehalose, D-melezitose, Gentiobiose, and D-turanose use it

In an embodiment of the present invention, it was confirmed that Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain isolated and identified from kimchi had alpha-amylase and alpha-glucosidase inhibitory activity. In addition, it was confirmed that the Lactobacillus plantarum LRCC5314 strain is a strain with high stability for 120 minutes or more even under artificial gastric juice conditions of pH 2.0, and has high stability for more than 12 hours under artificial bile solution conditions. strain was confirmed. In addition, it was confirmed that the colon has the ability to adhere to the epithelial cells Caco-2 cells.

One aspect of the present invention provides a probiotic composition comprising, as an active ingredient, the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture, or its lysate.

The Lactobacillus plantarum LRCC5314 strain deposited with the accession number KCCM12540P is as described above.

The term "culture" refers to a material containing a culture medium and/or a cultured strain, but is not limited thereto, but is a culture medium or strain itself cultured including the Lactobacillus plantarum LRCC5314 strain deposited with the above-mentioned accession number KCCM12540P. can The culture is not limited in its formulation, and may be, for example, liquid or solid.

In the present invention, the 'culture solution' refers to a cell culture solution or culture medium remaining after culturing the strain except for the strain.

The term "lysate" refers to a cell lysate obtained by disrupting the Lactobacillus plantarum LRCC5314 strain deposited under the accession number KCCM12540P by a chemical method or a physical method using a surfactant, etc.

The term "composition" is a concept that includes not only conventional food, functional food or dietary supplements, but also those approved as pharmaceuticals.

The carrier may be one or more selected from among diluents, lubricants, binders, disintegrants, sweeteners, stabilizers, and preservatives, and as additives, one or more selected from fragrances, vitamins, and antioxidants can be used by

In the present invention, all pharmaceutically acceptable carriers and additives can be used, and specifically, diluents include lactose monohydrate, trehalose, corn starch, and soybean oil. , microcrystalline cellulose or mannitol is preferred, the lubricant is preferably magnesium stearate or talc, and the binder is polyvinylpyrrolidone (PVP: polyvinyipyrolidone) Or hydroxypropyl cellulose (HPC: hydroxypropylcellulose) is preferably selected. In addition, the disintegrant is selected from carboxymethylcellulose calcium (Ca-CMC), sodium starch glycolate, polacrylin potassium or crospovidone (cross-linked polyvinylpyrrolidone). Preferably, the sweetener is selected from sucrose, fructose, sorbitol or aspartame, and the stabilizer is carboxymethylcellulose sodium (Na-CMC), beta-cyclodextrin (β-cyclodextrin), white wax (white bee's wax) or xanthan gum (xanthan gum), and the preservative is methyl p-hydroxy benzoate (methlparaben), propyl p-hydroxybenzoate (propylparaben), or potassium sorbate ( potassium sorbate) is preferred.

In addition, the Lactobacillus plantarum LRCC5314 strain of the present invention can be used as a seed for producing various lactic acid bacteria fermented milk and fermented products. may include tofu, soybean paste, cheonggukjang, jelly, kimchi, and the like, but is not necessarily limited thereto. The fermented milk and fermented products can be easily prepared by replacing only the strain with the lactic acid bacteria strain of the present invention in a conventional manufacturing method.

As described above, the composition of the present invention may be a food or food additive, and the food is preferably a fermented food. In addition, the composition of the present invention may be in powder or granular form, and the powder or granular form may be easily prepared by a conventional method in the art.

One aspect of the present invention provides a composition for lowering blood sugar comprising, as an active ingredient, the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, a culture thereof, or a lysate thereof.

The term "blood sugar" refers to glucose contained in the blood, and the blood glucose supplied to the human body enters the cells with the help of insulin to generate energy necessary to maintain normal functions. In a normal person, after a meal, blood sugar rises temporarily and then falls back to a normal level by insulin. and will be thrown out.

The term "lowering blood sugar" refers to lowering blood sugar, i.e., 'inhibition of rise in blood sugar', whereby alpha-glucosidase converts carbohydrates degraded by alpha-amylase to monosaccharides. Inhibition of the activity of these enzymes limits the sugar concentration after meals by delaying the hydrolysis and absorption of carbohydrates.

The Lactobacillus plantarum LRCC5314 strain of the present invention can lower blood sugar by controlling and regulating the digestion and absorption of sugar through alpha-amylase and alpha-glucosidase inhibitory action. can

In the present invention, the composition for lowering blood sugar may be embodied as a pharmaceutical composition, a food additive, a food composition (particularly a functional food composition), or a feed additive according to the purpose or aspect of use, and Lactobacillus as an active ingredient in the composition The content of the plantarum LRCC5314 strain may also be adjusted in various ranges depending on the specific form of the composition, the purpose of use, or the aspect.

