Lactobacillus fermentum strain and composition for preventing or treating metabolic diseases comprising same

Abstract

Provided are a novel Lactobacillus fermentum strain (accession number KCTC 14106BP) and a composition containing the same for preventing or treating metabolic diseases. The Lactobacillus fermentum strain of the present invention can inhibit fat accumulation in white adipose tissue and increase anti-inflammatory cells. In addition, Lactobacillus fermentum can effectively inhibit fat accumulation in brown adipose tissue and liver tissue. The Lactobacillus fermentum strain exhibits excellent blood glucose improving effects, particularly reducing fasting blood glucose levels. The Lactobacillus fermentum strain can effectively improve insulin resistance by improving glucose tolerance and increasing insulin sensitivity. Therefore, the Lactobacillus fermentum strain can be useful for preventing or treating metabolic diseases such as obesity.

Description

[Technical field]

[0001] The present disclosure relates to novel Lactobacillus fermentum strains and compositions comprising same for preventing or treating metabolic diseases. [Background technology]

[0002] As society develops, obesity appears as one of the serious diseases, and the World Health Organization (WHO) recognizes obesity as a disease that should be treated. Recently, a trend toward increasing prevalence of obesity has also been observed in the Republic of Korea due to a Westernized diet, and treatment and prevention for obesity have become an increasing concern. Obesity refers to a state of excessive accumulation of body fat resulting from an imbalance between food consumption and energy consumption. In addition, obesity is closely related to insulin resistance, glucose tolerance, hyperlipidemia, etc., and may be accompanied by metabolic diseases including cardiovascular disease, fatty liver disease, cancer, and diabetes as complications.

[0003] Recently, it has become known that inflammatory response in adipose tissue of obese patients increases, so obesity may be considered as low-grade systemic inflammation.In particular, it has been reported that inflammatory response increases due to inflammatory macrophages that increase in proportion to the size of adipose tissue, and inflammatory adipokines produced and secreted by adipose tissue in this condition are pathogenic factors of metabolic diseases such as cardiovascular disease and diabetes.Therefore, obesity, which increases these molecules, may be seen as the cause of almost all adult diseases.

[0004] Currently prescribed obesity drugs include Xenical (Roche Pharmaceuticals., Switzerland), Reductil (Abbott Co., USA) and Exolise (Arkopharma LLC, France). Obesity drugs are broadly classified into appetite suppressants, energy expenditure promoters, or fat absorption inhibitors, and most obesity drugs are appetite suppressants that suppress appetite by regulating neurotransmitters related to the hypothalamus. However, conventional obesity drugs have side effects such as cardiac disease, respiratory disease, and nervous system disease, and also have problems in that they have low in vivo durability. Therefore, there is a need to develop a safe and effective obesity treatment.

[0005] On the one hand, probiotics using safe microorganisms (e.g., lactic acid bacteria) to prevent or treat obesity have been actively studied. In particular, studies have shown that lactic acid bacteria exhibit effects such as maintaining normal intestinal flora, improving intestinal flora, antidiabetic and antihyperlipidemic effects, inhibiting carcinogenesis, inhibiting colon cancer, and non-specific activation of the host immune system.

[0006] Regarding lactic acid bacteria related to obesity prevention and treatment effects, Korean Patent No. 10-1494279 discloses Lactobacillus plantarum KY1032 strain (Accession No. KCCM-10430) having an inhibitory effect on adipocyte differentiation, Korean Patent No. 10-0996577 discloses Lactobacillus curvatus HY7601 (Accession No. KCTC 11456BP) having an obesity inhibitory effect, and Korean Patent No. 10-1394348 discloses Lactobacillus plantarum DSR920 strain (Accession No. KCCM 11210P) having an inhibitory effect on adipocyte differentiation, but none of these are mature enough to achieve commercial success.

[0007] Therefore, there is a need to continue to search for new strains with superior anti-obesity effects. Summary of the Invention [Problem to be solved by the invention]

[0008] Therefore, as a result of research to develop a new strain having excellent anti-obesity effects, the inventors discovered that Lactobacillus fermentum strain (accession number KCTC 14106BP) exhibits excellent anti-obesity effects, thereby completing the present invention. [Means for solving the problem]

[0009] One aspect of the present invention provides a Lactobacillus fermentum strain (Accession No. KCTC 14106BP).

