Application of bifidobacterium animalis subsp. Lactis or composition in preparation of product for preventing and / or treating metabolic diseases

Abstract

The invention relates to an application of a bifidobacterium animalis subsp. Lactis BGI-N3 (the preservation number is CCTCC (China Center For Type Culture Collection) NO: M20221585) or a composition containing the bifidobacterium animalis subsp. Lactis BGI-N3 as an active ingredient in the preparation of a product for preventing and / or treating metabolic diseases, and particularly relates to an application of the bifidobacterium animalis subsp. Lactis BGI-N3 or a composition containing the bifidobacterium animalis subsp. Lactis BGI-N3 as an active ingredient in the preparation of a product for preventing and / or treating metabolic diseases. The bifidobacterium animalis subsp. Lactis comprises a 16S rDNA (ribosomal deoxyribonucleic acid) sequence as shown in SEQ ID NO: 1. The animal bifidobacterium subsp. Lactis BGI-N3 provided by the embodiment of the invention has good safety and excellent gastrointestinal tract environment tolerance, and can effectively reduce the food calorie conversion rate, regulate the systemic chronic inflammation level and improve the fat accumulation condition so as to prevent and / or treat metabolic diseases, such as obesity.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, and in particular to an application of animal Bifidobacterium lactis subspecies or a composition containing the same as an effective ingredient in preparing a product for preventing and / or treating metabolic diseases. Background Art

[0002] Obesity is a chronic metabolic disease. In obese individuals caused by a high-calorie diet, adipose tissue releases a variety of cytokines and inflammatory mediators, which trigger and maintain a low-level systemic inflammatory state and lead to dysfunction of immune cells. The presence of chronic inflammation, in turn, affects the development and maintenance of obesity by affecting the differentiation and maturation of adipocytes. This interaction leads to a series of health problems such as heart disease, hypertension, diabetes, fatty liver and even cancer.

[0003] There are currently a variety of treatment and intervention strategies for metabolic diseases caused by high-calorie diets, especially obesity, such as drug therapy, surgical treatment, and diet and exercise intervention. Drug therapy can effectively treat obesity to a certain extent, but its applicable population and scenarios are strictly limited, and there are side effects such as indigestion, oily bowel movements, and nausea, and patients are often required to take medication for a long time. Weight loss surgery is only suitable for patients with severe obesity or complications, and patients need to continue to adjust their diet and lifestyle after surgery, otherwise the risk of weight rebound still exists. The method of diet and exercise intervention helps individuals gradually lose weight and relieve obesity and its related complications by adjusting eating habits, reducing the intake of high-calorie foods, and increasing physical exercise. However, this method is not easy to stick to. In summary, although the above methods provide a variety of solutions to the problem of obesity, each strategy has its inherent limitations.

[0004] Therefore, there is an urgent need for a safe and effective method to improve a series of chronic metabolic diseases (especially obesity) caused by a high-calorie diet. Summary of the invention

[0005] The present invention aims to provide an innovative solution to the above technical problems. To this end, the present invention proposes a use of animal Bifidobacterium lactis subsp. or a composition containing the same as an active ingredient in the preparation of a product for preventing and / or treating metabolic diseases.

[0006] The embodiment of the first aspect of the present invention proposes the use of an animal Bifidobacterium lactis subspecies or a composition containing the same as an effective ingredient in the preparation of a product for preventing and / or treating metabolic diseases, wherein the animal Bifidobacterium lactis subspecies comprises a 16S rDNA sequence as shown in SEQ ID NO: 1. The animal Bifidobacterium lactis subspecies reduces the level of serum inflammatory factors, regulates the body's immune and metabolic functions, and improves the food heat conversion rate, thereby improving abnormal conditions such as increased body fat and liver pathological improvement caused by a high-calorie diet, providing a scientific and effective choice for consumers seeking health management.

[0007] In some embodiments, the metabolic disease comprises at least one selected from the group consisting of obesity, type II diabetes, hyperlipidemia, non-alcoholic fatty liver disease, and metabolic syndrome.

[0008] In some embodiments, the metabolic disease is obesity.

[0009] In some embodiments, the composition regulates energy metabolism by regulating inflammation levels to prevent and / or treat the metabolic disease.

[0010] In some embodiments, the animal Bifidobacterium lactis subspecies is animal Bifidobacterium lactis subspecies BGI-N3 ( Bifidobacterium animalis subsp. lactis BGI-N3).