One aspect of the present invention provides a pharmaceutical composition for preventing or treating diabetes, comprising the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture or its lysate as an active ingredient.

The term "diabetes" is a chronic metabolic disease in which glucose is excreted in urine due to an abnormally high concentration of glucose in the blood due to a relative lack of insulin. That is, carbohydrates in the digestive system are decomposed by alpha-amylase and alpha-glucosidase, which are digestive enzymes, and are finally decomposed into glucose and fructose and absorbed in the small intestine. Accordingly, an excessive reaction of these enzymes causes a rapid production of sugar, which leads to diseases such as diabetes.

The Lactobacillus plantarum LRCC5314 strain of the present invention controls and regulates the digestion and absorption of sugar through an alpha-amylase and alpha-glucosidase inhibitory action to lower blood sugar, and ultimately has an antidiabetic effect through blood sugar control. Therefore, it can be usefully used as a pharmaceutical composition for preventing or treating diabetes.

Alpha-amylase is an enzyme that breaks down the α-D-(1,4)-glucan bond of carbohydrates. It breaks down carbohydrates in food and converts them into sugars that are easy to be absorbed into the body, a carbon and energy source for maintaining life. is used as Through this, the metabolic importance of starch-degrading enzymes such as alpha-amylase can be understood, but often an overreaction of starch-degrading enzymes causes rapid sugar production and consequent organic acid production, leading to diabetes, obesity, and acidosis. will cause disease Accordingly, when the activity of alpha-amylase is modulated, that is, inhibited, it is effective in treating diseases such as diabetes, obesity and acidosis.

Alpha-glucosidase has the function of cleaving the glycosidic bond between sugar and sugar, sugar and lipid, or between sugar and protein in polysaccharides in the cell, so when the enzyme reaction of glucosidase is inhibited, absorption of sugar in the small intestine is delayed. It is effective in treating diabetes.

The pharmaceutical composition can prevent or treat diseases regulated by alpha-amylase or alpha-glucosidase inhibitory activity. Since the strain of the present invention exhibits alpha-amylase or alpha-glucosidase inhibitory activity, it can exhibit an excellent therapeutic effect on diseases such as diabetes, Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain of the present invention, and its culture Water or a lysate thereof may be used as a pharmaceutical composition for preventing or treating diabetes.

The pharmaceutical composition of the present invention is safely absorbed into the body regardless of the intake amount. Dosage or intake is the type and content of the active ingredient and other ingredients contained in the pharmaceutical composition, the type of formulation and the patient's age, weight, general health, sex and diet, administration time, administration route and composition secretion rate, treatment It can be adjusted according to various factors including the duration and the drugs used at the same time. When the pharmaceutical composition of the present invention is administered to the human body, there is no fear of side effects compared to chemically synthesized foods and drugs containing alpha-amylase and alpha-glucosidase inhibitors. The food or pharmaceutical composition of the present invention may be formulated by including one or more pharmaceutical or food acceptable carriers in addition to the active ingredient. Pharmaceutical or food acceptable carriers may be used in a mixture of saline, sterile water, Ringer's solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components, and if necessary, antioxidants, buffers, Other common additives such as bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets. Furthermore, it can be preferably formulated according to each disease or component using an appropriate method in the art, or a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA.

Formulations of the pharmaceutical composition of the present invention may be granules, powders, granules, coated tablets, tablets, capsules, soups, extracts, suppositories, syrups, juices, suspensions, emulsions, sustained-release preparations of the active compound, etc. have.

In the pharmaceutical composition according to the present invention, the content of the novel lactic acid bacteria, that is, the Lactobacillus plantarum LRCC5314 strain, its culture, or its lysate, as an active ingredient in the pharmaceutical composition according to the present invention is not significantly limited, for example, 0.01 to 99% by weight based on the total weight of the composition, Preferably 0.5 to 50% by weight, more preferably 1 to 30% by weight. In addition, the pharmaceutical composition according to the present invention may further include an additive such as a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. Carriers, excipients and diluents that may be included in the pharmaceutical 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, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In addition, the pharmaceutical composition of the present invention is a Lactobacillus plantarum LRCC5314 strain, a culture thereof or a lysate thereof, in addition to diabetes, obesity, fatty liver, hyperlipidemia, cardiovascular disease, hypertension, arteriosclerosis, or a known effective having a therapeutic effect on metabolic syndrome It may contain one or more types of components.

The pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or a formulation for parenteral administration by a conventional method, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants, etc. It may be formulated using diluents or excipients.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the active ingredient, for example, starch, calcium carbonate, sucrose ), lactose or gelatin may be mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Liquid formulations for oral administration include suspensions, solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included in addition to commonly used simple diluents such as water and liquid paraffin. have.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used. Furthermore, it can be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (latest edition), Mack Publishing Company, Easton PA.

