Strain for improving bone health and application thereof

Abstract

The invention provides a bacterial strain for improving bone health and application of the bacterial strain. The preservation number of bifidobacterium animalis subsp. Lactis BL-11 is CGMCC (China General Microbiological Culture Collection Center) No.20847. The strain can promote osteoblasts and inhibit osteoclasts in a cell experiment; by constructing a mouse model with abnormal bone growth, the BL-11 probiotics have an obvious improvement effect on bone development indexes of the model mouse, promote bone formation and inhibit bone resorption; furthermore, the BL-11 probiotics can improve the ostealgia score, life quality and bone mineral density of people with osteoporosis or bone mass reduction. The results indicate the application potential of the probiotics in improvement of abnormal bone growth and prevention and treatment of osteoporosis.

Description

Technical Field

[0001] The present invention relates to the field of probiotics and human health, and in particular to a strain for improving bone health and application thereof. Background Art

[0002] Bones are important supporting structures of the human body, providing mechanical support, protecting internal organs, and participating in mineral storage and regulation. Maintaining bone health is essential for overall health. Bone health refers to the good state of strength, density and function of bone tissue. Bone metabolism is a dynamic equilibrium process, which is mainly regulated by osteoblasts and osteoclasts. Osteoblasts are responsible for the formation and mineralization of new bones, while osteoclasts remove aged or damaged bone tissue through bone resorption. The maintenance of bone health depends on the balance of osteogenesis and osteoclasts. When osteoclast activity exceeds osteogenic activity, bone loss and decreased bone density will occur. In addition, calcium, phosphorus, vitamin D, protein and various hormones (such as parathyroid hormone, estrogen, testosterone and growth hormone) have an important influence on the development and metabolism of bones.

[0003] Abnormal bone growth refers to an individual's bone growth rate that is lower than normal, resulting in a discrepancy between bone age and actual age. This situation may be caused by a variety of factors, including genetics, malnutrition, endocrine disorders, and chronic diseases. Currently, abnormal bone growth is an increasingly concerning health issue, especially in children and adolescents. Abnormal bone growth has significant epidemiological differences worldwide. There are large differences in the prevalence between developed and developing countries, which is closely related to nutritional status, medical level, and public health policies. In developing countries, malnutrition remains one of the main causes of abnormal bone growth, especially in areas lacking basic medical facilities. Therefore, the study of bone health and abnormal bone growth is not only of great scientific significance, but also helps to formulate public health policies and clinical treatment strategies to improve the quality of life of patients with bone diseases.

[0004] The current treatment and intervention strategies for abnormal bone growth mainly include several aspects: ① Nutritional supplementation and dietary balance. For abnormal bone growth caused by the lack of these nutrients, calcium and vitamin D supplementation is the basic strategy; ensure that the diet contains enough protein, vitamins and minerals, especially during the growth period of children and adolescents. ② Hormone drug treatment. For patients with growth hormone deficiency, recombinant human growth hormone (rhGH) treatment is the main intervention measure, which can significantly promote height growth and bone development. In addition, thyroid hormone replacement and sex hormone therapy are also used to promote bone growth and maturation. ③ Physical therapy and exercise. Moderate exercise, such as weight-bearing exercise and resistance training, helps to increase bone density and promote bone health. ④ Surgical intervention. For example, bone correction surgery and bone transplantation, regeneration technology, these means can provide new treatment options for dealing with severe bone deformities or functional disorders caused by abnormal bone growth.

[0005] In recent years, research on probiotics in the field of bone health has gradually emerged. Probiotics refer to active microorganisms that are beneficial to the host and mainly exist in the intestines. They have a positive impact on host health through multiple mechanisms, including promoting digestion, enhancing immune function, and potentially affecting bone health. Studies have found that probiotics can improve the intestinal environment and promote the absorption of vitamin D and calcium. Some specific probiotic strains (such as Lactobacillus and Bifidobacterium) can increase the efficiency of calcium absorption in the small intestine. Chronic inflammation is an important factor in bone loss and abnormal bone growth. Probiotics can reduce the levels of systemic inflammatory markers (such as tumor necrosis factor α) by regulating the intestinal microbiota. Reducing inflammatory responses not only helps maintain bone density, but also promotes normal bone development. In terms of immune regulation, probiotics can enhance the intestinal barrier function, prevent the invasion of pathogenic microorganisms, and thus reduce the overreaction of the immune system. This immunomodulatory effect has a positive effect on the prevention of chronic diseases such as inflammatory bowel disease, which are often associated with bone health problems. Additionally, although human studies are relatively sparse, preliminary findings suggest that probiotic supplementation may have a positive impact on bone health in children and older adults.

