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

Abstract

The present invention relates to a novel fermented lactobacillus strain (preservation number KCTC 14106BP) and a composition for preventing or treating metabolic diseases comprising the strain. The fermented lactobacillus of the present invention can inhibit fat accumulation and increase anti-inflammatory cells in white adipose tissue, and can effectively inhibit fat accumulation in brown adipose tissue and liver tissue. The fermented lactobacillus strain shows an excellent hypoglycemic effect, particularly can reduce fasting blood sugar level, and can effectively improve insulin resistance by improving glucose tolerance and increasing insulin sensitivity. Therefore, the fermented lactobacillus strain can be usefully used to prevent or treat metabolic diseases such as obesity.

Description

Technical Field

[0001] The present disclosure relates to a novel Lactobacillus fermentum strain and a composition comprising the strain for preventing or treating metabolic diseases. Background Art

[0002] As society develops, obesity has become a serious disease, and the World Health Organization (WHO) has recognized obesity as a disease that requires treatment. Recently, a trend of increasing obesity prevalence due to a Westernized diet has also been observed in Korea, and interest in the treatment and prevention of obesity has been growing. Obesity refers to a state of excessive accumulation of body fat due to an imbalance between food consumption and energy expenditure. 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 been known that inflammatory responses are increased in adipose tissue in obese patients, and obesity is sometimes considered a form of low-grade systemic inflammation. In particular, inflammatory macrophages, which increase in proportion to the size of adipose tissue, have been reported to increase inflammatory responses, and inflammatory adipokines produced and secreted by adipose tissue in this state are pathogenic factors for metabolic diseases such as cardiovascular disease and diabetes. Therefore, obesity, which increases these molecules, can be regarded as the cause of almost all adult diseases.

[0004] The obesity drugs currently prescribed include Xenical (Roche Pharmaceuticals., Switzerland), Reductil (Abbott Co., the U.S.) and Exolise (Arkopharma LLC, France) etc. Obesity drugs are mainly divided into appetite suppressants, energy consumption promoters or fat absorption inhibitors, and most obesity drugs are appetite suppressants, which suppress appetite by controlling neurotransmitters associated with the hypothalamus. However, conventional obesity drugs have side effects such as heart disease, respiratory diseases and nervous system diseases, and have the problem of low persistence in vivo. Therefore, it is necessary to develop safe and effective obesity treatment.

[0005] Meanwhile, probiotics using safe microorganisms such as lactic acid bacteria to prevent or treat obesity have been actively studied. In particular, studies have shown that lactic acid bacteria have effects such as maintaining normal intestinal flora, improving intestinal flora, anti-diabetic and anti-lipid effects, inhibiting carcinogenesis, inhibiting colon cancer, and nonspecific activity of the host immune system.

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

[0007] Therefore, there is a need to continue researching new strains with excellent anti-obesity effects. Summary of the invention

[0008] Technical issues

[0009] Therefore, the present inventors have studied and developed new strains having excellent anti-obesity effects, and found that the fermentation Lactobacillus strain (Accession No. KCTC 14106BP) exhibits excellent anti-obesity effects, thereby completing the present invention.

[0010] Solution

[0011] One aspect of the present invention provides a Lactobacillus fermentum strain (preservation number KCTC 14106BP).

[0012] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, wherein the pharmaceutical composition comprises a Lactobacillus fermentum strain (preservation number KCTC 14106BP) or a culture thereof as an active ingredient.

[0013] Another aspect of the present invention provides a food composition, comprising a Lactobacillus fermentum strain (preservation number KCTC 14106BP).

[0014] Another aspect of the present invention provides a feed composition for preventing or inhibiting metabolic diseases, the feed composition comprising a Lactobacillus fermentum strain (Deposit No. KCTC 14106BP) or a culture thereof.

[0015] Still another aspect of the present invention provides a method for preventing and treating metabolic diseases, the method comprising administering Lactobacillus fermentum strain (Deposit No. KCTC 14106BP) or a culture thereof to a subject.

