Composition containing Lactobacillus paracasei N1115 for improving bone health and application thereof

Abstract

The present invention belongs to the technical field of microorganisms and compositions thereof, and discloses a composition containing Lactobacillus paracasei N1115 for improving bone health and its application. The present invention first discovered that Lactobacillus paracasei N1115 has the effect of improving osteoporosis, and the composition of the bacteria, milk calcium and vitamin D in a specific ratio can synergistically improve the relevant characteristics of bone biomorphology, activate the relevant pathways of the body, and then promote the synthesis of bone marrow osteoblasts and regulate the activity of osteoclasts, thereby improving bone health. Formula powder, liquid products and postbiotic products containing Lactobacillus paracasei N1115 compositions, while improving osteoporosis, also have the advantages of high safety and no adverse reactions or dependence in long-term consumption. The present invention is suitable for preparing food or health food with improved bone health, especially for middle-aged and elderly people.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and compositions thereof, in particular to a composition containing Lactobacillus paracasei N1115 for improving bone health and application thereof. Background Art

[0002] With the advent of an aging society, bone health issues have become a common health issue faced by the world. Among bone diseases, osteoporosis, as a chronic degenerative disease, is high among the middle-aged and elderly population. According to statistics, the incidence of osteoporosis in women over 50 years old is 1 / 3 and that in men is 1 / 5, both higher than the incidence of breast cancer and prostate cancer. Osteoporotic fractures are expected to account for more than half of all fracture cases in the next 30 years. Osteoporosis leads to reduced bone mass, microstructural damage, reduced bone strength and density, and causes widespread bone brittleness and increased risk of fractures.

[0003] At present, drugs for the treatment of osteoporosis are developing rapidly, including bone resorption inhibitors such as bisphosphonates, calcitonin, and estrogen, and bone formation promoters such as parathyroid hormone analogs. However, some drugs have certain side effects, such as gastrointestinal adverse reactions and hypercalcemia. Therefore, for safety reasons, the use of the characteristics of specific microorganisms to regulate metabolic pathways related to bone health can alleviate the symptoms of osteoporosis, and long-term consumption of probiotic products that are beneficial to intestinal health will not have adverse effects.

[0004] Among the existing microorganisms and related composite products, screening out microbial strains that have the function of improving bone health and how to improve the effect of the strains have become key issues to be solved. In addition, since many probiotic products only have live bacteria with corresponding effects, and live bacteria have limited tolerance to the gastrointestinal environment, many probiotics cannot reach the intestine and are inactivated by digestive juices such as gastric juice, which is a problem that affects the application effect of probiotics. On the other hand, there are also problems such as high cost of preparing specific forms of products such as freeze-dried powder and poor portability of low-temperature liquid products. Summary of the invention

[0005] The present invention screens probiotic strains that can be used in food and health products, and unexpectedly finds that the bacteria and metabolites of Lactobacillus paracasei N1115 have the effect of improving bone health, especially improving osteoporosis by regulating metabolism and activating the body's immune pathways. The present invention further enhances the ability of Lactobacillus paracasei N1115 to improve bone health by using a specific composition.

[0006] Therefore, one object of the present invention is to provide a safe, long-term edible composition of Lactobacillus paracasei N1115, milk calcium and vitamin D without toxic side effects and its use in the preparation of a product with the function of improving bone health, so as to achieve synergistic enhancement among the components and enhance the effect of improving bone health; Another object of the present invention is to provide a formula powder, a liquid product and a postbiotic product for improving bone health containing a composition of Lactobacillus paracasei N1115. After inactivation, Lactobacillus paracasei N1115 still has the effect of improving bone health, solving the problems of high preparation cost and non-portability of live bacteria products, and also having the advantages of safety and no toxic side effects.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is: The invention provides a Lactobacillus paracasei N1115 composition for improving bone health. The composition comprises a Lactobacillus paracasei N1115 bacterial agent, milk calcium and vitamin D. The content ratio of the Lactobacillus paracasei N1115 bacterial agent, milk calcium and vitamin D is 0.2-20 mg: 1000-1700 mg: 5-10 μg.

