Bifidobacterium longum subsp.infantis capable of improving physiological activity function of vitamin D and repairing bone injury and application of bifidobacterium longum subsp.infantis

Abstract

The invention discloses bifidobacterium longum subsp.infantis capable of improving the physiological activity function of vitamin D and repairing bone injuries and application of the bifidobacterium longum subsp.infantis, and belongs to the technical field of biological medicine. The bifidobacterium longum subsp. Infantis CCFM1449 can significantly improve the total bone mineral density, cortical bone mineral density, femoral distal bone trabecula bone mineral density, bone trabecula thickness, bone trabecula connectivity, bone surface area, bone volume and bone volume fraction of an osteoporosis mouse, and reduce the bone trabecula separation of the osteoporosis mouse; the content of vitamin D metabolites in serum of the osteoporosis mice is increased, the content of serum calcium and alkaline phosphatase in the serum of the osteoporosis mice is reduced, and the level of osteocalcin and type I procollagen amino-terminated peptide in the serum is increased. Therefore, the bifidobacterium longum subsp. Infantis CCFM1449 has the effects of relieving bone calcium loss, maintaining bone health, repairing bone injury, increasing osteoblasts and reducing osteoporosis degree.

Description

Technical Field

[0001] The invention relates to a strain of Bifidobacterium longum infantis subspecies capable of improving the physiological activity function of vitamin D and repairing bone damage and application thereof, belonging to the technical field of biomedicine. Background Art

[0002] Osteoporosis is a systemic metabolic disease caused by various reasons, which leads to bone loss and reduction, changes in bone tissue microstructure, and increased bone brittleness, which easily causes fractures in patients. Osteoporosis is divided into two categories: primary osteoporosis and secondary osteoporosis. Primary osteoporosis is further divided into two categories: senile osteoporosis and postmenopausal osteoporosis.

[0003] Common symptoms of osteoporosis include pain, shortened stature, hunchback, fractures and decreased respiratory function. Calcium supplementation is a basic measure for the treatment of osteoporosis, but calcium supplementation alone is far from enough for the treatment of osteoporosis, and additional medications need to be used according to the patient's condition. At present, bisphosphonates, selective estrogen receptor modulators, estrogen and calcitonin are mainly used clinically to treat osteoporosis. These drugs can effectively promote the formation of osteoblasts and inhibit the formation of osteoclasts. However, bisphosphonates can cause severe bone, joint or muscle discomfort in some patients, and a small number of patients have an increased risk of jaw osteonecrosis when suffering from dental diseases or receiving invasive dental treatments; long-term use of alendronic acid is associated with the occurrence of subtrochanteric and femoral shaft fractures; the selective estrogen receptor modulator raloxifene can increase the risk of venous thromboembolism and stroke; estrogen can increase the risk of endometrial hyperplasia and cancer in patients with intact uterus, and the incidence of gallstone disease and venous thromboembolism is also increased by 2 to 3 times; calcitonin may cause allergic reactions, and severe patients may experience anaphylactic shock, and large-scale use of calcitonin can cause patients to have hypocalcemia, and some patients also experience joint pain and skeletal muscle pain. Therefore, it is urgent to find a drug or treatment method that can effectively relieve osteoporosis and that will not cause complications and side effects in patients with long-term use.

[0004] Vitamin D (VD) is an important element for maintaining bone physiological function and bone growth and development. It plays an important role in bone calcium absorption. It can absorb calcium from the intestine into the blood and improve the calcium absorption effect. At the same time, it can also help guide calcium from the blood to the bones, so that calcium in osteoblasts will not be lost excessively, avoiding the occurrence of osteoporosis and children's bone growth and development disorders. Secondly, it can also promote the formation of osteoblasts. The active form of VD 1,25 dihydroxyvitamin D (1,25 (OH) 2D) can also act as a direct agonist to bind to the vitamin D receptor (VDR) for regulation in the human body. Appropriate supplementation of VD for children and the elderly will have direct benefits to bones, such as improving the body's muscle strength and regulating the body's balance ability; it plays an important role in promoting human bone health. Improving the metabolic capacity of VD helps promote the absorption of calcium by bones, thereby preventing and / or treating osteoporosis. However, no safe products that can effectively prevent and / or treat osteoporosis by increasing the metabolic level of VD have been found. Summary of the invention