[0016] Effects of the Invention

[0017] The present disclosure relates to a novel Lactobacillus fermentum (preservation number KCTC 14106BP) strain and a composition for preventing or treating metabolic diseases comprising the strain. 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 sugar improvement effects, particularly can reduce fasting blood sugar 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 usefully used to prevent or treat metabolic diseases such as obesity. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The exemplary embodiments may be understood in more detail through the following description in conjunction with the accompanying drawings, in which:

[0019] Figures 1 to 3 It is a graph showing the body weight of mice fed a high-fat diet after oral administration of 16 lactic acid bacteria strains, respectively, compared with mice fed a high-fat diet alone.

[0020] Figure 4 The graph compares the sizes of adipocytes in white adipose tissue from mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with Lactobacillus fermentum (L.fermentum) GB103 strain.

[0021] Figure 5 The graph compares the number of M1 macrophages, M2 macrophages, and regulatory T (Treg) cells in white adipose tissue of mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with Lactobacillus fermentum GB103 strain.

[0022] Figure 6 The graph compares the sizes of fat cells in brown adipose tissue of mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermentation Lactobacillus GB103 strain.

[0023] Figure 7 The graph compares the liver weights of mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermentation Lactobacillus GB103 strain.

[0024] Figure 8The graph compares the degree of fat accumulation in liver tissue of mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermentation Lactobacillus GB103 strain.

[0025] Fig. 9 The graph compares the fasting blood glucose levels of mice fed a normal diet, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermented Lactobacillus GB103 strain to confirm the blood glucose lowering effect.

[0026] Fig.10 is a graph comparing blood sugar levels over time after administration of glucose in mice fed a normal chow, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermentation Lactobacillus GB103 strain to confirm the glucose tolerance improving effect; and

[0027] Fig.11 This graph compares blood sugar levels over time after insulin administration in mice fed a normal chow, mice fed a high-fat diet, mice fed a high-fat diet and orally administered with common lactic acid bacteria, and mice fed a high-fat diet and orally administered with the fermentation Lactobacillus GB103 strain to confirm the insulin resistance-improving effect. DETAILED DESCRIPTION

[0028] One aspect of the present invention provides a Lactobacillus fermentum strain (preservation number KCTC 14106BP).

[0029] Lactobacillus is an aerobic or facultative anaerobic Gram-positive rod widely distributed in nature. The genus Lactobacillus includes Lactobacillus fermentum, Lactobacillus sakei, etc. The inventors selected a new strain of Lactobacillus fermentum with excellent anti-obesity effect and named it "Lactobacillus fermentum GB103". The strain was deposited in the Korea Collection for Type Cultures, Korea Research Institute of Bioscience and Biotechnology, Korea Research Institute of Bioscience and Biotechnology on September 6, 2019 with the deposit number SD1342. The same strain was deposited in the Korea Collection for Type Cultures, Korea Research Institute of Bioscience and Biotechnology, Korea Research Institute of Bioscience and Biotechnology on January 14, 2020 with the deposit 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.

[0030] As used herein, the term "Lactobacillus fermentum GB103" may be interchangeably described as Lactobacillus fermentum GB103 or Lactobacillus fermentum strain (Deposit No. KCTC 14106BP).

[0031] Another aspect of the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, wherein the pharmaceutical composition comprises a Lactobacillus fermentum strain (preservation number KCTC 14106BP) or a culture thereof as an active ingredient.

[0032] Lactobacillus fermentum strain (preserving number KCTC 14106BP) is the same as above. In this case, the strain can be alive or dead, and preferably live strain. In addition, the culture of the strain may contain or may not contain the strain, preferably contains the strain.