[0008] The lactobacillus paracasei selected in the present invention ( Lactobacillus paracasei ) N1115 was isolated from traditional fermented dairy products in Inner Mongolia and was deposited in the General Microbiology Center of China Microbiological Culture Collection Committee on March 17, 2011. Its deposit number is: CGMCC No.4691, and it was disclosed in the Chinese patent application number CN201110357058.8 on January 23, 2013.

[0009] As a further limitation of the present invention, the Lactobacillus paracasei N1115 bacterial agent is a live bacterial agent or an inactivated bacterial agent.

[0010] The inactivated bacterium agent is prepared by inoculating Lactobacillus paracasei N1115 in a fermentation liquid containing skim milk, and then fermenting, centrifuging, inactivating and freeze-drying. The inactivation condition is 90-98° C. for 20-40 minutes. The amount of live bacteria after inactivation is less than 3000 CFU / g.

[0011] As a further limitation of the present invention, the viable bacterial count of the Lactobacillus paracasei N1115 bacterial agent or the viable bacterial count before inactivation thereof is ≥ 8×10 10 CFU / g.

[0012] As another limitation of the present invention, the improvement of bone health is achieved by improving osteoblast activity and regulating bone metabolism.

[0013] As a further limitation of the present invention, the regulating bone metabolism comprises upregulating the metabolism of prochondroitin, glycerophosphatidylcholine, sphingosine, butyrate and cystine.

[0014] The present invention also provides the use of any one of the above compositions in preparing a product having the function of improving bone health.

[0015] As a further limitation of the present invention, in every 100 g of the product having the function of improving bone health, the content of Lactobacillus paracasei N1115 bacterial agent is 0.2-20 mg, the content of milk calcium is 1000-1700 mg, and the content of vitamin D is 5-10 μg.

[0016] The invention provides a formula powder for improving bone health, comprising any one of the above-mentioned compositions containing Lactobacillus paracasei N1115 for improving bone health.

[0017] The present invention provides a liquid product for improving bone health, which is a liquid product prepared by dispersing any one of the above compositions containing Lactobacillus paracasei N1115 for improving bone health into water, edible oil or liquid milk.

[0018] The present invention provides a postbiotic product for improving bone health, comprising any one of the above compositions containing Lactobacillus paracasei N1115 for improving bone health, wherein the Lactobacillus paracasei N1115 bacterial agent in the composition is an inactivated bacterial agent.

[0019] The present invention also provides the use of any one of the above-mentioned postbiotic products in the preparation of a product having the function of improving bone health.

[0020] Preferably, the formula powder is formula milk powder.

[0021] Due to the adoption of the above technical solution, the present invention has the following technical advances compared with the prior art: The present invention discovers for the first time that Lactobacillus paracasei N1115 can improve bone health by improving osteoblast activity and regulating bone metabolism. It regulates the bone metabolic pathway by upregulating the metabolism of prochondroitin, glycerophosphatidylcholine, sphingosine, butyrate and cystine.

[0022] When Lactobacillus paracasei N1115 is combined with milk calcium and vitamin D to form a composition, the effect of improving bone health is more significant, which is better than using milk calcium and vitamin D alone, or using Lactobacillus paracasei N1115 alone. It can significantly improve the femoral bone density and bone microstructure destruction of mice, improve osteoclast activity, and effectively increase the bone fluorescence intensity of the dexamethasone-induced osteoporosis zebrafish model, thereby playing a role in improving osteoporosis.

[0023] In the composition of the present invention, milk calcium not only has high bioavailability but also has excellent sensory properties, and is an ideal natural food-grade calcium source; vitamin D has the effects of increasing intestinal calcium absorption, promoting bone mineralization, maintaining muscle strength, improving balance ability, and maintaining stable blood calcium and phosphorus levels. In addition, all three raw materials are food-grade raw materials, which are safer and more affordable than medicines, and will not cause adverse reactions or dependence after long-term consumption.

[0024] The present invention also found that after the inactivation of Lactobacillus paracasei N1115, it can still synergistically enhance the effect with milk calcium and vitamin D, and the effect of improving bone health is not affected by the inactivation of bacteria. Therefore, it is suitable for preparing postbiotic products, and there is no need to make products for low-temperature storage in order to retain activity, thereby reducing costs.