[0006] In summary, probiotics have potential benefits in promoting bone health and alleviating abnormal bone growth, and have broad application prospects in bone health. The present invention aims to screen and apply specific probiotic strains to provide a safe and effective probiotic that promotes osteogenesis and inhibits osteoclastogenesis, filling the gap in the prior art. Summary of the invention

[0007] The present invention provides a strain that can effectively improve bone health and its application. Specifically, the animal Bifidobacterium lactis subspecies BL-11 probiotics described in the present invention can promote osteoblasts and inhibit osteoclasts in cell experiments; by constructing a mouse model of abnormal bone growth, the BL-11 probiotics have a significant improvement effect on the bone development indicators of the model mice, promote bone formation, and inhibit bone resorption; further, the BL-11 probiotics can improve the bone pain score, quality of life and bone density of people with osteoporosis or osteopenia. These results suggest the potential application of the probiotics in improving abnormal bone growth and preventing and treating osteoporosis.

[0008] According to one aspect of the present invention, there is provided a Bifidobacterium animalis subsp. lactis, characterized in that the deposit number of the Bifidobacterium animalis subsp. lactis is CGMCC No.20847.

[0009] According to another aspect of the present invention, a preparation of Bifidobacterium animalis subspecies lactis is provided, the preparation containing the Bifidobacterium animalis subspecies lactis, and the preparation includes one or more of the fermentation broth, fermentation broth precipitate, fermentation broth supernatant, live bacteria, dead bacteria, freeze-dried powder, and lysate of the Bifidobacterium animalis subspecies lactis.

[0010] According to another aspect of the present invention, there is provided a bacterial preparation of Bifidobacterium lactis subsp. animalis, wherein the strain of Bifidobacterium lactis subsp. animalis is a solid preparation or a liquid preparation. Preferably, the Bifidobacterium lactis subsp. animalis or the preparation or the bacterial preparation is used in a product that promotes or differentiates osteoblasts.

[0011] According to another aspect of the present invention, there is provided a use of the Bifidobacterium animalis subsp. lactis or the preparation or the bacterial agent in a product having an inhibitory effect on osteoclasts.

[0012] According to another aspect of the present invention, there is provided the use of the animal Bifidobacterium lactis subsp. or the preparation or the bacterial agent in a product having one or more effects of improving bone volume percentage, trabecular number, trabecular thickness, and bone density.

[0013] According to another aspect of the present invention, there is provided a use of the Bifidobacterium lactis subsp. animalis or the preparation or the bacterial agent in the preparation of a product for treating and / or assisting in the promotion of bone formation, characterized in that the number of viable bacteria of the Bifidobacterium lactis subsp. animalis in the product is 1×10 3 -1×10 14 CFU / mL or 1×10 3 -1×10 14 CFU / g.

[0014] According to another aspect of the present invention, there is provided a use of the Bifidobacterium lactis subsp. animalis or the preparation or the bacterial agent in the preparation of a product for treating and / or assisting in inhibiting bone resorption, wherein the number of viable cells of the Bifidobacterium lactis subsp. animalis in the product is 1×10 3 -1×10 14 CFU / mL or 1×10 3 -1×10 14 CFU / g.

[0015] According to another aspect of the present invention, there is provided a use of the Bifidobacterium animalis subsp. lactis or the preparation or the bacterial agent in a product for preventing osteoporosis.