[0033] Based on the total weight of the composition, the composition comprises a therapeutically effective amount or a nutritionally effective concentration of an active ingredient, Lactobacillus fermentum strain (preservation number KCTC 14106BP) or a culture thereof, wherein the content thereof is 10 4 Up to 10 16 CFU / g, preferably 10 6 Up to 10 12 CFU / g, or including cultures with an equivalent amount of live strains. Generally speaking, for adult patients, 1×10 6 CFU / g or more of live strains, preferably 1×10 8 Up to 1×10 12 CFU / g of live strains.

[0034] As used herein, the term "metabolic disease", also referred to as metabolic syndrome, is a disease believed to be caused by insulin resistance and is a symptom of two or more abnormalities in cholesterol, blood pressure and blood sugar levels. Metabolic syndrome is a conceptualization of the phenomenon that the risk factors for various cardiovascular diseases and type 2 diabetes are clustered together as a disease group, and encompasses insulin resistance and all the complex and diverse metabolic abnormalities and clinical features associated with insulin resistance. Specifically, a metabolic disease can be any one selected from the group consisting of obesity, hypertension, arteriosclerosis, hyperlipidemia, fatty liver, nonalcoholic fatty liver disease, hyperinsulinemia, diabetes and insulin resistance syndrome.

[0035] As used herein, the term "obesity", also referred to as polyadiposity, is a disease in which excessive body fat is abnormally accumulated. Irregular eating habits, excessive food intake, lack of physical activity, endocrine system diseases, genetic factors, psychological factors, medication, etc. can all lead to obesity. In addition, obesity can increase the incidence of arteriosclerosis, cardiovascular disease (stroke and ischemic cardiovascular disease), hypertension, diabetes, hyperlipidemia, fatty liver, cholelithiasis, obstructive sleep apnea, menstrual disorders, polycystic ovarian disease, infertility, loss of libido, depression, degenerative arthritis, 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.

[0036] As used herein, the term "hypertension" refers to the phenomenon of increased perfusion blood pressure of blood flowing through arteries. If the systolic blood pressure reaches 140 mmHg and the diastolic blood pressure reaches above 90 mmHg, it can generally be diagnosed as hypertension. Hypertension has no obvious symptoms. There are primary (or idiopathic) hypertension of unknown cause, and secondary hypertension caused by kidney disease, endocrine disease, pregnancy poisoning, etc. Most cases of hypertension (90% to 95%) are primary hypertension, which is believed to be caused by genetic reasons as well as environmental factors (such as obesity, stress, alcohol and smoking).

[0037] As used herein, the term "atherosclerosis" is defined as a phenomenon in which the width of the arterial wall becomes narrower due to loss of elasticity of the arterial wall, abnormal tissue proliferation, and accumulation of fat in the inner layer of the arterial wall. Arteriosclerosis is a term for arterial lesions and is 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 artery sclerosis.

[0038] As used herein, the term "hyperlipidemia" is a disease caused by a large amount of lipids (such as triglycerides and cholesterol) in the blood due to the inability of lipid metabolism to proceed normally. Specifically, 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.

[0039] As used herein, the term "fatty liver" is a state in which fat is excessively accumulated in liver cells due to lipid metabolism disorder, and is defined as a condition in which fat reaches 5% or more of the 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 drinking and non-alcoholic fatty liver disease (NAFLD) which is not caused by alcohol.

[0040] As used herein, the term "non-alcoholic fatty liver disease" refers to a condition in which fatty liver is not caused by alcohol, and includes a range of liver damage processes, from simple fatty degeneration of the liver, non-alcoholic steatohepatitis (NASH) to cirrhosis. The causes of non-alcoholic fatty liver disease are: side effects of drugs such as antiarrhythmic agents, antiviral agents, steroids, or cytotoxic agents; excessive calorie consumption such as carbohydrates; obesity; diabetes; and some genetic causes.

[0041] As used herein, the term "hyperinsulinemia" is a condition in which there is an excess of insulin in the blood. Insulin is secreted by the pancreas and is a hormone that regulates blood sugar levels and promotes the flow of glucose into muscle and other peripheral tissues. Therefore, hyperinsulinemia may occur due to a disorder of the insulin-secreting organ, the pancreas, and is most often caused by insulin resistance.

