Bifidobacterium pseudocatenulatum and uses thereof

Abstract

The application discloses Bifidobacterium Pseudocatenulatum and application thereof, and the Bifidobacterium Pseudocatenulatum YSBP01CCTCC NO: M 20232067. Experiments prove that the Bifidobacterium Pseudocatenulatum can enhance the antioxidant capacity of Caenorhabditis elegans, prolong the life of the nematode, improve the activity and the reproductive capacity of the nematode, reduce fat accumulation, improve the mitochondrial membrane potential, and delay the aging of the body. The Bifidobacterium Pseudocatenulatum can also significantly inhibit the weight increase of a high-fat-diet mouse, reduce the TG level in serum of the mouse, significantly reduce the fasting blood glucose concentration, and effectively prevent and relieve obesity and type 2 diabetes.

Description

Technical Field

[0001] This invention belongs to the field of probiotic technology. More specifically, it relates to Bifidobacterium pseudobulbarum and its applications. Background Technology

[0002] With advancements in medical technology and increased life expectancy, aging and its associated chronic diseases have become the leading cause of death, posing a significant threat to the health of the elderly. Therefore, improving the quality of life for the elderly, increasing healthy lifespan, and reducing their disease burden remain substantial challenges. On the other hand, abundant material resources have led to a high prevalence of obesity in modern society, highlighting related problems such as increased incidence of metabolic diseases like diabetes, fatty liver disease, and cardiovascular disease, thus increasing mortality risk and threatening public health. Currently, reducing excessive fat accumulation and preventing or delaying adipose tissue dysfunction have been proven to be effective interventions for extending and improving lifespan.

[0003] Since Metchnikoff proposed his famous "yogurt for longevity" theory in 1907, numerous studies have shown that probiotics can delay aging by regulating the gut microbiota, increasing the abundance of beneficial bacteria, reducing the number of pathogenic bacteria, and improving the body's immunity and antioxidant capacity. They can also intervene in the pathogenesis of obesity and type 2 diabetes by regulating bile acid synthesis and fat and glucose metabolism. However, probiotics are specific at the genus, species, and strain levels, and different strains often exhibit significant differences in their probiotic functions and effects. Therefore, isolating and screening high-performance probiotic strains is crucial for the research and development of probiotics and related products.

[0004] Bifidobacterium pseudocatenulatum is prevalent in people of all ages and is the dominant Bifidobacterium in the gut of the Chinese population. Several strains of this species have been shown to have probiotic functions, including improving glucose and lipid metabolism, regulating immunity, and alleviating anxiety and depression. In the food industry, Bifidobacterium pseudocatenulatum demonstrates its potential as a food leavening agent by secreting phytase and β-glucosidase, thereby increasing the body's utilization of minerals and isoflavones in bread. Although several strains of this species possess probiotic functions, research on its anti-aging and antioxidant effects has not yet been reported. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a *Bifidobacterium pseudosporidis*.

[0006] A second objective of this invention is to provide the application of the aforementioned *Bifidobacterium pseudosporidis* in the preparation of antioxidant products.

[0007] A third objective of this invention is to provide the application of the aforementioned *Bifidobacterium pseudosporidis* in the preparation of anti-aging products.

[0008] A fourth objective of this invention is to provide the application of the aforementioned Bifidobacterium pseudosporidis in the preparation of products for treating obesity.

[0009] The fifth objective of this invention is to provide the application of the aforementioned Bifidobacterium pseudosporidis in the preparation of hypoglycemic products.

[0010] The technical solution of this invention is summarized as follows:

[0011] Bifidobacterium Pseudocatenulatum YSBP01CCTCC NO: M20232067.

[0012] The application of the above-mentioned Bifidobacterium pseudosporidis in the preparation of antioxidant products.

[0013] The application of the aforementioned Bifidobacterium pseudosporidis in the preparation of anti-aging products.

[0014] The application of the above-mentioned Bifidobacterium pseudosporidis in the preparation of products for treating obesity.

[0015] The application of the aforementioned Bifidobacterium pseudosporidis in the preparation of hypoglycemic products.