[0010] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a metabolic disease comprising, as an active ingredient, a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

[0011] Yet another aspect of the present invention provides a food composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (accession number KCTC 14106BP) or a culture thereof.

[0012] Yet another aspect of the present invention provides a feed composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

[0013] Yet another aspect of the present invention provides a method for preventing and treating metabolic disease comprising administering to a subject a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof. Effect of the Invention

[0014] The present disclosure relates to a novel Lactobacillus fermentum strain (Accession No. KCTC 14106BP) and a composition for preventing or treating metabolic diseases comprising the same. The Lactobacillus fermentum according to the present invention can inhibit fat accumulation in white adipose tissue and increase anti-inflammatory cells. In addition, the Lactobacillus fermentum according to the present invention can effectively inhibit fat accumulation in brown adipose tissue and liver tissue. The Lactobacillus fermentum strain can show excellent blood glucose improving effect, in particular, can reduce fasting blood glucose level. The Lactobacillus fermentum strain can effectively improve insulin resistance by improving glucose tolerance and increasing insulin sensitivity. Therefore, the Lactobacillus fermentum strain can be usefully used to prevent or treat metabolic diseases such as obesity.

[0015] The illustrative embodiments can be more fully understood from the following description taken in conjunction with the accompanying drawings. [Brief description of the drawings]

[0016] [Figure 1] FIG. 1 is a graph showing the body weight of mice fed a high-fat diet following oral administration of each of the 16 strains of lactic acid bacteria compared to mice fed only a high-fat diet. [Diagram 2] FIG. 1 is a graph showing the body weight of mice fed a high-fat diet following oral administration of each of the 16 strains of lactic acid bacteria compared to mice fed only a high-fat diet. [Diagram 3] FIG. 1 is a graph showing the body weight of mice fed a high-fat diet following oral administration of each of the 16 strains of lactic acid bacteria compared to mice fed only a high-fat diet. [Figure 4] For comparison, this figure compares the size of adipocytes in white adipose tissue from mice fed a normal chow diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered general lactic acid bacteria, and mice fed a high-fat diet and orally administered L. fermentum GB103 strain. [Diagram 5]1 is a graph comparing the numbers of M1 macrophages, M2 macrophages, and regulatory T (Treg) cells in white adipose tissue from mice fed a normal chow diet, mice fed a high-fat diet, mice fed a high-fat diet and oral administration of general lactic acid bacteria, and mice fed a high-fat diet and oral administration of L. fermentum GB103 strain. [Figure 6] These images compare the size of adipocytes in brown adipose tissue from mice fed a normal chow diet, mice fed a high-fat diet, mice fed a high-fat diet and oral administration of general lactic acid bacteria, and mice fed a high-fat diet and oral administration of L. fermentum strain GB103. [Figure 7] This is a graph comparing the liver weights of mice fed a normal solid feed diet, mice fed a high-fat diet, mice fed a high-fat diet and oral administration of general lactic acid bacteria, and mice fed a high-fat diet and oral administration of L. fermentum GB103 strain. [Figure 8] These images compare the degree of fat accumulation in liver tissue of mice fed a normal solid feed diet, mice fed a high-fat diet, mice fed a high-fat diet and oral administration of general lactic acid bacteria, and mice fed a high-fat diet and oral administration of L. fermentum GB103. [Figure 9] This is a graph comparing fasting blood glucose levels of mice fed a normal solid feed diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered general lactic acid bacteria, and mice fed a high-fat diet and orally administered L. fermentum GB103 strain to confirm the blood glucose lowering effect. [Figure 10] This is a graph comparing blood glucose levels over time after glucose administration in mice fed a normal solid feed diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered general lactic acid bacteria, and mice fed a high-fat diet and orally administered L. fermentum GB103 strain to confirm the effect of improving glucose tolerance. [Figure 11]This is a graph comparing blood glucose levels over time after insulin administration in mice fed a normal solid feed diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered general lactic acid bacteria, and mice fed a high-fat diet and orally administered L. fermentum GB103 strain to confirm the effect of improving insulin resistance.

[0017] [Best Mode for Carrying Out the Invention] One aspect of the present invention provides a Lactobacillus fermentum strain (Accession No. KCTC 14106BP).