[0042] As used herein, the term "diabetes" is a chronic disease characterized by a relative or absolute lack of insulin leading to glucose intolerance. Diabetes includes all types of diabetes, such as type 1 diabetes, type 2 diabetes and hereditary diabetes, but is not limited thereto. Type 1 diabetes is insulin-dependent diabetes, mainly caused by β-cell destruction. As used herein, the term "type 2 diabetes" is insulin-independent diabetes, which is caused by insulin resistance. Type 2 diabetes is caused by the fact that no increase in insulin is detected in muscle and adipose tissue, or even if insulin is detected, it does not act effectively.

[0043] As used herein, the term "insulin resistance" refers to a state in which cells do not respond to insulin, which lowers blood sugar levels, and therefore cannot burn glucose efficiently. When insulin resistance is high, the body thinks more insulin is needed and produces more insulin. This can lead to hyperinsulinemia, high blood pressure or dyslipidemia, as well as heart disease and diabetes.

[0044] As used herein, the term "insulin resistance syndrome" is a general term for diseases caused by insulin resistance. It is characterized by cellular resistance to the action of insulin, hyperinsulinemia and elevated very low-density lipoprotein (VLDL) and triglycerides, as well as decreased high-density lipoprotein (HDL), hypertension, etc., and is recognized as a risk factor for cardiovascular disease and type 2 diabetes.

[0045] According to an embodiment of the present invention, the strain can inhibit fat accumulation in white adipose tissue. Specifically, 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 decrease in the size of fat cells in the white adipose tissue of the mouse model. At the same time, when common lactic acid bacteria were orally administered to a mouse model fed a high-fat diet, the size of fat cells in the white adipose tissue of the mouse model did not decrease. Based on this, it was confirmed that the strain inhibited fat accumulation in white adipose tissue ( Figure 4 ).

[0046] According to an embodiment of the present invention, the strain can reduce metabolic inflammation (meta-inflammation) in white adipose tissue. As used herein, the term "metabolic inflammation" is a chronic and low-grade inflammation, referring to an inflammatory response that occurs when an excess of nutrients or metabolites are provided. In particular, it is known that chronic inflammatory responses caused by obesity play an important role in increasing insulin resistance and developing metabolic abnormalities.

[0047] In an 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 in the white adipose tissue of the mouse model, and a significant increase in M2 macrophages and Treg cells. At the same time, when common lactic acid bacteria were orally administered to a mouse model fed a high-fat diet, M1 macrophages in the white adipose tissue of the mouse model were significantly reduced, but M2 and Treg cells did not change. Based on this, it was confirmed that the strain inhibited the inflammatory response in white adipose tissue ( Figure 5 ).

[0048] According to an embodiment of the present invention, the strain can inhibit fat accumulation in brown adipose tissue. Specifically, 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 fat cells in the brown adipose tissue of the mouse model. At the same time, when common lactic acid bacteria were orally administered to a mouse model fed a high-fat diet, the size of fat cells in the brown adipose tissue of the mouse model did not decrease. Based on this, it is confirmed that the strain inhibits fat accumulation in brown adipose tissue ( Figure 6 ).

[0049] According to an embodiment of the present invention, the strain can reduce liver weight and inhibit fat accumulation in liver tissue. Specifically, 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 liver weight and fat accumulation in the mouse model's tissue. At the same time, when common lactic acid bacteria were orally administered to a mouse model fed a high-fat diet, a large amount of fat accumulated in the mouse model's liver tissue, 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 ( Figure 7 and Figure 8 ).

[0050] According to an embodiment of the present invention, the strain can reduce blood sugar levels. Preferably, the strain can reduce fasting blood sugar levels and improve glucose tolerance. Specifically, in an embodiment of the present invention, oral administration of the strain to a mouse model fed a high-fat diet results in a reduction in the fasting blood sugar level and the blood sugar level after administration of glucose in the mouse model. At the same time, when common lactic acid bacteria were orally administered to a mouse model fed a high-fat diet, neither the fasting blood sugar level nor the blood sugar level after administration of glucose was reduced. Based on this, it is confirmed that the strain reduces fasting blood sugar levels and improves glucose tolerance ( Fig. 9 and Fig.10 ).

