Probiotic preparation containing BL11 strain and capable of improving obesity-related metabolism and senescence indexes of middle-aged and elderly people and application of probiotic preparation

Abstract

The invention relates to a probiotic preparation containing a BL11 strain and capable of improving obesity-related metabolism and senescence indexes of middle-aged and elderly people and application of the probiotic preparation. The strain in the probiotic preparation is composed of a bifidobacterium longum subsp. Longum BL11 strain and a pediococcus acidilactici PA53 strain. The two strains can cooperate with each other and promote each other, and have a synergistic effect in the aspect of improving obesity-related metabolic indexes, which is specifically embodied in that: the weight increase and body fat rate of mice fed with high fat diet can be reduced; the low-density lipoprotein level, the total cholesterol level and the triglyceride level are reduced, and the high-density lipoprotein level is improved; fasting blood-glucose, glycosylated hemoglobin level and insulin resistance index are reduced; chronic inflammation is relieved; meanwhile, the composition also has excellent performance in the aspect of improving senescence-related indexes, the content of malondialdehyde in serum can be reduced, the activity of superoxide dismutase and glutathione peroxidase can be improved, and the activity of telomerase can be improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of probiotics, and relates to a probiotic preparation containing strain BL11 for improving middle-aged and elderly obesity-related metabolism and aging indexes and its application. Background Art

[0002] Metabolic syndrome (MetS) is a group of complex metabolic disorder syndromes with insulin resistance as the core pathophysiological basis, including central obesity, impaired glucose tolerance or type 2 diabetes, hypertension, dyslipidemia, etc. Metabolic syndrome not only significantly increases the incidence risks of cardiovascular diseases and type 2 diabetes, but also is closely related to various senile diseases such as cognitive dysfunction and non-alcoholic fatty liver.

[0003] In recent years, with the proposal and development of the "microbiota-gut-organ axis" theory, intestinal microbiota dysbiosis has been considered as one of the important environmental factors in the occurrence and development of metabolic syndrome. Metabolic syndrome patients generally have characteristic changes such as reduced intestinal microbiota diversity and disordered microbiota structure. This microbiota imbalance can affect host metabolism through multiple mechanisms: (1) Energy metabolism regulation disorder: The short-chain fatty acids produced by the fermentation of dietary fiber by the microbiota are reduced, resulting in weakened inhibition of hepatic gluconeogenesis and increased lipolysis in adipose tissue; (2) Intestinal barrier damage: The expression of tight junction proteins is down-regulated, increasing endotoxin translocation and inducing chronic low-grade inflammation; (3) Bile acid metabolism disorder: Affecting the farnesoid X receptor and G protein-coupled bile acid receptor 1 signaling pathways, interfering with the balance of glucose and lipid metabolism; (4) Brain-gut axis dysfunction: Affecting the secretion of metabolic regulatory peptides by enteroendocrine cells, resulting in abnormal appetite regulation and excessive energy intake.

[0004] Based on the close association between intestinal microbiota and metabolic syndrome, microecological intervention has become a current research hotspot. At present, the types of probiotic products for metabolic improvement are still few. Therefore, it is very meaningful to search for and develop more probiotic intervention strategies, especially probiotic combination strategies. Summary of the Invention

[0005] Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a probiotic preparation containing strain BL11 for improving middle-aged and elderly obesity-related metabolism and aging indexes and its application.

[0006] To achieve the purpose of this invention, the present invention adopts the following technical solutions: In the first aspect, the present invention provides a probiotic preparation containing strain BL11 for improving middle-aged and elderly obesity-related metabolism and aging indexes, and the strain in the probiotic preparation is Bifidobacterium longum subsp. with the preservation number of CGMCC No. 24412 Bifidobacterium longum subsp . longumBifidobacterium longum subsp. BL11 strain and Pediococcus acidilactici PA53 strain with the preservation number of CGMCC No. 18798 Pediococcus acidilactici It consists of PA53 strain. Among them, the Bifidobacterium longum subsp. BL11 strain involved in the present invention was isolated from the fecal sample of a healthy breastfed infant in Bama Town, Bama Yao Autonomous County, Hechi City, Guangxi Zhuang Autonomous Region, China.