[0025] The present invention is suitable for preparing formula powders, liquid products, postbiotic products and other foods or health foods that can improve bone health, and is particularly suitable for middle-aged and elderly people who are prone to osteoporosis.

[0026] The present invention is further described in detail below with reference to the accompanying drawings and specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Fluorescence microscope images of zebrafish vertebrae of different groups in Experimental Example 1 of the present invention; Figure 2 This is a graph showing the results of fluorescence intensity analysis of zebrafish vertebrae in different groups in Experimental Example 1 of the present invention, where “**” represents p<0.05 and “***” represents p<0.01; Figure 3 This is a graph showing the results of the bone tissue microstructure analysis experiment in Experimental Example 2 of the present invention, wherein: Figure 3 A is the reconstructed image of the femur. Figure 3 B is the statistical chart of bone density test results. Figure 3 C is the statistical chart of bone volume fraction results, Figure 3 D is the statistical chart of trabecular thickness results. Figure 3 E is the statistical chart of trabecular number results, Figure 3 F is the statistical graph of trabecular separation results, and the same letters in each group indicate p < 0.05; Figure 4 This is a graph showing the results of the osteoblast and osteoclast marker detection experiment in Experimental Example 2 of the present invention, wherein: Figure 4 A is the result of femoral tartrate-resistant acid phosphatase staining. Figure 4 B is the statistical result of the number of positive cells stained with tartrate-resistant acid phosphatase. Figure 4 C is a statistical graph of serum alkaline phosphatase test results, and groups marked with the same letters indicate p < 0.05; Figure 5 It is a Venn diagram of the intestinal flora structure in the microbiome analysis experiment in the effect test example 2 of the present invention; Figure 6 This is a UPGMA clustering tree / combination diagram of intestinal flora structure in the microbiome analysis experiment in effect test example 2 of the present invention; Figure 7 The relative abundance result diagram of representative differential bacteria at the genus level in the effect test example 2 of the present invention, wherein Duncanella genus ( Duncaniella ) relative abundance results, Muribaculum Relative abundance results of Lactobacillus genus ( Lactobacillus ) relative abundance results, Heterobacillus genus ( Allobaculum ) relative abundance results, Isoprevotella ( Alloprevotella ) relative abundance results, Bacteroides ( Bacteroides ) and the relative abundance results of other genus ( Alistipes ), "*" represents p<0.05, "**" represents p<0.01; "***" represents p<0.001, and "****" represents p<0.0001; Figure 8 This is a graph showing the results of the metabolomics analysis experiment related to bone health in Experimental Example 2 of the present invention, wherein: Figure 8 A is the heat map of differential metabolites between the Lp group and the GIOP group. Figure 8 B is the volcano plot of differential metabolites between the Lp group and the GIOP group. DETAILED DESCRIPTION

[0028] The present invention is further described in detail below by specific examples. It should be understood that the described examples are only used to explain the present invention, and are not intended to limit the present invention.

[0029] The freeze-dried bacterial powder of Lactobacillus paracasei N1115 used in the present invention has a viable cell count of 8×10 10 ~3.5×10 11 CFU / g range, specifically 8×10 10 , 1×10 11 , 2.5×10 11 , 3×10 11 and 3.5×10 11 Typical but non-limiting viable counts such as CFU / g; The calcium content in the milk mineral salt used in the present invention is expressed as Ca 2+ The content of the polyol is calculated to be in the range of 24 to 29 g / 100 g, and specifically may be 24, 25, 27 and 29, etc., which are typical but not limiting contents.

[0030] The lactobacillus paracasei selected in the present invention ( Lactobacillus paracasei ) N1115 was isolated from traditional fermented dairy products in Inner Mongolia and was deposited in the General Microbiology Center of China Microbiological Culture Collection Committee on March 17, 2011. Its deposit number is: CGMCC No.4691, and it was disclosed in the Chinese patent application number CN201110357058.8 on January 23, 2013.

