Bifidobacterium adolescentis for relieving estrogen-related metabolic disorder and obesity and application of bifidobacterium adolescentis

Abstract

The invention discloses bifidobacterium adolescentis for relieving estrogen-related metabolic disorder and obesity and application thereof, and belongs to the technical field of microorganisms. The bifidobacterium adolescentis CCFM1506 screened by the invention can effectively improve ovariectomized individual estrogen disorder symptoms, obviously slow down the weight increase speed and weight increment of estrogen disorder individuals, increase the expression quantity of serum estradiol, CYP11A1 and ERalpha in adrenal gland and the high-density lipoprotein content in serum, reduce the total cholesterol and glucose level and improve the ovariectomized ovariectomized ovariectomized ovariectomized ovariectomized ovariectomized ovariectomized ovariectomized ovariectomized abnormal glucose and lipid metabolism is adjusted; the invention has the advantages of simple preparation method and wide application prospect, can be used for preparing medicines for relieving or treating estrogen-related metabolic disorders and obesity, and can be used for preparing health-care products beneficial to controlling in-vivo fat and maintaining the health level of blood fat, and has obvious physiological effects and wide application prospects, and can be used for preparing the medicines for relieving or treating the estrogen-related metabolic disorders and obesity and preparing health-care products beneficial to controlling in-vivo fat and maintaining the health level of blood fat. A new thought and a new basis are provided for developing functional foods or medicines based on the probiotics.

Description

Technical Field

[0001] This invention relates to a strain of Bifidobacterium adolescentis that alleviates estrogen-related metabolic disorders and obesity, and its applications, belonging to the field of microbial technology. Background Technology

[0002] In the female physiological process, the homeostasis of estrogen levels plays a crucial role in maintaining host metabolism. As women age, ovarian function gradually declines; during the perimenopausal and postmenopausal periods, estrogen levels drop rapidly, causing dysfunction of the hypothalamus-pituitary-ovarian axis. This endocrine imbalance is a major contributing factor to a series of metabolic syndromes, including weight gain, visceral fat accumulation, abnormal bone metabolism, and insulin resistance.

[0003] Currently, clinical practice mainly involves hormone replacement therapy combined with selective estrogen receptor modulators to supplement estrogen. However, long-term use can lead to serious adverse reactions such as a 25% increased risk of breast cancer and a 30% increased risk of venous thrombosis, limiting its widespread application. Therefore, developing safe and effective non-hormone replacement methods has become a current research hotspot.

[0004] In recent years, research has revealed that gut microbiota participate in the metabolic regulation of host estrogen through pathways such as the gut-hepatic-estrogen cycle and the gut-brain-gonadal axis. Therefore, regulating the composition and function of the gut microbiota has become a promising strategy for estrogen metabolism intervention. Probiotics can indirectly or directly affect estrogen metabolism by regulating gut microbiota homeostasis, immune responses, and metabolic functions. In existing probiotic research and products, insufficient strain specificity is one of the main technical challenges in regulating estrogen disorders. Although many probiotic products on the market label themselves as containing "Bifidobacterium" or "Lactobacillus," the strains used often have not undergone functional screening targeting estrogen metabolic pathways and lack systematic verification of their mechanisms of action, often failing to effectively regulate estrogen metabolism. Existing studies have shown that only certain specific strains, such as *Lactobacillus plantarum* CCFM1180, have demonstrated the potential to participate in estrogen metabolism regulation by modulating β-glucuronidase activity or the enterohepatic circulation in animal experiments, exhibiting a certain ability to alleviate estrogen metabolism-related metabolic disorders and obesity. However, research on these functional strains remains fragmented, and their ability to alleviate estrogen metabolism-related metabolic disorders and obesity is limited. Therefore, there is an urgent need to develop new strains that can effectively intervene in estrogen disorders. Summary of the Invention

[0005] This invention provides a strain of Bifidobacterium adolescentis (CCFM1506), taxonomically named Bifidobacterium adolescentis, which was deposited on June 6, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No.: 66474), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0006] In one embodiment, the Bifidobacterium adolescentis CCFM1506 is derived from a fecal sample of a woman of childbearing age in Wuxi City, Jiangsu Province.

[0007] In one embodiment, the Bifidobacterium adolescentis CCFM1506 has the following characteristics:

[0008] (1) Significantly increases estradiol and high-density lipoprotein in mammalian serum.

[0009] (2) Significantly reduces total cholesterol and glucose in mammalian serum.

[0010] The present invention also provides a composition containing the aforementioned Bifidobacterium adolescentis CCFM1506.

[0011] In one embodiment, the composition includes, but is not limited to, microbial preparations.

[0012] In one embodiment, the amount of Bifidobacterium adolescentis CCFM1506 added to the microbial preparation is not less than 1×10⁻⁶. 8 CFU / g or 1×10 8 CFU / mL.