[0007] The present invention creatively develops a brand-new probiotic compounding method and a brand-new strategy for improving obesity-related metabolic indexes, that is, compounding and combining Bifidobacterium longum subsp. Bifidobacterium longum subsp . longum BL11 strain and Pediococcus acidilactici Pediococcus acidilactici PA53 strain, and it is found that the two can cooperate with each other, promote each other, and have a synergistic effect in improving obesity-related metabolic indexes, specifically reflected in: (1) It can reduce the weight gain and body fat rate of mice fed with high-fat diet, and reduce visceral fat; (2) It can reduce the levels of low-density lipoprotein, total cholesterol and triglyceride in mice fed with high-fat diet, and increase the level of high-density lipoprotein; (3) It can reduce the fasting blood glucose, glycated hemoglobin level and insulin resistance index of mice fed with high-fat diet; (4) Alleviate chronic inflammation; (5) Improve aging-related indexes.

[0008] Preferably, the viable count ratio of the Bifidobacterium longum subsp. Bifidobacterium longum subsp. longum BL11 strain to Pediococcus acidilactici Pediococcus acidilactici PA53 strain is (1-10):1, such as 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0009] Preferably, in the probiotic preparation, the total viable count is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, such as 1×10 9 CFU / g (CFU / mL), 2×10 9 CFU / g (CFU / mL), 5×10 9 CFU / g (CFU / mL), 8×10 9 CFU / g (CFU / mL), 1×10 10 CFU / g (CFU / mL), 5×10 10 CFU / g (CFU / mL), 1×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 1×10 13CFU / g (CFU / mL), etc. Other specific point values within this numerical range can be selected and will not be elaborated one by one here.

[0010] Preferably, the dosage form of the probiotic preparation includes solution, powder, capsule, tablet or granule.

[0011] Preferably, the probiotic preparation further contains a cell protectant, and the cell protectant includes any one or a combination of at least two of skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinylpyrrolidone, sucrose, lactose, trehalose, sorbitol or xylitol.

[0012] Preferably, the probiotic preparation further contains a functional adjuvant, and the functional adjuvant includes any one or a combination of at least two of fructooligosaccharide, galactooligosaccharide, xylooligosaccharide, isomaltooligosaccharide, soybean oligosaccharide, inulin, polydextrose, α-lactalbumin or lactoferrin.

[0013] Preferably, the probiotic preparation further contains excipients, and the excipients include any one or a combination of at least two of filler, binder, wetting agent, disintegrant, emulsifier, solubilizer, solubilizing agent, osmotic pressure regulator, colorant, pH regulator, antioxidant, bacteriostatic agent or buffer.

[0014] In a second aspect, the present invention provides the use of the probiotic preparation according to the first aspect in the preparation of a preparation for improving blood glucose metabolism and / or blood lipid metabolism.

[0015] In a third aspect, the present invention provides the use of the probiotic preparation according to the first aspect in the preparation of a preparation for preventing, improving or assisting in improving obesity-related metabolic syndrome.

[0016] In a fourth aspect, the present invention provides the use of the probiotic preparation according to the first aspect in the preparation of a nutritional supplement for maintaining metabolic health.

[0017] Compared with the prior art, the present invention has the following beneficial effects: The present invention creatively develops a brand-new probiotic compounding method and a brand-new strategy for improving obesity-related metabolic indicators, that is, Bifidobacterium longum subsp. Bifidobacterium longum subsp . longum BL11 strain and Pediococcus acidilactici Pediococcus acidilacticiWhen the BL11 strain and the PA53 strain are used in combination, it is found that they can cooperate with each other, promote each other, and have a synergistic effect on improving obesity-related metabolic indicators, specifically manifested in: (1) it can reduce the weight gain and body fat percentage of mice fed with a high-fat diet, and reduce visceral fat; (2) it can reduce the levels of low-density lipoprotein, total cholesterol and triglyceride in mice fed with a high-fat diet, and increase the level of high-density lipoprotein; (3) it can reduce the fasting blood glucose, glycated hemoglobin level and insulin resistance index of mice fed with a high-fat diet; (4) it can alleviate chronic inflammation; (5) it can improve aging-related indicators.