[0005] In order to solve the above problems, the present invention screened out a strain of Bifidobacterium longum subsp. infantis. This Bifidobacterium longum subsp. infantis has the effects of improving the physiological activity of vitamin D and alleviating osteoporosis, which is specifically embodied in: (1) significantly improving the level of vitamin D metabolites in the blood of osteoporotic mice; (2) significantly improving the bone-related indicators of osteoporotic mice; (3) significantly improving the bone health of osteoporotic mice. Therefore, Bifidobacterium longum subsp. infantis has great application prospects in the preparation of products for improving the physiological activity of vitamin D, preventing and / or treating osteoporosis, and / or preventing and / or treating bone loss.

[0006] The technical solution of the present invention is as follows:

[0007] The present invention provides a strain of Bifidobacterium longum subsp. infantis CCFM1449,

[0008] Bifidobacterium longum subsp. infantis CCFM1449

[0009] It was deposited in the Guangdong Provincial Microbiological Culture Collection Center on October 31, 2024, with the deposit number GDMCC No: 65384, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

[0010] In one embodiment of the present invention, the Bifidobacterium longum subsp. infantis CCFM1449 is isolated from human feces. The strain is sequenced and analyzed, and its 16SrRNA sequence is shown in SEQ ID NO.1. The sequence is compared in GenBank, and the result shows that the strain is Bifidobacterium longum subsp. infantis, and is named Bifidobacterium longum subsp. infantis CCFM1449.

[0011] In one embodiment of the present invention, the Bifidobacterium longum subsp. infantis CCFM1449 is a Gram-positive bacillus that does not produce spores and is arranged singly, in pairs or in a V-shaped pattern. After anaerobically cultured at 36° C. for 3 days on an MRS (lactic acid bacteria culture medium) plate, the colonies are milky white, round, convex, with a smooth surface and neat edges.

[0012] The present invention also provides a microbial preparation containing the Bifidobacterium longum subspecies infantis CCFM1449.

[0013] The invention also provides a product containing the microbial preparation.

[0014] In one embodiment of the present invention, the viable count of Bifidobacterium longum subsp. infantis CCFM1449 in the product is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0015] In one embodiment of the present invention, the product comprises food, medicine or health care product.

[0016] In one embodiment of the present invention, the food comprises a dairy product, a bean product or a fruit and vegetable product produced using a starter containing Bifidobacterium longum subsp. infantis CCFM1449; or the food comprises a starter containing Bifidobacterium longum subsp. infantis (Bifidobacterium longumsubsp.

[0017] infantis)CCFM1449 solid beverage.

[0018] In one embodiment of the present invention, the medicine contains Bifidobacterium longum subsp. infantis CCFM1449, a drug carrier and / or a pharmaceutical excipient.

[0019] In one embodiment of the present invention, the method for preparing the starter is as follows: Bifidobacterium longum subsp. infantis CCFM1449 is inoculated into a culture medium at an inoculum amount of 5 to 8% of the total mass of the culture medium, and cultured at 37° C. for 18 hours to obtain a culture solution; the culture solution is centrifuged to obtain bacterial cells; the bacterial cells are washed 2 to 4 times with a phosphate buffer having a pH of 7.2, and then resuspended with a freeze-drying protective agent containing 100 g / L of trehalose to obtain a resuspended liquid; the resuspended liquid is freeze-dried by a vacuum freezing method to obtain Bifidobacterium longum subsp. infantis.

[0020] infantis)CCFM1449 bacterial powder; the mass ratio of the lyophilized protective agent to the bacterial body is 2:1.

[0021] In one embodiment of the present invention, the culture medium comprises 87.7% water, 10% enzyme-hydrolyzed skim milk, 0.5% glucose, 1.5% tryptone and 0.3% dissolved yeast extract, accounting for 87.7% of the total mass of the culture medium.