[0051] According to an embodiment of the present invention, the strain can improve insulin resistance. Specifically, in an embodiment of the present invention, when the strain is orally administered to a mouse model fed a high-fat diet, the blood glucose level of the mouse model is significantly reduced after insulin administration. At the same time, when common lactic acid bacteria are orally administered to a mouse model fed a high-fat diet, neither the fasting blood glucose level nor the blood glucose level after insulin administration is reduced. Based on this, it is confirmed that the strain improves insulin resistance by increasing insulin sensitivity ( Fig.11 ).

[0052] The composition may further comprise a cryoprotectant or excipient. Specifically, the cryoprotectant may be one or more selected from the group consisting of glycerol, trehalose, maltodextrin, skimmed milk powder and starch. In addition, the excipient may be one or more selected from the group consisting of glucose, dextrin and skimmed milk powder.

[0053] Based on the gross weight of the composition, the composition can include 0.01wt% to 20wt% or 0.01wt% to 10wt% of a cryoprotectant, specifically, the composition can include 5wt% to 20wt% of glycerol, 2wt% to 10wt% of trehalose, 2wt% to 10wt% of maltodextrin, 0.5wt% to 2wt% of skim milk powder and 0.1wt% to 1wt% of starch. In addition, based on the gross weight of the composition, the composition can include 75wt% to 95wt% or 85wt% to 95wt% of an excipient.

[0054] Another aspect of the present invention provides a food composition, comprising a Lactobacillus fermentum strain (preservation number KCTC 14106BP).

[0055] The Lactobacillus fermentum strain (Accession No. KCTC 14106BP) is the same as that described above.

[0056] The food composition includes all forms, such as functional food, nutritional supplement, health food, food additives, and the types of food compositions can be prepared into various forms according to conventional methods known in the art.

[0057] In the case where 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. Usually, when preparing food or beverages, the addition amount of the active ingredient in the raw material composition containing the strain can be 0.0001wt% to 1wt%, particularly 0.001wt% to 0.1wt%.

[0058] Another aspect of the present invention provides a feed composition for preventing or inhibiting metabolic diseases, the feed composition comprising a Lactobacillus fermentum strain (Deposit No. KCTC 14106BP) or a culture thereof.

[0059] The Lactobacillus fermentum strain (Accession No. KCTC 14106BP) is the same as that described above.

[0060] The feed composition for preventing or improving metabolic diseases can be prepared according to various methods for preparing feed compositions known in the art by adding Lactobacillus fermentum strain (Deposit No. KCTC 14106BP) in an appropriate effective concentration range.

[0061] Still another aspect of the present invention provides a method for preventing and treating metabolic diseases, the method comprising administering Lactobacillus fermentum strain (Deposit No. KCTC 14106BP) or a culture thereof to a subject.

[0062] The object may suffer from metabolic diseases. In addition, the object may be a mammal, preferably a human. In this case, the fermented lactobacillus strain (preserving number KCTC 14106BP) is the same as described above. In addition, the route of administration, dosage and frequency of administration of the strain or its culture may be administered to the object in various ways and amounts according to the patient's condition and the presence or absence of side effects, and the best method of administration, dosage and frequency of administration may be suitably selected by those skilled in the art in an appropriate range. In addition, the type of metabolic diseases is as described above.

[0063] Another aspect of the present invention provides use of a Lactobacillus fermentum strain (preservation number KCTC 14106BP) or a culture thereof for treating metabolic diseases.

[0064] In this case, the Lactobacillus fermentum strain (Accession No. KCTC 14106BP) is the same as that described above. In addition, the types of metabolic diseases are as described above. DETAILED DESCRIPTION

[0066] The present invention will be described in more detail below by the following examples. However, the following examples are only used to illustrate the present invention, and the scope of the present invention is not limited by the following examples.