[0013] In one embodiment, the microbial preparation is a solid or liquid preparation.

[0014] The present invention also provides the use of the above-mentioned Bifidobacterium adolescentis CCFM1506, or the above-mentioned microbial preparation, in the preparation of drugs for alleviating diseases related to estrogen metabolism disorders and / or increasing estrogen levels in the peripheral blood of women.

[0015] In one embodiment, the amount of Bifidobacterium adolescentis (CCFM1506) added to the drug is not less than 1×10⁻⁶. 8CFU / g or 1×10 8 CFU / mL.

[0016] The present invention also provides the application of the aforementioned Bifidobacterium adolescentis CCFM1506 in the preparation of products for relieving obesity, wherein the relieving of obesity includes, but is not limited to, one or more of the following functions: controlling body fat and maintaining healthy blood lipid levels.

[0017] In one embodiment, the amount of Bifidobacterium adolescentis (Bifidobacterium adolescentis) CCFM1506 added to the product is not less than 1×10⁻⁶. 8 CFU / g or 1×10 8 CFU / mL.

[0018] In one embodiment, the product is food, medicine, or health product.

[0019] In one embodiment, the food is a dairy product, soy product, or fruit and vegetable product produced using Bifidobacterium adolescentis CCFM1506 or a fermentation agent of the above-mentioned microbial preparation.

[0020] In one embodiment, the dairy products include fermented milk, flavored fermented milk, fermented milk beverages, cream, cheese, milk-containing beverages, or milk powder; the soy products include soy milk and soy milk powder; and the fruit and vegetable products include fruit and vegetable products made from at least one of cabbage, white radish, cucumber, beet, yellow peach, or bayberry products.

[0021] In one embodiment, the food is a fermented food, including solid food, liquid food, or semi-solid food.

[0022] In one embodiment, the food is a beverage or snack containing Bifidobacterium adolescentis CCFM1506 or the above-mentioned microbial preparation.

[0023] In one embodiment, the drug contains Bifidobacterium adolescentis CCFM1506, as well as a drug carrier and / or pharmaceutical excipients.

[0024] In one embodiment, the carrier includes one or more of the following commonly used in medicine: fillers, adhesives, wetting agents, disintegrants, lubricants, and flavoring agents.

[0025] In one embodiment, the dosage form of the drug is granules, capsules, tablets, pills, or oral liquid.

[0026] This invention also provides the application of the above-mentioned Bifidobacterium adolescentis CCFM1506 in the preparation of a drug or functional food having at least one of the following functions:

[0027] (a) It can reduce the amount of weight gain in mammals and decrease the proportion of visceral fat;

[0028] (b) Increase serum estradiol levels in mammals;

[0029] (c) Increase the expression of CYP11A1 and ERα in mammalian adrenal tissue.

[0030] (d) Increases high-density lipoprotein (HDL-C) in mammalian serum;

[0031] (e) Reduces total cholesterol (TC) and glucose (Glu) in mammalian serum;

[0032] (f) Increase the content of SCFA in the intestinal environment of mammals.

[0033] The present invention also provides a composition containing live bacteria of the above-mentioned Bifidobacterium adolescentis CCFM1506.

[0034] In one embodiment, the number of viable Bifidobacterium adolescentis in the composition is ≥1×10⁻⁶. 8 CFU / g or 1×10 8 CFU / mL.

[0035] The present invention also provides a probiotic preparation containing Bifidobacterium adolescentis CCFM1506, wherein the probiotic preparation is a powder obtained by drying a bacterial solution containing Bifidobacterium adolescentis CCFM1506.

[0036] In one embodiment, the drying refers to vacuum freeze drying.

[0037] In one embodiment, the viable count of Bifidobacterium adolescentis CCFM1506 in the bacterial agent is ≥1×10⁻⁶. 8 CFU / g.

[0038] Beneficial effects

[0039] The Bifidobacterium adolescentis GDMCC No: 66474 screened in this invention has a significant effect in alleviating or treating estrogen-related metabolic disorders and obesity, specifically including:

[0040] (1) Significantly inhibited the rate of weight gain and weight increment in ovariectomized rats;

[0041] (2) Significantly increased serum estradiol in ovariectomized rats and increased the expression levels of CYP11A1 and ERα in adrenal tissue;

[0042] (3) It can increase high-density lipoprotein (HDL-C) in the serum of ovariectomized rats and decrease total cholesterol (TC) and glucose (Glu) in the serum of ovariectomized rats, which helps to alleviate metabolic disorders such as obesity, Cushing's syndrome and fatty liver.