[0018] The BL11 strain involved in the present invention is classified and named as Bifidobacterium longum subsp. longum Bifidobacterium longum subsp. longum , the preservation unit is the General Microbiological Center of the China Committee for Culture Collection of Microorganisms. The preservation time is February 21, 2022, and the preservation number is CGMCC No. 24412. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0019] The PA53 strain involved in the present invention is classified and named as Pediococcus acidilactici. The preservation unit is the General Microbiological Center of the China Committee for Culture Collection of Microorganisms. The preservation time is November 4, 2019, and the preservation number is CGMCC No. 18798. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Description of the Drawings

[0020] Figure 1 It is a statistical result graph of the weight growth rate of mice in each group; Figure 2 It is a statistical result graph of the body fat percentage of mice in each group; Figure 3 It is a statistical result graph of the fasting blood glucose level of mice in each group; Figure 4 It is a statistical result graph of the glycated hemoglobin level of mice in each group; Figure 5 It is a statistical result graph of the insulin level of mice in each group; Figure 6 It is a statistical result graph of the area under the oral glucose tolerance curve of mice in each group; Figure 7 It is a statistical result graph of the serum total cholesterol level of mice in each group; Figure 8 It is a statistical result graph of the serum triglyceride level of mice in each group; Figure 9 It is a statistical result graph of the serum low-density lipoprotein level of mice in each group; Figure 10It is a statistical result graph of the serum high-density lipoprotein levels of each group of mice; Figure 11 It is a statistical result graph of the serum inflammatory factor TNF-α levels of each group of mice; Figure 12 It is a statistical result graph of the serum inflammatory factor IL-6 levels of each group of mice; Figure 13 It is a statistical result graph of the triglyceride levels in the liver tissues of each group of mice; Figure 14 It is a statistical result graph of the malondialdehyde content in the liver tissues of each group of mice; Figure 15 It is a statistical result graph of the superoxide dismutase activity in the liver tissues of each group of mice; Figure 16 It is a statistical result graph of the glutathione peroxidase activity in the liver tissues of each group of mice; Figure 17 It is a statistical result graph of the telomerase activity of each group of mice; In the figure, represents a significant difference compared with the control group ( p <0.001); * represents a significant difference compared with the model group ( p <0.05); ** represents a significant difference compared with the model group ( p <0.01); *** represents a significant difference compared with the model group ( p <0.001). Detailed implementation manners

[0021] The technical solutions of the present invention will be further described below through specific implementation manners. Those skilled in the art should understand that the embodiments are only for helping to understand the present invention and should not be regarded as specific limitations on the present invention.

[0022] The culture media involved in the following embodiments are as follows: MRS culture medium (g / L): peptone 10 g / L, beef extract 10 g / L, glucose 15 g / L, lactose 15 g / L, yeast powder 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 80 1 mL / L, cysteine hydrochloride 0.5 g / L.

[0023] The taxonomic name of the BL11 strain involved in the following embodiments is Bifidobacterium longum subsp. Bifidobacterium longum subsp . longum , the preservation time is February 21, 2022, and the preservation number is CGMCC No. 24412.

[0024] The taxonomic naming of the PA53 strain involved in the following examples is Pediococcus acidilactici Pediococcus acidilactici , the preservation time was November 4, 2019, and the preservation number was CGMCC No. 18798.

[0025] The bacterial suspension involved in the following examples: The required strain was inoculated into MRS liquid medium and cultured at 37°C for 24 h for activation to obtain an activated solution; the activated solution was inoculated into MRS liquid medium at an inoculation amount of 3% (v / v) and cultured at 37°C for 24 h to obtain a bacterial solution; the bacterial solution was centrifuged at 6000 g for 6 min, and the cells were resuspended with PBS to obtain the bacterial suspension. Example

[0026] This example explored the ability of probiotic preparations to improve relevant metabolic indexes in high-fat model mice: (1) Test animals: C57BL / 6J male mice (12 months old) (80 mice). These mice were raised in a controlled environment with the room temperature maintained at 22 ± 2°C, the humidity at 55% ± 5%, following a 12 h light / dark cycle, and with free access to food and water.

[0027] (2) Animal grouping: After 1 week of adaptive feeding of the above mice, 80 mice were randomly divided into 8 groups (10 mice in each group): control group, model group, BL11 intervention group, PA53 intervention group, ATCC8042 intervention group (commercial Pediococcus acidilactici intervention group), BL11 + PA53 composite group 1 (the ratio of viable bacteria numbers was 1:1), BL11 + PA53 composite group 2 (the ratio of viable bacteria numbers was 10:1), BL11 + ATCC8042 composite group (the ratio of viable bacteria numbers was 1:1).

