Bifidobacterium longum subsp. longum C-5 with body weight regulation efficacy and its application

Abstract

The present invention relates to Bifidobacterium longum subsp. longum C-5 with the efficacy of regulating body weight and its application. The taxonomic naming of the Bifidobacterium longum subsp. longum C-5 is Bifidobacterium longum subsp. longum , and the preservation number is GDMCC No: 64542, and the preservation date is April 22, 2024. This strain can improve the efficacy of obesity, hyperlipidemia, or inflammatory bowel disease caused by a high-fat diet through various action mechanisms such as regulating blood glucose and lipid levels, improving oxidative stress response, regulating adipose tissue metabolism, and alleviating liver injury or colonic tissue inflammation, significantly improving the symptoms of metabolic disorders, and providing coping strategies and reference basis for the prevention and treatment of obesity.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial culture, and relates to a Bifidobacterium longum subsp. longum C-5 with weight-regulating efficacy and its application. Background Art

[0002] Obesity is generally accompanied by symptoms such as disorders in lipid metabolism levels in the body, increases in serum total cholesterol and triglyceride levels, etc. Among them, lipid metabolism disorders will further exacerbate the body's inflammatory response, promote the secretion of inflammatory factors such as TNF-α and IL-6, and form a chronic low-grade inflammatory state. This state not only damages the metabolic balance state but may also have a serious impact on the liver. Research shows that the liver, as the core organ of lipid metabolism, is vulnerable to the effects of obesity and lipid metabolism disorders and develops into diseases such as fatty liver or non-alcoholic fatty liver disease. These pathological changes not only exacerbate the degree of metabolic disorders but also further increase the risk of complications such as cardiovascular diseases.

[0003] Probiotics can improve the intestinal microecological balance of the host, enhance the body's health level, and affect the health and metabolism of the host by regulating the composition and abundance of the intestinal microbiota, participating in immune regulation, or improving gastrointestinal physiological functions. The intestinal flora diversity of obese individuals is usually low, and the intestinal flora can affect and regulate body weight by regulating the intestinal barrier function and reducing the secretion of inflammatory factors. Probiotics can regulate the composition of native symbiotic bacteria or directly colonize in the intestine to increase the abundance of the intestinal microbiota with bile salt hydrolase ability, thereby increasing the concentration of unconjugated bile acids, affecting lipid metabolism, and alleviating various diseases caused by obesity. Therefore, how to provide a probiotic product with weight-regulating efficacy to relieve the related symptoms caused by obesity, hyperlipidemia, or other inflammatory bowel diseases, so as to regulate the body's lipid metabolism and improve the body's health level has important significance and application value. Summary of the Invention

[0004] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a Bifidobacterium longum subsp. longum C-5 with weight-regulating efficacy and its application.

[0005] To achieve the purpose of this invention, the following technical solutions are adopted by the present invention:

[0006] In the first aspect, the present invention provides a Bifidobacterium longum subsp. longum C-5 with weight-regulating efficacy, and the taxonomic nomenclature of the Bifidobacterium longum subsp. longum C-5 is Bifidobacterium longum subsp. longum , and the preservation number is GDMCC No: 64542, and the preservation date is April 22, 2024.

[0007] A novel Bifidobacterium longum subsp. longum strain with weight-regulating efficacy was isolated from the intestine of a healthy human body and preserved. It was named Bifidobacterium longum subsp. longum C-5 strain. This strain has the efficacy of regulating body weight, and can also regulate blood glucose and lipid levels, improve oxidative stress response, regulate adipose tissue metabolism, relieve liver injury and colonic tissue inflammation, etc. Therefore, this Bifidobacterium longum subsp. longum C-5 strain can be used to prepare agents with the efficacy of preventing, improving or treating hyperlipidemia or inflammatory bowel disease caused by a high-fat diet.

[0008] In a second aspect, the present invention provides a culture of Bifidobacterium longum subsp. longum C-5 as described in the first aspect, and the culture is prepared by the following method: inoculating Bifidobacterium longum subsp. longum C-5 into a culture medium and culturing at 35-38 °C for 22-26 h.