[0022] In one embodiment of the present invention, the pH of the culture medium is 6.8.

[0023] The present invention also provides the use of the Bifidobacterium longum infantis subspecies CCFM1449 or the microbial preparation in preparing medicines for preventing and / or treating osteoporosis.

[0024] In one embodiment of the present invention, the prevention and / or treatment of osteoporosis comprises at least one of the following:

[0025] (1) Improve the level of vitamin D metabolism in osteoporotic individuals;

[0026] (2) Improving bone-related indicators of individuals with osteoporosis; the bone-related indicators include total bone density, cortical bone density, distal femoral trabecular bone density, trabecular separation, trabecular thickness, trabecular number, trabecular connectivity, bone surface area, bone volume and bone volume fraction.

[0027] In one embodiment of the present invention, the viable count of Bifidobacterium longum subspecies infantis CCFM1449 in the drug is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0028] The present invention also provides the use of the Bifidobacterium longum infantis subspecies CCFM1449 in preparing a health product that helps to improve bone density.

[0029] In one embodiment of the present invention, in the health product, the viable count of Bifidobacterium longum subspecies infantis CCFM1449 is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0030] Beneficial effects:

[0031] The present invention screened out a strain of Bifidobacterium longum subsp.

[0032] infantis)CCFM1449, this Bifidobacterium longum subsp.

[0033] infantis)CCFM1449 has the function of improving the physiological activity of vitamin D and alleviating osteoporosis, which is specifically reflected in:

[0034] (1) Significantly increased total bone density, cortical bone density, distal femoral trabecular bone density, trabecular thickness, trabecular number, trabecular connectivity, bone surface area, bone volume, and bone volume fraction in osteoporotic mice;

[0035] (2) Significantly reduced trabecular separation in osteoporotic mice;

[0036] (3) Increase the levels of vitamin D metabolites 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D in the serum of osteoporotic mice;

[0037] (4) Reduce the levels of serum calcium and alkaline phosphatase in the serum of osteoporotic mice, and increase the levels of serum osteocalcin and type I procollagen amino-terminal propeptide.

[0038] Therefore, Bifidobacterium longum subsp. infantis CCFM1449 has great application prospects in the preparation of products for preventing and / or treating influenza.

[0039] Biomaterial Deposit

[0040] A strain of Bifidobacterium longum subsp. infantis CCFM1449, taxonomically named Bifidobacterium longum subsp. infantis, was deposited in the Guangdong Provincial Microbiological Culture Collection on October 31, 2024, with the deposit number GDMCC No: 65384, and the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 : The content of 25-hydroxyvitamin D in serum of osteoporotic mice in different groups;

[0042] Figure 2 : The content of 1,25-dihydroxyvitamin D in serum of osteoporotic mice in different groups;

[0043] Figure 3 : The serum calcium content in the serum of osteoporotic mice in different groups;

[0044] Figure 4 : The levels of alkaline phosphatase in serum of osteoporotic mice in different groups;

[0045] Figure 5 : The content of osteocalcin in serum of osteoporotic mice in different groups;

[0046] Figure 6 : The content of amino-terminal propeptide of type Ⅰ procollagen in serum of osteoporotic mice in different groups;

[0047] Figure 7 : Total bone density levels of osteoporotic mice in different groups;

[0048] Figure 8 :Cortical bone density levels in different groups of osteoporotic mice;

[0049] Fig. 9 :Bone mineral density of distal femur trabeculae in different groups of osteoporotic mice;

[0050] Fig.10 :The level of trabecular separation in different groups of osteoporotic mice;

[0051] Fig.11 :The trabecular thickness levels of osteoporotic mice in different groups;

[0052] Fig.12 :The number of trabecular bones in different groups of osteoporotic mice;

[0053] Fig.13:The level of trabecular connectivity in different groups of osteoporotic mice;

[0054] Fig.14 :Bone surface area levels of osteoporotic mice in different groups;

[0055] Fig.15 :Bone volume levels of osteoporotic mice in different groups;

[0056] Fig.16 : Bone volume fraction levels in different groups of osteoporotic mice. DETAILED DESCRIPTION

[0057] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0058] The culture medium involved in the following examples is as follows:

[0059] MRS solid culture medium: peptone 10g / L, beef extract powder 5g / L, yeast extract powder 4g / L, glucose 20g / L, sodium acetate 5g / L, dipotassium hydrogen phosphate 2g / L, triammonium citrate 2g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, Tween 80 1g / L, agar 15g / L, cysteine ​​salt 0.5g / L, pH 6.8.