[0018] Lactobacillus is an aerobic or facultative anaerobic Gram-positive rod-shaped bacterium that is widely distributed in nature. The genus Lactobacillus includes L. fermentum, L. sakei, etc. The present inventors selected a novel Lactobacillus fermentum strain with excellent anti-obesity effects, which they named "Lactobacillus fermentum GB103". The strain was deposited at the Korea Collection for Type Cultures, Korea Research Institute of Bioscience and Biotechnology on September 6, 2019 under the accession number SD1342. The strain was deposited at the Korea Collection for Type Cultures, Korea Research Institute of Bioscience and Biotechnology on January 14, 2020 under the accession number KCTC 14106BP. In addition, the strain belongs to a probiotic strain, is harmless to the human body, and can be used without side effects.

[0019] As used herein, the term "Lactobacillus fermentum GB103" is interchangeably referred to as L. fermentum GB103 or Lactobacillus fermentum strain (Accession No. KCTC 14106BP).

[0020] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating a metabolic disease comprising, as an active ingredient, a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

[0021] The Lactobacillus fermentum strain (Accession No. KCTC 14106BP) is the same as above. In this case, the strain may be live or dead, with live strains being preferred. In addition, the culture of the strain may or may not contain the strain, with strains being preferred.

[0022] The composition comprises a therapeutically effective amount, or a nutritionally effective concentration, of the active ingredient Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof, based on the total weight of the composition, wherein the active ingredient is 10 4 ~10 16 CFU / g, preferably 10 6 ~10 12 CFU / g content or cultures with an equivalent number of viable strains. Generally, for adult patients, 1 × 10 6 Live strains of at least 1 × 10 CFU / g, preferably 8 ~1×10 12 The CFU / g of live strain may be administered in a single dose or in several doses.

[0023] As used herein, the term "metabolic disease" is also called metabolic syndrome, and is a disease assumed to be caused by insulin resistance, and is a condition with abnormalities in two or more of cholesterol, blood pressure, and blood glucose levels.Metabolic syndrome is a conceptualization of the phenomenon in which various cardiovascular disease and type 2 diabetes risk factors are clustered together as a disease group, and includes insulin resistance and all of the complex and various metabolic abnormalities and clinical features related to insulin resistance.In particular, the metabolic disease can be any one selected from the group consisting of obesity, hypertension, arteriosclerosis, hyperlipidemia, fatty liver, non-alcoholic fatty liver disease, hyperinsulinemia, diabetes, and insulin resistance syndrome.

[0024] As used herein, the term "obesity" is also known as adiposity, and is a disease in which excess body fat is abnormally accumulated. Irregular eating habits, excessive food intake, lack of physical activity, endocrine system disease, genetic factors, psychological factors, drugs, etc. can cause obesity. In addition, obesity increases the incidence of arteriosclerosis, cardiovascular disease (stroke and ischemic cardiovascular disease), hypertension, diabetes, hyperlipidemia, fatty liver, cholelithiasis, obstructive sleep apnea, menstrual irregularities, polycystic ovarian disease, infertility, libido loss, depression, osteoarthritis, gout, etc. Obesity can be simple obesity, symptomatic obesity, childhood obesity, adult obesity, cell proliferation obesity, cell hypertrophy obesity, upper body obesity, lower body obesity, visceral fat obesity, or subcutaneous fat obesity.

[0025] As used herein, the term "hypertension" refers to the phenomenon of increased perfusion pressure of blood flowing through an artery. Hypertension can generally be diagnosed when the systolic blood pressure reaches 140 mmHg and the diastolic blood pressure reaches 90 mmHg or higher. Hypertension exists as primary (or essential) hypertension, which has no noticeable symptoms and no known cause, and secondary hypertension, which is caused by renal disease, endocrine disease, pregnancy toxemia, etc. Most cases of hypertension (90-95%) are primary hypertension, which is assumed to be caused by genetic causes combined with environmental factors such as obesity, stress, alcohol, and smoking.

[0026] As used herein, the term "arteriosclerosis" is defined as the phenomenon of narrowing of the arterial wall due to loss of elasticity of the arterial wall, proliferation of abnormal tissue, and accumulation of fat inside the intima of the arterial wall.Arteriosclerosis refers to the pathological changes of arteries, and is a term named according to the organ affected by arteriosclerosis.For example, arteriosclerosis includes, but is not limited to, cerebral infarction caused by arteriosclerosis, and myocardial infarction caused by coronary atherosclerosis.

[0027] As used herein, the term "hyperlipidemia" refers to a disease caused by a large amount of lipids, such as triglycerides and cholesterol, in the blood due to improper lipid metabolism.In particular, hyperlipidemia refers to a state in which lipid components, such as triglycerides, LDL cholesterol, and free fatty acids, in the blood increase.Hyperlipidemia includes, but is not limited to, hypercholesterolemia or hypertriglyceridemia.