[0009] Among them, the above "35-38 °C" can be, for example, 35 °C, 35.5 °C, 36 °C, 36.5 °C, 37 °C, 37.5 °C, 38 °C, etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0010] The above "22-26 h" can be, for example, 22 h, 22.5 h, 23 h, 23.5 h, 24 h, 24.5 h, 25 h, 25.5 h, 26 h, etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0011] In a third aspect, the present invention provides a probiotic agent with weight-regulating efficacy, and the strains in the probiotic agent with weight-regulating efficacy include the Bifidobacterium longum subsp. longum C-5 strain described in the first aspect.

[0012] Preferably, in the probiotic agent, the viable count of Bifidobacterium longum subsp. longum C-5 is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, such as 1×10 9 CFU / mL (CFU / g), 2×10 9 CFU / mL (CFU / g), 5×10 9 CFU / mL (CFU / g), 8×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), 5×10 10 CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0013] Preferably, the strains in the probiotic agent with weight-regulating efficacy further include Lactobacillus gasseri Lactobacillus gasseri CKCC 1913, with the preservation number of CGMCC No. 23175 and the preservation date of August 23, 2021.

[0014] The present invention also creatively discovers that the above-mentioned Lactobacillus gasseri Lactobacillus gasseri strain CKCC 1913 can be compounded with Bifidobacterium longum subsp. Bifidobacterium longum subsp. longum C-5 strain for regulating body weight and improving the body's metabolic level, and has more excellent effects than single bacterial agents or other compounding methods, indicating that CKCC 1913 bacteria and C-5 bacteria have a synergistic effect in regulating and controlling body weight, regulating blood sugar and blood lipids, and relieving liver injury and colon tissue inflammation.

[0015] Preferably, the ratio of the viable count of Bifidobacterium longum subsp. C-5 to CKCC 1913 bacteria is 1:10 - 10:1. For example, it can be 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 5:1, 6:1, 8:1, 10:1, etc. Other specific point values within this numerical range can be selected and will not be elaborated here one by one.

[0016] Preferably, the dosage form of the probiotic agent includes solution, freeze-dried powder, capsule, tablet or granule.

[0017] Preferably, the dosage form of the probiotic agent is a solution, which is prepared by the following method: inoculating C-5 strain and CKCC1913 strain into a culture medium respectively, and sequentially performing activation and fermentation culture to obtain a fermentation broth; centrifuging the fermentation broth respectively, and resuspending with a solvent to obtain a C-5 bacterial suspension and a CKCC 1913 bacterial suspension, and mixing the C-5 bacterial suspension and the CKCC 1913 bacterial suspension according to the viable count ratio to obtain the probiotic agent;

[0018] Preferably, the dosage form of the probiotic agent is a freeze-dried powder, which is prepared by the following method:

[0019] Inoculating C-5 strain and CKCC 1913 strain into a culture medium respectively, and sequentially performing activation and fermentation culture to obtain a fermentation broth; centrifuging the fermentation broth respectively, mixing with a cryoprotectant and then performing freeze-drying to obtain C-5 bacterial powder and CKCC 1913 bacterial powder, and mixing the C-5 bacterial powder and the CKCC 1913 bacterial powder according to the viable count ratio to obtain the probiotic agent.

[0020] Preferably, the probiotic agent further includes a cryoprotectant;

[0021] The protective agent includes any one or a combination of at least two of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinylpyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol.

[0022] Fourthly, the present invention provides an application of the Bifidobacterium longum subsp. longum C-5 as described in the first aspect, or the culture as described in the second aspect, or the probiotic agent as described in the third aspect in the preparation of a preparation for regulating blood glucose and blood lipid levels.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention has isolated and preserved a new Bifidobacterium longum subsp. longum strain with weight-regulating efficacy from the intestinal tract of healthy humans, named Bifidobacterium longum subsp. longum C-5 strain. This strain has the efficacy of regulating body weight, and can also regulate blood glucose and blood lipid levels, improve oxidative stress response, regulate adipose tissue metabolism, relieve liver injury and colon tissue inflammation, etc. Therefore, this Bifidobacterium longum subsp. longum C-5 strain can be used to prepare preparations with the function of preventing, improving or treating obesity, hyperlipidemia or inflammatory bowel disease caused by high-fat diet.