[0031] The experimental animals of the effect test example 1 of the present invention are 3-day-old wild-type AB strain zebrafish, which are bred and provided by Junlebao Fish Farming Center with a license number of SYXK (Zhejiang) 2022-0004. They are raised in fish farming water at 28°C, and the water quality conditions are as follows: 200 mg of instant sea salt is added to every 1 L of reverse osmosis water, the conductivity is 450~550 μS / cm; pH is 6.5~8.5; hardness is 50~100 mg / L of CaCO3.

[0032] The experimental animals of effect test example 2 of the present invention are 12-week-old C57BL / 6J male mice, which were purchased from Jiangsu Huachuang Xinnuo Co., Ltd. and are specific pathogen-free experimental animals with a license number of SCXK (Su) 2020-0009. Example 1

[0033] This embodiment provides a composition containing Lactobacillus paracasei N1115 for improving bone health. The product is composed of 0.5 mg of Lactobacillus paracasei N1115 bacterial agent, 1200 mg of milk calcium (in the form of Ca 2+ The Lactobacillus paracasei N1115 bacterial agent was a live bacterial agent with a live bacterial count of 3.4×10 11 CFU / g, milk calcium is expressed in terms of calcium content 2+ The milk mineral salt is provided in an amount of 25 g / 100 g, and the vitamin D is provided by a vitamin D powder having a vitamin D content of 2.25 mg / g. Example 2

[0034] This embodiment provides a composition containing Lactobacillus paracasei N1115 for improving bone health. The product is composed of 0.5 mg of Lactobacillus paracasei N1115 bacterial agent, 1200 mg of milk calcium (in the form of Ca 2+ The Lactobacillus paracasei N1115 inoculant was an inactivated inoculant, and the number of live bacteria before inactivation was 3.4×10 11 CFU / g, milk calcium is expressed in terms of calcium content 2+ The milk mineral salt is provided in an amount of 25 g / 100 g, and the vitamin D is provided by a vitamin D powder having a vitamin D content of 2.25 mg / g.

[0035] The inactivated bacteria agent is prepared by inoculating Lactobacillus paracasei N1115 in a fermentation liquid containing skim milk, and then fermenting, centrifuging, inactivating and freeze-drying. The inactivation condition is to maintain at 96°C for 20 minutes, and the amount of live bacteria after inactivation is less than 3000 CFU / g. Example 3

[0036] This embodiment provides a composition containing Lactobacillus paracasei N1115 for improving bone health. The product is composed of 0.5 mg of Lactobacillus paracasei N1115 bacterial agent, 1200 mg of milk calcium (in the form of Ca 2+ The Lactobacillus paracasei N1115 bacterial agent was a live bacterial agent with a live bacterial count of 3.4×10 11 CFU / g, milk calcium is expressed in terms of calcium content 2+ The vitamin D is provided in the form of 25 g / 100 g milk mineral salts (ignoring impurities).

[0037] In the composition containing Lactobacillus paracasei N1115 for improving bone health, the amount of live bacteria of the Lactobacillus paracasei N1115 bacterial agent or the amount of live bacteria before inactivation is ≥8×10 10 CFU / g, has been proven to improve bone health by improving osteoblast activity and regulating bone metabolism, including upregulating the metabolism of prochondroitin, glycerophosphatidylcholine, sphingosine, butyrate and cystine.

[0038] In order to investigate whether Lactobacillus paracasei N1115, milk calcium and vitamin D have a synergistic effect in improving osteoporosis, the present invention conducts the following experiments: Effect Test Example 1

[0039] (1.1) Zebrafish validation experimental method: 240 wild-type AB zebrafish of 3 dpf were randomly divided into eight groups, each with 30 zebrafish. The eight groups were raised in a 28°C incubator. The eight groups were respectively counted as normal group, model group, alendronate sodium tablet group, milk mineral salt + VD group, N1115 group, commercially available Lactobacillus paracasei group (viable bacteria count was 3.4×10 11 CFU / g), milk mineral salt + VD + N1115 group and milk mineral salt + VD + inactivated bactericide group; Except for the normal group, the other seven groups were treated with an equal amount of dexamethasone in a water-soluble manner to treat zebrafish, and a zebrafish osteoporosis model was established. The dosage ratio of Lactobacillus paracasei N1115 bacterial agent, milk mineral salts and vitamin D in Example 1 was converted to zebrafish, and the dosage ratio of Lactobacillus paracasei N1115 inactivated bacterial agent, milk mineral salts and vitamin D in Example 2 was converted to zebrafish. Based on the principle of the controlled variable method, the corresponding water-soluble samples of each group were administered, as shown in Table 1; Table 1 Statistics of administration methods and dosages in different groups

