A probiotic agent for promoting height development containing Lactobacillus casei LC89 strain and application thereof

Abstract

The present invention relates to a probiotic agent for promoting height development containing Lactobacillus casei LC89 strain and its application. Lactobacillus cheese LC89 strain and Bifidobacterium longum subsp. infantis Bifidobacterium longum subsp.infantis The probiotics are composed of BI03 strains. In terms of growth and development indicators, they can significantly increase the weight and length of mice; in terms of bone development indicators, they can significantly increase the bone density and bone length of mice; in terms of biochemical indicators, they can significantly increase the levels of growth hormone and insulin-like growth factor-1 in mice.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotics and relates to a probiotic agent for promoting height development containing a Lactobacillus casei LC89 strain and an application thereof. Background Art

[0002] Probiotics are active microorganisms that are beneficial to the health of the host. By regulating the intestinal microecological system, they can improve children's nutrient absorption, enhance immune function and regulate the endocrine system. Specifically, probiotics can promote the absorption of key nutrients such as calcium, iron, and vitamins, which are essential for bone health and overall growth and development. In addition, probiotics can also affect the secretion of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) by regulating the gut-brain axis mechanism. GH promotes the growth of long bones by stimulating the proliferation and differentiation of chondrocytes, which is particularly important during the rapid growth period of children and adolescents; IGF-1 stimulates the activity of osteoblasts, promotes the production and mineralization of bone matrix, helps increase the length and density of bones, and thus promotes the growth and development of children.

[0003] In summary, probiotics can comprehensively promote children's growth and development through multiple mechanisms such as improving nutrient absorption, enhancing immune function, and regulating hormone secretion. Therefore, it is very meaningful to find and develop more probiotic intervention strategies, especially probiotic combination strategies. Summary of the invention

[0004] In view of the deficiencies in the prior art, the object of the present invention is to provide a probiotic for promoting height development containing Lactobacillus casei LC89 strain and an application thereof, and specifically to provide a probiotic for promoting height development containing Lactobacillus casei LC89 strain and an application thereof in the preparation of a product for promoting height development.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a probiotic for promoting height development containing Lactobacillus casei LC89 strain, wherein the strains in the probiotic are composed of Lactobacillus casei LC89 strain with a preservation number of CGMCC No.15409 and Bifidobacterium longum subsp. infantis BI03 strain with a preservation number of CGMCC No.24473.

[0007] The present invention creatively develops a new probiotic compounding method and a new strategy for promoting children's height development, that is, Lactobacillus casei LC89 strain and Bifidobacterium longum subsp. infantis BI03 strain are compounded and used together. It is found that the two can cooperate with each other, promote each other, and synergize in promoting height development. Animal test results show that: (1) in terms of growth and development indicators, the weight and body length of mice can be significantly increased; (2) in terms of bone development indicators, the bone density and bone length of mice can be significantly improved; (3) in terms of biochemical indicators, the levels of growth hormone and insulin-like growth factor-1 of mice can be significantly improved.

[0008] When the amount of bacteria used is the same, the combination of the two bacteria significantly improves the above-mentioned effects compared with a single LC89 strain or a single BI03 strain. Therefore, the probiotic agent provides a new strategy for promoting growth and development. Since Lactobacillus casei and Bifidobacterium longum infantis subspecies are both probiotics, they are highly safe and not prone to dependence when used to prepare related efficacy products.

[0009] Preferably, the ratio of the viable counts of the Lactobacillus casei LC89 strain to the Bifidobacterium longum subsp. infantis BI03 strain is (1-5):1, for example, 1:1, 2:1, 3:1, 4:1, 5:1, etc. Other specific point values ​​within the above numerical range can be selected and will not be described one by one here.

[0010] Preferably, in the probiotic agent, the total number of viable bacteria is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, for example 1×10 9 CFU / mL (CFU / g), 5×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), 5×10 10 CFU / mL (CFU / g), 1×10 11 CFU / mL (CFU / g), 1×10 12 CFU / mL (CFU / g), 1×10 13 CFU / mL (CFU / g), etc.; other specific point values ​​within this numerical range can be selected and will not be described here one by one.

[0011] Preferably, the dosage form of the probiotic includes solution, lyophilized powder, capsule, tablet or granule.

[0012] Preferably, the probiotic is in the form of a solution, which is prepared by the following method:

[0013] The LC89 strain and the BI03 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged and resuspended with a solvent to obtain an LC89 bacterial suspension and a BI03 bacterial suspension; the LC89 bacterial suspension and the BI03 bacterial suspension are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

[0014] Preferably, the probiotic is in the form of a freeze-dried powder, which is prepared by the following method:

[0015] The LC89 strain and the BI03 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and freeze-dried to obtain LC89 bacterial powder and BI03 bacterial powder; the LC89 bacterial powder and the BI03 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

[0016] Preferably, the protective agent is selected from any one or a combination of at least two of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol.

