Composition of probiotic preparation of bifidobacterium longum infantis and medicinal and edible mixture and application thereof in regulating blood lipid, blood sugar, protecting cardiovascular and cerebrovascular system and liver and gall

Abstract

The present application belongs to the technical field of probiotics, and particularly relates to a composition of a postbiotic preparation of Bifidobacterium longum subsp. infantis and a medicinal and edible mixture and application thereof in regulating blood lipids, blood sugar, protecting cardiovascular and cerebrovascular systems and liver and gallbladder. The composition of the present application comprises a postbiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and a medicinal and edible mixture. The present application can effectively alleviate sugar and lipid metabolism disorder induced by high-fat and high-cholesterol diet, and in combination with the medicinal and edible mixture, natural plant components with metabolic regulation and cardiovascular and cerebrovascular protection effects are used as adjuvant therapy, so that the sugar and lipid metabolism disorder caused by high-fat and high-cholesterol diet can be better alleviated, liver cells can be protected, and chronic inflammatory reaction can be relieved, thereby having important application prospects for preventing and treating hyperlipidemia and relieving cardiovascular and cerebrovascular damage caused by metabolic dysfunction.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of probiotics, and specifically relates to a composition of a postbiotic preparation of Bifidobacterium longum subsp. infantis and a homoeopathy mixture and application thereof in regulating blood lipid, blood sugar, protecting cardiovascular system and liver and gallbladder. BACKGROUND

[0002] With the change of modern dietary structure and lifestyle, sugar and lipid metabolism dysfunction has become an important threat to global health. Long-term high-fat and high-cholesterol diet leads to lipid metabolism disorder and insulin sensitivity reduction in the body, directly causing metabolic abnormalities such as hyperlipidemia and hyperglycemia. These metabolic imbalances not only significantly increase the risk of atherosclerosis, coronary heart disease and hypertension and other cardiovascular diseases, but also exacerbate vascular sclerosis and myocardial damage through mechanisms such as chronic inflammation and endothelial damage. In addition, sugar and lipid metabolism dysfunction is also closely related to cerebrovascular diseases, including cerebral ischemic injury, neuroinflammatory response and cognitive function decline. Chronic low-grade inflammation and oxidative stress caused by hyperlipidemia and metabolic disorders not only lead to fibrosis of the cardiovascular system, vascular endothelial dysfunction, but also cause damage to the central nervous system, manifested as decreased learning ability, memory impairment and increased risk of cognitive degenerative diseases.

[0003] In recent years, there has been increasing attention to the prevention and treatment of metabolic disorder-related cardiovascular and cerebrovascular diseases through improving the balance of intestinal microecology. However, current research on the improvement of sugar and lipid metabolism dysfunction and cardiovascular and cerebrovascular damage induced by high-fat and high-cholesterol diet by probiotic postbiotics still needs further study. SUMMARY

[0004] The present application aims to provide a composition of a postbiotic preparation of Bifidobacterium longum subsp. infantis and a homoeopathy mixture and application thereof in regulating blood lipid, blood sugar, protecting cardiovascular system and liver and gallbladder, which has the effects of improving sugar and lipid metabolism and protecting cardiovascular system.

[0005] The application further provides a composition of a probiotic preparation of Bifidobacterium longum subsp. Infantis and a medicinal and edible mixture, which comprises a probiotic preparation of Bifidobacterium longum subsp. Infantis NKU FB3-14 and a medicinal and edible mixture; the Bifidobacterium longum subsp. Infantis NKU FB3-14 has a preservation number of CGMCC No. 25762 and is preserved in the China General Microbiological Culture Collection Center on September 21, 2022; the probiotic preparation of the Bifidobacterium longum subsp. Infantis NKU FB3-14 comprises heat-inactivated bacteria, heat-inactivated bacteria in bacteria liquid or heat-inactivated supernatant in bacteria liquid of the Bifidobacterium longum subsp. Infantis NKU FB3-14; the concentration of the Bifidobacterium longum subsp. Infantis NKU FB3-14 in the bacteria liquid of the Bifidobacterium longum subsp. Infantis NKU FB3-14 is 1x10 8 ~ 3x10 9 CFU / mL; the medicinal and edible mixture comprises the following raw materials in parts by mass: 10-20 parts of ginseng, 10-20 parts of longan meat, 10-20 parts of Poria cocos, 10-20 parts of lotus leaf, 5-15 parts of yam, 5-15 parts of coix seed, 5-15 parts of polygonatum, 5-15 parts of wheat, 5-15 parts of orange peel, 5-15 parts of white lentil and 4-6 parts of hawthorn.

[0006] Preferably, the probiotic preparation of the Bifidobacterium longum subsp. Infantis NKU FB3-14 comprises heat-inactivated bacteria in the bacteria liquid of the Bifidobacterium longum subsp. Infantis NKU FB3-14; the heat-inactivated bacteria are further vacuum freeze-dried; the concentration of the Bifidobacterium longum subsp. Infantis NKU FB3-14 in the bacteria liquid of the Bifidobacterium longum subsp. Infantis NKU FB3-14 is 2x10 9 ~ 3x10 9 CFU / mL.