[0040] After 96 hours of treatment at 28°C, samples were collected and stained with alizarin red. Ten zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. NIS-ElementsD3.20 advanced image processing software was used to analyze and collect data. The fluorescence intensity of the zebrafish spine was analyzed, and the anti-osteoporosis efficacy of the sample was evaluated based on the statistical analysis results of this indicator. The statistical processing results are expressed as mean±SE. SPSS26.0 software was used for statistical analysis, and p<0.05 indicated that the difference was statistically significant. This experiment analyzed the zebrafish spine. The fluorescence microscope images of the zebrafish spine in different groups are shown in the following figure. Figure 1 The results of the fluorescence intensity analysis of zebrafish vertebrae in different groups are shown in Figure 2 shown.

[0041] (1.2) Zebrafish validation experiment results: Figure 2 It can be seen that the fluorescence intensity of the model group was significantly lower than that of the normal group (p<0.001), indicating that dexamethasone caused increased calcium ion loss, leading to the formation of osteoporosis, and the model was successfully established.

[0042] Alendronate sodium tablets are a bone resorption inhibitor for the treatment and prevention of osteoporosis. They have a strong affinity with hydroxyapatite in the bone and play an anti-bone resorption role by inhibiting the activity of osteoclasts. Compared with the model group, the fluorescence intensity of this group was significantly enhanced (p<0.001), indicating that drug intervention effectively improved the symptoms of osteoporosis. The fluorescence intensity of the milk mineral salt + VD group, N1115 group and milk mineral salt + VD + N1115 group also increased significantly (p<0.001), indicating that all three groups have the effect of improving osteoporosis; the fluorescence intensity of the milk mineral salt + VD + N1115 group was significantly higher than that of the milk mineral salt + VD group (p<0.01) and the N1115 group (p<0.001), indicating that the combined consumption of milk mineral salt, VD and Lactobacillus paracasei N1115 can synergize and improve osteoporosis more significantly. The fluorescence intensity of the milk mineral salt + VD + inactivated bacteria group was also higher than that of the milk mineral salt + VD group and the N1115 group, indicating that the combination of inactivated bacteria, milk mineral salt and VD also has a more significant effect on improving osteoporosis. There was no significant difference between the commercially available Lactobacillus paracasei group and the model group, indicating that the strain has no function in improving osteoporosis, and its fluorescence intensity was significantly lower than that of the N1115 group (p<0.001). Effect Test Example 2

[0043] 72 12-week-old C57BL / 6J male mice purchased from Jiangsu Huachuang Xinnuo Co., Ltd. were randomly divided into 6 groups, with 12 mice in each group. The osteoporosis model construction and drug administration of each group are as follows: For ease of presentation, calcium carbonate is denoted as CCa, milk mineral salt (milk calcium) is denoted as MCa, vitamin D3 is denoted as VD, and Lactobacillus casei N1115 is denoted as Lp.

[0044] A. CON group: normal drinking water, intramuscular injection and oral gavage of equal volume of normal saline; B. GIOP group: dexamethasone (5 mg / kg / week) was injected intramuscularly and an equal amount of normal saline was administered intragastrically; C. CCa group: dexamethasone (5 mg / kg / week) intramuscular injection and calcium carbonate (0.27 g / kg / day) gavage; D. Lp group: intramuscular injection of dexamethasone (5 mg / kg / week), Lactobacillus casei N1115 (1×10 9 CFU / day) by intragastric administration; E. MCa+ Lp group: intramuscular injection of dexamethasone (5 mg / kg / week), milk mineral salts (0.44 g / kg / day) and Lactobacillus casei N1115 (1×10 9 CFU / day) by intragastric administration; F. MCa+ VD+Lp group: intramuscular injection of dexamethasone (5 mg / kg / week), milk mineral salts (0.44 g / kg / day), vitamin D3 (0.7 μg / kg / day) and Lactobacillus casei N1115 (1×10 9 CFU / day) by intragastric administration. This group was based on the dosage of the formula in Example 3 converted to mice.