[0043] (4) Compared with the model group, the content of acetic acid, propionic acid, butyric acid, valeric acid and isovaleric acid in feces after intervention with Bifidobacterium adolescentis CCFM1506 was increased by 49.32%, 39.96% and 125.43% respectively, which is a significant effect.

[0044] The Bifidobacterium adolescentis CCFM1506 strain of this invention, compared with estrogen drugs, has a more prominent effect on peripheral blood estrogen, total cholesterol (TC), glucose (Glu), and short-chain fatty acids. Compared with the effect of strain GDMCC NO:61634 after normalization, it significantly reduced the weight gain and visceral fat ratio in mammals by 48.90% and 24.56%, respectively, increased estrogen by 9.33%, significantly increased the estrogen-related gene CYP11A1 in adrenal tissue by 10.15%, and significantly increased the butyrate content in feces by 47.05%, showing a superior effect in alleviating metabolic disorders and obesity.

[0045] The strains screened in this invention can be used to prepare drugs to alleviate or treat estrogen-related metabolic disorders and obesity. They can also be used to prepare health products or foods that help control body fat and maintain healthy blood lipid levels, thus exerting their effects widely and having very valuable application prospects.

[0046] Preservation of biological materials

[0047] Bifidobacterium adolescentis (CCFM1506), taxonomically named Bifidobacterium adolescentis, was deposited on June 6, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No.: 66474), located at 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0048] Figure 1 The effects of Bifidobacterium adolescentis CCFM1506 on body weight gain and visceral fat percentage in ovariectomized rats.

[0049] Figure 2 The effect of Bifidobacterium adolescentis CCFM1506 on serum estrogen levels in ovariectomized rats.

[0050] Figure 3 This is a schematic diagram showing the changes in CYP11A1 and ERα in the adrenal tissue of ovariectomized rats after intervention with Bifidobacterium adolescentis CCFM1506.

[0051] Figure 4 The effect of Bifidobacterium adolescentis CCFM1506 on high-density lipoprotein in the serum of ovariectomized rats.

[0052] Figure 5 This diagram illustrates the changes in total cholesterol and glucose levels in the serum of ovariectomized rats after intervention with Bifidobacterium adolescentis CCFM1506.

[0053] Figure 6 This diagram illustrates the changes in short-chain fatty acids in the feces of ovariectomized rats after intervention with Bifidobacterium adolescentis CCFM1506. Detailed Implementation

[0054] To make the above-mentioned objectives, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to specific examples.

[0055] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0056] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in different places in this specification does not necessarily refer to the same embodiment, nor is it a single or selective embodiment that excludes other embodiments. To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to specific examples.

[0057] The female SD rats used in the following examples were purchased from Spiford (Beijing) Biotechnology Co., Ltd.

[0058] The GDMCC NO:61634 strain involved in the following examples has been disclosed in patent application with publication number CN113249256A.

[0059] The culture media involved in the following examples are as follows:

[0060] MRS liquid culture medium: 10g beef extract; 10g tryptone; 5g yeast extract; 20g glucose; 5g anhydrous sodium acetate; 0.1g MgSO4·7H2O; 0.05g MnSO4·H2O; 2g diammonium citrate; 2.6g K2HPO4·3H2O; 1mL Tween 80; 0.8g L-cysteine ​​hydrochloride; adjust pH to 6.8±0.2; bring volume to 1L. Autoclave at 115℃ for 20min.

[0061] MRS solid medium: 2% agar powder is added to the MRS liquid medium.

[0062] MRS liquid medium with mass percentage (0.05%-0.1%) cysteine: Add 0.08% cysteine ​​hydrochloride to the MRS liquid medium.

[0063] The preparation of Bifidobacterium adolescentis suspension involved in the following examples

[0064] Bifidobacterium adolescentis was inoculated into MRS solid medium and cultured at 37°C for 72 h to obtain single colonies. The prepared single colonies were then inoculated into MRS liquid medium and cultured at 37°C for 24 h to activate them.

[0065] After activation for three generations, the bacterial culture was inoculated into 1 L of MRS liquid medium at an inoculum size of 2% (v / v). After shaking and mixing, the culture was incubated at 37°C for 20 h in an anaerobic incubator. After centrifugation at 8000 g / min and 4°C for 15 min, the supernatant was removed, and the culture was washed three times with sterile physiological saline containing 0.05%-0.1% L-cysteine ​​hydrochloride. After centrifugation under the same conditions and removal of supernatant, the culture was resuspended in 60% glycerol to obtain the bacterial culture before gavage. The culture was then stored in a refrigerator for later use.

[0066] Before conducting animal experiments, the bacterial suspension was taken out, centrifuged at 6000 r / min for 5 min, washed 3 times with sterile physiological saline, resuspended in physiological saline, shaken evenly, and the number of viable bacteria at the beginning and after one week of storage was determined by plate pour method.