[0028] As used herein, the term "fatty liver" refers to the state of excessive accumulation of fat in liver cells caused by lipid metabolism disorder, and is defined as the case where fat reaches 5% or more of liver weight. This causes various diseases such as angina pectoris, myocardial infarction, stroke, arteriosclerosis, fatty liver, pancreatitis, etc. Fatty liver is divided into alcoholic fatty liver caused by alcohol consumption and non-alcoholic fatty liver disease (NAFLD) that is not caused by alcohol.

[0029] As used herein, the term "non-alcoholic fatty liver disease" refers to the case where fatty liver is not caused by alcohol, and includes a series of liver damage processes ranging from simple steatosis in liver to non-alcoholic steatohepatitis (NASH) to cirrhosis.The causes of non-alcoholic fatty liver disease are side effects of drugs such as antiarrhythmic drugs, antiviral drugs, steroids, or cytotoxic drugs; excessive calorie consumption such as carbohydrates; obesity; diabetes; and some genetic causes.

[0030] As used herein, the term "hyperinsulinemia" refers to a disease in which an excess of insulin is present in the blood. Insulin is a hormone secreted by the pancreas that controls blood sugar levels and promotes glucose influx into muscles and other peripheral tissues. Thus, hyperinsulinemia can occur due to disorders of the pancreas, the insulin-secreting organ, and is most commonly caused by insulin resistance.

[0031] As used herein, the term "diabetes" refers to a chronic disease characterized by a relative or absolute deficiency of insulin resulting in glucose intolerance. Diabetes includes all types of diabetes, and may be, for example, but not limited to, type 1 diabetes, type 2 diabetes, and hereditary diabetes. Type 1 diabetes is insulin-dependent diabetes that results mainly from the destruction of β-cells. As used herein, the term "type 2 diabetes" refers to non-insulin-dependent diabetes, which is caused by insulin resistance. Type 2 diabetes is caused because no increase in insulin is detected in muscle and fat tissue, or even if an increase in insulin is detected, the action of insulin does not occur effectively.

[0032] As used herein, the term "insulin resistance" refers to a state in which cells do not respond to insulin to lower blood glucose levels, and therefore cannot burn glucose effectively. When insulin resistance is high, the body thinks it needs more insulin and produces more insulin. This results in hyperinsulinemia, hypertension, or dyslipidemia, as well as heart disease and diabetes.

[0033] As used herein, the term "insulin resistance syndrome" is a collective term for diseases caused by insulin resistance, which is characterized by cellular resistance to insulin action, hyperinsulinemia and increased very low density lipoprotein (VLDL) and triglycerides, and decreased high density lipoprotein (HDL), hypertension, etc., and is recognized as a risk factor for cardiovascular disease and type 2 diabetes.

[0034] According to an embodiment of the present invention, the strain can inhibit fat accumulation in white adipose tissue. In particular, in an embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet resulted in a significant reduction in the size of adipocytes in the white adipose tissue of the mouse model. Meanwhile, when general lactic acid bacteria was orally administered to a mouse model fed a high-fat diet, the size of adipocytes in the white adipose tissue of the mouse model did not decrease. Based on this, it was confirmed that the strain inhibits fat accumulation in white adipose tissue (Figure 4).

[0035] According to an embodiment of the present invention, the strain can reduce meta-inflammation in white adipose tissue. As used herein, the term "meta-inflammation" refers to chronic and low-grade inflammation, which is an inflammatory response that occurs when an excessive amount of nutrients or metabolites are provided. In particular, it is known that the chronic inflammatory response caused by obesity plays an important role in the process of increasing insulin resistance and developing metabolic disorders.

[0036] In one embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet resulted in a decrease in M1 macrophages and a significant increase in M2 macrophages and Treg cells in the white adipose tissue of the mouse model. Meanwhile, when general lactobacillus was orally administered to a mouse model fed a high-fat diet, M1 macrophages in the white adipose tissue of the mouse model were significantly decreased, but M2 and Treg cells were unchanged. Based on this, it was confirmed that the strain inhibited inflammatory responses in white adipose tissue (Figure 5).