[0045] Six groups of mice were subjected to bone tissue microstructure analysis, osteoblast and osteoclast marker detection, microbiome analysis, and bone health-related metabolomics analysis. The significance between groups in each group of results was marked with letters. The details are as follows: (2.1) Experimental methods for bone tissue microstructure analysis:

[0046] The femurs of mice in each group were collected, and the distal femur structure was scanned by micro-CT. The bone morphological parameters such as mouse bone density (BMD), bone volume fraction (BV / TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.sp) were statistically analyzed, and corresponding graphs were drawn to compare the differences in the microstructure of bone tissue of mice in each group, and to analyze the effect of the Lactobacillus paracasei N1115 composition of the present invention on improving bone health. (2.2) Experimental results of bone tissue microstructure analysis:

[0047] The results are as follows Figure 3 As shown in the figure, the reconstruction image of the femur D by micro-CT scanning is shown in the figure Figure 3A, shows that in the femoral microstructure of osteoporotic mice, the trabeculae change from plate-shaped to rod-shaped, and the cortical bone is thin, showing the characteristics of osteoporosis. Compared with normal mice, the femoral bone mineral density (BMD) of GIOP mice is significantly reduced, such as Figure 3 B; Bone volume fraction (BV / TV) is significantly reduced, such as Figure 3 C; Trabecular thickness (Tb.Th) was significantly reduced, such as Figure 3 D; The number of trabecular bone (Tb.N) is significantly reduced, such as Figure 3 E; Trabecular separation (Tb.sp) increased significantly, such as Figure 3 F, showing characteristics of osteoporosis; after CCa intervention, BMD, BV / TV, Tb.Th and Tb.sp showed an improving trend, but there was no significant difference; after Lp and Lp+MCa intervention, BMD, BV / TV and Tb.sp improved significantly, and the other parameters showed an improving trend, but there was no significant difference; after Lp+MCa+VD intervention, BMD, BV / TV, Tb.Th, Tb.N and Tb.sp were all significantly improved.

[0048] The above results indicate that Lp, MCa + Lp and MCa + VD + Lp can significantly improve the femoral bone density and bone microarchitecture destruction in GIOP mice, and the improvement effect is more obvious after the additional addition of Lp. (2.3) Experimental methods for detecting osteoblast and osteoclast markers:

[0049] Femurs of mice in each group were collected, decalcified and sliced ​​for tartrate-resistant acid phosphatase (TRAP) staining, and the histopathological morphology was observed and analyzed under an optical microscope, and the number of TRAP-positive cells was counted to analyze the activity of osteoclasts. Serum of mice in each group was collected, and alkaline phosphatase (ALP) was detected using a biochemical kit, and quantitative analysis was performed using an enzyme-labeled instrument to analyze the activity of osteoblasts. (2.4) Results of osteoblast and osteoclast marker detection experiments:

[0050] The results are as follows Figure 4 As shown in the figure, compared with the control group, the number of TRAP-positive cells near the trabeculae of GIOP mice (the part pointed by the arrow) increased significantly, that is, the activity of osteoclasts increased, and different intervention substances improved this phenomenon to varying degrees, such as Figure 4 A. The number of TRAP-positive cells in tissues was counted and statistically analyzed, and it was found that Lp, MCa+Lp, and MCa+VD+Lp could significantly reduce the number of TRAP-positive cells in femoral tissues of GIOP mice, that is, significantly improve the activity of osteoclasts in tissues, such as Figure 4B. ALP is an exoenzyme produced and secreted by osteoblasts, which can effectively show the activity of osteoblasts in the body. The serum ALP content of GIOP mice was significantly lower than that of normal mice, that is, the activity of osteoblasts in the body was reduced. The intervention of different components showed an improvement effect on the ALP content of GIOP mice, but there was no statistical difference. At the same time, after adding VD and Lp, the improvement effect on the activity of osteoblasts in GIOP mice was more obvious. Figure 4 C.