[0016] According to another aspect of the present invention, there is provided a method for preparing a drug for treating and / or assisting in the treatment of osteoblast-promoting, osteoclast-inhibiting diseases and osteoporosis, wherein the drug comprises a therapeutically effective amount of the animal Bifidobacterium lactis subspecies or the bacterial agent. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The effect of animal Bifidobacterium lactis subspecies BL-11 of the present invention on osteoblasts and osteoclasts under in vitro conditions is shown. (A) Osteoblast activity; (B) Osteoblast differentiation; (C) Osteoclast viability; (D) Osteoclast differentiation. ns>0.05, *p<0.05, **p<0.01, ***p<0.001.

[0018] Figure 2 The micro-CT representative images of the femur of mice treated with animal Bifidobacterium lactis subspecies BL-11 in the present invention are shown. MNC, fecal bacteria group of children with normal bone transplant development (Model normal control); MNB, fecal bacteria group of children with normal bone transplant development MNB (Model normal BL-11) gavaged with BL-11 probiotics; MC, fecal bacteria group of children with abnormal bone transplant growth (Model control); MCB, fecal bacteria group of children with abnormal bone transplant growth (Model controlBL-11) gavaged with BL-11 probiotics.

[0019] Figure 3The femur CT index results of mice treated with animal Bifidobacterium lactis subspecies BL-11 in the present invention are shown. (A) Bone volume percentage BV / TV%; (B) Trabecular number (Tb.N); (C) Trabecular thickness (Tb.Th); (D) Bone density (BMD); (E) Trabecular separation (Tb.Sp). n=10, ns>0.05, *p<0.05, **p<0.01, ***p<0.001.

[0020] Figure 4 The contents of bone development-related indicators in mice treated with animal Bifidobacterium lactis subspecies BL-11 of the present invention are shown. (A) Serum P1NP content; (B) Serum CTX1 content. n=10, ns>0.05, *p<0.05, **p<0.01, ***p<0.001.

[0021] Figure 5 The gene expression in the femur of mice after treatment with the animal Bifidobacterium lactis subsp. BL-11 of the present invention is shown. (A) Relative level of femur ALP mRNA; (B) Relative level of femur TRAP mRNA. n=10, ns>0.05, *p<0.05, **p<0.01, ***p<0.001.

[0022] Figure 6 The results of osteoblastic and osteoclastogenic staining of the femur of mice treated with animal Bifidobacterium lactis subspecies BL-11 in the present invention are shown. (A) Representative images of Alizarin Red S (indicating osteoblastic nodules) and TRAP staining (TRAP, tartrate-resistant acid phosphatase, osteoclast marker enzyme); (BC) Statistical results of the proportion of positive areas of Alizarin Red S and TRAP staining. n=10, ns>0.05, *p<0.05, **p<0.01, ***p<0.001. Example

[0023] Example 1: Cultivation and sample preparation of Bifidobacterium animalis subspecies lactis BL-11

[0024] The probiotic strain BL-11 described in the present invention is isolated from breast milk of healthy women and has been preserved in the General Microbiology Center of China Microbiological Culture Collection Administration on October 10, 2020. The classification name is Bifidobacterium lactis, and the preservation number is CGMCC No. 20847. The original species name is Bifidobacterium lactis, and the species name has now been changed to Bifidobacterium anlmalis subsp. lactis (refer to the announcement issued by the National Health Commission on the update of the "List of Bacteria that can be used in Food" and the "List of Bacteria that can be used in Infant Food").

[0025] In the experiment, the frozen BL-11 strain was activated and subcultured for two generations, then inoculated into MRS liquid culture medium at 2-4%, cultured continuously for 16h-24h, and centrifuged (6000r / min, 10min, 4°C). The obtained supernatant was the fermentation supernatant of the BL-11 strain. The bacterial sludge was washed twice with PBS, and then the bacterial count was adjusted to 5×10 with sterile PBS solution. 9 CFU / mL was used for subsequent animal experiments.

[0026] Example 2: Effects of Bifidobacterium animalis subsp. lactis BL-11 on osteoblasts and osteoclasts

[0027] This example is used to illustrate the promoting effect of animal Bifidobacterium lactis subspecies BL-11 on osteoblasts and the inhibiting effect on osteoclasts.