[0037] According to an embodiment of the present invention, the strain can inhibit fat accumulation in brown adipose tissue. In particular, in an embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet resulted in a significant reduction in the size of adipocytes in the brown adipose tissue of the mouse model. Meanwhile, when general lactic acid bacteria was orally administered to a mouse model fed a high-fat diet, the size of adipocytes in the brown adipose tissue of the mouse model did not decrease. Based on this, it was confirmed that the strain inhibits fat accumulation in brown adipose tissue (Figure 6).

[0038] According to an embodiment of the present invention, the strain can reduce liver weight and inhibit fat accumulation in liver tissue. In particular, in an embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet resulted in reduced liver weight and fat accumulation in the tissue of the mouse model. Meanwhile, when general lactic acid bacteria was orally administered to a mouse model fed a high-fat diet, a lot of fat accumulated in the liver tissue of the mouse model, and there was no change in liver weight. Based on this, it was confirmed that the strain reduced liver weight and inhibited fat accumulation in tissue (Figures 7 and 8).

[0039] According to an embodiment of the present invention, the strain can reduce blood glucose levels. Preferably, the strain can reduce fasting blood glucose levels and improve glucose tolerance. In particular, in an embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet resulted in a reduction in fasting blood glucose levels and blood glucose levels after administration of glucose in the mouse model. Meanwhile, when general lactobacillus was orally administered to a mouse model fed a high-fat diet, fasting blood glucose levels and blood glucose levels after administration of glucose were not reduced. Based on this, it was confirmed that the strain reduced fasting blood glucose levels and improved glucose tolerance (Figures 9 and 10).

[0040] According to an embodiment of the present invention, the strain can improve insulin resistance. In particular, in an embodiment of the present invention, when the strain is orally administered to a mouse model fed a high-fat diet, blood glucose levels are significantly reduced in the mouse model after insulin administration. Meanwhile, when general lactobacillus is orally administered to a mouse model fed a high-fat diet, fasting blood glucose levels and blood glucose levels after insulin administration are not reduced. Based on this, it was confirmed that the strain improves insulin resistance by increasing insulin sensitivity (FIG. 11).

[0041] The composition may further comprise a cryoprotectant or excipient. In particular, the cryoprotectant may be one or more selected from the group consisting of glycerol, trehalose, maltodextrin, skim milk powder, and starch. In addition, the excipient may be one or more selected from the group consisting of glucose, dextrin, and skim milk powder.

[0042] The composition may comprise 0.01wt%-20wt% or 0.01wt%-10wt% cryoprotectant based on the total weight of the composition, specifically, the composition may comprise 5wt%-20wt% glycerol, 2wt%-10wt% trehalose, 2wt%-10wt% maltodextrin, 0.5wt%-2wt% skim milk powder, and 0.1wt%-1wt% starch. Additionally, the composition may comprise 75wt%-95wt% or 85wt%-95wt% excipient based on the total weight of the composition.

[0043] Yet another aspect of the present invention provides a food composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (accession number KCTC 14106BP) or a culture thereof.

[0044] The Lactobacillus fermentum strain (accession number KCTC 14106BP) was the same as described above.

[0045] In this case, the metabolic disease is as described above, and in particular the strain may be a food composition for preventing or inhibiting obesity.

[0046] The food compositions include all forms of functional foods, dietary supplements, health foods, food additives, etc., and the types of food compositions can be prepared in various forms according to conventional methods known in the art.

[0047] When the strain is used as a food additive, the strain can be added as it is or can be used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment). In general, when preparing a food or beverage, the active ingredient can be added in an amount of 0.0001wt% to 1wt%, specifically 0.001wt% to 0.1wt%, in the raw material composition containing the strain. However, in the case of long-term intake for health and hygiene or health management purposes, the amount may be below the above range.

[0048] Yet another aspect of the present invention provides a feed composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

[0049] The Lactobacillus fermentum strain (accession number KCTC 14106BP) was the same as described above.

[0050] A feed composition for preventing or ameliorating metabolic diseases can be prepared by adding a suitable effective concentration range of Lactobacillus fermentum strain (accession number KCTC 14106BP) according to various methods for preparing feed compositions known in the art.

[0051] Yet another aspect of the present invention provides a method for preventing and treating metabolic disease comprising administering to a subject a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

[0052] The subject may have a metabolic disease. In addition, the subject may be a mammal, preferably a human. In this case, the Lactobacillus fermentum strain (accession number KCTC 14106BP) is the same as described above. In addition, the administration route, dosage, and administration frequency of the strain or its culture may be administered to the subject in various ways and amounts depending on the patient's condition and the presence or absence of side effects, and the optimal administration method, dosage, and administration frequency can be appropriately selected within an appropriate range by a person skilled in the art. In addition, the type of metabolic disease is as described above.