[0025] The taxonomic nomenclature of the C-5 strain involved in the present invention is Bifidobacterium longum subsp. longum , the preservation unit is the Guangdong Provincial Microbial Culture Collection Center, the preservation number is GDMCC No: 64542, the preservation date is April 22, 2024, and the preservation address is the 5th floor of Building 59, No. 100, Xianlie Middle Road, Guangzhou;

[0026] The taxonomic nomenclature of the CKCC 1913 strain involved in the present invention is Lactobacillus gasseri Lactobacillus gasseri , the preservation unit is the General Microbiology Center of the China Microbial Culture Collection Management Committee, the preservation number is CGMCC No. 23175, the preservation date is August 23, 2021, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a statistical result chart of the body weight and inguinal fat content levels of each group of mice;

[0028] Figure 2 It is a statistical result chart of the content levels of ALT, TC, GLU, and NEFA in the serum of each group of mice;

[0029] Figure 3 It is a statistical result chart of the immune and oxidative stress levels of each group of mice;

[0030] Figure 4It is a statistical result graph of the metabolic levels of adipose tissues in each group of mice;

[0031] Figure 5 It is a statistical result graph of the content levels of TC, TG, and FAS in the liver tissues of each group of mice;

[0032] Figure 6 It is a statistical result graph of the metabolic levels of short-chain fatty acids in the feces of each group of mice;

[0033] Figure 7 It is a pathological observation result graph of liver, adipose, and colon tissue sections of each group of mice. Detailed implementation manners

[0034] The technical solutions of the present invention will be further described below through specific implementation manners. Those skilled in the art should understand that the described embodiments are only for helping to understand the present invention and should not be regarded as specific limitations to the present invention.

[0035] ① The taxonomic naming of the C-5 strain involved below is Bifidobacterium longum subsp. longum , and the preservation number is GDMCC No: 64542;

[0036] ② The taxonomic naming of the CKCC 1913 strain involved below is Lactobacillus gasseri Lactobacillus gasseri , and the preservation number is CGMCC No. 23175;

[0037] ③ The CICC 24935 involved below is the Bifidobacterium longum subsp. longum CICC 24935 strain.

[0038] The culture media and their formulas involved below are:

[0039] MRS liquid medium: Beef extract 10.0 g, glucose 20.0 g, peptone 10.0 g, yeast extract 5.0 g, sodium chloride 5.0 g, diammonium hydrogen citrate 2.0 g, dipotassium hydrogen phosphate 2.0 g, magnesium sulfate 0.2 g, manganese sulfate 0.05 g, Tween-80 1.0 g, made up to 1000 mL with distilled water, pH 6.2 - 6.4.

[0040] MRS solid medium: Beef extract 10.0 g, glucose 20.0 g, peptone 10.0 g, yeast extract 5.0 g, sodium chloride 5.0 g, diammonium hydrogen citrate 2.0 g, dipotassium hydrogen phosphate 2.0 g, magnesium sulfate 0.2 g, manganese sulfate 0.05 g, Tween-80 1.0 g, 2% agar powder, made up to 1000 mL with distilled water, pH 6.2 - 6.4.

[0041] The main components of the mouse food formula involved below are (by mass percentage):

[0042] Regular feed: corn 43.8%, secondary flour 12%, wheat 17%, salted soybean meal 7%, Peruvian fish meal 4%, and American muscle 5%; the main nutritional components are: crude protein 19.54%, crude fat 4.43%, crude fiber 2.14%, calcium 1.36%, and phosphorus 0.86%.

[0043] High-fat feed: maintenance basal diet 48.1%, lard 15%, sucrose 20%, casein 10%, experimental animal premix 2%; the main nutritional components are: protein 19.379%, fat 17.133%, crude fiber 3.321%, ash 5.3114%, calcium 1.26735%, and phosphorus 0.82223%.

[0044] The C57BL / 6J male mice (8 weeks old) involved below were purchased from Beijing SPF Biotechnology Co., Ltd.