[0028] (1) Osteoblast proliferation and differentiation assay

[0029] Mouse embryonic osteoblast MC3T3-E1 cells (purchased from Shanghai Fuheng Biotechnology Co., Ltd.) were inoculated in α-MEM medium (containing 10% FBS and 1% penicillin-streptomycin) and cultured at 37°C and 5% CO2 until 80% confluence. Digest, count and re-inoculate in 96-well plates at an appropriate cell density per well. After cell attachment, the culture medium was removed and α-MEM medium containing different concentrations (v / v%) of probiotic supernatant (such as 0%, 5%, 10% and 20%) was added. The group without probiotic supernatant was set as a blank control.

[0030] ①Cell activity detection:

[0031] After 2 days of culture, the proliferation of cells was detected using the MTT kit. According to the experimental instructions, MTT solution (10 μL per well) was added and incubated at 37°C for 4 hours. DMSO (100 μL per well) was added and shaken for 10 minutes to dissolve the formed crystals. The absorbance (OD value) was measured at 570 nm on an ELISA reader.

[0032] ②Cell differentiation detection:

[0033] Mouse MC3T3-E1 cell osteogenic differentiation medium (Saiye.MUXMT-90021) was selected for osteogenic induction culture. After 2-3 weeks of culture, the formation of mineralized nodules was detected using an Alizarin Red staining kit. Specifically, after terminating the culture, the culture medium was discarded, the cells were rinsed 3 times with PBS, fixed with 4% paraformaldehyde for 30 minutes, and then rinsed with PBS 3 times, and Alizarin Red dye was added for staining for 10 minutes. After rinsing with PBS, the Alizarin Red dye in the cells was dissolved with a solvent (acetic acid: glycerol = 5: 1), and the cell debris was removed by centrifugation at 3000g for 5 minutes. An appropriate volume of supernatant was taken and the absorbance value (OD value) at 405nm was detected by an ELISA instrument.

[0034] (2) Osteoclast experiment

[0035] Mouse mononuclear macrophages (RAW264.7) are one of the most commonly used in vitro models for osteoclast research. In this example, RAW264.7 cells were inoculated in DMEM medium (containing 10% FBS and 1% penicillin-streptomycin) and cultured to 80% confluence at 37°C and 5% CO2. Digest, count and re-inoculate in 24-well plates or 96-well plates at an appropriate cell density per well. After the cells adhere, the culture medium is removed and DMEM culture medium containing different concentrations (v / v) of probiotic supernatant (such as 0%, 5%, 10%, 20%) is added. A control group (without probiotic supernatant) and an experimental group (different concentrations of probiotic supernatant) are set up.

[0036] ①Cell activity detection:

[0037] After 2 days of culture, the cell activity was detected using an MTT kit. MTT solution (10 μL per well) was added and incubated at 37°C for 4 hours. DMSO (100 μL per well) was added and shaken for 10 minutes to dissolve the formed crystals. The absorbance (OD value) was measured at 570 nm on an ELISA reader.

[0038] ②Osteoclast differentiation detection:

[0039] RANKL (receptor activator of nuclear factor κB ligand, 100 ng / mL) was used to induce RAW264.7 cells to differentiate into osteoclasts. During this period, the culture medium was changed every 3 days and RANKL was supplemented. After 5-7 days of culture, the activity of osteoclasts was detected using an acid phosphatase (TRAP) staining kit (Kamiya tartrate-resistant acid phosphatase TRAP staining kit). Specifically, after terminating the culture, the culture medium was discarded, the cells were rinsed 3 times with PBS, fixed with 4% paraformaldehyde for 30 minutes, and then rinsed with PBS 3 times, and then TRAP staining solution was added according to the kit instructions, and incubated at room temperature for 1 hour. The staining solution was removed, washed 3 times with PBS, and then dissolved with glacial acetic acid for 10 minutes, and the absorbance (OD value) was measured at 550nm on an ELISA reader.