[0067] The results showed that the initial viable count was 3.15 × 10⁻⁶. 9 CFU / mL, viable bacterial count after 1 week was 2.73 × 10⁻⁶. 9 The CFU / mL value did not change in magnitude, indicating that freezing the bacterial culture would not affect the experiment and it can be used for animal experiments.

[0068] The detection methods involved in the following embodiments are as follows:

[0069] The expression levels of CYP11A1 and ERα were detected as follows:

[0070] The expression levels of CYP11A1 and ERα genes were determined using real-time quantitative polymerase chain reaction (qRT-PCR). First, RNA was extracted from fresh tissue, using the following method:

[0071] 0.2g of fresh adrenal tissue obtained from rat dissection was repeatedly ground in a mortar (180℃, 4h high-temperature enzyme inactivation) with liquid nitrogen. Then, 1mL of TrizoL reagent was added to the mortar, and grinding continued until the liquid was basically clear. The mixture was then collected into a 1.5mL enzyme-free centrifuge tube and allowed to stand at room temperature for 15min. 200μL of chloroform solution was added to the centrifuge tube, and the mixture was gently shaken for 15s. The mixture was allowed to stand at room temperature for 10min, and then centrifuged at 4℃ and 12000r / min for 15min. 600μL of the colorless upper aqueous phase was transferred to another enzyme-free centrifuge tube, and 500μL of isopropanol was added. Invert the tube to mix thoroughly, let stand at room temperature for 10 min, then centrifuge at 12000 rpm for 10 min at 4 °C. Discard the supernatant, leaving the white precipitate formed at the bottom of the centrifuge tube containing RNA. Add 1 mL of 75% ethanol solution prepared with DEPC water, vortex to resuspend, centrifuge at 7500 rpm for 5 min at 4 °C, discard the supernatant, and allow to evaporate and dry at room temperature. Add 30 μL of RNase-free water to the dried RNA. After the RNA dissolves, determine the RNA concentration and purity using Nanodrop, and assess the RNA quality by agarose gel electrophoresis. Using the extracted total RNA as a template, reverse transcribe cDNA according to the instructions of the HiScript III All-in-one RT SuperMix Perfect for qPCR Reverse Transcription Kit from Novizan Biotechnology Co., Ltd., and store at -20 °C. Primers for rat CYP11A1 and ERα protein genes and the internal reference gene GADPH gene are shown in Table 1.

[0072] Table 1. Sequences of rat CYP11A1 protein gene and internal reference β-actin primers

[0073]

[0074] qRT-PCR reaction system and conditions:

[0075] use The CFX96TM real-time quantitative PCR instrument was used for PCR amplification and the fluorescence signal was read.

[0076] The c-kit gene qRT-PCR reaction system is as follows:

[0077]

[0078] The qRT-PCR reaction conditions for the c-kit gene are as follows:

[0079] The temperature was set at 95℃ for 30 seconds, then at 95℃ for 10 seconds, followed by 60℃ for 30 seconds, for a total of 40 cycles. The β-Actin gene was used as an internal reference gene, and the results were analyzed using CFX96Manager software.

[0080] The method for detecting the concentration of short-chain fatty acids in fecal contents involved in the following examples is as follows:

[0081] The feces collected before the end of the experiment were freeze-dried, the dry weight of the feces was calculated, and the samples were stored at -80℃. The specific method is as follows:

[0082] 20 mg of feces was weighed and resuspended in 500 μL of saturated NaCl solution. 20 μL of 10% H₂SO₄ solution was added. 1 mL of anhydrous diethyl ether was added, and the mixture was shaken thoroughly to extract short-chain fatty acids. The extract was then centrifuged at 12000 rpm at 4℃ for 15 min. The upper ether phase was collected and dried with 0.25 g of anhydrous Na₂SO₄. After standing for 30 min, the extract was centrifuged at 12000 rpm at 4℃ for 5 min. The upper ether phase was then collected, and the content of short-chain fatty acids in the lyophilized rat feces was determined by GC-MS. An Rtx-Wax column (30 m long, 25 μm inner diameter) was used. He was used as the carrier gas at a flow rate of 2 mL / min. The injection volume was 1 μL. The temperature was increased to 140℃ at 7.5℃ / min, then increased to 200℃ at 60℃ / min and held for 3 min. The ionization temperature was 20℃. The analysis was performed in full scan mode. A standard curve was obtained using the external standard method to calculate the concentrations of various short-chain fatty acids.

[0083] Example 1: Isolation of Bifidobacterium adolescentis CCFM1506

[0084] 1. Isolation and screening of Bifidobacterium strains:

[0085] (1) Fecal samples were collected from women of childbearing age in Wuxi City, Jiangsu Province using a disposable sterile fecal collection device. The fecal samples were enriched in an anaerobic incubator (N2:CO2:H2 = 80:10:10) in MRS liquid medium containing 5-10 g / L fructooligosaccharides and 0.5-1 g / L cysteine. Single colonies were selected for purification culture.