The pharmaceutical composition of the present invention may be administered orally or parenterally to mammals including humans according to a desired method, and parenteral administration methods include external application to the skin, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, and intramuscular administration. There is an intravenous injection or an intrathoracic injection injection method.

The dosage of the pharmaceutical composition of the present invention is not particularly limited as long as it is a pharmaceutically effective amount, and the range may vary depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of disease. Varies. A typical daily dosage of the pharmaceutical composition of the present invention is not particularly limited, and may be administered once a day or divided into several doses.

One aspect of the present invention provides a food composition for preventing or improving diabetes, comprising the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture or its lysate as an active ingredient.

The Lactobacillus plantarum LRCC5314 strain of the present invention controls and regulates the digestion and absorption of sugar through an alpha-amylase and alpha-glucosidase inhibitory action to lower blood sugar, and ultimately has an antidiabetic effect through blood sugar control. Therefore, it can be usefully used as a food composition for preventing or improving diabetes.

The "food composition" may include any food in a conventional sense, and may be used interchangeably with terms known in the art, such as functional food and health functional food.

As used herein, the term “functional food” refers to food manufactured and processed using raw materials or ingredients useful for the human body according to Health Functional Food Act No. 6727, and “functionality” refers to the structure of the human body. And it means ingestion for the purpose of obtaining useful effects for health purposes such as regulating nutrients for function or physiological action.

In addition, the term "health functional food" as used in the present invention refers to a food manufactured and processed by extracting, concentrating, refining, mixing, etc., a specific ingredient as a raw material or a specific ingredient contained in a food raw material for the purpose of health supplementation, It refers to a food designed and processed to sufficiently exert biological control functions, such as biological defense, regulation of biological rhythm, prevention and recovery of disease, etc. with respect to the living body by the above ingredients. It can perform functions related to recovery, etc.

There is no limitation on the kind of food in which the composition of the present invention can be used. In addition, the composition comprising the Lactobacillus plantarum LRCC5314 strain of the present invention, a culture thereof, or a lysate thereof as an active ingredient may be prepared by mixing appropriate other auxiliary ingredients that may be contained in food and known additives according to the selection of those skilled in the art. have. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes, and the like, and it can be prepared by adding the Lactobacillus plantarum LRCC5314 strain according to the present invention, a culture thereof, or a lysate thereof as a main component to juice, tea, jelly, juice, and the like.

In addition, foods that can be applied to the present invention include, for example, special nutritional foods (eg, formula milk, infant food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (eg, ramen, noodles, etc.), health supplements , Seasoned foods (eg soy sauce, soybean paste, red pepper paste, mixed soy sauce, etc.), sauces, sweets (eg snacks), dairy products (eg fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi, pickles, etc.) ), beverages (eg, fruit, vegetable beverages, soy milk, fermented beverages, etc.), and natural seasonings (eg, ramen soup, etc.).

When the health functional food composition of the present invention is used in the form of a beverage, it may contain various sweeteners, flavoring agents, or natural carbohydrates as additional ingredients, as in a conventional beverage. In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin , alcohol, a carbonation agent used in carbonated beverages, and the like. In addition, it may contain the pulp for the production of natural fruit juice, fruit juice beverage and vegetable beverage.

One aspect of the present invention provides an anti-inflammatory pharmaceutical composition comprising the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture or its lysate as an active ingredient.

The term "anti-inflammatory" refers to the action of inhibiting inflammation, and the regulation of the inflammatory response is known to be very complex, which is known to enhance the in vivo repair system and reduce damage. However, if the inflammatory response continues due to repeated tissue damage or regeneration, ROS and RNS are overproduced in inflammation-related cells, resulting in permanent gene modification. As such, ROS and RNS are deeply related to the inflammatory response that regulates the actions of various cells in vivo. During the inflammatory process, large amounts of proinflammatory cytokines, nitric oxide (NO), and prostaglandin E2 (PGE2) are released from inducible nitric oxide synthase (iNOS) and cyclooxygenase. Produced by cyclooxygenase-2 (COX-2). Inflammation is a cause of various inflammatory diseases, and the composition comprising the Lactobacillus plantarum LRCC5314 strain of the present invention, a culture thereof, or a lysate thereof may have a preventive and ameliorating effect on various inflammatory diseases through anti-inflammatory action.