[0053] Yet another aspect of the present invention provides use of a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof for treating a metabolic disease.

[0054] In this case, the Lactobacillus fermentum strain (accession number KCTC 14106BP) was the same as that described above. In addition, the type of metabolic disease was as described above. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0055] The present invention will now be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited to the following examples. EXAMPLES

[0056] Example 1. Screening of strains with anti-obesity effects Among 16 strains of lactobacilli purchased from Mediogen Co., lactobacilli with anti-obesity efficacy were selected using a mouse model. In particular, C57BL / 6 mice fed a 60% high-fat diet (HFD) and then orally administered each lactobacillus were used as the experimental group. Mice fed only a 60% high-fat diet were used as the control group. In this case, each of the lactobacilli was daily administered at 5 × 10 9 CFU / mouse were orally administered. The anti-obesity effect was determined based on the body weight difference between the experimental and control groups.

[0057] As a result, Lactobacillus fermentum GB103 strain was the most effective in inhibiting weight gain. In this case, there were some L. fermentum lactic acid bacteria that did not show anti-obesity effects (Figures 1 to 3). Based on this, it was confirmed that Lactobacillus fermentum GB103 strain shows anti-obesity effects among L. fermentum lactic acid bacteria.

[0058] Example 2. Confirmation of the lipid accumulation inhibitory effect of L. fermentum GB103 strain in white adipose tissue A mouse model was used to confirm the fat accumulation inhibitory effect of L. fermentum GB103 (accession number KCTC 14106BP) strain in white adipose tissue. In detail, C57BL / 6 mice fed with a 60% high-fat diet (HFD) and then orally administered with L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as the negative control group, and mice fed with a 60% high-fat diet and orally administered with general lactic acid bacteria (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed with a normal solid diet (NCD) were used as the normal group. In this case, general lactic acid bacteria purchased from Mediogen Co. or L. fermentum GB103 strain were added at 5 × 10 9 CFU / mouse were orally administered daily.

[0059] After the administration of each test substance is completed, white adipose tissue is separated by autopsy. The separated white adipose tissue is fixed with 10% neutral buffered formalin. Then, paraffin sections are prepared and stained with hematoxylin and eosin (H&E), and then cell size is observed.

[0060] The results showed that the size of adipocytes in the negative and positive control groups fed a high-fat diet increased compared to the normal group, while the size of adipocytes in the experimental group orally administered L. fermentum GB103 strain was significantly decreased (Figure 4). Based on this, it was confirmed that L. fermentum GB103 strain effectively inhibits fat accumulation in white adipose tissue.

[0061] Example 3. Confirmation of immune cell regulation effect of L. fermentum GB103 strain in white adipose tissue A mouse model was used to confirm the changes in immune cells caused by administration of L. fermentum GB103 strain in white adipose tissue. In detail, C57BL / 6 mice fed a 60% (HFD) diet and then orally administered L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as the negative control group, and mice fed a 60% high-fat diet and oral administration of general Lactobacillus casei (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed a normal solid diet (NCD) were used as the normal group. In this case, general Lactobacillus casei or L. fermentum GB103 strain purchased from Mediogen Co. were added at 5 × 10 9 CFU / mouse were orally administered daily.

[0062] After the administration of each test substance was completed, immune cells were isolated from the epididymal white adipocyte tissue of the mice in each group by autopsy, and the cells were stained with specific antibodies and then analyzed by flow cytometry. The adipose tissue was minced with sharp scissors and incubated with 2 mg / mL collagen Type II (Worthington) at 37°C for 45 minutes, and then the stromal vascular fraction (SVF) was isolated by centrifugation. After washing with FACS buffer, it was then blocked at 4°C for 10 minutes to inhibit non-specific binding. Anti-MHCII (Biolegend, CA, USA), anti-F4 / 80 (Biolegend, CA, USA), anti-CD206 (Biolegend, CA, USA), anti-CD11c (Biolegend, CA, USA), and anti-CD11b (Biolegend, CA, USA) antibodies were added to M1 / M2 macrophages, reacted at 4°C for 30 minutes, and then washed with FACS buffer. Treg cells were subjected to the same procedure as above using anti-TCRβ (Biolegend, CA, USA) and anti-CD4 (Biolegend, CA, USA) antibodies, then treated with EBIOSCIENCE™ Foxp3 / Transcription Factor Staining Buffer for fixation and permeabilization, and stained with anti-Foxp3 antibody (eBioscience, CA, USA). Among MHCII+F4 / 80+ macrophages, CD11b+CD206+ cells were analyzed in M2 macrophages, and CD11b+CD11c+ cells were analyzed in M1 macrophages. Treg cells were analyzed in TCRβ+CD4+Foxp3+ cells.