[0086] 2. Molecular biological identification of Bifidobacteria

[0087] (1) Take 1 mL of the bacterial cells screened in step 1 and activated for 3 generations (cultured for 12-48h) for bacterial identification, centrifuge at 6000r / min for 3min, discard the supernatant to obtain the bacterial cells.

[0088] (2) After adding 1 mL of sterile water and washing the bacterial cells by blowing, centrifuge at 10000 r / min for 1 min, discard the supernatant to obtain the bacterial cells, add 500 μL of sterile water to resuspend them, and use them as a template for bacterial culture.

[0089] (3) 16S rDNA PCR system:

[0090] The bacterial 16S rDNA PCR reaction system (20 μL) contained: 27F, 0.5 μL; 1492R, 0.5 μL; Taq enzyme, 1 μL; template, 1 μL; and ddH2O, 8 μL.

[0091] PCR conditions: 94℃ for 5 min; 94℃ for 30 s; 55℃ for 30 s; 72℃ for 2 min; 72℃ for 10 min; step 2-4 30×; 12℃ for 2 min.

[0092] (4) Prepare 1% agarose gel, then mix the PCR product with 10000× Loading buffer, load 2μL, run at 120V for 30min, and then perform gel imaging.

[0093] (5) The PCR product of 16S rDNA was sequenced and analyzed. The sequencing result is shown in SEQ ID NO.1. The obtained sequence was searched and compared with GenBank using BLAST. The results showed that the nucleic acid sequence similarity with Bifidobacterium adolescentis was 99.79%. It was named Bifidobacterium adolescentis CCFM1506 and stored at -80℃ with 60% (v / v) glycerol for later use.

[0094] Example 2: Effects of Bifidobacterium adolescentis CCFM1506 on body weight and visceral fat ratio in ovariectomized rats

[0095] (1) Preparation of Bifidobacterium adolescentis CCFM1506 bacterial suspension

[0096] After the Bifidobacterium adolescentis CCFM1506 strain was taken out of the -80℃ freezer, it was streaked on MRS solid medium and cultured at 37℃ for 48h. Single colonies were picked and cultured on MRS liquid medium at 37℃ for 24h to prepare seed culture.

[0097] The prepared seed culture was inoculated into a new MRS liquid medium at an inoculation rate of 2% (v / v) and cultured at 37°C for 24 h. The culture was repeated for one generation in the same manner to prepare the fermentation broth of Bifidobacterium adolescentis CCFM1506.

[0098] The prepared Bifidobacterium adolescentis CCFM1506 fermentation broth was then centrifuged at 6000 r / min and 4℃ for 5 min, and then resuspended with 60% (v / v) glycerol to obtain a bacterial suspension. The suspension was stored in a refrigerator and centrifuged before being used in animal experiments, and then resuspended with physiological saline.

[0099] (2) Thirty healthy male SD rats aged 3 months were taken and allowed to acclimatize to the environment for 1 week. They were then randomly divided into 5 groups:

[0100] Six rats were included in each of the following groups: sham-operated group, ovariectomized model group, estradiol group (treatment group), Bifidobacterium adolescentis intervention group (CCFM1506), and Lactobacillus plantarum CCFM1180 group. Except for the sham-operated group, which only underwent ovarian exposure, all other groups had both ovaries removed. All rats rested for two weeks post-surgery. Then, starting on day 15 post-surgery, the sham-operated group and the ovariectomized model group received physiological saline; the intervention group received a gavage concentration of 5 × 10⁻⁶. 9 The bacterial suspension of CFU / mL was administered by gavage starting at 9:00 AM every day, 0.2 mL each time; the estradiol drug group was given estradiol at a dose of 28 μg / kg / day to each rat.

[0101] The grouping and treatment methods of the experimental animals are shown in Table 2.

[0102] Table 2 Grouping of experimental animals

[0103]

[0104] Rats' body weight was recorded weekly at the start of the intervention. At the end of the experiment, fresh feces were collected and frozen at -80°C. At the end of the experiment, rats were fasted but allowed free water for 12 hours. After anesthesia with isoflurane, blood was collected from the abdominal aorta, and the rats were euthanized by cervical dislocation. Blood samples were centrifuged at 3500 rpm for 15 minutes, and the supernatant was collected and frozen at -80°C for blood parameter analysis. Adrenal gland tissue was quickly removed, rinsed in pre-cooled physiological saline, and blood was collected. This blood was then flash-frozen in liquid nitrogen and transferred to -80°C for subsequent analysis of relevant parameters in the adrenal gland homogenate.