[0011] In some embodiments, the composition comprises an effective amount of the Bifidobacterium animalis subsp. lactis and a pharmaceutically or food acceptable excipient.

[0012] In some embodiments, the composition further comprises other components for preventing and / or treating metabolic diseases.

[0013] In some embodiments, the 3 CFU / mL to 1.0×10 11 CFU / mL Bifidobacterium animalis subsp. lactis The composition was used for the dosage of BGI-N3.

[0014] In some embodiments, the composition comprises a dose of Bifidobacterium animalis subsp. lactis greater than 1.0×10 5 CFU / mL.

[0015] In some embodiments, the product is a medicine, a health food or a functional food.

[0016] The embodiments of the present invention achieve the following beneficial effects: (1) Animal Bifidobacterium lactis subspecies BGI-N3 has a long history of application in the food field, has good safety, and is suitable for a wide range of people. The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiment of the present invention has no potential harm to the body and has good safety. At the same time, compared with drug intervention and high-calorie diet, the use of probiotic preparations is less likely to cause side effects. The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiment of the present invention is suitable for long-term use, avoiding the drug dependence and side effects caused by traditional drug intervention. Therefore, it is suitable for preventing and / or treating metabolic diseases (especially obesity) and liver pathological changes caused by them. In addition, the supplementation of probiotics helps maintain the balance of intestinal microecology, which can bring long-term benefits to intestinal health and reduce the risk of obesity.

[0017] (2) The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiments of the present invention has excellent tolerance to the gastrointestinal environment, ensuring that its active ingredients can safely reach the intestine and exert their effects, and is therefore suitable for preventing and / or treating metabolic diseases, especially obesity.

[0018] (3) The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiment of the present invention can reduce the level of serum inflammatory factors, regulate the body's immune and metabolic functions, and improve the food calorie conversion rate, thereby achieving the purpose of improving abnormal conditions such as increased body fat and liver pathological improvement caused by a high-calorie diet, and ultimately achieving multi-dimensional effective prevention or improvement (treatment) of metabolic diseases (especially obesity). BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the present invention or related technologies, the following briefly introduces the drawings required for use in the embodiments or related technical descriptions. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0020] Figure 1 Schematic diagram of food calorie conversion rate results of the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention (* indicates p<0.05 relative to the model control group; ** indicates p<0.01 relative to the model control group; *** indicates p<0.001 relative to the model control group; tested by ONE-WAY ANOVA); Figure 2Schematic diagram of free fatty acid level determination results of the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiment of the present invention (* indicates p<0.05 relative to the model control group; ** indicates p<0.01 relative to the model control group; *** indicates p<0.001 relative to the model control group; tested by ONE-WAY ANOVA); Figure 3 Schematic diagram of the measurement results of perirenal fat and peritesticular fat weight in the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiment of the present invention (* indicates p<0.05 relative to the model control group; ** indicates p<0.01 relative to the model control group; *** indicates p<0.001 relative to the model control group; tested by ONE-WAY ANOVA); Figure 4 Schematic diagram of the results of measuring serum TNF-α levels of systemic inflammatory markers in the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention (* indicates p<0.05 relative to the model control group; ** indicates p<0.01 relative to the model control group; *** indicates p<0.001 relative to the model control group; tested by ONE-WAYANOVA); Figure 5 Schematic diagram of the results of measuring serum CRP levels of systemic inflammatory markers in the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention (* indicates p<0.05 relative to the model control group; ** indicates p<0.01 relative to the model control group; *** indicates p<0.001 relative to the model control group; tested by ONE-WAYANOVA); Figure 6 Graph showing the results of liver tissue pathology analysis in the normal control group, model control group, and BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention. DETAILED DESCRIPTION

[0021] The present invention is further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art and are not intended to limit the present invention in any way.

[0022] The present invention is made based on the following understanding: In obese individuals caused by a high-calorie diet, adipose tissue releases a variety of cytokines and inflammatory mediators, which can trigger and maintain a low-level systemic inflammatory state and lead to dysfunction of immune cells. The presence of chronic inflammation, in turn, affects the development and maintenance of obesity by affecting the differentiation and maturation of adipocytes. This interaction leads to a series of health problems such as heart disease, hypertension, diabetes, fatty liver and even cancer. Specifically, a high-calorie diet changes the composition of intestinal microorganisms, increases the abundance of harmful bacteria, reduces the abundance of beneficial bacteria, and destroys the integrity and function of the intestinal barrier, thereby causing chronic inflammation; accordingly, chronic inflammation damages cells, thereby affecting mitochondrial function and energy metabolism, further exacerbating obesity symptoms.