[0063] The results showed that the amount of inflammatory M1 macrophages in the negative and positive control mice fed a high-fat diet was higher than that in the normal group, while the amount of M1 macrophages in the experimental mice orally administered L. fermentum GB103 strain was significantly lower than that in the negative control group. It was also confirmed that the amount of anti-inflammatory M2 macrophages and Treg cells in the negative and positive control mice fed a high-fat diet was lower than that in the normal group, while the amount of M2 macrophages and Treg cells in the experimental mice orally administered L. fermentum GB103 strain was significantly increased compared to the negative control group. Based on this, the immune cell regulating effect of L. fermentum GB103 strain in white adipose tissue was confirmed (Figure 5).

[0064] Example 4. Confirmation of the lipid accumulation inhibitory effect of L. fermentum GB103 strain in brown adipose tissue Using a mouse model, the fat accumulation inhibitory effect of L. fermentum GB103 strain in brown adipose tissue was confirmed. In detail, C57BL / 6 mice fed a 60% (HFD) diet and then orally administered L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as the negative control group, and mice fed a 60% high-fat diet and orally administered general lactic acid bacteria (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed a normal solid diet (NCD) were used as the normal group. In this case, general lactic acid bacteria purchased from Mediogen Co. or L. fermentum GB103 strain were added at 5 × 10 9 CFU / mouse were orally administered daily.

[0065] After the administration of each test substance is completed, the brown adipose tissue is separated from the mice in each group by autopsy.The separated brown adipose tissue is fixed with 10% neutral buffered formalin.Then, paraffin sections are prepared and stained with H&E, and then the cell size is observed.

[0066] The results showed that the size of adipocytes in the negative and positive control mice fed a high-fat diet increased compared to the normal group, while the size of adipocytes in the experimental group orally administered with L. fermentum GB103 strain was significantly decreased (Figure 6). Based on this, it was confirmed that L. fermentum GB103 strain effectively inhibits fat accumulation in brown adipose tissue.

[0067] Example 5. Confirmation of lipid accumulation inhibitory effect of L. fermentum GB103 strain in liver tissue A mouse model was used to confirm the lipid accumulation inhibitory effect of L. fermentum GB103 strain in liver tissue. In detail, C57BL / 6 mice fed a 60% (HFD) diet and then orally administered L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as the negative control group, and mice fed a 60% high-fat diet and orally administered general lactic acid bacteria (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed a normal solid diet (NCD) were used as the normal group. In this case, general lactic acid bacteria purchased from Mediogen Co. or L. fermentum GB103 strain were added at 5 × 10 9 CFU / mouse were orally administered daily.

[0068] After the administration of each test substance was completed, the liver was excised by autopsy, and the liver weight was measured to compare and evaluate the liver weight between groups. The excised liver was fixed with 10% buffered formalin, and then paraffin sections were prepared and stained with H&E, and then the cell size was observed.

[0069] The results showed that the liver weights of the negative and positive control mice fed a high-fat diet were increased compared to the normal group, while the liver weights of the experimental mice orally administered L. fermentum GB103 strain were significantly decreased compared to the negative control group (Figure 7). Furthermore, compared to the normal group, fat accumulation in liver tissues was increased in the negative and positive control mice fed a high-fat diet. However, almost no fat was accumulated in the liver tissues of the experimental mice orally administered L. fermentum GB103 strain, and the fat accumulation level was similar to that of the negative control group (Figure 8). Based on this, it was confirmed that L. fermentum GB103 strain effectively inhibited liver weight and fat accumulation in liver tissues.