[0067] Example 1. Screening of strains with anti-obesity effects

[0068] A mouse model was used to select lactic acid bacteria with anti-obesity effects from 16 lactic acid bacteria purchased from Mediogen Co., Specifically, C57BL / 6 mice fed with a 60% high fat diet (HFD) and then orally administered with each lactic acid bacteria were used as the experimental group. Mice fed only with a 60% high fat diet were used as the control group. In this case, each mouse was fed 5×10 9 Each lactic acid bacteria was orally administered by CFU. The anti-obesity effect was determined based on the difference in body weight between the experimental group and the control group.

[0069] As a result, the Lactobacillus fermentum GB103 strain was the most effective in suppressing weight gain. In this case, some Lactobacillus fermentum lactic acid bacteria did not show anti-obesity effects ( Figures 1 to 3 ). Based on this, it was confirmed that the fermentation Lactobacillus GB103 strain exhibited anti-obesity effects among the fermentation Lactobacillus lactic acid bacteria.

[0070] Example 2. Confirmation of the fat accumulation inhibitory effect of Lactobacillus fermentum GB103 strain in white adipose tissue

[0071] A mouse model was used to confirm the fat accumulation inhibitory effect of Lactobacillus fermentum GB103 (Accession No. KCTC 14106BP) strain in white adipose tissue. Specifically, C57BL / 6 mice fed with a 60% high-fat diet (HFD) and then orally administered with Lactobacillus fermentum GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as a negative control group, and mice fed a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal chow diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9 Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0072] After the administration of each test substance was completed, white adipose tissue was isolated by autopsy. The isolated white adipose tissue was fixed with 10% neutral buffered formalin. Then, paraffin sections were prepared, stained with hematoxylin and eosin (H&E), and then the cell size was observed.

[0073] The results showed that the adipocyte size of the negative control group and the positive control group fed with a high-fat diet increased compared with the normal group, while the adipocyte size of the experimental group orally administered with the Lactobacillus fermentum GB103 strain was significantly reduced ( Figure 4 ). Based on this, it was confirmed that the fermentation Lactobacillus GB103 strain effectively inhibits fat accumulation in white adipose tissue.

[0074] Example 3. Confirmation of the immune cell regulatory effect of Lactobacillus fermentum GB103 strain in white adipose tissue

[0075] A mouse model was used to confirm that the administration of Lactobacillus fermentum GB103 strain resulted in changes in immune cells in white adipose tissue. Specifically, C57BL / 6 mice fed with 60% (HFD) and then orally administered with Lactobacillus fermentum GB103 strain were used as the experimental group. In addition, mice fed only a 60% high-fat diet were used as a negative control group, and mice fed a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9 Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0076] After the administration of each test substance, immune cells were isolated from the epididymal white adipocyte tissue of each group of mice by necropsy, the cells were stained with specific antibodies, and then analyzed by flow cytometry. The adipose tissue was minced with curved scissors, incubated with 2 mg / mL type II collagen (Worthington) at 37°C for 45 minutes, and then the stromal vascular fraction (SVF) was separated by centrifuge. It was washed with FACS buffer and then blocked at 4°C for 10 minutes to inhibit nonspecific 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 and 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 and then analyzed by eBioscience TMFoxp3 / transcription factor staining buffer was used for fixation and permeability to stain with anti-Foxp3 antibody (eBioscience, CA, USA). In MHCII+F4 / 80+ macrophages, CD11b+CD206+ cells were analyzed with M2 macrophages, and CD11b+CD11c+ cells were analyzed with M1 macrophages. Treg cells were analyzed with TCRβ+CD4+Foxp3+.