[0023] There are currently a variety of treatment interventions for metabolic diseases caused by high-calorie diets, especially obesity, such as drug therapy, surgical treatment, and diet and exercise intervention. In terms of drug therapy, drugs approved for long-term weight management include orlistat, phentermine, naltrexone-bupropion compound preparations, etc. They work through different mechanisms, such as inhibiting fat absorption, reducing hunger, and controlling appetite. However, on the one hand, this method has adverse drug effects that cannot be ignored. For example, orlistat may cause discomfort such as indigestion and oily bowel movements, while phentermine and naltrexone-bupropion compound preparations may cause side effects such as palpitations, hypertension, nausea, and dry mouth; on the other hand, this method requires patients to take drugs for a long time. Once the drug is interrupted, the weight is likely to rise rapidly, and it also puts pressure on the patient's liver metabolism. The scope of use of this drug is also severely limited to its clear user population requirements. Bariatric surgery is only suitable for patients with severe obesity or complications. It is not the most appropriate solution for individuals with mild to moderate obesity. And after surgery, patients need to continue to adjust their diet and lifestyle, otherwise the risk of weight rebound still exists. The method of diet and exercise intervention is to adjust eating habits, reduce the intake of high-calorie foods and increase physical exercise, so as to help individuals gradually lose weight and alleviate obesity and its related complications. Obviously, the success rate of this method is very low. In summary, although the above methods provide diverse solutions to the problem of obesity, each strategy has its inherent limitations. In this context, active probiotics, as a natural therapy for regulating the balance of intestinal flora, have shown significant potential in combating weight gain and lipid accumulation caused by a high-fat diet.

[0024] Active probiotics have potential in combating weight gain and lipid accumulation caused by a high-calorie diet. They can effectively improve the symptoms of obesity and related complications by regulating intestinal flora. These probiotics provide a new therapy for the prevention and / or treatment of metabolic diseases caused by a high-calorie diet by regulating the balance of intestinal flora, enhancing intestinal barrier function, improving the body's antioxidant capacity, and reducing inflammatory responses in the body. However, the current research results on intestinal flora are inconsistent. As for the various types of lactobacilli and bifidobacteria that have received more attention, there is strain specificity. Different strains may have differences in colony morphology, growth characteristics, metabolites, physiological functions, etc. Even different strains of the same species have differences in genetic and metabolic characteristics that affect the final functional effect of preventing and / or treating metabolic diseases. Therefore, for patients with metabolic diseases, it is necessary to select probiotics that can effectively prevent and treat metabolic diseases (such as obesity) based on scientific research at the strain level, and have good safety and excellent gastrointestinal environmental tolerance.

[0025] In the present embodiment, "probiotics" should be understood as "live microorganisms that have a beneficial effect on the health of the host by improving the intestinal microbial environment of the host in the gastrointestinal tract of animals including humans". When live microorganisms with probiotic activity are provided in the form of dried cells or fermentation products in the form of a single or composite strain, the probiotics can have a beneficial effect on the intestinal flora of the host.

[0026] According to the above practical needs, the first aspect of the embodiments of the present invention provides the use of animal Bifidobacterium lactis subsp. or a composition containing it as an active ingredient in the preparation of a product for preventing and / or treating metabolic diseases, wherein the animal Bifidobacterium lactis subsp. comprises a 16S rDNA sequence as shown in SEQ ID NO: 1.

[0027]

[0028] In some embodiments, the metabolic disease is selected from at least one of the group consisting of obesity, type II diabetes, hyperlipidemia, non-alcoholic fatty liver disease, and metabolic syndrome.

[0029] In some embodiments, the metabolic disease is obesity.

[0030] In some embodiments, the composition regulates energy metabolism by regulating inflammation levels to prevent and / or treat the metabolic disease.

[0031] In some embodiments, the composition modulates the inflammation level by modulating tumor necrosis factor-alpha (TNF-α) and C-reactive protein (CRP) levels.

[0032] In some embodiments, the animal Bifidobacterium lactis subspecies is animal Bifidobacterium lactis subspecies BGI-N3 ( Bifidobacterium animalis subsp. lactis BGI-N3).