[0016] The present invention has the following beneficial effects:

[0017] Experiments have shown that *Bifidobacterium pseudodocatenulatum* (YSBP01CCTCCNO: M 20232067) of this invention can enhance the antioxidant capacity of *C. elegans*, prolong the lifespan of nematodes, improve their motility and reproductive capacity, reduce fat accumulation, increase mitochondrial membrane potential, and delay aging. Furthermore, it significantly inhibits weight gain in mice fed a high-fat diet, reduces serum triglyceride levels, and significantly lowers fasting blood glucose concentration, effectively preventing and alleviating obesity and type 2 diabetes. Attached Figure Description

[0018] Figure 1 The effect of YSBP01 on the lifespan of Caenorhabditis elegans N2;

[0019] Figure 2 Changes in motility (A) and fertility (B) of YSBP01 after feeding it with Caenorhabditis elegans;

[0020] Figure 3 The effect of feeding YSBP01 on the fat content of nematodes;

[0021] Figure 4The effect of feeding YSBP01 on the mitochondrial membrane potential of nematodes;

[0022] Figure 5 The antioxidant properties of YSBP01 (A: MDA content; B: SOD activity; C: GSH content; D: CAT activity);

[0023] Figure 6 The effects of YSBP01 on body weight (A, B) and serum triglycerides (C) in mice fed a high-fat diet;

[0024] Figure 7 The effect of YSBP01 on fasting blood glucose levels in type 2 diabetic mice. Detailed Implementation

[0025] Bifidobacterium Pseudocatenulatum YSBP01 was deposited on October 30, 2023, at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, with accession number CCTCC NO: M 20232067.

[0026] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0027] The reagents or formulations used in the following examples:

[0028] NGM solid medium: 3g sodium chloride, 2.5g tryptone, 17g agar powder, 25mL potassium phosphate buffer (1mol / L), 1mL 1M CaCl2 aqueous solution, 1mL 1M MgSO4 aqueous solution; add distilled water to a final volume of 1000mL, autoclave at 121℃ for 20min. After the temperature drops to approximately 60℃, add 1mL of 5mg / mL cholesterol solution (95% ethanol aqueous solution as solvent) under aseptic conditions, shake well, and pour into plates.

[0029] LB liquid medium: 5g yeast extract, 10g tryptone, 10g NaCl, add distilled water to a final volume of 1000mL, autoclave at 121℃ for 20min.

[0030] MRS liquid medium for 0.5% L-cysteine ​​hydrochloride: MRS broth (52.2 g / L, manufacturer: Shandong Top Biotechnology Co., Ltd.), 0.5% L-cysteine ​​hydrochloride anhydrous, add distilled water to a final volume of 1 L, autoclave at 121℃ for 20 min.

[0031] MRS solid medium of 0.5% L-cysteine ​​hydrochloride: Add 2% agar to MRS liquid medium of 0.5% L-cysteine ​​hydrochloride.

[0032] Wild-type Caenorhabditis elegans N2 and Escherichia coli OP50 were purchased from Fujian Shangyuan Biotechnology Co., Ltd.

[0033] The kits for measuring malondialdehyde (MDA) content, glutathione (GSH) content, and catalase (CAT) activity were purchased from Nanjing Jiancheng Biotechnology, and the kit for measuring superoxide dismutase (SOD) activity was purchased from Beyotime Biotechnology.

[0034] M9 buffer: Weigh 6g disodium hydrogen phosphate, 3g potassium dihydrogen phosphate, 5g sodium chloride, and 1mL 1mol / L magnesium sulfate buffer. Add distilled water to bring the volume to 1000mL and autoclave at 121℃ for 20min.

[0035] Lysis buffer: Weigh 0.1g sodium hydroxide, add 500μL sodium hypochlorite solution, and after heat dissipation, add 4mL M9 buffer to dissolve and mix well. The lysis buffer should be prepared fresh for use.