[0040] According to the results of in vitro cell experiments ( Figure 1 ), after the fermentation supernatant of the animal Bifidobacterium lactis subspecies BL-11 probiotic in the present invention was used to treat MC3T3-E1 osteoblasts, the cell viability showed a significant upward trend ( Figure 1 A), and it increases with the increase of fermentation liquid addition (v / v). Specifically, the average relative activity is about 148.46% when the addition amount is 5%, 163.57% when the addition amount is 10%, and as high as 176.62% when the addition amount is 20%. It can be seen that the addition amount of 20% (v / v) has the best effect on promoting the growth of osteoblasts. During the MC3T3-E1 induced differentiation process, the mineralized nodules after BL-11 probiotic treatment increased significantly (Alizarin red staining, Figure 1 B), the highest value was reached when the addition amount was 20%, and the OD405nm reading at this time was about 2 times that of the untreated group, indicating that the probiotics can promote bone formation. In terms of osteoclasts, the cell viability of osteoclast RAW264.7 was detected ( Figure 1 C) and differentiation ( Figure 1 D), it can be found that the animal Bifidobacterium lactis subspecies BL-11 in the present invention can inhibit the proliferation of osteoclasts RAW264.7 and reduce its activity. The inhibitory effect is significantly reduced with the increase of the fermentation liquid addition amount (v / v). The average relative activity value is reduced to 82.06% at 5% addition, 78.37% at 10%, and 67.28% at 20%. Consistently, the test results of BL-11 on the inhibition of osteoclast differentiation also show that the OD550nm reading can be reduced to about half of the untreated cell group at 20% addition.

[0041] Example 3: Bifidobacterium animalis subsp. lactis BL-11 improves bone formation and bone resorption in mice with abnormal bone growth

[0042] This example is used to illustrate the beneficial effects of BL-11 animal Bifidobacterium on bone metabolism in mice with abnormal bone growth.

[0043] ① Experimental and testing methods:

[0044] Children who met the criteria for abnormal bone growth were selected from the pediatric endocrinology clinic, and normally developing children whose physical examination indicators were normal and whose age, gender, parents' height, family living standards and other conditions matched those of children with abnormal growth were selected from nearby primary schools. Fresh stool specimens were collected, the fecal residue was filtered out, and the microorganisms in the feces were obtained by high-speed centrifugation. Fecal bacteria freeze-dried powder was prepared using vacuum freeze-drying technology.

[0045] Forty 3-week-old clean wild-type C57BL / 6 mice (purchased from Vital River) were purchased and first fed for 3 days in an SPF environment with constant temperature and humidity and alternating light and dark. Then, they were treated with broad-spectrum antibiotics for 5 days of bowel cleansing. Finally, they were randomly divided into 4 groups: MNC (Model normal control) group of children with normal bone transplantation, MNB (Model normal BL-11) group of children with normal bone transplantation and gavage of BL-11 probiotics, MC (Model control) group of children with abnormal bone transplantation, and MCB (Model control BL-11) group of children with abnormal bone transplantation and gavage of BL-11 probiotics, with 10 mice in each group. All groups of mice were gavaged with the corresponding fecal bacteria in the morning and BL-11 probiotics in the afternoon (the number of viable bacteria was 5*10^9 CFU). The experiment lasted for a total of 6 weeks.

[0046] After the experiment, serum samples were collected before the mice were killed for subsequent detection of osteoblastic and osteoclast-related indicators. After the mice were killed, the femur tissue of the left hind limb was removed from the excess skin and muscle and other soft tissues, and the bone tissue for subsequent mRNA detection was retained. The femur was scanned using a Skyscan1276 micro CT instrument (rukermicroCT, Kontich, Belgium). After the scan, the image was reconstructed using NRecon software, and the CTAn program was used to analyze parameters such as bone volume percentage (BV / TV%), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp) and bone density (BMD). In order to perform histological staining analysis, the part below the knee joint of the right hind limb of the mouse fixed in formalin solution was removed from the excess skin and muscle and other soft tissues, and the bone tissue for subsequent mRNA detection was retained. The remaining bone tissue was placed in a 10% EDTA decalcification solution for decalcification for 4 weeks, and then the decalcified bone tissue was embedded in paraffin and sliced ​​for subsequent Alizarin Red S and TRAP staining.

[0047] ②Experimental results:

[0048] The femur specimens of mice in each group were scanned by micro-CT. Figure 2 CT showed that the bone structure of the MC group mice was relatively loose, and the bone reticular structure of the MC mice (MCB) treated with BL-11 strain became denser; the effect of gavage of BL-11 on the bone structure of normal mice was not obvious.