[0033] The animal Bifidobacterium lactis subspecies provided in the embodiment of the present invention belongs to the genus Bifidobacterium ( Bifidobacterium ), which was named Bifidobacterium animalis subsp. lactis BGI-N3 ( Bifidobacterium animalis subsp. lactis BGI-N3); the deposit registration number is CCTCC NO:M20221585, the deposit institution is: China Center for Type Culture Collection; the deposit unit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province; the postal code is 430072; the deposit time is October 17, 2022.

[0034] In an embodiment of the present invention, the animal Bifidobacterium lactis subspecies BGI-N3 having the sequence shown in SEQ ID NO: 1 can be understood by those skilled in the art as the original strain. It is understandable that strains such as animal Bifidobacterium lactis subspecies BGI-N3 can undergo spontaneous mutations or be artificially cultured to form variant strains, such as the deletion of nucleotides, the addition of nucleotides, or the replacement of nucleotides. The "variant strain" has a highly identical gene sequence and extremely similar biological functions with the "animal Bifidobacterium lactis subspecies BGI-N3", and the mutated gene will not substantially affect the conserved sequence of animal Bifidobacterium lactis subspecies BGI-N3, and thus will not affect the genetic stability of animal Bifidobacterium lactis subspecies BGI-N3. More specifically, the "variant strain" is also a strain of the BGI-N3 species, showing the physiological activity and functional characteristics of the BGI-N3 species.

[0035] The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiment of the present invention has no potential harm to the body and has good safety; at the same time, compared with drug intervention of high-calorie diet, the use of probiotic preparations is less likely to cause side effects. The animal Bifidobacterium lactis subspecies BGI-N3 provided in the embodiment of the present invention is suitable for long-term use, avoiding the drug dependence and side effects caused by traditional drug intervention, and is therefore suitable for preventing and / or treating metabolic diseases (especially obesity) and liver pathological changes caused by them; and the supplementation of probiotics helps to maintain the balance of intestinal microecology, which can bring long-term benefits to intestinal health and reduce the risk of obesity. In addition, animal Bifidobacterium lactis subspecies BGI-N3 has excellent gastrointestinal environmental tolerance, can reduce the level of serum inflammatory factors, regulate the body's immune and metabolic functions, and improve the food heat conversion rate, thereby achieving the purpose of improving abnormal conditions such as increased body fat and liver pathological improvement caused by high-calorie diet, and ultimately achieving multi-dimensional effective prevention or improvement (treatment) of metabolic diseases (especially obesity).

[0036] In some embodiments, the composition comprises an effective amount of the Bifidobacterium animalis subsp. lactis and a pharmaceutically or food acceptable excipient.

[0037] In some embodiments, the composition further comprises other components for preventing and / or treating metabolic diseases.

[0038] In some embodiments, the composition further comprises prebiotics and / or probiotics.

[0039] In some embodiments, the composition is administered orally or rectally.

[0040] In some embodiments, the composition is in the form of a powder, granules, capsule, tablet, solution, suspension, syrup, ointment, plaster, or suppository.

[0041] In some embodiments, the pharmaceutically acceptable excipient is, for example, a carrier, an excipient, a diluent, an adjuvant, a vehicle, an excipient, a carrier or a combination thereof, and / or other substances having a concentration that does not affect the activity of the microorganism, and the present invention is not limited thereto.

[0042] The term "pharmaceutically acceptable carrier" refers to any preparation or carrier medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects on the host or patient. Representative carriers include water, oil, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, viscosity enhancers, transdermal enhancers, etc. Their preparations are well known to those skilled in the art of cosmetics or topical drugs. For additional information about the carrier, reference can be made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0043] In some embodiments, based on the total volume or total weight of the composition, the composition may contain 1.0×10 3 CFU / mL to 1.0×10 11 CFU / mL Bifidobacterium animalis subsp. lactis The composition is used in a dosage of BGI-N3. In some embodiments, the composition comprises 1.0×10 4 CFU / mL to 1.0×10 10 CFU / mL Bifidobacterium animalis subsp. lactis In some embodiments, 1.0×10 5 CFU / mL to 1.0×10 9 CFU / mL Bifidobacterium animalis subsp. lactis The composition was used for the dosage of BGI-N3.