In one embodiment of the present invention, when Raw 264.7 cells are treated with Lactobacillus plantarum LRCC5314 strain culture medium, lactobacilli in LPS-treated Raw 264.7 cells compared to the control induced by LPS, without affecting cell survival It was confirmed that when the Bacillus plantarum LRCC5314 strain culture was treated, the level of NO production, which is an important mediator in the progression of inflammation, was inhibited. That is, it was confirmed that the Lactobacillus plantarum LRCC5314 strain had an anti-inflammatory effect by significantly reducing the amount of NO production in the inflammatory cells without causing cytotoxicity. Therefore, the anti-inflammatory effect of the present invention may be achieved by inhibiting NO production.

Since the strain of the present invention exhibits NO production inhibitory ability and can exhibit excellent therapeutic effects on inflammatory diseases, the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain of the present invention, its culture or its lysate is an anti-inflammatory pharmaceutical composition, That is, it can be used as a pharmaceutical composition for preventing or treating inflammatory diseases.

The 'inflammatory disease' is a generic term for diseases whose main lesion is inflammation, and is not limited thereto, but allergic asthma, allergic rhinitis, allergic mucositis, urticaria and anaphylax. diseases, myopathy including systemic sclerosis, dermatomyositis and inclusion body myositis, arthritis, atopic dermatitis, psoriasis, asthma, multiple sclerosis, ssRNA and dsRNA viral infections, sepsis, polychondritis , scleroderma, eczema, gout, periodontal disease, Behcet's syndrome, edema, vasculitis, Kawasaki disease, diabetic retinitis, autoimmune pancreatitis, vasculitis, glomerulonephritis, acute and chronic bronchitis, and influenza infection.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable” means exhibiting non-toxic properties to cells or humans exposed to the composition. The carrier may be used without limitation as long as it is known in the art, such as buffers, preservatives, softening agents, solubilizers, isotonic agents, stabilizers, bases, excipients, lubricants, and the like.

In addition, the pharmaceutical composition of the present invention can be used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. have. Furthermore, it can be used in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel, or external preparation for skin in the form of a gel.

On the other hand, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "administration" of the present invention means introducing a predetermined substance to an individual by an appropriate method, and the administration route of the composition may be administered through any general route as long as it can reach a target tissue. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, may be administered intrarectally, but is not limited thereto.

The term “individual” refers to all animals, including humans, rats, mice, and livestock. Preferably, it may be a mammal including a human.

The term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level is determined by the sex, age, and weight of the patient. , health condition, disease type, severity, drug activity, drug sensitivity, administration method, administration time, administration route, and excretion rate, treatment period, factors including drugs used in combination or concurrently, and other medical fields. It can be readily determined by one of ordinary skill in the art according to known factors.

One aspect of the present invention provides an anti-inflammatory food composition comprising the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture or its lysate as an active ingredient.

As for the anti-inflammatory as described above, when the composition comprising the Lactobacillus plantarum LRCC5314 strain of the present invention, its culture or its lysate as an active ingredient is used as a food composition, the Lactobacillus plantarum LRCC5314 strain itself It can be added or used together with other foods or food ingredients, and can be appropriately used according to a conventional method. The mixing amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment), and may further include a food pharmaceutically acceptable food supplement additive. Since the composition of the present invention uses novel lactic acid bacteria derived from kimchi, a traditional fermented food, as an active ingredient, there is no problem in terms of stability, so there is no significant limitation on the mixing amount.

There is no limitation on the kind of food in which the composition of the present invention can be used. Examples of food include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes. And, it can be prepared by adding the Lactobacillus plantarum LRCC5314 strain according to the present invention, a culture thereof or a lysate thereof as a main component to juice, tea, jelly, juice, and the like.

One aspect of the present invention provides a food composition for relieving stress, comprising the Lactobacillus plantarum LRCC5314 ( Lactobacillus plantarum LRCC5314) strain deposited with accession number KCCM12540P, its culture or its lysate as an active ingredient.

The term “stress” refers to a detrimental response or change to an organism caused by an external stimulus. "Stress relief" can be used interchangeably with 'anti-stress', and is a term that collectively represents a resistance action against stress, for example, a preventive effect, a stress reducing effect, or various symptoms caused by stress (eg, depression). , cognitive function or learning ability decrease, headache, indigestion, fatigue, rough skin, neurosis, gastric ulcer, etc.)

The food composition for relieving stress of the present invention can suppress cortisol hypersecretion.

In one embodiment of the present invention, H295R cells, which are human adrenocortical cells, are treated with LPS, which is known as a stress-inducing substance, and treated with Lactobacillus plantarum LRCC5314 strain. As a positive control, compared to H295R cells treated only with LPS. In the case of H295R cells treated with the Lactobacillus plantarum LRCC5314 strain, the cortisol secretion was reduced by more than 3 times, it was found that the antistress effect of the Lactobacillus plantarum LRCC5314 strain was excellent.