[0077] The results showed that the number of inflammatory M1 macrophages in the negative and positive control group mice fed a high-fat diet was higher than that in the normal group, while the number of M1 macrophages in the experimental group mice orally administered with the fermented Lactobacillus GB103 strain was significantly lower than that in the negative control group. It was also confirmed that the number of anti-inflammatory M2 macrophages and Treg cells in the negative and positive control group mice fed a high-fat diet was lower than that in the normal group, while the number of M2 macrophages and Treg cells in the experimental group mice orally administered with the fermented Lactobacillus GB103 strain was significantly increased compared with the negative control group. Based on this, the immune cell regulatory effect of the fermented Lactobacillus GB103 strain in white adipose tissue was confirmed ( Figure 5 ).

[0078] Example 4. Confirmation of the inhibitory effect of Lactobacillus fermentum GB103 on fat accumulation in brown adipose tissue

[0079] A mouse model was used to confirm the fat accumulation inhibitory effect of Lactobacillus fermentum GB103 strain in brown adipose tissue. Specifically, C57BL / 6 mice fed with 60% (HFD) and then orally administered with Lactobacillus fermentum GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as a negative control group, and mice fed a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9 Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0080] After the administration of each test substance was completed, brown adipose tissue was isolated from each group of mice by necropsy. The isolated brown adipose tissue was fixed with 10% neutral buffered formalin. Then, paraffin sections were prepared, stained with H&E, and then the cell size was observed.

[0081] The results showed that the adipocyte size of the negative control group and the positive control group mice fed with a high-fat diet increased compared with the normal group, while the adipocyte size of the experimental group orally administered with the Lactobacillus fermentum GB103 strain was significantly reduced ( Figure 6). Based on this, it was confirmed that the fermentation Lactobacillus GB103 strain effectively inhibits fat accumulation in brown adipose tissue.

[0082] Example 5. Confirmation of the inhibitory effect of Lactobacillus fermentum GB103 on fat accumulation in liver tissue

[0083] A mouse model was used to confirm the fat accumulation inhibitory effect of fermented Lactobacillus GB103 strain in liver tissue. Specifically, C57BL / 6 mice fed with 60% (HFD) and then orally administered with fermented Lactobacillus GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as a negative control group, and mice fed a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9 Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0084] After administration of each test substance, the liver was removed by autopsy and the liver weight was measured to compare and evaluate the liver weight among the groups. The extracted liver was fixed with 10% buffered formalin, paraffin sections were prepared, stained with H&E, and the cell size was observed.

[0085] The results showed that the liver weight of the negative control group and the positive control group mice fed with a high-fat diet increased compared with the normal group, while the liver weight of the experimental group mice orally administered with the fermentation Lactobacillus GB103 strain was significantly reduced compared with the negative control group ( Figure 7 In addition, compared with the normal group, the fat accumulation in the liver tissue of the negative and positive control group mice fed with a high-fat diet increased, however, the fat accumulation in the liver tissue of the experimental group mice orally administered with the Lactobacillus fermentum GB103 strain was very small, and the fat accumulation level was similar to that of the negative control group ( Figure 8 ). Based on this, it was confirmed that the Lactobacillus fermentum GB103 strain effectively inhibited liver weight and fat accumulation in liver tissue.

[0086] Example 6. Confirmation of the fasting blood sugar lowering and glucose tolerance improving effects of Lactobacillus fermentum GB103 strain

[0087] A glucose tolerance test (GTT) was performed to confirm the glucose tolerance improving effect of the fermented Lactobacillus GB103 strain. First, C57BL / 6 mice fed with 60% (HFD) and then orally administered with the fermented Lactobacillus GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as a negative control group, and mice fed with a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9 Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0088] For the glucose tolerance test, mice were fasted for 16 hours or longer, and then a glucose solution was injected intraperitoneally at a dose of 1 g / kg. Then, blood was collected from the tail vein of the mice after 0, 30, 60, 90, and 120 minutes, and blood glucose was measured using a glucometer. In this case, blood glucose at 0 minutes refers to fasting blood glucose.