[0045] The preparation method of the bacterial suspension and bacterial powder involved below is: after activating the strain, inoculate it into the culture medium for cultivation to obtain the culture solution; centrifuge the culture solution, resuspend the bacterial cells to obtain the bacterial suspension, or further add a cryoprotectant for freeze-drying to prepare a freeze-dried bacterial powder product. Example 1

[0046] In this example, a strain of Bifidobacterium longum subsp. with body weight regulation efficacy was isolated and screened, and the steps are as follows:

[0047] Select samples isolated from the healthy human intestine, dilute them 10-fold with normal saline with a mass concentration of 0.9% for 3 times, coat them on the solid medium, and after culturing at 37°C for 48 h, pick out colonies with different morphologies and streak-purify them on the surface of the modified MRS solid medium. Pick out single colonies and expand the culture in a liquid medium at 37°C, and then preserve them with glycerol with a mass concentration of 35%. Example 2

[0048] In this example, morphological identification and 16S rRNA molecular biology identification were carried out on the strain screened in Example 1, and the steps are as follows:

[0049] (1) Morphological identification:

[0050] Inoculate the strain into the MRS medium, anaerobically culture it at 37°C for 48 h, and then observe it under a microscope. After smear and Gram staining, microscopic examination shows that: the Gram staining is positive, the strain morphology is multi-shaped, and it presents as a milky white, smooth surface, convex, and neatly edged circular colony on the MRS medium.

[0051] (2)16S rRNA molecular biological identification:

[0052] Take out the strain stored at -80°C, inoculate it into a centrifuge tube containing 20 mL of MRS liquid medium at a ratio of 2% (v / v), culture it at 37°C for 24 h, then perform centrifugal separation, centrifuge at 8000 rpm for 10 min, discard the supernatant, and collect the bacterial cells. Extract the genomic DNA of the strain, add universal bacterial primers for PCR amplification, and send the amplified product to a sequencing company for sequencing and identification. After sequencing and analysis, the 16S rRNA sequence of this strain is shown as SEQ ID No:1. Compare the sequenced sequence with the nucleic acid sequences in GeneBank, and the result shows that the strain is *Bifidobacterium longum* subsp. *longum*.

[0053] SEQ ID No:1:

[0054]

[0055] Based on the results of 16S rRNA molecular biological identification and morphological identification in Example 2, it was confirmed that the strain belongs to Bifidobacterium longum subsp. longum, and was named Bifidobacterium longum subsp. longum Bifidobacterium longum subsp. longum Strain C-5.

[0056] Test Example 1

[0057] This test example explored the effects of strain C-5 on various symptom indexes of obese mice. The specific steps were as follows:

[0058] (1) Experimental animals: 96 male C57BL / 6J mice at 8 weeks old were adaptively fed with ordinary feed in an animal house (room temperature: 22±2°C, humidity: 55%±10%, lighting 12 h) for one week (free diet and water).

[0059] (2) Grouping and intervention methods: After adaptive feeding, the mice were randomly divided into 8 groups: Group S1 (C-5 bacterial suspension, high-fat diet), Group S2 (CKCC 1913 bacterial suspension, high-fat diet), Group S3 (C-5 bacterial suspension + CKCC 1913 bacterial suspension, viable bacteria number ratio 10:1, high-fat diet), Group S4 (C-5 bacterial suspension + CKCC 1913 bacterial suspension, viable bacteria number ratio 1:1, high-fat diet), Group S5 (C-5 bacterial suspension + CKCC 1913 bacterial suspension, viable bacteria number ratio 1:10, high-fat diet), Group S6 (CICC 24935 bacterial suspension + CKCC 1913 bacterial suspension, viable bacteria number ratio 10:1, high-fat diet), Group S7 was the model group (sterile PBS solution, high-fat diet), and Group S8 was the blank group (sterile PBS solution, ordinary feed). Each group had 12 mice. The total viable bacteria in each of Groups S1-S6 were all 10 9 CFU. Each group was gavaged once a day, and the gavage volume of the bacterial suspension / sterile PBS solution was 0.2 mL each time; during the test period, the mice in each group were allowed free diet and water.