[0060] MRS liquid culture medium: peptone 10g / L, beef extract powder 5g / L, yeast extract powder 4g / L, glucose 20g / L, sodium acetate 5g / L, dipotassium hydrogen phosphate 2g / L, triammonium citrate 2g / L, magnesium sulfate 0.2g / L, manganese sulfate 0.05g / L, Tween 80 1g / L, cysteine ​​salt 0.5g / L, pH 6.8.

[0061] The detection methods involved in the following embodiments are as follows:

[0062] Detection method of viable bacteria count: adopt the national standard "GB 4789.35-2016 National Food Safety Standard Food Microbiology Detection Lactic Acid Bacteria Detection".

[0063] The present invention will be further described below in conjunction with specific embodiments.

[0064] Example 1: Screening and identification of Bifidobacterium longum subsp. infantis

[0065] 1. Screening

[0066] Human feces were used as samples. After pretreatment, the samples were stored in a -80°C refrigerator in about 20% glycerol. After being taken out and thawed, 0.5 mL of the sample was mixed and added to 4.5 mL. Gradient dilutions were performed with 0.9% saline containing 0.05% cysteine. Appropriate gradient dilutions were selected and spread on MRS plates with 0.05% cysteine. The plates were cultured at 37°C for 48 hours. Typical colonies were picked and streaked on MRS plates for purification. Single colonies were picked and transferred to liquid MRS culture medium (containing 0.05% cysteine) for bacterial enrichment and stored in 30% glycerol to obtain strain CCFM1449.

[0067] 2. Identification

[0068] The genome of CCFM1449 was extracted, and the 16S rDNA of CCFM1449 was amplified and sequenced (Shanghai Sangon Biotechnology Co., Ltd.). The sequence was compared in GenBank, and the results showed that the strain was Bifidobacterium longum subsp. infantis, and was named Bifidobacterium longum subsp. infantis CCFM1449.

[0069] Example 2: Cultivation of Bifidobacterium longum subsp. infantis

[0070] Bifidobacterium longum subsp. infantis CCFM1449 was inoculated into MRS solid medium (containing 0.05% cysteine) and cultured at 37°C for 48h, and its colonies were observed and its bacteria were observed under a microscope, and it was found that it was a Gram-positive bacillus, non-spore-producing, single, paired or V-shaped arrangement. After anaerobic culture at 36°C on MRS (lactic acid bacteria culture medium) plates for 3 days, the colonies were milky white, round, convex, smooth on the surface, and neat on the edges.

[0071] Bifidobacterium longum subsp. infantis CCFM1449 was inoculated into MRS liquid culture medium (containing 0.05% cysteine) and cultured at 37°C for 48 hours. A growth curve was prepared, and it was found that it reached a stable period after 12 hours of culture at 37°C. At the same time, it was observed that it was a heterotypic fermentation and could produce acid and gas from glucose.

[0072] Bifidobacterium longum subsp. infantis CCFM1449 was inoculated into MRS liquid culture medium (containing 0.05% cysteine) and cultured at 10, 15, 20, 25, 30, 35, 40, 45, and 50°C for 48 h, and then its growth was observed. It grew well at 20-35°C and was still able to grow at 45°C, but hardly grew at 15°C or below or at 50°C.