[0105] Effects of Bifidobacterium adolescentis CCFM1506 on rat weight gain rate and weight gain, as follows: Figure 1 A. Compared with ovariectomized rats, *Bifidobacterium adolescentis* CCFM1506 and estradiol reduced the weight gain in ovariectomized rats (weight gain in the ovariectomized group: 89.46±15.85g; weight gain in the *Bifidobacterium adolescentis* CCFM1506 group: 30.79±12.31g, reduction: 65.58%; weight gain in the estradiol group: 32.20±2.07g, reduction: 64.01%). Simultaneously, *Lactobacillus plantarum*, administered via gavage to the ovariectomized rat group, also significantly reduced the weight gain in ovariectomized rats during the intervention period (60.25±9.38g, reduction: 32.65%). These results indicate that *Bifidobacterium adolescentis* CCFM1506 has a significant improving effect in preventing and reducing weight gain due to decreased estrogen levels.

[0106] Meanwhile, compared with ovariectomized rats, Bifidobacterium adolescentis CCFM1506 reduced the proportion of visceral fat in ovariectomized rats. Figure 1 (B) Compared with ovariectomized rats, Bifidobacterium adolescentis CCFM1506 reduced the proportion of visceral fat in ovariectomized rats by 46.05%. In contrast, compared with the ovariectomized group, the sham-operated group, Lactobacillus plantarum CCFM1180, and estrogen reduced the proportion of visceral fat in ovariectomized rats by 38.05%, 27.64%, and 30.85%, respectively. This indicates that the Bifidobacterium adolescentis CCFM1506 group has a superior ability to reduce the proportion of visceral fat compared to the sham-operated group, Lactobacillus plantarum CCFM1180, and estrogen group, and can prevent diseases such as increased visceral fat and central obesity caused by decreased estrogen levels.

[0107] The results of this embodiment indicate that Bifidobacterium adolescentis CCFM1506 has a significant effect in alleviating estrogen-related metabolic disorders and obesity.

[0108] Example 3: Bifidobacterium adolescentis (CCFM1506) increases estrogen levels

[0109] The experimental animals were grouped and treated in the same way as in Example 2, and the serum estradiol level was measured according to the kit instructions.

[0110] like Figure 2 As shown, compared with the ovariectomized rat model group, *Bifidobacterium adolescentis* CCFM1506 increased serum estradiol levels by 47.64%, restoring serum estradiol levels in ovariectomized rats to levels similar to those in the sham-operated group (*Bifidobacterium adolescentis* CCFM1506 group: 414.98±38.55 ng / L; sham-operated group: 406.29±25.19 ng / L). Similarly, intervention with *Lactobacillus plantarum* CCFM1180 and estrogen also partially restored serum estradiol levels in ovariectomized rats (*Lactobacillus plantarum* CCFM1180: 377.39±35.13 ng / L; estrogen group: 380.73±40.16 ng / L), increasing levels by 35.04% and 35.04%, respectively. The results of this example indicate that intervention with *Bifidobacterium adolescentis* CCFM1506 can compensate for the decrease in estrogen levels caused by reduced ovarian function.

[0111] Example 4: Bifidobacterium adolescentis CCFM1506 increases the expression levels of CYP11A1 and ERα genes in the adrenal tissue of ovariectomized rats.

[0112] The grouping, modeling, and treatment methods for SD rats were the same as in Example 2. The expression levels of CYP11A1 and ERα genes were determined using real-time quantitative polymerase chain reaction (qRT-PCR).

[0113] CYP11A1 catalyzes the conversion of cholesterol to pregnenolone in the adrenal glands, and indirectly regulates estrogen levels by generating androgen precursors which are then converted to estrogen by aromatase in peripheral tissues. Abnormal CYP11A1 activity can lead to an imbalance in the adrenal-derived androgen / estrogen ratio, affecting reproductive and metabolic homeostasis. Experimental results are as follows: Figure 3 As shown in Figure A, gavage administration of *Bifidobacterium adolescentis* CCFM1506 significantly increased the mRNA level of CYP11A1 in the adrenal tissue of ovariectomized rats, reaching 2.17±0.63 compared to the ovariectomized group, which was superior to the 1.97±0.42 in the *Lactobacillus plantarum* CCFM1180 group. The increased CYP11A1 expression level suggests that the pathway for cholesterol to estrogen conversion is activated; *Bifidobacterium adolescentis* CCFM1506 may promote estrogen synthesis by increasing CYP11A1 expression in adrenal tissue.