[0028] (3) Animal modeling and intervention methods: The control group was fed with normal feed + normal purified water, the model group was fed with high-fat feed containing 60% fat + normal purified water, and each probiotic intervention group was fed with high-fat feed containing 60% fat + the bacterial suspension of each group (the total amount of bacteria given to each group was 10 9 CFU / day / mouse), and the intervention was continuous for 12 weeks.

[0029] (4) Index analysis: (4.1) Effects on body weight growth rate and body fat rate: After the intervention, the body weights of the mice in each group were weighed, and the body weight growth rate (%) of the mice was calculated; the visceral fat content of the mice was measured by nuclear magnetic resonance (MRI), and the body fat rate (%) of the mice was calculated. The statistical results of the averages of each group are as Figure 1 and Figure 2 shown.

[0030] As shown by Figure 1and Figure 2 It can be seen that, compared with the control group, the body weight growth rate and body fat rate of the mice in the model group were significantly increased. After probiotic intervention, different degrees of reversal effects occurred, and the reversal effects of the BL11+PA53 composite groups 1 and 2 were better, which could significantly reduce the body weight growth rate and body fat rate of the mice in the model group.

[0031] (4.2) Effects on blood glucose indexes: After the intervention, blood was collected from the tail vein of the mice to measure the fasting blood glucose level, glycated hemoglobin (HbA1c) level and insulin level of the mice, and the insulin resistance index (HOMA-IR) was calculated. The results are as Figures 3 - 5 shown.

[0032] At the same time, the oral glucose tolerance test of the mice was carried out. The glucose dosage for the glucose tolerance test of the mice was 2 g per kilogram of body weight (2 g / kg), and a 20% glucose solution was prepared with normal saline; the mice were fasted for 16 hours, and the mice maintained normal drinking water during this period; before starting the glucose tolerance test, the body weight of each mouse was weighed, blood was taken from the tip of the mouse's tail, and the fasting blood glucose was measured with a blood glucose meter, and the measured value was regarded as the blood glucose value at 0 min; glucose was intragastrically administered and timing started from the moment it was completed. The blood glucose values of each mouse were measured at 15, 30, 60, 90 and 120 min, and the area under the oral glucose tolerance curve (OGTT AUC) was calculated. The statistical results of the average values of each group are as Figure 6 shown.

[0033] From Figures 3 - 6 it can be seen that, compared with the control group, the fasting blood glucose level, HbA1c level and HOMA-IR of the mice in the model group were significantly increased, showing obvious glucose metabolism disorders. After probiotic intervention, different degrees of reversal effects occurred, and the reversal effects of the BL11+PA53 composite groups 1 and 2 were better, which could more significantly reduce the fasting blood glucose level, HbA1c level and HOMA-IR of the mice in the model group, and the OGTT results were close to the normal level, indicating that the BL11 strain and the PA53 strain had synergistic effects in improving glucose metabolism disorders.

[0034] (4.3) Effects on blood lipid indexes: After the intervention, the mice were sacrificed, blood was collected from the eyeballs, and after centrifugation at 1200×g for 10 min, the serum was extracted and stored at -80°C. An enzyme-linked immunosorbent assay kit was used to detect the levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), and high-density lipoprotein (HDL-C) in the serum. The statistical results of the average values of each group are as Figures 7 - 10 shown.

[0035] From Figures 7 - 10It can be seen that compared with the control group, the levels of serum total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL-C) in the model group of mice were significantly increased, while the level of high-density lipoprotein (HDL-C) was significantly decreased, showing obvious lipid metabolism disorders. After probiotic intervention, varying degrees of reversal effects occurred, and the reversal effects of the BL11+PA53 composite groups 1 and 2 were better, superior to those of the BL11 intervention group and the PA53 intervention group, indicating that the BL11 strain and the PA53 strain had synergistic effects in improving lipid metabolism disorders.

[0036] (4.4) Effects on serum inflammatory factors: After the intervention ended, the mice were sacrificed, blood was collected from the eye sockets, and after centrifugation at 1200×g for 10 min, the serum was extracted and stored at -80°C. An enzyme-linked immunosorbent assay kit was used to detect the levels of the inflammatory factors TNF-α and IL-6 in the serum. The statistical results of the averages of each group are as Figures 11 - 12 shown.