[0036] Example 1: Isolation and Identification of Strains

[0037] 1. Isolation of strains

[0038] Fecal samples from 50-year-old male volunteers in Shaoxing, China, were serially diluted using sterile, anaerobic saline solution to obtain concentration gradients of 10-10. -1 Up to 10 -11 Dilute the bacterial solution within a certain range. Take 10... -9 10 -10 10 -11 Three gradients of bacterial suspensions were plated (MRS solid medium containing 0.5% L-cysteine ​​hydrochloride). After even spreading, the plates were inverted and placed in anaerobic culture bags for incubation. The growth time of the strain was approximately 24-48 hours. Single colonies suspected to be *Bifidobacterium pseudomicrobium* were selected and isolated from the colonies and transferred to a new medium (MRS solid medium containing 0.5% L-cysteine ​​hydrochloride) for further incubation using the streak method. Slightly transparent, small single colonies were picked, and a small amount was gently scraped from the surface using a 10 μL pipette tip. The colony was then thoroughly mixed with 10 μL of pure water, and 3 μL was used for PCR amplification.

[0039] 2. Molecular biological identification

[0040] (1) Take 3 μL of the bacterial culture obtained in step 1 and prepare the PCR amplification system and run the PCR amplification according to the following system (see Table 1 and Table 2).

[0041] Table 1: PCR reaction system

[0042]

[0043] Table 2: PCR reaction procedure

[0044]

[0045] SEQ ID NO.1 (upstream primer): 27f(5′-AGAGTTTGATCCTGGCTCAG-3′)

[0046] SEQ ID NO.2 (downstream primer): Nucleotide sequence listed: 1492r(5′-TACGGTTACCTTGTTACGACTT-3′)

[0047] (2) Prepare 1.5% agarose gel by adding 10 μL of Ultra GelRed (10000×) nucleic acid dye to every 100 mL.

[0048] (3) Add 5 μL of the above PCR reaction solution to each well. Use DL2000 marker as a band size control. The band size is about 327 bp, which can be preliminarily identified as Bifidobacterium pseudosporidis.

[0049] (4) Samples initially identified as Bifidobacterium pseudosporidis were further subjected to PCR amplification using universal primers (SEQ ID NO.1, SEQ ID NO.2). The amplified PCR liquid was sent to Beijing Qingke Biotechnology Co., Ltd. (hereinafter referred to as Qingke Biotechnology) for sequencing. After the sequencing results were obtained, the sequence was compared on the website https: / / blast.ncbi.nlm.nih.gov / Blast.cgi to obtain the comparison results.

[0050] (5) After comparison, the samples identified as *Bifidobacterium pseudocatenulatum* were expanded and cultured in MRS liquid medium containing 0.5% L-cysteine ​​hydrochloride for 48 h. 100 μL of the bacterial culture was then amplified again by PCR. After amplification, the samples were sent to Qingke Biotechnology for sequencing, while the remaining bacterial cultures were cultured further. The 16S rDNA sequence (SEQ ID No. 3) obtained from sequencing was compared with the NCBI BLAST sequence. The similarity to *Bifidobacterium pseudocatenulatum* in Genebank was greater than 99%, confirming that the strain was indeed *Bifidobacterium pseudocatenulatum*. If identified as *Bifidobacterium unimodalum* after secondary sequencing, it is a pure *Bifidobacterium unimodalum* (named YSBP01). The bacterial culture was centrifuged, the supernatant was discarded, and cryopreservation buffer (bacterial culture to cryopreservation buffer volume ratio 1:1) was added (the cryopreservation buffer is a 60% glycerol aqueous solution). After aliquoting, it was stored at -80°C. It is deposited at the China Center for Type Culture Collection.

[0051] Example 2: Tolerability evaluation of YSBP01

[0052] Bifidobacterium Pseudocatenulatum YSBP01CCTCC NO: M20232067, abbreviated as YSBP01.