[0007] Preferably, the mass ratio of the probiotic preparation of the Bifidobacterium longum subsp. Infantis NKU FB3-14 and the medicinal and edible mixture is (2-4):1.

[0008] The application further provides application of the composition described in the above scheme in the preparation of food.

[0009] Preferably, the food comprises food that helps improve sugar and lipid metabolism.

[0010] The sugar and lipid metabolism comprises sugar and lipid metabolism caused by high-fat and high-cholesterol diet.

[0011] Preferably, the improvement of glycolipid metabolism comprises one or more of the following: reducing body weight, controlling body weight gain, reducing blood glucose level, reducing triglyceride level, reducing cholesterol level, reducing appetite, improving food energy conversion efficiency, reducing organ weight, reducing epididymal fat content, inhibiting intestinal absorption of fat, inhibiting intestinal absorption of cholesterol, reducing adipocyte area, increasing serum leptin level, protecting hepatocytes, regulating bile acid metabolism, inhibiting bile acid reabsorption, and improving intestinal flora.

[0012] Preferably, the organ comprises liver and / or epididymis.

[0013] The present application also provides use of the composition described in the above-mentioned scheme in the preparation of a drug for preventing, treating and / or alleviating a disease, which comprises one or more of the following: obesity, glycolipid metabolism dysfunction, intestinal inflammation, intestinal flora imbalance, hyperlipidemia, and cardiovascular and cerebrovascular diseases.

[0014] Preferably, the prevention, treatment and / or alleviation of cardiovascular and cerebrovascular diseases comprises one or more of the following: reducing cardiovascular risk factor level, increasing serum NO level, reducing cardiovascular and cerebrovascular injury, alleviating impaired vascular function, promoting vasodilation and blood flow improvement, inhibiting myocardial cell fibrosis, protecting the nervous system, repairing the blood-brain barrier, and improving brain inflammation.

[0015] Beneficial effects:

[0016] The application provides a composition of a probiotic preparation of Bifidobacterium longum subsp. Infantis and a medicinal and edible mixture, and the composition comprises a probiotic preparation of Bifidobacterium longum subsp. Infantis NKUFB3-14 and a medicinal and edible mixture; the preservation number of the Bifidobacterium longum subsp. Infantis NKUFB3-14 is CGMCC No. 25762; the medicinal and edible mixture comprises the following raw materials in parts by mass: 10-20 parts of ginseng, 10-20 parts of longan meat, 10-20 parts of Poria cocos, 10-20 parts of lotus leaf, 5-15 parts of yam, 5-15 parts of coix seed, 5-15 parts of polygonatum, 5-15 parts of malt, 5-15 parts of orange peel, 5-15 parts of white lentil and 4-6 parts of hawthorn. The composition of the application comprises a probiotic preparation of Bifidobacterium longum subsp. Infantis NKUFB3-14 and a medicinal and edible mixture. The application can effectively relieve sugar and lipid metabolism disorder induced by high-fat and high-cholesterol diet, and the combination of the medicinal and edible mixture and the natural plant components with metabolic regulation and cardiovascular protection can better relieve sugar and lipid metabolism disorder caused by high-fat and high-cholesterol diet, relieve chronic inflammatory reaction, and thus has important application prospects for preventing and treating hyperlipidemia and relieving cardiovascular damage caused by metabolic dysfunction. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed in the embodiments.

[0018] Figure 1 A result graph of the influence of different forms of the probiotic preparation of Bifidobacterium longum subsp. Infantis NKUFB3-14 on the body weight of model mice;

[0019] Figure 2 A result graph of the influence of different forms of the probiotic preparation of Bifidobacterium longum subsp. Infantis NKUFB3-14 on the glucose (A), total cholesterol (B) and triglyceride (C) in the serum of model mice;

[0020] Figure 3 A result graph of the combination rate percentage of the probiotic preparation and the medicinal and edible mixture with taurocholate (A) and glycocholate (B) at different compounding ratios;

[0021] Figure 4 A result graph of the influence of the probiotic preparation of Bifidobacterium longum subsp. Infantis NKUFB3-14 and the medicinal and edible mixture composition on the body weight of model mice;

[0022] Figure 5The figure shows the effects of a combination of a postbiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and a mixture of food and medicine homologous ingredients on daily food intake (A), average energy intake (B), and food utilization (C) in model mice.

[0023] Figure 6 The effect of the combination of Bifidobacterium longum infant subspecies NKU FB3-14 postbiotic preparation and food-medicine homology mixture on liver weight (A) and epididymal fat weight (B) in model mice is shown in the figure.

[0024] Figure 7 The figure shows the effects of a combination of a postbiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and a mixture of food and medicine homologous ingredients on serum glucose, total cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) in model mice.