[0017] In a second aspect, the present invention provides use of the probiotic agent described in the first aspect in preparing a product for promoting height development.

[0018] Preferably, the product further comprises a functional adjuvant, and the functional adjuvant is selected from any one or a combination of at least two of fructooligosaccharides, galacto-oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, inulin, polydextrose, α-lactalbumin or lactoferrin.

[0019] Preferably, the product further comprises excipients, which are selected from any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators or antioxidants.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention creatively develops a new probiotic compounding method and a new strategy for promoting children's height development, that is, Lactobacillus casei LC89 strain and Bifidobacterium longum subsp. infantis BI03 strain are compounded and used together. It is found that the two can cooperate with each other, promote each other, and synergize in promoting height development. Animal test results show that: (1) in terms of growth and development indicators, the weight and body length of mice can be significantly increased; (2) in terms of bone development indicators, the bone density and bone length of mice can be significantly improved; (3) in terms of biochemical indicators, the levels of growth hormone and insulin-like growth factor-1 of mice can be significantly improved.

[0022] When the amount of bacteria used is the same, the combination of the two bacteria significantly improves the above-mentioned effects compared with a single LC89 strain or a single BI03 strain. Therefore, the probiotic agent provides a new strategy for promoting growth and development. Since Lactobacillus casei and Bifidobacterium longum infantis subspecies are both probiotics, they are highly safe and not prone to dependence when used to prepare related efficacy products.

[0023] The LC89 strain involved in the present invention is classified and named as Lactobacillus casei, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration Committee, the preservation time is March 5, 2018, the preservation number is CGMCC No.15409, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0024] The BI03 strain involved in the present invention is classified and named as Bifidobacterium longum subsp. infantis, and the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration. The preservation time is March 7, 2022, the preservation number is CGMCC No.24473, and the address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is the statistical result of bone density of each group of mice;

[0026] Figure 2 is the statistical result graph of bone length of each group of mice;

[0027] Figure 3 is a statistical result graph of the insulin-like growth factor-1 level in the serum of each group of mice;

[0028] Figure 4This is a statistical graph of the growth hormone levels in the serum of each group of mice. DETAILED DESCRIPTION

[0029] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0030] The culture medium formula involved in the following examples is as follows:

[0031] MRS medium: peptone 10g / L, beef extract 10g / L, glucose 20g / L, sodium acetate 2g / L, yeast powder 5g / L, diammonium hydrogen citrate 2g / L, K2PO4·3H2O 2.6g / L, MgSO4·7H2O 0.1g / L, MnSO4 0.05g / L, Tween 801mL / L, cysteine ​​hydrochloride 0.5g / L.

[0032] The LC89 strain involved in the following examples is classified and named as Lactobacillus casei, the preservation time is March 5, 2018, and the preservation number is CGMCC No.15409.

[0033] The BI03 strain involved in the following examples is classified and named Bifidobacterium longum subsp. infantis, with a preservation date of March 7, 2022 and a preservation number of CGMCC No.24473.

[0034] The following method for preparing a bacterial suspension involves: inoculating the desired strain into a liquid culture medium, culturing at 37°C for 24 hours for activation, and continuously activating twice to obtain an activated solution; inoculating the activated solution into a liquid culture medium at an inoculum rate of 5% (v / v), culturing at 37°C for 24 hours to obtain a bacterial solution; centrifuging the bacterial solution at 4°C and 5000 rpm for 10 minutes, filtering to obtain bacterial bodies, and resuspending the bacterial bodies in a PBS solution to obtain the product.

[0035] Example

[0036] This example explores the ability of probiotics to promote the growth and development of mice:

[0037] (1) Experimental animals: healthy SPF 3-week-old BALB / c male mice (purchased from Shanghai Experimental Animal Center). These mice were housed in a controlled environment with room temperature maintained at 20-24°C, humidity at 50-60%, and a 12-h light / dark cycle. They were allowed to eat and drink as they pleased.

[0038] (2) Animal grouping: After one week of adaptive feeding as described above, 70 mice were randomly divided into seven groups (10 mice in each group): control group (CON), LC89 strain group (LC89 group), BI03 strain group (BI03 group), composite bacteria group 1 (LC89+BI03 group, the ratio of live bacteria was 1:1), composite bacteria group 2 (LC89+BI03 group, the ratio of live bacteria was 5:1), composite bacteria group 3 (LC89+ATCC15707 group, the ratio of live bacteria was 1:1), and composite bacteria group 4 (ATCC393+BI03 group, the ratio of live bacteria was 1:1).

[0039] (3) Intervention methods:

[0040] Each probiotic intervention group was gavaged with 1×10 9 The mice were given 300 μL of the corresponding single bacterial suspension or compound bacterial suspension at 100 CFU / mL and were allowed to drink distilled water freely. The control group was gavaged with 300 μL of normal saline per mouse every day from day 1 to day 28 of the formal experiment and were allowed to drink distilled water freely.