[0053] 1. Evaluation of YSBP01 artificial gastric juice tolerance performance

[0054] (1) Test methods

[0055] Frozen YSBP01 was inoculated at a 1% inoculum into MRS liquid medium containing 0.5% L-cysteine ​​hydrochloride and cultured anaerobically at 37°C. After 24 hours of culture, the bacterial culture was transferred at a 5% inoculum into fresh MRS liquid medium containing 0.5% L-cysteine ​​hydrochloride and cultured anaerobically at 37°C. After 18 hours of culture, 5 ml of bacterial culture was collected under aseptic conditions and centrifuged at 8000 rpm for 5 min to separate the bacterial cells. The bacterial precipitate was resuspended in physiological saline and thoroughly mixed, then centrifuged at 8000 rpm for 5 min to collect the bacterial cells. YSBP01 was resuspended in equal volumes of artificial gastric fluid at pH=2 and pH=3 and thoroughly mixed, then incubated anaerobically at 37°C. Bacterial cultures were serially diluted 10-fold with physiological saline at 0 h, 1.5 h, and 3 h of incubation, and plated at appropriate dilutions. Two dilutions were selected for each sample, and two plates were prepared for each dilution. After spreading, the plates were placed upside down in a 37°C incubator for anaerobic culture. After 1-2 days of culture, the growth status of the colonies on the plates was observed, the number of colonies was recorded, and the survival rate was calculated.

[0056]

[0057] Where A0 is the number of colonies after 0 hours of incubation, and An is the number of colonies after n hours of incubation.

[0058] (2) Test Results

[0059] The survival rate of YSBP01 in artificial gastric fluid is shown in Table 3.

[0060] Table 3: Statistics on Tolerance to Artificial Gastric Fluid in YSBP01

[0061]

[0062] 2. Evaluation of the tolerance performance of YSBP01 artificial intestinal fluid

[0063] (1) Test methods

[0064] YSBP01 was resuspended in an equal volume of artificial intestinal fluid and thoroughly mixed, then incubated anaerobically at 37°C. After 0h, 4h, and 8h of incubation, the bacterial suspensions were serially diluted 10-fold with physiological saline, and appropriate dilutions were selected for plating. Two dilutions were selected for each sample, and two plates were prepared for each dilution. The plated plates were inverted and anaerobically cultured at 37°C for 1-2 days. The growth status of the colonies on the plates was observed, the colony count was recorded, and the survival rate was calculated.

[0065]

[0066] Where A0 is the number of colonies after 0 hours of incubation, and An is the number of colonies after n hours of incubation.

[0067] (2) Test Results

[0068] The survival of YSBP01 in artificial intestinal fluid is shown in Table 4.

[0069] Table 4: Statistics on tolerance to artificial intestinal fluid in YSBP01

[0070]

[0071] Example 3: Lifetime test of YSBP01 on wild-type Caenorhabditis elegans N2 (hereinafter referred to as nematode)

[0072] 1. Activation of microbial strains

[0073] Take 200 μL of YSBP01 that has been frozen at -80℃, inoculate it into 10 mL of MRS liquid medium containing 0.5% L-cysteine ​​hydrochloride, place it in an anaerobic bag, and anaerobic culture at 37℃ for 24–48 h to activate the strain.

[0074] 2. Cultivation of Nematodes

[0075] Escherichia coli OP50 (OD=0.5) was spread on NGM solid medium as food for nematodes. Then, nematodes were picked and placed on the surface of NGM solid medium and placed in a biochemical incubator at 20°C for culture. During the culture period, the nematodes were transferred to fresh NGM plates coated with Escherichia coli OP50 every day (cultured to L4 stage).

[0076] 3. Synchronization of nematodes

[0077] Collect L4 stage nematodes into centrifuge tubes, add 1 mL of M9 buffer to wash away excess E. coli OP50, allow to stand and precipitate, then remove the supernatant. Repeat washing 3 times. Then add 1 mL of lysis buffer and shake well, remove the supernatant, and then centrifuge with M9 buffer (3000 r / min, 1 min) and wash 3 times. Remove the supernatant and retain the eggs. Transfer the eggs to a new NGM plate and incubate in a 20℃ biochemical incubator. After about 16-18 hours, they will grow into L1 stage larvae.

[0078] 4. Determination of nematode lifespan

[0079] YSBP01 was cultured in MRS liquid medium containing 0.5% L-cysteine ​​hydrochloride to the logarithmic growth phase.