[0044] In some embodiments, the composition comprises a dose of Bifidobacterium animalis subsp. lactis greater than 1.0×10 5 CFU / mL. It is understood that probiotics are prepared into bacterial powder through conventional processes in the microbial industry such as high-density fermentation, filtration centrifugation, freeze drying or spray drying, in which the number of viable bacteria is 1.0×10 4 to 1.0×10 12 CFU / g. The bacterial powder can be prepared into the above-mentioned various forms when applied. In some embodiments, the bacterial powder is dissolved in water to obtain different bacterial concentrations (e.g., greater than 1.0×10 5 CFU / mL).

[0045] In some embodiments, the product is a medicine, a health food or a functional food.

[0046] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0047] Unless otherwise specified, the quantitative tests in the following examples were performed three times and the results were averaged.

[0048] Example 1 Isolation, identification and preservation of Bifidobacterium animalis subspecies lactis BGI-N3 In this example, Bifidobacterium animalis subspecies lactis BGI-N3 was isolated, identified and preserved.

[0049] (1) Isolation of strains One sample of traditional Chinese farm fermented yogurt was collected and placed in an anaerobic chamber. 0.2 g of the sample was mixed in 1 mL of sterile phosphate buffered saline (PBS, item number B640011, purchased from Shanghai Biotech Co., Ltd.), and then graded diluted. 4 Take 100ul from the sample dilution of 100 times, spread it on TPY agar medium (Cat. No. 027350, Guangdong Huankai Microbiology Technology Co., Ltd.), and culture it anaerobically at 37℃ for 48 to 72h. Then select a single colony for streaking isolation to obtain an isolated pure culture strain.

[0050] (2) Identification of strains The isolated pure culture strain was extracted using a bacterial genomic DNA extraction kit (Cat. No. DP302, purchased from Tiangen Biochemical Technology Beijing Co., Ltd.). The 16S rDNA gene was amplified by PCR using the extracted genomic DNA as a template, wherein the sequences of the primers used are shown in SEQ ID NO: 2 and SEQ ID NO: 3 (SEQ ID NO: 2, 27F-AGAGTTTGATCATGGCTCAG, SEQ ID NO: 3, 1492R-TAGGGTTACCTTGTTACGACTT), and the PCR amplification program is shown in Table 1, with 30 cycles. The PCR amplification product was sequenced using the Sanger method to obtain the 16S rDNA sequence of the strain (SEQID NO: 1). By comparing the NCBI database, it was found that the strain with the highest homology was Bifidobacterium animalis subsp. lactis , the similarity was 100.00%, and it was named Bifidobacterium animalis subsp. lactis BGI-N3 ( Bifidobacterium animalis subsp. lactis BGI-N3).

[0051] Table 1 PCR amplification procedure

[0052] (3) Preserved strains BGI-N3( Bifidobacterium animalis subsp. lactis The strain was preserved at BGI-N3), the preservation registration number is CCTCC NO:M20221585, the preservation institution is: China Center for Type Culture Collection; the preservation unit address is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province; the postal code is 430072; the preservation time is October 17, 2022.

[0053] Example 2 Gastrointestinal tolerance of Bifidobacterium animalis subspecies lactis BGI-N3 In this example, the intestinal environment tolerance of animal Bifidobacterium lactis subspecies BGI-N3 was evaluated by simulating the human gastrointestinal environment.

[0054] Activate the strain and count it. Place the bacterial liquid in artificial gastric juice (pH=3.0), artificial intestinal juice (pH=6.8), and TPY liquid medium containing 0.3% bile salt (Cat. No. 027312, purchased from Guangdong Huankai Microbiological Technology Co., Ltd.), and incubate at 37°C for 2 hours. The initial viable count value is regarded as the survival rate of 100%. The viable count of each group was performed at the completion of the experiment. The survival rate (%) = 100% × the number of viable bacteria after 2 hours / the initial number of viable bacteria.

[0055] Table 2 Gastrointestinal environmental tolerance of Bifidobacterium animalis subsp. lactis BGI-N3

[0056] The gastrointestinal environment tolerance of animal Bifidobacterium lactis subspecies BGI-N3 is shown in Table 2. The survival rate of animal Bifidobacterium lactis subspecies BGI-N3 after 2 hours under gastric acid conditions is (91.1±5.5)%, the survival rate after 2 hours under simulated small intestinal fluid conditions is (94.9%±2.8)%, and the survival rate of the strain under 0.3% bile salt concentration conditions is (94.9%±5.1)%. The above results show that the animal Bifidobacterium lactis subspecies BGI-N3 in the embodiment of the present invention has excellent tolerance to gastric acid, intestinal fluid and bile salt in the gastrointestinal system, and can survive in the gastrointestinal tract, which is a prerequisite for its prebiotic effect.