[0041] (4) Testing indicators:

[0042] (4.1) Measurement of mouse weight and length:

[0043] During the experiment, the weight and body length (nose tip to anus) of mice were measured every week. The results are shown in Table 1 (weight, unit: g) and Table 2 (body length, unit: cm) (W0, W1, W2, W3, and W4 in the table represent before the experiment, after the first week of the experiment, after the second week, after the third week, and after the fourth week of the experiment, respectively). Compared with the control group, the weight and body length of mice in each probiotic intervention group increased significantly, especially the LC89 and BI03 combination group performed better, with a higher weight and body length than other probiotic groups throughout the intervention period.

[0044] Table 1

[0045]

[0046] Table 2

[0047]

[0048] (4.2) Bone density and bone length measurement:

[0049] After the experiment, the mice were killed, and the femurs and tibias on both sides were quickly peeled off, and the attached soft tissues were removed. The right femur was measured for bone density using a dual-energy X-ray absorptiometry. The right tibia was measured for the distance between the proximal and distal articular surfaces of the tibia using a vernier caliper, i.e., the tibia bone length.

[0050] The statistical results of bone density of mice in each group are shown in Figure 1As shown in Figure 2, the bone density of mice in each probiotic intervention group was significantly higher than that in the control group, among which the LC89 and BI03 combination group had the best effect; similarly, the statistical results of the tibia bone length of mice in each group are shown in Figure 2. Figure 2 As shown, the tibia length of mice in each probiotic intervention group was significantly higher than that in the control group, especially the LC89 and BI03 combination group performed better. In summary, the probiotic agent involved in the present invention has a significant promoting effect on bone growth and bone density enhancement in mice.

[0051] (4.3) Serological index testing:

[0052] After the experiment, the mice were killed, and serum was prepared from blood samples collected by cardiac puncture using serum separation tubes, and tested using mouse insulin-like growth factor 1 (IGF-1) ELISA research kit and mouse growth hormone (GH) ELISA research kit. The results were as follows: Figure 3 and Figure 4 As shown, the levels of IGF-1 and GH in the serum of mice were increased in each probiotic intervention group compared with the control group, among which the combination of LC89 and BI03 had the most significant effect, indicating that the probiotic agent involved in the present invention can regulate the levels of insulin-like growth factor 1 and growth hormone in serum, thereby promoting growth and development.

[0053] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0054] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0055] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A probiotic for promoting height development containing Lactobacillus casei LC89 strain, characterized in that: The strain in the probiotic is Lactobacillus casei with a preservation number of CGMCC No.15409 Lactobacillus casei LC89 strain and Bifidobacterium longum subsp. infantis with a deposit number of CGMCC No. 24473 Bifidobacterium longumsubsp. infantile BI03 strain composition.

2. The probiotic according to claim 1, characterized in that Lactobacillus casei Lactobacillus casei Bifidobacterium longum subsp. infantis Bifidobacterium longumsubsp. infantis The ratio of the number of viable cells of the BI03 strain was (1-5):

1.

3. The probiotic according to claim 1, characterized in that In the probiotics, the total number of viable bacteria is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

4. The probiotic according to claim 1, characterized in that The dosage form of the probiotics includes solution, lyophilized powder, capsule, tablet or granule.

5. The probiotic according to claim 4, characterized in that The dosage form of the probiotic is a solution, which is prepared by the following method: The LC89 strain and the BI03 strain were inoculated into the culture medium respectively for activation and fermentation culture in sequence to obtain fermentation broth; the fermentation broth was centrifuged and resuspended with a solvent to obtain LC89 bacterial suspension and BI03 bacterial suspension; The LC89 bacterial suspension and the BI03 bacterial suspension are mixed according to the ratio of viable bacteria count to obtain the probiotic agent.

6. The probiotic according to claim 4, characterized in that The dosage form of the probiotic is a freeze-dried powder, which is prepared by the following method: The LC89 strain and the BI03 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain a fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and freeze-dried to obtain LC89 bacterial powder and BI03 bacterial powder; the LC89 bacterial powder and the BI03 bacterial powder are mixed according to the ratio of the number of live bacteria to obtain the probiotic agent.

7. The probiotic according to claim 6, characterized in that The protective agent is selected from any one of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinyl pyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol, or a combination of at least two thereof.

8. Use of the probiotic according to any one of claims 1 to 7 in the preparation of a product for promoting height development.

9. The use according to claim 8, characterized in that: The product also includes a functional additive, which is selected from any one of fructooligosaccharides, galacto-oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, inulin, polydextrose, α-lactalbumin or lactoferrin, or a combination of at least two thereof.

10. The use according to claim 8, characterized in that: The product also includes auxiliary materials, which are selected from any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators or antioxidants.

Images