There is no particular limitation on the type of the food composition. Examples of foods to which the composition of the present invention can be added include health functional foods such as beverages, gums, vitamin complexes, and drinks, and include all food compositions in a conventional sense.

The food composition includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, It may contain a protective colloidal thickener, a pH adjuster, a stabilizer, a preservative, glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like.

The composition of the present invention is a probiotic composition, pharmaceutical composition or food because the composition comprising the Lactobacillus plantarum LRCC5314 strain, a culture thereof or a lysate thereof, as an active ingredient, is a novel lactic acid bacterium derived from kimchi, a traditional fermented food. Even when used as a composition, side effects may be less than that of general synthetic compounds, so it can be safely included and usefully used.

Various aspects of the present invention are as follows.

One aspect of the present invention is a Lactobacillus plantarum LRCC 5314 (Lactobacillus plantarum LRCC 5314) strain having a hypoglycemic, antidiabetic, anti-inflammatory or stress-relieving effect, wherein the strain is a heat-treated dead cell.

In one aspect of the present invention, the strain is deposited with accession number KCCM12540P, provides a strain.

In one aspect of the present invention, the strain is isolated from kimchi, and provides a strain comprising the 16S rDNA base sequence of SEQ ID NO: 1.

In one aspect of the present invention, the strain modulates inflammatory factors of TNF-a, IL-6 or CCL-2, or the strain modulates GLUT4, Adiponectin or Leptin's blood glucose-related genes, or the strain is NPY or It provides a strain that expresses Y2R stress improvement related gene, or the strain regulates excessive secretion of cortisol in the blood.

Another aspect of the present invention is the strain according to any one of the aspects of the present invention; lysate thereof; its culture; its ferment; and extracts of the strains, lysates, cultures or ferments; It provides a microbial preparation comprising one or more of.

In another aspect of the present invention, the strain provides a dead cell, microbial preparation.

Another aspect of the present invention provides a probiotic composition comprising, as an active ingredient, the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a composition for lowering blood sugar comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating diabetes, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

Another aspect of the present invention provides a food composition for preventing or improving diabetes, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

Another aspect of the present invention provides an anti-inflammatory pharmaceutical composition comprising, as an active ingredient, the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof.

Another aspect of the present invention provides an anti-inflammatory food composition comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient.

Another aspect of the present invention provides a food composition for improving cortisol hypersecretion, comprising the heat-treated Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) strain of the one aspect of the present invention, a culture thereof, or a lysate thereof as an active ingredient do.

The strain of the present invention may be a Tyndalated Lactobacillus plantarum LRCC5314 strain.

In one embodiment, the "tindalhwa" is a method of killing live lactic acid bacteria.

In one embodiment, the method for killing may be a strain that has undergone a process of lyophilization after heat-treating the activated live cells at a temperature of 90 to 100° C. for 10 to 50 minutes.

Hereinafter, the configuration and effect of the present invention will be described in more detail through Examples, Preparation Examples and Experimental Examples. These Examples, Preparation Examples and Experimental Examples are only for illustrating the present invention, and the scope of the present invention is not limited by these Examples, Preparation Examples and Experimental Examples.

Example 1 - Strain Identification and Tyndallization

The inventors of the present invention collected traditional kimchi, added sterile distilled water to the collected sample to make a diluted solution, and then homogenized the inside of the sample with a grinding homogenizer (Stomacher, Pro-Media SH-001, ELMEX). After diluting the dilution step by step with sterile saline, 0.1 ml was taken. This was smeared on MRS solid medium supplemented with 0.002% by weight of BCP (Bromocresol purple) and 1.5% by weight of agar, and cultured for 48 hours in an incubator maintained at 37°C. After culturing, colonies showing yellow rings were selected and subcultured 2-3 times in the same medium for each individual to purely isolate each strain. Separated strains were inoculated into MRS broth for each individual, cultured at 37°C for 48 hours, and then used while refrigerated at 4°C.

The nucleotide sequence of the 16s rDNA gene of the strain is the same as SEQ ID NO: 1. As a result of the nucleotide sequence analysis, it was confirmed that the strain has a maximum of 99% homology with the Lactobacillus sp. strain, and the present inventors identified the strain as a new Lactobacillus plantarum strain, named LRCC5314 to the Korea Microbial Conservation Center 2019 It was deposited on June 20, 2016 (Accession No. KCCM12540P).

Tyndalization of lactic acid bacteria is a method of killing live lactic acid bacteria, and after culturing the strain in the maximum activated state by passing the third passage, heat killed cells were prepared at 95° C. for 30 minutes and used after freeze-drying.