[0073] Bifidobacterium longum subsp. infantis CCFM1449 was inoculated into MRS liquid medium (containing 0.05% cysteine) and cultured at 37°C for 48 h, then transferred into fresh MRS medium (containing 0.05% cysteine), cultured under the same conditions for 30 h, centrifuged at 6000g for 15 min, washed with 0.9% saline and centrifuged again at 6000g for 10 min to obtain the cells, which were resuspended in 30% sucrose solution and frozen at -80°C for use.

[0074] Example 3: Effect of Bifidobacterium longum subsp. infantis on the levels of vitamin D metabolites in osteoporotic mice

[0075] 3-4 weeks old SPF grade C57BL / 6J male mice were divided into 3 groups, namely blank group (CON), model group (MOD), positive control group (VD) and CCFM1449 experimental group (intervention group), 8 mice in each group. The animals were raised in the Experimental Animal Center of Jiangnan University, fed with ordinary feed, with a constant temperature of 21-26℃, humidity of 40-70%, noise less than or equal to 60dB, and animal illumination of 15-20LX (all animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Management Committee of Jiangnan University).

[0076] The experimental period was 42 days in total. The model was established on the 7th day. Except for the blank group, the other mice were gavaged with 200 μL of 90 mg / kg BW / d retinoic acid once a day to induce osteoporosis for three weeks. After the model was established, the positive control group was gavaged with 200 μL of 0.06 μg / kg BW / d VD solution once a day, and the intervention group was gavaged with 0.2 mL of 1×10 viable bacteria once a day. 9 CFU / mL bacterial suspension, the blank group and the model group were only gavaged with an equal amount of sterile saline as a control, and all groups had free access to water and food for two weeks until the mice were killed on the 42nd day.

[0077] The experimental animal groups and treatment methods are shown in Table 1:

[0078] Table 1 Experimental animal groups

[0079]

[0080]

[0081] After killing the mice, blood samples were collected and centrifuged to separate serum. The serum was tested using the Shanghai ELISA Mouse 25-hydroxyvitamin D ELISA Kit (ml038442) and Mouse 1,25-hydroxyvitamin D ELISA Kit (ml062981). Figure 1 , Figure 2 It can be seen that osteoporosis leads to a decrease in the level of vitamin D metabolites in the blood of mice; the positive control group significantly increased the level of vitamin D metabolites, and the levels of 25-hydroxyvitamin D and 1,25-hydroxyvitamin D increased by 42.27% and 27.90% respectively compared with the model group. The CCFM1449 experimental group significantly increased the level of vitamin D metabolites in osteoporotic mice compared with the model group, and the levels of metabolites 25-hydroxyvitamin D and 1,25-hydroxyvitamin D increased by 16.53% and 22.79% respectively compared with the model group, reaching the same level as the blank group.

[0082] The above experimental results show that Bifidobacterium longum subspecies infantis CCFM1449 can significantly increase the level of vitamin D metabolites in osteoporotic mice.

[0083] Example 4: Effect of Bifidobacterium longum subsp. infantis CCFM1449 on serum calcium levels in the serum of osteoporotic mice The mouse grouping and modeling methods were the same as in Example 3.

[0084] On the 42nd day, the mice were killed, blood samples were taken, serum was separated by centrifugation, and serum calcium level was tested using Nanjing Jiancheng Serum Calcium Kit (C004-2-1). Figure 3 .

[0085] Depend on Figure 3 It can be seen that the serum calcium level of osteoporosis model mice was significantly higher than that of blank group mice. The positive control group reduced the serum calcium level by 18.02% compared with the model group. The serum calcium level of osteoporosis mice in the CCFM1449 experimental group was significantly reduced by 20.81% compared with the model group, and returned to normal levels, indicating that Bifidobacterium longum infantis subspecies CCFM1449 played a role in alleviating bone calcium loss, and the effect was better than VD.

[0086] Example 5: Effect of Bifidobacterium longum subspecies infantis CCFM1449 on serum alkaline phosphatase levels in osteoporotic mice

[0087] The mouse grouping and modeling methods were the same as in Example 3.

[0088] On the 42nd day, the mice were killed, blood samples were collected, serum was separated by centrifugation, and serum alkaline phosphatase AKP level was detected using Nanjing Jiancheng serum alkaline phosphatase kit (A059-2-2). Figure 4 .