[0049] Furthermore, the relevant indicators were analyzed ( Figure 3AE), compared with the MNC group, the bone volume percentage (BV / TV%), trabecular number (Tb.N), trabecular thickness (Tb.Th) and bone density (BMD) of the MC group were significantly reduced, and the trabecular separation (Tb.Sp) was significantly increased; specifically, the bone volume percentage (BV / TV%), trabecular number (Tb.N), trabecular thickness (Tb.Th) and bone density (BMD) of the fecal bacteria group (MC) of children with abnormal bone growth were 71.4%, 68.5%, 91.5% and 80.2% of those of the fecal bacteria group (MNC) of children with normal bone development, respectively; the trabecular separation (Tb.Sp) value of the fecal bacteria group (MC) of children with abnormal bone growth was 108.9% of that of the fecal bacteria group (MNC) of children with normal bone development. These statistical results show that the MC group successfully simulated the pathological state of abnormal bone growth in young children. After intervention with BL-11 probiotics, bone parameters such as bone volume percentage (BV / TV%), trabecular number (Tb.N), trabecular thickness (Tb.Th) and bone mineral density (BMD) were significantly improved and approached the index values ​​of normal mice, which were 90.7%, 88.6%, 97.3% and 87.2% of the fecal bacteria group of children with normal bone development (MNC), respectively. In the fecal bacteria group of children with transplanted growth abnormalities (MCB) gavaged with BL-11 probiotics, the above indicators were increased by 26.9%, 29.2%, 6.3% and 8.6% respectively compared with those in the fecal bacteria group of children with abnormal bone growth (MC); the trabecular separation (Tb.Sp) was reduced by about 9.6% compared with the fecal bacteria group of children with abnormal bone growth (MC).

[0050] In order to explore whether there are similar trends in serum osteoblastic and osteoclastogenic indicators in mice, serum indicators related to osteoblastic (P1NP) and osteoclastogenic (CTX1) were tested. The results showed that compared with the MNC group, the content of osteoblastic indicator P1NP in the MC group was significantly reduced (0.89 vs 2.98 ng / ml), and the content of osteoclastogenic indicator CTX1 was significantly increased (0.61 vs 0.57 ng / ml). After BL-11 intervention, the levels of P1NP increased to 2.14 ng / ml, and CTX1 decreased to 0.43 ng / ml ( Figure 4 AB).

[0051] According to the mRNA detection results of osteoblasts and osteoclasts in femoral tissue ( Figure 5AB), compared with the normal bone development group (MNC), the relative expression level of osteogenesis-related genes (ALP) in the abnormal bone growth MC group was significantly downregulated (1vs 0.46), and the expression of osteoclast-related indicators (TRAP) was significantly upregulated (1vs 1.62). After BL-11 probiotic intervention, the expression of ALP increased significantly (1.44vs 0.46), and the gene of TRAP decreased significantly (1.04vs 1.62), which was consistent with the trend of the above-mentioned bone formation and bone resorption results.

[0052] The results of osteoblast- and osteoclast-specific staining of femoral tissue sections showed that ( Figure 6 AC), compared with the MNC group, the MC group had disordered trabecular structure and decreased number ( Figure 6 A), bone mineral deposition is significantly reduced ( Figure 6 B), osteoclasts increased significantly ( Figure 6 C); after BL-11 intervention, these abnormalities were improved and closer to normal.

[0053] Example 4: Animal Bifidobacterium lactis subspecies BL-11 improves bone health in osteoporotic patients

[0054] This example is used to evaluate the effect of animal Bifidobacterium lactis subsp. BL-11 in improving bone density, bone metabolism markers and overall bone health in osteoporosis patients.