[0051] The above results show that the Lactobacillus paracasei N1115 composition for improving bone health of the present invention can significantly improve osteoblast activity and osteoblast activity. (2.5) Experimental methods for microbiome analysis:

[0052] Fecal samples of each group of mice were taken for 16S rRNA analysis, the intestinal microbial genera and relative abundance of each group of mice were counted, and Venn diagrams and UPGMA clustering trees / and abundance combination diagrams were drawn to analyze the effects of the Lactobacillus paracasei N1115 composition for improving bone health of the present invention on beneficial and harmful intestinal bacteria. (2.6) Microbiome analysis experimental results:

[0053] The results are as follows Figures 5 to 7 There were significant differences between the mice in the GIOP model group and the mice in the Lp group. Specifically, compared with the GIOP group, the intervention with Lp and its combination significantly affected the relative abundance of at least 59 bacterial species in the intestines of GIOP mice, such as Figure 5 From the UPGMA cluster tree analysis, it can be observed that the intestinal flora structure of GIOP mice is obviously clustered, and there is a clear separation trend from the CON control group and the Lp and its combination intervention group, indicating that GIOP can significantly change the overall structure of the intestinal flora of mice, and Lp and its combination can reverse this situation. Figure 6 .

[0054] like Figure 7 The relative abundance of representative differential bacteria in the intestinal flora of the Lp group, GIOP model group and CON control group was further analyzed. The results showed that there were significant differences in the intestinal flora of the Lp and GIOP groups at the genus level; among them, Duncanella ( Duncaniella ), Lactobacillus spp. Lactobacillus ), Heterobacillus spp. Allobaculum ), Isoprevotella spp. Alloprevotella )-based probiotics were upregulated; Muribaculum , Bacteroides Bacteroides ), another genus ( Alistipes ) were downregulated.

[0055] The above results show that the Lp and its composition of the present invention can significantly improve the abundance of beneficial bacteria in the intestinal flora and reduce the abundance of harmful bacteria. (2.7) Experimental methods for bone health-related metabolomics analysis:

[0056] Fecal samples from each group of mice were taken for non-targeted metabolite analysis to explore the effects of Lp and its combination on the metabolic status of the intestinal microbial environment of GIOP mice. (2.8) Results of bone health-related metabolomics analysis experiments:

[0057] The results showed that there were differences in 396 key metabolites related to bone health between the Lp group and the GIOP group; there were differences in 195 key metabolites related to bone health between the MCa+Lp group and the GIOP group; there were differences in 343 metabolites between the Ca+VD+Lp group and the GIOP group. These metabolites are enriched in pathways related to amino acid biosynthesis, aminoacyl-tRNA biosynthesis, tryptophan metabolism, sphingolipid metabolism, and cofactor biosynthesis. In particular, there are relatively abundant differences in the differences in metabolites between the Lp group and the GIOP group, such as Figure 8 , among which the more abundant metabolites were related to the development of OP or bone health, as shown in Table 2 .

[0058] Table 2 Metabolite difference results

[0059] These results suggest that Lp and its combination can significantly affect the relative contents of intestinal microbial metabolites in GIOP mice, some of which are closely related to bone health. Embodiment 4-6

[0060] Examples 4 to 6 respectively provide a composition product having the function of improving bone health. The difference is that the types or amounts of raw materials in each example are different. The specific raw material ratios of different examples are shown in Table 3.

[0061] Table 3 Raw material ratios in Examples 4 to 6

[0062] The preparation method of the composition products in Examples 4 to 6 is to weigh the raw materials according to the ratio table, compound them, and mix them evenly according to the existing technology in the art.

[0063] In every 100g of the composition product with the function of improving bone health, the content of Lactobacillus paracasei N1115 bacterial agent is 0.2~20mg, the content of milk calcium is 1000~1700mg, and the content of vitamin D is 5~10μg. Example 7

[0064] In view of the effect of improving osteoporosis, Lactobacillus paracasei N1115 can be used as a raw material for food or health food for improving bone health. In this embodiment, the composition of Example 1 is used as one of the raw materials to prepare a formula powder for improving bone health.