[0037] From Figures 11 - 12 the data results, it can be seen that compared with the control group, the levels of the serum inflammatory factors TNF-α and IL-6 in the model group of mice were significantly increased. After probiotic intervention, the levels of the inflammatory factors TNF-α and IL-6 decreased to varying degrees, and the intervention effects of the BL11+PA53 composite groups 1 and 2 were better, superior to those of the BL11 intervention group and the PA53 intervention group, indicating that the BL11 strain and the PA53 strain had synergistic effects in this efficacy aspect.

[0038] (4.5) Effects on the content of triglyceride (TG) in liver tissue: After the intervention ended, the mice were sacrificed, and liver tissue homogenates of the mice were taken to detect the content of triglyceride (TG) in the supernatant of the homogenates. The statistical results of the averages of each group are as Figure 13 shown.

[0039] From Figure 13 the data results, it can be seen that compared with the control group, the content of triglyceride (TG) in the liver tissue of the model group of mice was significantly increased. After probiotic intervention, the level of triglyceride (TG) decreased to varying degrees, and the intervention effects of the BL11+PA53 composite groups 1 and 2 were better, superior to those of the BL11 intervention group and the PA53 intervention group, indicating that the BL11 strain and the PA53 strain had synergistic effects in this efficacy aspect.

[0040] (4.6) Effects on aging-related indicators: After the intervention ended, liver tissue of the mice was taken to detect the oxidative stress indicators (content of malondialdehyde (MDA), activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px)) and telomerase activity in the supernatant of the homogenates. The statistical results of the averages of each group are asFigures 14 - 17 as shown

[0041] It can be seen from Figures 14 - 17 that compared with the control group, the serum MDA content of the mice in the model group was significantly increased, while the activities of SOD, GSH-Px and telomerase were significantly decreased. After probiotic intervention, the above indexes were improved to varying degrees, and the intervention effects of the BL11+PA53 composite groups 1 and 2 were better than those of the BL11 intervention group and the PA53 intervention group, indicating that the BL11 strain and the PA53 strain have synergistic effects in improving the aging-related indexes.

[0042] The applicant declares that the technical solution of the present invention is illustrated by the above embodiments, but the present invention is not limited to the above embodiments, that is, it does not mean that the present invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, the equivalent substitution of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

[0043] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various 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.

[0044] In addition, it should be noted that in the above specific embodiments, the various specific technical features described can be combined in any suitable way without conflict. To avoid unnecessary repetition, the present invention will not separately describe various possible combination methods.

Claims

1. A probiotic preparation containing BL11 strain for improving obesity-related metabolism and aging indicators in the middle-aged and elderly, characterized in that: The strain in the probiotic preparation is Bifidobacterium longum subspecies longum with a deposit number of CGMCC No. 24412. Bifidobacterium longum subsp . longum BL11 strain and Pediococcus acidilactici with a deposit number of CGMCC No. 18798 Pediococcus acidilactici PA53 strain composition.

2. The probiotic preparation according to claim 1, characterized in that Bifidobacterium longum subsp. longum Bifidobacterium longum subsp. longum BL11 strain and Pediococcus acidilactici Pediococcus acidilactici The ratio of the number of viable bacteria of the PA53 strain is (1-10):

1.

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

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

5. The probiotic preparation according to claim 1, characterized in that The probiotic preparation also contains a bacterial protective agent, which includes 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.

6. The probiotic preparation according to claim 1, characterized in that The probiotic preparation also contains functional additives, which include any one of fructooligosaccharides, galacto-oligosaccharides, xylooligosaccharides, isomaltooligosaccharides, soybean oligosaccharides, inulin, polydextrose, α-lactalbumin or lactoferrin, or a combination of at least two of them.

7. The probiotic preparation according to claim 1, characterized in that The probiotic preparation also contains excipients, which include any one of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents or buffers, or a combination of at least two of them.

8. Use of the probiotic preparation according to any one of claims 1 to 7 in the preparation of a preparation for improving blood sugar metabolism and / or blood lipid metabolism.

9. Use of the probiotic preparation according to any one of claims 1 to 7 in the preparation of a preparation for preventing, improving or assisting in improving obesity-related metabolic syndrome.

10. Use of the probiotic preparation according to any one of claims 1 to 7 in the preparation of a nutritional supplement for maintaining metabolic health.

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