[0089] Depend on Figure 4 It can be seen that the serum alkaline phosphatase level of osteoporosis model mice was significantly higher than that of normal mice. The positive control group significantly reduced the serum alkaline phosphatase level of osteoporosis mice by 15.83%. Compared with the model group, the serum alkaline phosphatase level of osteoporosis mice in the CCFM1449 experimental group was significantly reduced by 13.59%, returning to normal levels, indicating that Bifidobacterium longum infantis subspecies CCFM1449 plays a role similar to VD in maintaining bone health, repairing bone damage, and reducing the degree of osteoporosis.

[0090] Example 6: Effect of Bifidobacterium longum subsp. infantis CCFM1449 on serum osteocalcin levels in osteoporotic mice The mouse grouping and modeling methods were the same as in Example 3.

[0091] On the 42nd day, the mice were killed, blood samples were collected, serum was separated by centrifugation, and osteocalcin (OC) levels were detected using the Elabscience Mouse Osteocalcin (OC / BGP) ELISA Kit (E-EL-M0864). Figure 5 .

[0092] Depend on Figure 5 It can be seen that the serum osteocalcin level of osteoporosis model mice (4.054ng / mL) was lower than that of normal mice (9.735ng / mL), the positive control group significantly increased the serum osteocalcin level of mice by about 134.09%, and the CCFM1449 experimental group significantly increased the serum osteocalcin level of osteoporosis mice by 366.22% compared with the model group, indicating that Bifidobacterium longum infantis subspecies CCFM1449 plays a role in enhancing osteoblast function, repairing bone damage and osteoporosis, and the effect is better than VD.

[0093] Example 7: Effect of Bifidobacterium longum subspecies infantis CCFM1449 on the level of amino-terminal propeptide of type I procollagen in serum of osteoporotic mice

[0094] The mouse grouping and modeling methods were the same as in Example 3.

[0095] On the 42nd day, the mice were killed, and blood samples were collected from the mice. Serum was separated by centrifugation, and the serum level of type I procollagen amino-terminal propeptide (PⅠNP) was detected using the Elabscience mouse type I procollagen amino-terminal propeptide (PⅠNP) enzyme-linked immunosorbent assay kit (E-EL-M0233). Figure 6 .

[0096] Depend on Figure 6 It can be seen that the level of serum type I procollagen amino-terminal propeptide (1046pg / mL) in osteoporosis model mice was lower than that in the blank group mice (1723pg / mL), the positive control group significantly increased the mouse serum PIPN level by about 51.53%, and the CCFM1449 experimental group significantly increased the level of serum type I procollagen amino-terminal propeptide in osteoporosis mice by 324.99% compared with the model group, indicating that Bifidobacterium longum infantis subspecies CCFM1449 plays a role in improving osteoblast synthesis, promoting new bone formation, repairing bone damage and osteoporosis, and the effect is better than VD.

[0097] Example 8: Effects of Bifidobacterium longum subspecies infantis CCFM1449 on bone-related indices in osteoporotic mice

[0098] The mouse grouping and modeling method were the same as in Example 3. The mice were killed on the 42nd day, and the femurs of the mice were taken for detection of bone-related indicators using Micro-CT. The observation parameters included total bone density (Bone Mean, BM), cortical bone density (Cortex Mean, CM), distal femoral trabecular bone density (Trabeculae Mean, TM), trabecular bone separation (trabecular bone separation, Tb.Sp), trabecular bone thickness (trabecular bone thickness, Tb.Th), trabecular bone number (trabecular bone number, Tb.N), trabecular bone connectivity (Conn.D), bone surface area (BS), bone volume (BV) and bone volume fraction (bone volume / total volume, BV / TV). The results are shown in Figure 7 to Figure 16 .