[0060] (3) The test was carried out for a total of 6 weeks (including the first week of adaptive feeding and five weeks of gavage treatment). After the end of the sixth week, the mice were sacrificed, and serum, adipose tissue, visceral tissue and other samples were collected for detection of relevant indexes.

[0061] (4) Detection and analysis of relevant indexes

[0062] (a) Body weight and fat content

[0063] The body weight of each mouse in each group was measured in the sixth week of the test; after the end of the last week of the test, the mice were sacrificed, and the inguinal fat was collected to detect the fat content. The detection and statistical results of the body weight and inguinal fat content of the mice are as Figure 1 shown.

[0064] Hyperlipidemia was induced by feeding mice a high-fat diet. Then, by monitoring the weight changes and body fat content of the mice, the effects of probiotics on weight loss and body fat reduction could be visually evaluated. As can be seen from the figure, during the test period, the weight of the mice in S7 (model group) was higher than that in S8 (control group). At the end of the test, the weight (184.44 g) and inguinal fat content (1.353 g) of the mice in S7 (model group) were significantly higher than the body weight (146.95 g) and inguinal fat content (0.675 g) of the mice in S8 (control group). The inguinal fat proved that the obesity model of mice was successfully established.

[0065] After intragastric administration of C-5 and CKCC 1913 bacterial suspensions, the weight gain of obese mice could be slowed down, and the C-5 bacterial suspension had the best effect. In addition, it was unexpectedly found that when C-5 and CKCC 1913 bacteria were used in combination, the effect of slowing down the weight gain and body fat accumulation of obese mice was better.

[0066] (b)Blood glucose and lipid levels

[0067] Mouse serum was taken and centrifuged at 571 g for 15 min at 4°C. The content levels of alanine aminotransferase (ALT), total cholesterol (TC), glucose (GLU), and non-esterified fatty acids (NEFA) in the serum were measured with reference to the ELISA kit instructions. The test results are as Figure 2 shown.

[0068] Obesity caused by continuous intake of high-fat foods can lead to abnormalities in related blood indicators. These abnormal changes in blood lipid and blood glucose levels are directly related to the incidence and mortality of coronary heart disease and other atherosclerotic diseases. Therefore, long-term high-fat diet may disrupt the normal balance of blood components, thereby increasing the risk of cardiovascular diseases.

[0069] From the statistical results in the figure, it can be seen that at the end of the test, the content levels of TC (16.92 mmol / L), ALT (154.1 U / L), NEFA (3.085 mmol / L), and GLU (8.437 mmol / L) in the mice of S7 (model group) were significantly higher than those in the mice of S8 (control group); after intervention with C-5 and CKCC 1913 bacterial suspensions, the blood lipid and blood glucose indicators could be effectively regulated and optimized, and the C-5 bacterial suspension had the best effect. The levels of each indicator in the mice of S1 group showed an obvious downward trend, and the effect was better than that of CICC 24935 bacteria; in addition, it was unexpectedly found that when C-5 and CKCC 1913 bacteria were used in combination, the related effect was better, which could effectively improve the related blood lipid and blood glucose indicators, thereby helping to reduce the incidence probability of coronary heart disease and other atherosclerotic diseases.

[0070] (c)Immune and oxidative stress levels

[0071] Mouse serum was taken and centrifuged at 571 g for 15 min at 4°C. The content level of tumor necrosis factor-α (TNF-α) in the serum was measured with reference to the ELISA kit instructions.

[0072] Mouse liver tissues were collected for homogenization. The content levels of malondialdehyde (MDA) and superoxide dismutase (SOD) were detected with reference to the kit instructions. The results are as Figure 3 shown.

[0073] Obesity caused by continuous intake of high-fat foods promotes adipose tissue peroxidation and inflammation, leading to an increase in the levels of TNF-α in the serum and MDA in the liver, and a decrease in the activity of SOD in the liver. This not only exacerbates insulin resistance but also accelerates the occurrence of diseases such as atherosclerosis. After intervention with C-5 and CKCC 1913 bacterial suspensions, the levels of TNF-α inflammatory factors in the serum decreased to varying degrees compared with the model group. They have a certain ability to inhibit lipid peroxidation, with the MDA level lower than that of the model group and the SOD activity higher than that of the model group. It can be seen that the two bacteria can inhibit the peroxidation reaction of the liver and relieve the oxidative stress reaction of the liver to varying degrees, and the effect of C-5 is the best. At the same time, when C-5 and CKCC 1913 are used in combination, the effects in anti-inflammatory and antioxidant aspects are better.