[0057] Example 3: In vivo effectiveness study of Bifidobacterium animalis subspecies lactis BGI-N3 in improving high-calorie diet In this example, the in vivo effectiveness study of animal Bifidobacterium lactis subspecies BGI-N3 in improving the adverse effects of a high-calorie diet was completed by comparing the results of food calorie conversion rate, free fatty acids (FFA), perirenal fat and peritesticular fat weight, and levels of inflammatory markers serum TNF-α and CRP in each treatment group and the results of liver tissue pathology analysis in a total of 5 groups of rats, including a normal control group, a model control group, and BGI-N3 low-dose, medium-dose, and high-dose intervention groups.

[0058] (1) A total of 50 6-week-old SPF SD male rats were cultured for a 7-day adaptation period and then randomly divided into 5 groups according to their body weight: normal control group (experimental group A), model control group (experimental group B), BGI-N3 low-dose intervention group (experimental group C), BGI-N3 medium-dose intervention group (experimental group D), and BGI-N3 high-dose intervention group (experimental group E). Experimental group A was fed with maintenance feed, and the remaining groups were fed with high-calorie feed. Experimental groups C and E were gavaged with BGI-N3 bacterial suspension once a day, with a dose of 1.0×10 5 CFU / d (BGI-N3 low-dose intervention group), 1.0×10 7 CFU / d (BGI-N3 medium-dose intervention group), 1.0×10 9 CFU / d (BGI-N3 high-dose intervention group), experimental group AB was gavaged with the same amount of sterile saline daily. The composition of the maintenance feed used in the experiment was in accordance with the provisions of "GB 14924.3-2010 Nutrient composition of compound feed for experimental animals", and its caloric value was 3.87kcal / g; the high-calorie feed was based on 57.6% maintenance feed, with 20.0% sucrose, 15.0% lard, 5% casein, 1.2% cholesterol, 0.6% calcium hydrogen phosphate, 0.4% stone powder and 0.2% sodium cholate, and its caloric value was 4.69kcal / g.

[0059] (2) The experiment lasted for a total of 35 days, and the body weight and food intake of the rats were recorded every week. After 35 days, 2 mL of blood was collected from the abdominal aorta. The blood was allowed to stand at room temperature for 30 minutes and centrifuged at 3000 rpm / min for 10 minutes to separate the serum. The serum was used to detect the levels of TNF-α, CRP, and FFA using a TNF-α detection kit (purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., item number H052-1-2), a CRP detection kit (purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd., item number H126-1-2), and a FFA detection kit (purchased from Shanghai ELISA Biotechnology Co., Ltd., item number ml092765). At the same time, the rats were anesthetized with 1% sodium pentobarbital, and the perirenal fat and epididymal fat tissue were weighed. The rat liver was fixed and paraffin-sectioned, and then Masson staining and HE staining were performed to observe the tissue pathological changes.

[0060] The experimental results are shown in Figures 1 - 6 In the chart, "*" indicates that there is a difference between this group and the model control group (p < 0.05), "**" indicates that there is a significant difference between this group and the model control group (p < 0.01), and "***" indicates that there is an extremely significant difference between this group and the model control group (p < 0.001).

[0061] Figure 1 It is a schematic diagram of the food heat conversion rate results of the normal control group, model control group, BGI-N3 low dose, medium dose, and high dose intervention group in the embodiment of the present invention. The food heat conversion rate of the model control group that only takes in a high-calorie diet is 20.25±2.60%, while the food heat conversion rate of the animal Bifidobacterium lactis subspecies BGI-N3 taken in different doses while feeding a high-calorie diet can reach a minimum of 14.80±0.98%, that is, the use of BGI-N3 reduces the heat conversion by 26.91% at most. The above results show that animal Bifidobacterium lactis subspecies BGI-N3 can significantly inhibit the conversion of intake calories into body weight.