[0099] Depend on Figure 7 to Figure 16It can be seen that compared with the blank group, the BM, TM, Tb.N, Conn.D, BS and BV of the osteoporosis model group mice were significantly decreased, which were reduced by 3.1%, 5.27%, 10.61%, 25.64%, 42.60%, 22.33%, 23.93% and 18.18%, respectively, while Tb.Sp was significantly increased by 18.58%; after VD treatment, the BM, TM, Tb.Th, Tb.N and Conn.D of the positive control group were increased by 1.68%, 3.66%, 5.82%, 40.66% and 8.31% respectively compared with the model group, Tb.Sp was reduced by 20.77%, while BS, BV and BV / TV did not increase significantly. After oral administration of Bifidobacterium longum infantis subsp. CCFM1449, the BM, CM, TM, Tb.Th, Tb.N, Conn.D, BS, BV and BV / TV of mice increased significantly by 3.19%, 4.29%, 2.73%, 16.18%, 11.99%, 109.03%, 17.48%, 29.54% and 46.98%, respectively, while Tb.Sp decreased significantly by 35.50%. It can be seen that Bifidobacterium adolescentis CCFM1449 can significantly increase the total bone density, cortical bone density, distal femoral trabecular bone density, trabecular thickness, trabecular connectivity, bone surface area, bone volume and bone volume fraction of osteoporotic mice, and significantly reduce the trabecular separation of osteoporotic mice.

[0100] Compared with oral administration of VD, oral administration of Bifidobacterium longum infantis subspecies CCFM1449 can significantly increase trabecular thickness (increased by 1.10 times), trabecular connectivity (increased by 1.93 times), bone surface area (increased by 1.24 times), bone volume (increased by 1.35 times), and bone volume fraction (increased by 1.52 times). These results indicate that intervention with Bifidobacterium longum infantis subspecies CCFM1449 has the efficacy of treating osteoporosis, and the effect is better than that of VD.

[0101] Example 9: Preparation of a product containing Bifidobacterium longum subsp. infantis CCFM1449

[0102] Bifidobacterium longum infantis subspecies CCFM1449 was used as the key ingredient, galacto-oligosaccharide and curcumin were added as prebiotics, and xylitol, maltodextrin and passion fruit juice powder were used as fillers. After mixing, sieving, homogenizing and drying, a 2g probiotic powder was prepared (the number of viable bacteria of Bifidobacterium longum infantis subspecies CCFM1449 was not less than 1×10 6 CFU / g), and when using, just dissolve it in water and eat it.

[0103] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A strain of Bifidobacterium longum subsp. infantis CCFM1449, characterized in that: The Bifidobacterium longum infantis subspecies CCFM1449 was deposited in the Guangdong Provincial Microbiological Culture Collection Center on October 31, 2024, with the collection number GDMCC No: 65384, and the collection address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou.

2. A microbial preparation containing Bifidobacterium longum subspecies infantis CCFM1449 according to claim 1.

3. A product containing the microbial preparation according to claim 2.

4. The product according to claim 3, characterized in that The products include food, medicine or health products.

5. The product according to claim 4, characterized in that The food includes fermented food; the medicine also contains a drug carrier and / or a pharmaceutical excipient.

6. Use of the Bifidobacterium longum subspecies infantis CCFM1449 described in claim 1 or the microbial preparation described in claim 2 in the preparation of a medicine for preventing and / or treating osteoporosis.

7. The use according to claim 6, characterized in that The prevention and / or treatment of osteoporosis comprises at least one of the following: (1) Improve the level of vitamin D metabolism in osteoporotic individuals; (2) Improving bone-related indicators of individuals with osteoporosis; the bone-related indicators include total bone density, cortical bone density, distal femoral trabecular bone density, trabecular separation, trabecular thickness, trabecular number, trabecular connectivity, bone surface area, bone volume and bone volume fraction.

8. The use according to claim 6 or 7, characterized in that In the medicine, the viable count of Bifidobacterium longum infantis subspecies CCFM1449 is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

9. Use of the Bifidobacterium longum subspecies infantis CCFM1449 according to claim 1 in the preparation of a health product that helps to improve bone density.

10. The use according to claim 9, characterized in that In the health product, the viable count of Bifidobacterium longum infantis subspecies CCFM1449 is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

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