Example 2 - Mouse administration experiment

(1) Acute toxicity test of heat-treated lactic acid bacteria LRCC5314 strain

To confirm the acute toxicity of heat-treated lactic acid bacteria, the experimental animals were administered orally daily for 12 weeks using different concentrations of the LRCC314 strain using 5-week-old C57BL/6J mice. Mortality and liver tissue of mice for different concentrations of LRCC5314 strain were observed during the administration period.

The experimental results were as shown in FIG. 1 .

The concentration of the LRCC5314 strain was 1x10 ⁸ cells/g, and as a result of oral administration, no mice died, and there was no significant difference compared with the control group in the H&E staining of liver tissue. Therefore, it was judged that there was no toxicity of the LRCC5314 strain, and animal experiments were continued.

(2) Experimental animals and groups

Using 5-week-old C57BL/6J mice, the experiment was conducted by dividing the mice into 2 groups by 10 mice per group. The first group was a control group treated with a high-fat diet (containing 30% saturated fat), and the second group was an experimental group, which was administered with LRCC5314 heat-treated lactic acid bacteria (1x10 ⁸ cells/g) at the same time as a high-fat diet, the third group were divided into groups administered with live LRCC5314 lactic acid bacteria (1x10 ⁸ cells/g) at the same time as a high-fat diet.

(3) Weight change measurement

Body weight was measured once a week at the same time every week after the initial measurement on the day of the start of the experiment. Body weight was measured by using an electronic scale in batches before morning feeding, and in order to minimize the weight error due to the movement of the mouse during measurement, the mouse was placed on a plastic bowl, and the weight that appeared when the mouse reached a stable state was recorded. .

The experimental results were as shown in FIG. 2 .

In this experiment, weight gain of all mice appeared from 1 week after high-fat feed, and weight loss was observed in the group administered with LRCC5314 heat-treated lactic acid bacteria (1x10 ⁸ cells/g) compared to the high-fat control group. Conversely, the group administered with LRCC5314 live lactic acid bacteria (1x10 ⁸ cells/g) showed a tendency to slightly increase body weight.

(4) Fasting glucose level (mg/dl)

Blood glucose was measured using a blood glucose tester by collecting blood from the tail vein after fasting for 12 hours or more once a week.

The experimental results were as shown in FIG. 3 .

On average, it was measured to be significantly higher in the high-fat diet group, and especially at week 12, the high-fat diet group had the highest value at 208.2 mg/dl. On the other hand, the group treated with LRCC5314 heat-treated lactic acid bacteria showed a level of 147.2 mg/dl, and the blood glucose level was 29% lower than that of the high-fat diet group, respectively.

(5) intraperitoneal glucose tolerance test (IPGTT) and insulin tolerance test (ITT)

To measure changes in glucose metabolism, LRCC5314 heat-treated lactic acid bacteria were administered to diabetic mice for 13 weeks, and then glucose tolerance test (IPGTT) and insulin tolerance test (ITT) were performed.

In the intraperitoneal glucose tolerance test, after fasting for more than 12 hours on the 13th week of the experiment, fasting blood glucose was measured, and 5% glucose solution was dissolved in distilled water at a volume of 2 g/kg body weight and injected intraperitoneally. After 30 minutes and 60 minutes after , after 90 minutes and after 120 minutes, blood glucose was measured by collecting tail vein blood from mice.

In addition, ITT was performed after fasting the mice for 12 hours 3 days before autopsy, measuring blood glucose, subcutaneously administering 5 unit/kg insulin, and intraperitoneally administering a glucose solution at a dose of 2 g/kg body weight 30 minutes after 30 minutes. After minutes, after 60 minutes, after 90 minutes, and after 120 minutes, blood glucose was measured by collecting tail vein blood from the mice.

Experimental results were as shown in FIGS. 4 and 5 .

As shown in FIGS. 4 and 5 , as a result of the glucose tolerance test, it was confirmed that the LRCC5314 heat-treated lactobacillus group significantly improved and rapidly decreased the glucose tolerance compared to the high-fat diet group. Also, in the insulin tolerability test, it was observed that the absorption (sensitivity) of blood glucose by insulin increased significantly and the fastest.

(6) Inflammation regulation-related gene expression level analysis

After 12 weeks of animal experimentation, blood was taken under anesthesia (Urethane 1.7 mg/kg) and perfused with PBS containing 0.1% diethylpyrocarbonate through the abdominal aorta, and then sacrificed. After that, the liver was taken and immediately stored at -70°C. In order to confirm the inflammatory control effect of LRCC 5314 heat-treated lactic acid bacteria, the mRNA expression levels of inflammatory cytokines TNF-a, IL-6, and CCL-2 factors in liver samples were confirmed by Real-Time PCR.