[0062] Figure 2 The figure is a schematic diagram of the free fatty acid level determination results of the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiment of the present invention. It can be seen that compared with the model control group without any intervention, the use of BGI-N3 significantly reduced the free fatty acid level in rats fed a high-calorie diet (p < 0.001), which indicates that the use of animal Bifidobacterium lactis subspecies BGI-N3 can effectively prevent and improve the increase in blood lipid levels caused by a high-calorie diet.

[0063] Figure 3The results of the measurement of perirenal fat and peritesticular fat weight in the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention are shown in the figure, which shows the accumulation of the main body fat in rats under a high-calorie diet. It can be seen that the total weight of perirenal fat and peritesticular fat weight in the model control group rats that only take a high-calorie diet is 18.28±4.96 g; while the fat weight of each intervention group fed with animal Bifidobacterium lactis subspecies BGI-N3 at the same time is as low as 10.909±4.098 g, which shows that the use of animal Bifidobacterium lactis subspecies BGI-N3 can effectively improve the local fat increase caused by a high-calorie diet.

[0064] Figure 4 and Figure 5 The results of measuring serum TNF-α and CRP levels of systemic inflammatory markers in the normal control group, model control group, BGI-N3 low-dose, medium-dose, and high-dose intervention groups in the embodiments of the present invention are shown in Figure 1. It can be seen that feeding animals with Bifidobacterium lactis subspecies BGI-N3 while on a high-calorie diet can significantly prevent and improve the occurrence of systemic chronic inflammation caused by a high-calorie diet.

[0065] Figure 6 It is the result of liver tissue pathological analysis of normal control group, model control group, BGI-N3 low dose, medium dose and high dose intervention group in the embodiment of the present invention. In the results of liver Masson staining, the fibrosis of the liver tissue of rats under the intervention of each dosage group of animal Bifidobacterium lactis subspecies was significantly alleviated compared with the model control group. In the results of liver HE staining, the degree of fatty degeneration of the liver of rats in each dosage group of animal Bifidobacterium lactis subspecies was significantly improved compared with the model control group. The above results illustrate that animal Bifidobacterium lactis subspecies can significantly improve the effect of fatty degeneration of rat liver and liver tissue fibrosis induced by a high-calorie diet.

[0066] The results of Example 3 comprehensively show that the animal Bifidobacterium lactis subspecies BGI-N3 in the examples of the present invention can effectively reduce the level of serum inflammatory factors, regulate the body's immune and metabolic functions, and improve the food calorie conversion rate, thereby achieving the purpose of improving abnormal conditions such as increased body fat and liver pathological improvement caused by a high-calorie diet, and ultimately achieving effective prevention or improvement (treatment) of metabolic diseases (especially obesity) in multiple dimensions.

[0067] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0068] It should be noted that, in this article, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the term "comprises" or any other variant thereof is intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0069] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. Use of Bifidobacterium animalis subsp. lactis or a composition containing it as an active ingredient in the preparation of a product for preventing and / or treating metabolic diseases, The Bifidobacterium animalis subsp. lactis comprises the 16S rDNA sequence shown in SEQ ID NO:

1.

2. The use according to claim 1, characterized in that: The metabolic disease includes at least one selected from the group consisting of obesity, type II diabetes, hyperlipidemia, non-alcoholic fatty liver disease and metabolic syndrome.

3. The use according to claim 1, characterized in that: The metabolic disease is obesity.

4. The use according to claim 1, characterized in that: The composition regulates energy metabolism by regulating inflammation levels, thereby preventing and / or treating the metabolic disease.

5. The use according to claim 1, characterized in that: The animal Bifidobacterium lactis subspecies is animal Bifidobacterium lactis subspecies BGI-N3 (with a preservation number of CCTCC NO: M20221585) Bifidobacterium animalis subsp. lactis BGI-N3).

6. The use according to claim 1, characterized in that: The composition comprises an effective amount of the Bifidobacterium animalis subspecies lactis and a pharmaceutically or food acceptable auxiliary material.

7. The use according to claim 6, characterized in that: The composition may further comprise other components for preventing and / or treating metabolic diseases.

8. The use according to claim 1, characterized in that: To include 1.0×10 3 CFU / mL to 1.0×10 11 CFU / mL Bifidobacterium animalis subsp. lactis The composition was used for the dosage of BGI-N3.

9. The use according to claim 1, characterized in that: The composition contains more than 1.0×10 5 CFU / mL.

10. The use according to claim 1, characterized in that: The product is a medicine, a health food or a functional food.

Images