[0074] (d)Adipose tissue metabolic level

[0075] White adipose tissues (epididymal fat, perirenal fat, and inguinal fat) were collected. After homogenization and grinding, the indexes of resistin and leptin were tested with reference to the kit instructions. The results are as Figure 4 shown.

[0076] Continuous high-fat diet usually leads to an increase in resistin levels, which may interfere with the insulin signaling pathway, reduce the sensitivity of mice to insulin, and thus trigger metabolic problems such as hyperglycemia and insulin resistance. Leptin can improve insulin resistance and fat metabolism, and thus relieve related symptoms.

[0077] As can be seen from the figure, the resistin (15.584 μg / L) in the adipose tissue of S7 (model group) mice was significantly higher than that in the control group (13.921 μg / L), and the leptin (6.297 μg / L) was significantly lower than that in the control group (7.662 μg / L); after intervention with C-5 and CKCC 1913 bacterial suspensions, the resistin and leptin in adipose tissue decreased and increased to varying degrees respectively, and the effect of C-5 was better. It can be seen that C-5 has a good effect on improving insulin resistance and fat metabolism, and has the effect of regulating the metabolism of adipose tissue in high-fat diet; at the same time, when C-5 and CKCC 1913 are used in combination, the effect is better.

[0078] (e) Level of liver injury

[0079] Mouse liver tissues were collected for homogenization, and the content levels of total cholesterol (TC), triglyceride (TG) and fatty acid synthase (FAS) were detected according to the kit instructions. The results are as Figure 5 shown.

[0080] Continuous high-fat diet is likely to lead to a large amount of cholesterol synthesis in the liver. Excessive accumulation of cholesterol in the liver will cause fatty liver, resulting in symptoms such as blood viscosity and atherosclerosis; fatty acid synthase (FAS) is synthesized in the liver and can be used as a potential therapeutic target for fatty liver and dyslipidemia.

[0081] After intragastric administration of C-5 and CKCC 1913, the levels of the three liver-related indicators all decreased to varying degrees, and the effect of C-5 was the best, indicating that C-5 has the effect of improving blood lipid levels and liver health status, and helps to reduce the related metabolic burden; at the same time, when C-5 and CKCC 1913 are used in combination, the effect is better.

[0082] (f) Short-chain fatty acid metabolism level

[0083] In the sixth week of the experiment, feces of mice in each group were collected. After freeze-drying the feces of mice, short-chain fatty acids (SCFAs) were extracted, and the levels of short-chain fatty acids (propionic acid, valeric acid, isobutyric acid) in fecal samples of each group were quantitatively analyzed by gas chromatography. The results are as Figure 6 shown.

[0084] Short-chain fatty acids (propionic acid, valeric acid, isobutyric acid) play important roles in intestinal microbial metabolism and the regulation of metabolic health, and can exert effects such as anti-inflammation, anti-bacterial, and regulation of intestinal integrity. After intragastric administration of C-5 and CKCC 1913, the content levels of the three short-chain fatty acids in the feces of mice increased to varying degrees, and C-5 had the best effect, indicating that C-5 has the ability to regulate short-chain fatty acid metabolism under a high-fat diet, which helps improve overall metabolic health; further, when C-5 and CKCC 1913 are used in combination, the related regulatory ability is better.

[0085] (g) Histopathological observation of liver, adipose, and colon tissue sections

[0086] Liver, adipose, and colon tissues of groups S1, S7, and S8 were collected for H&E and Oil Red O staining sections, and the pathological changes in the morphology of the section tissues or cells were observed through an optical microscope. The results are as Figure 7 shown.