[0070] Example 6: Confirmation of the fasting blood glucose lowering and glucose tolerance improving effects of L. fermentum GB103 strain A glucose tolerance test (GTT) was performed to confirm the glucose tolerance improving effect of L. fermentum GB103 strain. First, C57BL / 6 mice fed a 60% (HFD) and then orally administered L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as the negative control group, and mice fed a 60% high-fat diet and orally administered general lactic acid bacteria (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed a normal solid diet (NCD) were used as the normal group. In this case, general lactic acid bacteria purchased from Mediogen Co. or L. fermentum GB103 strain were used at 5 × 10 9 CFU / mouse were orally administered daily.

[0071] To carry out glucose tolerance test, mice were fasted for more than 16 hours, and then glucose solution was injected intraperitoneally at a dose of 1g / kg. Blood was then collected from the tail vein of the mice at 0, 30, 60, 90, and 120 minutes, and blood glucose was measured using a glucometer. In this case, blood glucose at 0 minutes represents fasting blood glucose.

[0072] The results showed that fasting blood glucose levels were significantly increased in the negative and positive control mice fed a high-fat diet compared to the normal group, while the fasting blood glucose levels of the experimental mice orally administered L. fermentum GB103 strain were significantly decreased compared to the negative control group (Figure 9).

[0073] In addition, it was confirmed that the blood glucose levels after glucose administration in the negative and positive control mice fed a high-fat diet were higher than those in the normal group, while the blood glucose levels of the experimental mice orally administered L. fermentum GB103 strain were significantly lower than those of the negative control group (Figure 10). Based on this, it was confirmed that L. fermentum GB103 strain exhibits the effects of lowering fasting blood glucose and improving glucose tolerance.

[0074] Example 7. Confirmation of the effect of L. fermentum GB103 strain in improving insulin resistance An insulin tolerance test (ITT) was performed to confirm the insulin resistance-improving effect of L. fermentum GB103 strain. First, C57BL / 6 mice fed a 60% (HFD) diet and then orally administered L. fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as the negative control group, and mice fed a 60% high-fat diet and orally administered general lactic acid bacteria (L. plantarum MG5120) were used as the positive control group. Furthermore, mice fed a normal solid diet (NCD) were used as the normal group. In this case, general lactic acid bacteria purchased from Mediogen Co. or L. fermentum GB103 strain were used at 5 × 10 9 CFU / mouse were orally administered daily.

[0075] To perform the insulin tolerance test, the mice were fasted for 4.5 hours, and then insulin solution was administered intraperitoneally at a dose of 1 U / kg. Blood was collected from the tail vein of the mice at 0, 30, 60, 90 and 120 minutes, and then blood glucose was measured by a glucometer.

[0076] The results confirmed that the blood glucose levels after insulin administration in the negative and positive control mice fed a high-fat diet were higher than those in the normal group, while the blood glucose levels of the experimental mice orally administered L. fermentum GB103 strain were significantly lower than those of the negative control group (Figure 11). Based on this, it was confirmed that L. fermentum GB103 strain effectively improves insulin resistance by increasing insulin sensitivity. [Accession number]

[0077] <Accession number> Depository institution: Korean Collection for Type Cultures (KCTC), Korea Research Institute of Bioscience and Biotechnology Accession number: KCTC14106BP Date of deposit: 20200114

Claims

1. Lactobacillus fermentum strain (accession number KCTC 14106BP).

2. A pharmaceutical composition for preventing or treating a metabolic disease, comprising as an active ingredient a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

3. 3. The pharmaceutical composition for preventing or treating a metabolic disease according to claim 2, wherein the metabolic disease is any one selected from the group consisting of obesity, hypertension, arteriosclerosis, hyperlipidemia, fatty liver, non-alcoholic fatty liver disease, hyperinsulinemia, diabetes mellitus, and insulin resistance syndrome.

4. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein said strain inhibits fat accumulation in white adipose tissue.

5. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein said strain reduces metabolic inflammation in white adipose tissue.

6. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein said strain inhibits fat accumulation in brown adipose tissue.

7. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein said strain inhibits fat accumulation in liver tissue.

8. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein said strain reduces blood glucose levels.

9. The pharmaceutical composition for preventing or treating a metabolic disease according to claim 2, wherein said strain reduces insulin resistance.

10. A food composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

11. A feed composition for preventing or inhibiting a metabolic disease, comprising a Lactobacillus fermentum strain (Accession No. KCTC 14106BP) or a culture thereof.

12. Use of a Lactobacillus fermentum strain (accession number KCTC 14106BP) or a culture thereof for the manufacture of a pharmaceutical composition for preventing or treating a metabolic disease.

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