[0114] Bifidobacterium adolescentis CCFM1506 significantly increased the mRNA level of ERα in the adrenal tissue of ovariectomized mice, reaching 1.908±0.21 compared to the ovariectomized group, which was close to the 2.03±0.70 level in the Lactobacillus plantarum CCFM1180 group. Figure 3 (B) Increased ERα expression levels indicate that ERα is activated due to elevated estrogen levels, and estrogen's ability to combat obesity requires ERα activation. Bifidobacterium adolescentis CCFM1506 can exert an effective anti-obesity effect by increasing ERα expression in abdominal adipose tissue, thereby mitigating metabolic disorders caused by estrogen withdrawal.

[0115] Example 5: Bifidobacterium adolescentis CCFM1506 increases high-density lipoprotein cholesterol (HDL-C) levels.

[0116] The experimental animals were grouped and treated in the same way as in Example 2. The serum high-density lipoprotein (HDL-C) was measured according to the method of the biochemical analyzer.

[0117] like Figure 4As shown, compared with the ovariectomized rat model group, *Bifidobacterium adolescentis* CCFM1506 increased the serum HDL-C level in rats by 37.67%, which was superior to the sham-operated group (29.81%, P=0.0235), *Lactobacillus plantarum* CCFM1180 (22.11%, P=0.0697), and the estrogen group (11.55%, P=0.5258). Intervention with *Bifidobacterium adolescentis* CCFM1506 increased HDL-C levels in ovariectomized rats, thereby enhancing cholesterol reverse transport capacity, providing raw materials for estrogen synthesis, and indirectly supporting estrogen production in tissues such as fat and adrenal glands after ovariectomy.

[0118] Example 6: Bifidobacterium adolescentis CCFM1506 significantly reduced total cholesterol (TC) and glucose (Glu) levels.

[0119] The experimental animals were grouped and treated in the same way as in Example 2. Serum total cholesterol (TC) and glucose (Glu) were measured using a biochemical analyzer.

[0120] like Figure 5 As shown in Figure A, compared with the ovariectomized rat model group, Bifidobacterium adolescentis CCFM1506 (concentration 1.88±0.44 mmol / L) reduced the serum TC level in rats by 31.38% relative to the ovariectomized group (concentration 2.74±0.35 mmol / L). Its ability to reduce serum total cholesterol was similar to that of Lactobacillus plantarum CCFM1180 group (concentration 1.98±0.69 mmol / L, reduction: 27.73%), sham-operated group (concentration 1.96±0.32 mmol / L, reduction: 28.47%), and estrogen group (concentration 2.02±0.48 mmol / L, reduction: 26.27%), thereby reducing the total cholesterol content in rats, which is beneficial for the prevention and treatment of metabolic disorders and obesity after estrogen withdrawal.

[0121] Compared with the ovariectomized rat model group, Bifidobacterium adolescentis CCFM1506 reduced the serum Glu level in rats by 28.11% ( ). Figure 5 B) The ability of Bifidobacterium adolescentis CCFM1506 to reduce serum total cholesterol was superior to that of the sham-operated group (reduced by 16.41%), Lactobacillus plantarum CCFM1180 (reduced by 16.70%), and the estrogen group (reduced by 19.88%), thereby reducing glucose levels in rats. Since serum glucose levels are negatively correlated with estrogen synthesis, the reduction in serum glucose levels in ovariectomized rats by Bifidobacterium adolescentis CCFM1506 intervention may be beneficial to estrogen synthesis.

[0122] Example 7: Bifidobacterium adolescentis CCFM1506 increases the content of short-chain fatty acids in rat feces.

[0123] The grouping, modeling, and treatment methods for SD rats were the same as in Example 2.

[0124] Feces collected before the end of the experiment were freeze-dried, and the content of short-chain fatty acids in the freeze-dried rat feces was determined by GC-MS. The concentrations of various short-chain fatty acids were then calculated.

[0125] Experimental results are as follows Figure 6 As shown, the levels of acetic acid, propionic acid, and butyric acid in the feces of ovariectomized rats were all lower than those in the sham-operated group. In the estradiol group, the acetic acid content was 21.06±2.30 μmol / g, the propionic acid content was 7.01±1.45 μmol / g, and the butyric acid content was 8.09±1.89 μmol / g, representing decreases of 8.47%, 16.59%, and 59.81% respectively compared to the ovariectomized group (P < 0.05). 乙酸 =0.8442, P 丙酸 =0.6164, P 丁酸 =0.2625). However, after gavage administration of Bifidobacterium adolescentis CCFM1506, the contents of acetic acid, propionic acid, and butyric acid were 35.24±4.84μmol / g, 11.76±2.11μmol / g, and 45.38±13.69μmol / g, respectively, which were significantly higher than those in the ovariectomized group, increasing by 49.32%, 39.96%, and 125.43%, respectively (P = 0.2625). 乙酸 =0.0047, P 丙酸 =0.0365, P 丁酸 =0.0041), and the improvement effect was better than that of the estradiol group, increasing by 60.08%, 56.55%, and 185.24% respectively compared with the estradiol group. Meanwhile, after gavage administration of *Lactobacillus plantarum* CCFM1180, the contents of acetic acid, propionic acid, and butyric acid were 33.52±7.23 μmol / g, 12.70±1.91 μmol / g, and 30.86±12.29 μmol / g, respectively, which were significantly higher than those of the ovariectomized group, increasing by 42.03%, 51.15%, and 53.30% respectively (P = 0.0041). 乙酸 =0.0169, P 丙酸 =0.0060, P 丁酸 =0.3594). This indicates that Bifidobacterium adolescentis CCFM1506 significantly increases the content of acetic acid and butyric acid in feces compared to Lactobacillus plantarum CCFM1180, while Lactobacillus plantarum is superior to Bifidobacterium adolescentis CCFM1506 in increasing the content of propionic acid.