[0055] Volunteers for this population trial were recruited according to the following inclusion and exclusion criteria: inclusion criteria: ① Age: women between 50 and 70 years old, ② bone density test showed osteoporosis or osteopenia, ③ no serious underlying diseases (such as diabetes, heart disease, etc.). Exclusion criteria: people using osteoporosis-related drugs (such as bisphosphonates). People who have recently taken antibiotics or other drugs that affect the intestinal flora. 20 participants who met the criteria were selected and divided into 2 groups (10 in the probiotic group and 10 in the placebo group). Probiotic group: took capsules containing 1 billion CFU of the target probiotics every day, and placebo group: took placebo capsules with the same appearance. The entire trial lasted for 3 months. Intervention measurement indicators included: (1) Subjective questionnaire evaluation: ① Bone pain score (0-10 points), ② Quality of life questionnaire (including dimensions such as mobility, pain, and fatigue). (2) Bone density measurement: The test is performed once before and after each intervention. The T value is often used to represent the patient's bone density in osteoporosis. A T value of less than -2.5 indicates osteoporosis; a T value of -1.0 to -2.5 indicates osteopenia and a high risk of fracture.

[0056] The results showed that (Table 1) the subjective pain score of the probiotic intervention group decreased, and the quality of life questionnaire score increased; compared with the baseline bone density T value (-2.33±0.59), the bone density increased significantly (-2.25±0.27). There was no significant improvement in the subjective pain score and quality of life questionnaire score of the placebo group; the bone density T value was -2.36±0.32, and the baseline value was -2.35±0.19, which showed a downward trend, which is consistent with the law of gradual loss of bone mass with age.

[0057] Table 1 Statistics of indicators of the test population

[0058]

[0059] In summary, the results of in vitro cell and animal experiments show that the animal Bifidobacterium lactis subspecies BL-11 probiotic strain can promote bone development and inhibit bone resorption by promoting osteoblastic development and inhibiting osteoclast proliferation and differentiation. Population experiments show that the intake of this probiotic can improve the bone density and bone metabolism of osteoporosis patients to a certain extent, and has a potential adjuvant therapeutic effect. Therefore, the BL-11 probiotic of the present invention has great development potential and application prospects in improving bone development and promoting related products such as bone health, especially for specific populations such as children with abnormal bone growth, osteoporosis women caused by aging, etc.

Claims

1. A Bifidobacterium animalis subsp. lactis, characterized in that: The preservation number of the animal Bifidobacterium lactis subspecies is CGMCC No.20847.

2. A preparation of Bifidobacterium animalis subsp. lactis, characterized in that The preparation contains the animal Bifidobacterium lactis subsp. lactis according to claim 1, and comprises one or more of the fermentation broth, fermentation broth precipitate, fermentation broth supernatant, live bacteria, dead bacteria, lyophilized powder, and lysate of the animal Bifidobacterium lactis subsp. lactis.

3. A bacterial preparation of Bifidobacterium animalis subsp. lactis, comprising the strain of Bifidobacterium animalis subsp. lactis according to claim 1, wherein the bacterial preparation is a solid preparation or a liquid preparation.

4. Use of the animal Bifidobacterium lactis subsp. lactis according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in a product having an effect of promoting or differentiating osteoblasts.

5. Use of the animal Bifidobacterium lactis subsp. lactis according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in a product having an inhibitory effect on osteoclasts.

6. Use of the animal Bifidobacterium lactis subsp. lactis according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in a product having one or more effects of improving bone volume percentage, trabecular number, trabecular thickness, and bone density.

7. Use of the animal Bifidobacterium lactis subsp. according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in the preparation of a product for treating and / or assisting in the promotion of bone formation, characterized in that: The viable count of animal Bifidobacterium lactis subspecies in the product is 1×10 3 -1×10 14 CFU / mL or 1×10 3 -1×10 14 CFU / g.

8. Use of the animal Bifidobacterium lactis subsp. according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in the preparation of a product for treating and / or assisting in inhibiting bone resorption, characterized in that: The viable count of animal Bifidobacterium lactis subspecies in the product is 1×10 3 -1×10 14 CFU / mL or 1×10 3 -1×10 14 CFU / g.

9. Use of the animal Bifidobacterium lactis subsp. lactis according to claim 1, the preparation according to claim 2, or the bacterial agent according to claim 3 in a product for preventing osteoporosis.

10. A method for preparing a drug for treating and / or assisting in treating osteoblast-promoting, osteoclast-inhibiting diseases and osteoporosis, characterized in that: The medicine comprises a therapeutically effective amount of the animal Bifidobacterium lactis subspecies according to claim 1 or the bacterial agent according to claim 2.

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