Specifically, total RNA from liver was isolated using Trizol (Invitrogen, Carlsbad, CA, USA), the concentration of RNA isolated at 260 nm was measured using a spectrophotometer, and absorbance measured at 260 nm and 280 nm. RNA purity was evaluated by the ratio of 1 μg of the isolated RNA was subjected to a reverse transcription reaction using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Carlsbad, CA, USA), and then,

The mRNA expression of the control group 18S and the inflammatory cytokines TNF-a, IL-6 (interleukin-6), and CCL-2 was analyzed with each primer (Bioneer, Daejeon, Korea) and the SYBR Green System (7300 Real-Time). PCR System, Applied Biosystems) was used.

The experimental results were as shown in FIG. 6 .

As shown in FIG. 6 , it was confirmed that the expression levels of all three inflammatory factors were significantly lowered in the LRCC5314 heat-treated lactic acid bacteria-treated group compared to the high-fat diet group.

(7) Analysis of gene expression level related to blood sugar control

To confirm the blood glucose control effect of LRCC 5314 heat-treated lactic acid bacteria, mRNA expression levels of GLUT4, Adiponectin, and Leptin factors in liver samples were confirmed by Real-Time RT qPCR. It proceeded in the same manner as described above.

The experimental results were as shown in FIG. 7 .

As shown in FIG. 7 , it was confirmed that the LRCC5314 heat-treated lactic acid bacteria-treated group significantly increased the expression of GLUT4 protein compared to the high-fat diet group, thereby promoting glucose absorption and contributing to blood sugar control, and remarkably inhibiting both Adiponectin and Leptin factors. It was confirmed that not only blood sugar control but also fat synthesis was controlled.

(8) Stress improvement related gene expression level analysis

In order to confirm the blood glucose and stress control effect of LRCC 5314 heat-treated lactic acid bacteria, mRNA expression levels of NPY and Y2R factors in liver samples were confirmed by Real-Time RT qPCR. It proceeded in the same manner as described above.

The experimental results were as shown in FIG. 8 .

As shown in FIG. 8 , it was confirmed that the LRCC5314 heat-treated lactic acid bacteria-administered group significantly reduced the expression of NPY and Y2R factors compared to the high-fat diet group, thereby regulating stress. In addition, when LRCC 5314 heat-treated lactic acid bacteria was administered, it was confirmed that the level of corticosteron (Cortisol) in the blood was also greatly reduced.

Preparation Example 1: Preparation of a pharmaceutical composition

Lactobacillus plantarum LRCC5314 (Lactobacillus plantarum LRCC5314) culture in the preparation of the following pharmaceutical composition can be replaced with the Lactobacillus plantarum LRCC5314 strain itself or a lysate thereof.

Preparation Example 1-1: Preparation of powder

Example 1 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients were mixed and filled in an airtight cloth to prepare a powder.

Preparation Example 1-2: Preparation of tablets

Example 1 10 mg

Corn Starch 100 mg

Lactose 100 mg

Magne stearate 2 mg

After mixing the above ingredients, tablets were prepared by tableting according to a conventional method for manufacturing tablets.

Preparation Example 1-3: Preparation of capsules

Example 1 10 mg

3 mg of crystalline cellulose

Lactose 15 mg

0.2 mg of magnesium stearate

After mixing the above ingredients, the capsules were prepared by filling in gelatin capsules according to a conventional manufacturing method of capsules.

Preparation Example 1-4: Preparation of a ring

Example 1 10 mg

Lactose 150 mg

Glycerin 100 mg

xylitol 50 mg

After mixing the above components, it was prepared so as to be 4 g per ring according to a conventional method.

Preparation Example 1-5: Preparation of granules

Example 1 15 mg

Soybean Extract 50 mg

glucose 200 mg

Starch 600 mg

After mixing the above components, 100 mg of 30% ethanol was added and dried at 60° C. to form granules, and then filled in a bag.

Preparation Example 1-6: Preparation of injection

Example 1 10 mg

Sodium metabisulfite 3.0 mg

Methylparaben 0.8 mg

Propylparaben 0.1 mg

Appropriate amount of sterile distilled water for injection

After mixing the above ingredients, 2 ml of them were filled in an ampoule and sterilized to prepare an injection.

Preparation Example 2: Preparation of a food composition

Preparation Example 2-1: Preparation of wheat flour food

0.5 parts by weight of Example 1 was added to 100 parts by weight of wheat flour, and breads, cakes, cookies, crackers and noodles were prepared using this mixture.

Preparation Example 2-2: Preparation of dairy products

0.5 parts by weight of Example 1 was added to 100 parts by weight of milk, and various dairy products such as butter and ice cream were prepared using the milk.