[0126] Short-chain fatty acids (SCFAs) positively regulate estrogen synthesis pathways by upregulating aromatase expression and modulating gut microbiota metabolism and host epigenetic modifications. Therefore, Bifidobacterium adolescentis CCFM1506 plays an important role in increasing the content of SCFAs in feces, mainly by increasing the content of acetic acid, butyric acid, and propionic acid in feces to alleviate estrogen-related metabolic disorders and obesity.

[0127] Example 8: Comparison of Bifidobacterium adolescentis CCFM1506 and Lactobacillus plantarum GDMCC NO:61634

[0128] The results of Bifidobacterium adolescentis CCFM1506 and GDMCC NO:61634 in animal experiments were normalized and compared with their model group (the modeling and animal treatment methods were the same as in Example 2, and the butyric acid determination method was exactly the same). The results are shown in Table 3.

[0129] Table 3 Comparison of results between Bifidobacterium adolescentis CCFM1506 and Lactobacillus plantarum GDMCC NO:61634

[0130]

[0131] Note: The rate of change refers to the percentage improvement in effect of the CCFM1506 group compared to the GDMCC NO:61634 group.

[0132] The results showed that, compared with GDMCC NO:61634, *Bifidobacterium adolescentis* CCFM1506 exhibited significantly lower weight gain and visceral fat percentage, significantly increased expression levels of CYP11A1 and ERα in adrenal tissue, and significantly higher fecal butyrate content. This suggests that strain CCFM1506 may inhibit fat accumulation by altering host metabolic pathways through regulation of host estrogen levels. The significant increase in butyrate suggests that strain CCFM1506 may promote butyrate production, which is considered to play an important role in maintaining gut health, promoting anti-inflammatory responses, and enhancing immune function. These results indicate that strain CCFM1506 may play a positive role in influencing host metabolism, particularly in weight control.

[0133] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A strain of Bifidobacterium adolescentis (CCFM1506), characterized in that, The Bifidobacterium adolescentis was deposited at the Guangdong Provincial Center for Microbial Culture Collection on June 6, 2025, with accession number GDMCC No: 66474.

2. A composition containing the live strain of Bifidobacterium adolescentis as described in claim 1.

3. The composition according to claim 2, characterized in that, Including but not limited to microbial preparations.

4. Food, medicine or health product containing the Bifidobacterium adolescentis as described in claim 1.

5. The use of the Bifidobacterium adolescentis according to claim 1 in the preparation of a medicament for alleviating diseases related to estrogen metabolism disorders and / or increasing estrogen levels in peripheral blood.

6. The application according to claim 5, characterized in that, The estrogen metabolism disorder-related diseases include, but are not limited to: obesity, diabetes, and metabolic syndrome.

7. The application of the Bifidobacterium adolescentis CCFM1506 according to claim 1 in the preparation of products for relieving obesity, characterized in that, The measures to alleviate obesity include, but are not limited to, controlling body fat and / or maintaining healthy blood lipid levels.

8. The application according to claim 7, characterized in that, The product is food, medicine, or health product; The food products are dairy products, soy products, or fruit and vegetable products containing the Bifidobacterium adolescentis CCFM1506.

9. The application according to claim 7, characterized in that, The drug contains the Bifidobacterium adolescentis as described in claim 1, as well as a drug carrier and / or pharmaceutical excipients.

10. The use of the Bifidobacterium adolescentis according to claim 1 in the preparation of a drug or functional food having at least one of the following functions: (a) Reduces the rate of weight gain in mammals and decreases the proportion of visceral fat; (b) Increase serum estradiol levels in mammals; (c) Increase the expression of CYP11A1 and ERα in mammalian adrenal tissue; (d) Increases high-density lipoprotein in mammalian serum; (e) Reduces total cholesterol and glucose in mammalian serum; (f) Increase the content of short-chain fatty acids in the intestinal environment of mammals.

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