[0087] It can be seen from the staining results in the figure that in the liver staining model, compared with S7 (model group), the positive area of S1 group (C-5 intervention group) was significantly reduced, indicating that C-5 has a good protective effect on liver damage caused by a high-fat diet and reduces the degree of liver pathological damage; in the observation of adipose sections, the adipose in S8 (control group) was dense and uniform, the adipose area of S7 (model group) was larger, while the adipose area of S1 group (C-5 intervention group) showed a shrinking trend, indicating that C-5 helps reduce adipose accumulation and has a certain improvement effect on obesity or fatty liver.

[0088] In S7 (model group), obvious folds were formed in the mucosal layer of the intestinal tissue. The mucosal epithelium was composed of simple columnar epithelium and goblet cells. A large number of intestinal glands were contained in the lamina propria. These intestinal glands were densely arranged, containing a large number of goblet cells and a small number of columnar cells. The muscularis mucosa separated the lamina propria from the submucosa. The submucosa was loose connective tissue, and focal aggregation of lymphocytes was visible; in contrast, the intestinal tissue of S1 group (C-5 intervention group) did not show the above abnormal manifestations, showing an obvious improvement effect, indicating that C-5 intervention treatment has a positive effect on maintaining intestinal health and reducing inflammatory responses.

[0089] In summary, the present invention isolated and preserved a new Bifidobacterium longum subsp. with the efficacy of body weight regulation, and named it Bifidobacterium longum subsp. Bifidobacterium longum subsp. longumThe C-5 strain has the efficacy of body weight regulation, and can also regulate blood glucose and lipid levels, improve oxidative stress response, regulate adipose tissue metabolism, relieve liver injury and colonic tissue inflammation, etc. Therefore, this Bifidobacterium longum subsp. C-5 strain can be used to prepare products with the effects of preventing, improving or treating obesity, hyperlipidemia or inflammatory bowel disease caused by a high-fat diet, etc.

[0090] The applicant declares that the technical solutions of the present invention are illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0091] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0092] In addition, it should be noted that, in the case of no contradiction, the various specific technical features described in the above specific embodiments can be combined in any suitable way. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A probiotic agent having the effect of reducing weight, characterized in that: The strains in the probiotic with weight loss efficacy are composed of Bifidobacterium longum subspecies longum C-5 and Lactobacillus gasseri CKCC 1913; Wherein, the classification of the Bifidobacterium longum subspecies longum C-5 is named Bifidobacterium longum subsp. longum , the deposit number is GDMCC No: 64542, and the deposit date is April 22, 2024; The classification of the Lactobacillus gasseri CKCC 1913 is named Lactobacillus gasseri Lactobacillus gasseri The deposit number is CGMCC No. 23175, and the deposit date is August 23, 2021.

2. The probiotic agent with weight loss effect as claimed in claim 1, characterized in that: In the probiotic, the viable count of the Bifidobacterium longum subspecies longum C-5 is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

3. The probiotic agent with weight loss effect as claimed in claim 1, characterized in that: The ratio of the viable counts of Bifidobacterium longum subspecies longum C-5 to Lactobacillus gasseri CKCC 1913 is 1:10-10:

1.

4. The probiotic agent having weight loss efficacy according to claim 1, characterized in that: The dosage form of the probiotics includes solution, lyophilized powder, capsule, tablet or granule.

5. The probiotic agent having weight loss efficacy according to claim 4, characterized in that: The dosage form of the probiotic is a solution, which is prepared by the following method: The C-5 strain and the CKCC 1913 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broths are centrifuged respectively, and resuspended with a solvent to obtain a C-5 bacterial suspension and a CKCC 1913 bacterial suspension; the C-5 bacterial suspension and the CKCC 1913 bacterial suspension are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent; Alternatively, the probiotic is in the form of a freeze-dried powder, which is prepared by the following method: The C-5 strain and the CKCC 1913 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and freeze-dried to obtain C-5 bacterial powder and CKCC 1913 bacterial powder; the C-5 bacterial powder and the CKCC 1913 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

6. The probiotic agent having weight loss efficacy according to claim 1, characterized in that: The probiotics also include a protective agent; The protective agent includes any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

7. Use of the probiotic according to any one of claims 1 to 6 in the preparation of a preparation for lowering blood sugar and blood lipid levels.

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