[0089] The results showed that the fasting blood glucose levels of the negative and positive control mice fed a high-fat diet increased significantly compared with the normal group. At the same time, the fasting blood glucose levels of the experimental group mice orally administered with the fermentation Lactobacillus GB103 strain were significantly reduced compared with the negative control group ( Fig. 9 ).

[0090] In addition, it was confirmed that after the administration of glucose, the blood glucose levels of the negative and positive control group mice fed with a high-fat diet were higher than those of the normal group, while the blood glucose levels of the experimental group mice orally administered with the fermentation Lactobacillus GB103 strain were significantly lower than those of the negative control group ( Fig.10 ). Based on this, it was confirmed that the fermentation Lactobacillus GB103 strain showed fasting blood sugar lowering and glucose tolerance improving effects.

[0091] Example 7. Confirmation of the insulin resistance improving effect of Lactobacillus fermentum GB103 strain

[0092] An insulin tolerance test (ITT) was performed to confirm the insulin resistance improving effect of the fermented Lactobacillus GB103 strain. First, C57BL / 6 mice fed with 60% (HFD) and then orally administered with the fermented Lactobacillus GB103 strain were used as the experimental group. In addition, mice fed only with a 60% high-fat diet were used as a negative control group, and mice fed with a 60% high-fat diet and orally administered with common lactic acid bacteria (Lactobacillus plantarum MG5120) were used as a positive control group. In addition, mice fed a normal diet (NCD) were used as a normal group. In this case, each mouse was fed 5×10 9Common lactic acid bacteria purchased from Mediogen Co., or Lactobacillus fermentum GB103 strain was orally administered CFU.

[0093] For the insulin tolerance test, mice were fasted for 4.5 hours and then insulin solution was intraperitoneally administered at a dose of 1 U / kg. Blood was collected from the tail vein of the mice after 0, 30, 60, 90 and 120 minutes, and blood glucose was measured using a glucometer.

[0094] The results confirmed that after the administration of insulin, the blood glucose levels of the negative and positive control mice fed a high-fat diet were higher than those of the normal group, while the blood glucose levels of the experimental group mice orally administered with the fermentation Lactobacillus GB103 strain were significantly lower than those of the negative control group ( Fig.11 ). Based on this, it was confirmed that the fermentation Lactobacillus GB103 strain effectively improved insulin resistance by increasing insulin sensitivity.

[0095] <Accession Number>

[0096] Name of depository institution: Korea Center for Typical Culture Collection (KCTC), Korea Institute of Biotechnology

[0097] Accession number: KCTC14106BP

[0098] Date of preservation: 20200114.

Claims

1. A Lactobacillus Fermentum strain with a preservation number of KCTC 14106BP.

2. A pharmaceutical composition for preventing or treating metabolic diseases, comprising a Lactobacillus fermentum strain with a deposit number of KCTC14106BP or a culture containing the strain as an active ingredient, wherein: The metabolic disease is any one selected from the group consisting of obesity, hyperlipidemia, fatty liver, diabetes and insulin resistance syndrome.

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

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

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

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

7. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein: The strain reduces blood sugar levels.

8. The pharmaceutical composition for preventing or treating metabolic diseases according to claim 2, wherein: The strain reduces insulin resistance.

9. A food composition comprising the fermentation Lactobacillus strain with a preservation number of KCTC 14106BP.

10. A feed composition for preventing or inhibiting metabolic diseases, the feed composition comprising a fermented Lactobacillus strain with a preservation number of KCTC14106BP or a culture containing the strain, wherein: The metabolic disease is any one selected from the group consisting of obesity, hyperlipidemia, fatty liver, diabetes and insulin resistance syndrome.

11. Use of the fermented Lactobacillus strain with the deposit number of KCTC 14106BP or a culture containing the strain in the preparation of a medicament for preventing or inhibiting metabolic diseases, wherein: The metabolic disease is any one selected from the group consisting of obesity, hyperlipidemia, fatty liver, diabetes and insulin resistance syndrome.

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