Preparation Example 2-3: Preparation of wire

Brown rice, barley, glutinous rice, and barley radish were pregelatinized by a known method and dried, and then roasted and prepared as a powder having a particle size of 60 mesh with a grinder.

Black soybeans, black sesame, and perilla were also steamed and dried by a known method, and then roasted and prepared into powder having a particle size of 60 mesh with a grinder.

It was prepared by mixing the grains, seeds and Example 1 prepared above in the following ratio.

Grains (30 parts by weight of brown rice, 17 parts by weight of barley, 20 parts by weight of barley),

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame),

Example 1 (1 part by weight),

Reishi (0.5 parts by weight),

Rehmannia (0.5 parts by weight).

Preparation Example 2-4: Preparation of health drinks

After instant sterilization by homogeneously mixing 1 g of Example 1 with auxiliary materials such as high fructose (0.5 g), oligosaccharide (4 g), sugar (2 g), salt (0.5 g), and water (77 g) This was prepared by packaging in small packaging containers such as glass bottles and plastic bottles.

Preparation Example 2-5: Preparation of vegetable juice

Vegetable juice was prepared by adding 2 g of Example 1 to 1,000 ml of tomato or carrot juice.

Preparation Example 2-6: Preparation of fruit juice

Fruit juice was prepared by adding 1 g of Example 1 to 1,000 ml of apple or grape juice.

Korea Microorganism Conservation Center (Overseas) KCCM12540P 20190620

<110> LOTTE FOODS CO.,LTD <120> Lactobacillus plantarum LRCC5314 having an effect of blood glucose lowering-activity and controlling stress and its application <130> DP-2021-0184 <160> 1 <170> KoPatentIn 3.0 <210> 1 <211> 960 <212> DNA <213> Artificial Sequence <220> <223> LRCC5314_16S rDNA <400> 1 tgtccggatt tattgggcgt aaagcgagcg caggcggttt tttaagtctg atgtgaaagc 60 cttcggctca accgaagaag tgcatcggaa actgggaaac ttgagtgcag aagaggacag 120 tggaactcca tgtgtagcgg tgaaatgcgt agatatatgg aagaacacca gtggcgaagg 180 cggctgtctg gtctgtaact gacgctgagg ctcgaaagta tgggtagcaa acaggattag 240 ataccctggt agtccatacc gtaaacgatg aatgctaagt gttggagggt ttccgccctt 300 cagtgctgca gctaacgcat taagcattcc gcctggggag tacggccgca aggctgaaac 360 tcaaaggaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgaagctac 420 gcgaagaacc ttaccaggtc ttgacatact atgcaaatct aagagattag acgttccctt 480 cgggggacatg gatacaggtg gtgcatggtt gtcgtcagct cgtgtcgtga gatgttgggt 540 taagtcccgc aacgagcgca acccttatta tcagttgcca gcattaagtt gggcactctg 600 gtgagactgc cggtgacaaa ccggaggaag gtggggatga cgtcaaatca tcatgcccct 660 tatgacctgg gctacacacg tgctacaatg gatggtacaa cgagttgcga actcgcgaga 720 gtaagctaat ctcttaaagc cattctcagt tcggattgta ggctgcaact cgcctacat 780 aagtcggaat cgctagtaat cgcggatcag catgccgcgg tgaatacgtt ccggggcctt 840 gtacacaccg cccgtcacac catgagagtt tgtaacaccc aaagtcggtg gggtaacctt 900 ttaggaacca gccgcctaag gtgggacaga tgattagggt gaagtcgtaa caagggtaac 960 960

Claims (7)

As a Lactobacillus plantarum LRCC 5314 ( Lactobacillus plantarum LRCC 5314) strain having a hypoglycemic, antidiabetic, anti-inflammatory or stress-relieving effect,
The strain is a heat-treated dead cell, strain.
According to claim 1,
The strain modulates inflammatory factors of TNF-a, IL-6 or CCL-2,
The strain regulates blood sugar-related genes of GLUT4, Adiponectin or Leptin,
The strain expresses a stress improvement related gene of NPY or Y2R, or
The strain is a strain that regulates excessive secretion of cortisol in the blood.
delete A composition for lowering blood sugar comprising the heat-treated Lactobacillus plantarum LRCC5314 strain of claim 1 as an active ingredient.
A food composition for preventing or improving diabetes, comprising the heat-treated Lactobacillus plantarum LRCC5314 strain of claim 1 as an active ingredient.
A food composition for anti-inflammatory, comprising the heat-treated Lactobacillus plantarum LRCC5314 strain of claim 1 as an active ingredient.
A food composition for improving cortisol hypersecretion, comprising the heat-treated Lactobacillus plantarum LRCC5314 strain of claim 1 as an active ingredient.

Images