[0080] Escherichia coli OP50 was cultured in LB liquid medium to the logarithmic growth phase;

[0081] Collect YSBP01 cells and Escherichia coli OP50 cells by centrifugation at 8000 r / min for 10 min at 4℃.

[0082] Ninety L1-stage nematodes were selected and placed on NGM plates containing YSBP01 and E. coli OP50, respectively, and incubated at 20°C in a biochemical incubator. Every other day, all surviving nematodes were transferred to fresh NGM plates coated with the same bacterial suspension to prevent newly hatched nematodes from affecting the experimental results. The experiment continued until all nematodes died, and nematode lifespan curves were plotted.

[0083] 5. Test Results

[0084] The results are as follows Figure 1 As shown, compared with the average lifespan (13 days) and longest lifespan (20 days) of OP50-fed E. coli, the average lifespan of YSBP01 was significantly extended by 35.4% (p<0.05), and the longest lifespan was extended by 6 days, indicating that YSBP01 can significantly extend the lifespan of nematodes.

[0085] Example 4: Effects of YSBP01 on the motility and reproductive capacity of nematodes

[0086] 1. Test Methods

[0087] (1) Exercise capacity is one of the key indicators of health. Exercise status and exercise capacity will gradually deteriorate and weaken with the aging of the body. Nematodes were cultured using strain YSBP01, and the culture method was the same as in Example 3. Escherichia coli OP50 was fed only as a control group. On the 3rd day, 20 nematodes were randomly selected and placed on fresh NGM plates. After the nematodes moved freely for 30 seconds, the frequency of body bending and swaying of the nematodes within 20 seconds was observed using a stereomicroscope. One body bending refers to the body bending from one direction to another and then returning to the original direction.

[0088] (2) The anti-aging drugs or strains selected should not impair the body's reproductive capacity, which is one of the most critical indicators of health. Therefore, the effect of YSBP01 on the reproductive capacity of nematodes was verified using the number of offspring as an indicator. Nematodes were cultured with the test strain according to the method described in Example 3 until the L4 stage. Two nematodes were randomly selected and placed on NGM plates coated with YSBP01 and NGM plates coated with Escherichia coli OP50, respectively, with 5 replicates in each group. During the nematode oviposition period, the oviposition adults were transferred to new NGM plates with the same bacterial solution every 1 day. The old plates were then placed in a 20°C biochemical incubator for further culture until the eggs hatched. The total number of larvae hatched on the plates was counted until the nematode eggs had finished hatching.

[0089] 2. Experimental Results

[0090] Compared with the E. coli OP50 control group, the frequency of body bending and swaying of nematodes in the YSBP01 group increased by 18.0% and 39.8% on days 3 and 12, respectively. Figure 2 A, p<0.05), the total number of eggs laid increased by 12.2% ( Figure 2 B, p<0.05). This indicates that feeding YSBP01 can significantly enhance the motility and reproductive capacity of nematodes, and has good anti-aging properties.

[0091] Example 5: Effects of YSBP01 on lipid content and mitochondrial membrane potential in nematodes

[0092] 1. Test Methods

[0093] (1) Nematodes were cultured using YSBP01, following the same culture method as in Example 3. Escherichia coli OP50 was used as a control group. After culturing for 3 days in the L4 stage, 20 nematodes were randomly selected and their fat content was measured using Oil Red O staining. The nematodes were rinsed three times with M9 buffer, and the pharynx was sealed with 25 mM levamisole hydrochloride. 200 μL of 4% paraformaldehyde solution was added and allowed to stand for 20 min. The solution was then removed, and the nematodes were subjected to three freeze-thaw cycles in liquid nitrogen. Finally, they were stained with 60% isopropanol Oil Red O (10 ml 60% isopropanol + 0.05 g Oil Red O). After 5 h, excess dye was rinsed off with M9 buffer, and the nematodes were observed using an optical microscope.