[0065] The specific preparation method is as follows: skim milk powder, whole milk powder, inulin, desalted whey powder, lecithin, the composition of Example 1, magnesium sulfate and ferrous sulfate are weighed in a weight ratio of 30:20:15:15:10:4.8:2.7:2.5, and the above raw materials are subjected to a dry-wet composite process to obtain a formula milk powder suitable for consumption by middle-aged and elderly people to prevent or improve osteoporosis.

[0066] In other embodiments, the composition of Example 1 is replaced by the composition of Example 2 to 6, and the formula powder having the function of preventing or improving osteoporosis is prepared according to the above method. Example 8

[0067] This example is a liquid product for improving bone health, which is prepared by dispersing the composition of Example 1 into liquid milk.

[0068] In other embodiments, the compositions of Examples 2 to 6 are used instead of the composition of Example 1 and are dispersed in water, edible oil or liquid milk, respectively, to prepare a liquid product for improving bone health. Example 9

[0069] This embodiment is a postbiotic preparation for improving bone health, which comprises the composition of Example 2, and further comprises maltodextrin and a sweetener.

[0070] In other embodiments, the Lactobacillus paracasei N1115 bacterial agent in the compositions of Examples 1 and 3 to 6 is replaced with an inactivated bacterial agent, and maltodextrin and a sweetener are added to prepare a postbiotic product.

[0071] The inactivated bacteria agent is prepared by inoculating Lactobacillus paracasei N1115 in a fermentation liquid containing skim milk, and then fermenting, centrifuging, inactivating, and freeze-drying. The inactivation conditions are 90-98°C for 20-40 minutes, specifically 90°C for 40 minutes, 93°C for 35 minutes, 98°C for 40 minutes, and 96°C for 20 minutes. The amount of live bacteria after inactivation is less than 3000 CFU / g.

[0072] It should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the above embodiments, those skilled in the art can still modify the technical solutions described in the above embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the scope of protection of the claims of the present invention.

Claims

1. A composition containing Lactobacillus paracasei N1115 for improving bone health, characterized in that: The composition comprises a bacterial agent of Lactobacillus paracasei N1115, milk calcium and vitamin D, wherein the content ratio of the bacterial agent of Lactobacillus paracasei N1115, milk calcium and vitamin D is 0.2-20 mg: 1000-1700 mg: 5-10 μg.

2. The composition containing Lactobacillus paracasei N1115 for improving bone health according to claim 1, characterized in that The Lactobacillus paracasei N1115 bacterial agent is a live bacterial agent or an inactivated bacterial agent.

3. The composition containing Lactobacillus paracasei N1115 for improving bone health according to claim 2, characterized in that, The live bacteria amount of the Lactobacillus paracasei N1115 bacterial agent or the live bacteria amount before inactivation is ≥ 8×10 10 CFU / g.

4. The composition containing Lactobacillus paracasei N1115 for improving bone health according to any one of claims 1 to 3, characterized in that The improvement of bone health is achieved by improving osteoblast activity and regulating bone metabolism.

5. The composition containing Lactobacillus paracasei N1115 for improving bone health according to claim 4, characterized in that: The regulation of bone metabolism includes upregulating the metabolism of prochondroitin, glycerophosphatidylcholine, sphingosine, butyrate and cystine.

6. Use of the composition containing Lactobacillus paracasei N1115 for improving bone health according to any one of claims 1 to 5 in preparing a product having the function of improving bone health.

7. A formula powder for improving bone health, characterized in that: The invention comprises the composition containing Lactobacillus paracasei N1115 for improving bone health according to any one of claims 1 to 5.

8. A liquid product for improving bone health, characterized in that: The invention is a liquid product obtained by dispersing the composition containing Lactobacillus paracasei N1115 for improving bone health as claimed in any one of claims 1 to 5 into water, edible oil or liquid milk.

9. A postbiotic product for improving bone health, characterized in that: The invention comprises the composition containing Lactobacillus paracasei N1115 for improving bone health according to any one of claims 1 to 5, wherein the Lactobacillus paracasei N1115 bacterial agent in the composition is an inactivated bacterial agent.

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