[0094] (2) Caenorhabditis elegans was cultured using YSBP01, and the culture method was the same as in Example 3. Escherichia coli OP50 was used as the control group. After culturing for 3 days in the L4 stage, 20 nematodes were randomly selected and their mitochondrial membrane potential was measured using JC-1 dye. The nematodes were incubated with freshly prepared JC-1 dye solution in the dark for 2 hours. The dye on the surface of the nematodes was washed off with M9 buffer. The red fluorescence (emission wavelength about 590 nm) and green fluorescence (emission wavelength 529 nm) were observed under a fluorescence microscope, and the images were taken and recorded. The fluorescence images were quantitatively calculated using ImageJ software.

[0095] 2. Test Results

[0096] Compared with the Escherichia coli OP50 control group, the fat content of nematodes fed with YSBP01 was significantly reduced. Figure 3 ImageJ software analysis revealed that the red light density of nematodes' lipids was significantly reduced after YSBP01 interference, decreasing by 38.6%. Figure 3 (p<0.05). Compared with OP50, JC-1 staining showed enhanced red fluorescence and decreased green fluorescence. Figure 4 ), and the red-green fluorescence ratio increased by 90.0% ( Figure 4 (p<0.05) indicates that YSBP01 significantly reduced fat deposition in nematodes, thereby affecting nematode fat metabolism, slowing the decline in mitochondrial membrane potential, and reducing mitochondrial oxidative damage.

[0097] Example 6: Effect of YSBP01 on the antioxidant capacity of nematodes

[0098] 1. Test Methods

[0099] Nematodes were cultured using strain YSBP01, following the same cultivation method as in Example 3. A control group fed only with *E. coli* OP50 was used. After culturing for another 3 days in the L4 stage, the nematodes were collected, washed three times with M9 buffer, and allowed to settle naturally. The nematodes were then homogenized in a homogenizer on ice, centrifuged at 4000 rpm at 4°C, and the supernatant was collected as the protein homogenate. Malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, glutathione (GSH) content, and catalase (CAT) activity were measured according to the kit instructions.

[0100] 2. Test Results

[0101] Compared with the control group of Escherichia coli OP50, the MDA content in nematodes fed with YSBP01 was significantly reduced by 38.0%. Figure 5 A, p<0.05), SOD activity was significantly increased by 58.8% ( Figure 5 B, p<0.05), the GSH content increased significantly by 59.5% ( Figure 5 C, p<0.05), CAT activity was significantly increased by 34.5% ( Figure 5 D, p<0.05), the above results indicate that YSBP01 can enhance the antioxidant capacity of nematodes by increasing SOD activity, CAT activity and GSH content and reducing MDA content in the body.

[0102] Example 7: Analysis of the effect of YSBP01 on obesity in mice on a high-fat diet

[0103] 1. Test Methods

[0104] (1) Analysis of the effect of YSBP01 on the body weight of mice on a high-fat diet

[0105] Six-week-old male C57BL / 6J mice were acclimatized for one week, weighed, labeled, and randomly divided into three groups: blank control group (NC), model control group (HFD), and YSBP01 intervention group (YSBP01). The grouping and treatment methods of the experimental animals are shown in Table 5.

[0106] Table 5 Grouping of experimental animals

[0107]

[0108] All mice were weighed at a fixed time each week, and their high-fat diet was changed every 48 hours with the weight of feed given and leftover recorded.

[0109] (2) Analysis of serum triglycerides (TG) in mice fed a high-fat diet using YSBP01

[0110] After intraperitoneal anesthesia for 3-5 minutes, blood was collected from the orbital vein of mice in each group. After standing at room temperature for 30 minutes, the blood was centrifuged at 3000 rpm for 10 minutes at 4°C. The supernatant was collected, aliquoted into centrifuge tubes, and stored at -80°C. The TG content in the serum was analyzed using a fully automated analyzer.

[0111] 2. Test Results

[0112] The mice were weighed and their weights recorded weekly during the experiment, and the results were as follows: Figure 6 A and Figure 6 As shown in Figure B, the HFD group mice showed significant weight gain, indicating successful model establishment. In the YSBP01 group, after intervention with *Bifidobacterium pseudomicrobium* (YSBP01), the weight of mice on a high-fat diet significantly decreased. Compared to the model control group, the weight gain in the YSBP01 group was significantly lower, indicating that *Bifidobacterium pseudomicrobium* (YSBP01) significantly inhibited the weight gain induced by a high-fat diet in mice. Figure 6 As shown in Figure C, compared with the blank control group, the serum TG content of mice in the HFD group was significantly increased (p<0.01). After gavage administration of Bifidobacterium pseudosporidis YSBP01, the TG content in the YSBP01 intervention group was significantly decreased compared with the HFD group (p<0.01), indicating that YSBP01 has a good regulatory effect on triglycerides.

[0113] Example 8: YSBP01 can reduce fasting blood glucose levels in type 2 diabetic mice.

[0114] 1. Test Methods

[0115] Six-week-old healthy male C57BL / 6J mice were used and acclimatized to their environment for one week. They were then randomly divided into four groups: blank control group (NC), model control group (DM), metformin control group (MET), and YSBP01 intervention group (YSBP01). The dose of bacterial suspension administered by gavage was 1.0 × 10⁻⁶. 9 CFU / mL. The grouping and treatment methods for experimental animals are shown in Table 6.

[0116] Table 6 Grouping of experimental animals

[0117]

[0118] Except for the blank control group, after 8 weeks of high-sugar, high-fat diet intervention, mice were intraperitoneally injected with 40 mg / kg / day of STZ (STZ powder dissolved in 0.1 mol / L sodium citrate buffer (pH 4.5) and the NC group was simultaneously administered an equal volume of 0.1 mol / L sodium citrate buffer. Injections were repeated for 5 consecutive days, and blood glucose was measured every 3 days after the injection. Mice with random blood glucose >16.7 mmol / L, accompanied by polydipsia, polyuria, and polyphagia, were considered diabetic mice.

[0119] Before the end of the 10th week of the experiment, the blood glucose homeostasis of mice was evaluated by oral glucose tolerance test (OGTT). The specific procedure was as follows: after fasting for 16 hours, but without the restriction of water, mice were given 30% glucose solution by gavage at a dose of 2 g / kg. Blood was collected from the tail vein before gavage (0 min) and at 15, 30, 60, 90 and 120 min after gavage, and the blood glucose value of the tail tip of the mice was measured using a Roche fully automated blood glucose meter.

[0120] 2. Test Results

[0121] Experimental results are as follows Figure 7 As shown, fasting blood glucose levels were significantly elevated in the DM group mice, while gavage administration of YSBP01 significantly reduced fasting blood glucose levels in the model mice, approaching those of the NC group. Its ability to reduce fasting blood glucose levels in mice was similar to that in the MET group. This indicates that YSBP01 has the effect of improving glycemic homeostasis and increasing oral glucose tolerance in mice fed a high-fat diet.

[0122] In summary, the YSBP01 strain provided by this invention can prolong the lifespan of nematodes, enhance their mobility and reproductive capacity, reduce fat accumulation, increase the production of SOD, GSH, CAT, etc., and reduce the content of MDA in the body. It also significantly inhibits the weight gain of mice on a high-fat diet, reduces the level of serum TG in mice, and significantly reduces fasting blood glucose concentration. It has antioxidant, fat-reducing, and anti-aging effects, and can effectively prevent and alleviate obesity and type 2 diabetes. It is a probiotic.

[0123] The product is a health food, medicine, feed or additive, and its formulation is in the form of liquid, jelly, stick, powder, tablet, granule or capsule.

[0124] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. Bifidobacterium Pseudocatenulatum YSBP01, with accession number CCTCCNO: M20232067.

2. The application of Bifidobacterium pseudosporidis YSBP01 as described in claim 1 in the preparation of antioxidant products.

3. The application of Bifidobacterium pseudosporidis YSBP01 as described in claim 1 in the preparation of anti-aging products.

4. The use of Bifidobacterium pseudosporidis YSBP01 as described in claim 1 in the preparation of products for treating obesity.

5. The application of Bifidobacterium pseudosporidis YSBP01 as described in claim 1 in the preparation of hypoglycemic products.

Images