[0025] Figure 8 Figure showing the effect of a combination of postbiotic preparations and food-medicine homologous mixtures of Bifidobacterium longum infant subspecies NKU FB3-14 on the intestinal fat absorption capacity of model mice.

[0026] Figure 9 The results show the effects of the combination of Bifidobacterium longum infant subspecies NKU FB3-14 postbiotic preparation and food-medicine homology mixture on the epididymal fat morphology (A-F) (H&E stained pathological sections) and adipocyte area (E) of model mice.

[0027] Figure 10 The effect of a combination of postbiotic preparations and food-medicine homologous mixtures of Bifidobacterium longum infantis subsp. NKU FB3-14 on serum leptin levels in model mice is shown in the figure.

[0028] Figure 11 The effect of the combination of postbiotic preparation and food-medicine homology mixture of Bifidobacterium longum infant subspecies NKU FB3-14 on the liver morphology (A-E) (H&E stained pathological sections) of model mice;

[0029] Figure 12 The effect of a combination of postbiotic preparations and food-medicine homologous mixtures of Bifidobacterium longum subsp. infantis NKU FB3-14 on lipid deposition in the liver of model mice (A-E) (Oil Red stained pathological sections);

[0030] Figure 13 The figure shows the effect of a combination of postbiotic preparations and food-medicine homologous mixtures of Bifidobacterium longum infant subsp. NKU FB3-14 on the levels of ALT(A) and AST(B) in the liver tissue of model mice.

[0031] Figure 14Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on cardiovascular risk factors of model mice;

[0032] Figure 15 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on serum nitric oxide (NO) level of model mice;

[0033] Figure 16 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on myocardial cell morphology (A-E) (H&E staining pathological section) of model mice;

[0034] Figure 17 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on myocardial cell fibrosis (A-E) (Masson staining pathological section) of model mice;

[0035] Figure 18 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on brain Nissl body injury (A-E) (Nissl staining pathological section) of model mice;

[0036] Figure 19 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on brain inflammatory factors IL-1β (A) and TNF-α (B) levels of model mice;

[0037] Figure 20 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on intestinal flora (A-C) of model mice;

[0038] Figure 21 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on intestinal flora functional clustering of model mice;

[0039] Figure 22 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on liver bile acid level (A) and bile acid synthesis (B) of model mice;

[0040] Figure 23 Figure for the effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation combined with medicinal and edible mixture composition on ileum (A) and serum (B) bile acid levels of model mice;

[0041] Figure 24 The combination of the long Bifidobacterium infantis NKUFB3-14 probiotic preparation and the medicinal and edible mixture composition before and after the main components are deleted on the percentage of the binding rate of taurine cholate (A) and glycine cholate (B) is shown in the result chart;

[0042] Figure 25 The combination of different long Bifidobacterium infantis probiotic preparations and medicinal and edible mixture compositions on the percentage of the binding rate of taurine cholate (A) and glycine cholate (B) is shown in the result chart. DETAILED DESCRIPTION

[0043] The present application provides a long Bifidobacterium infantis probiotic preparation and medicinal and edible mixture composition, including long Bifidobacterium infantis NKUFB3-14 probiotic preparation and medicinal and edible mixture; the preservation number of the long Bifidobacterium infantis NKUFB3-14 is CGMCC No. 25762; the long Bifidobacterium infantis NKUFB3-14 probiotic preparation includes heat-inactivated bacteria liquid of long Bifidobacterium infantis NKUFB3-14, heat-inactivated bacteria of bacteria liquid or heat-inactivated supernatant of bacteria liquid; the concentration of long Bifidobacterium infantis NKUFB3-14 in the bacteria liquid of long Bifidobacterium infantis NKUFB3-14 is 1x10 8 ~ 3x10 9 CFU / mL; according to mass parts, the medicinal and edible mixture includes the following raw materials: 10-20 parts of ginseng, 10-20 parts of longan meat, 10-20 parts of poria cocos, 10-20 parts of lotus leaf, 5-15 parts of yam, 5-15 parts of coix seed, 5-15 parts of polygonatum, 5-15 parts of wheat, 5-15 parts of orange peel, 5-15 parts of white kidney bean and 4-6 parts of hawthorn.

[0044] In the specific implementation process of the present application, the composition is composed of long Bifidobacterium infantis NKUFB3-14 probiotic preparation and medicinal and edible mixture.

[0045] In the present application, the long Bifidobacterium infantis NKUFB3-14 has been disclosed in Chinese patent CN116286551A, and it is proved by experiments that the long Bifidobacterium infantis NKUFB3-14 is a safe strain and can be used for human and animal bodies.

[0046] In the implementation of the present application, the probiotic preparation of B. longum subsp. infantis NKU FB3-14 includes heat-inactivated material of bacterial bodies in the bacterial liquid of B. longum subsp. infantis NKU FB3-14, which can better control weight gain and more effectively regulate glycolipid-related indicators in serum; the heat-inactivated material is further vacuum freeze-dried; the vacuum degree of the vacuum freeze-drying is 10-100 Pa; the temperature of the vacuum freeze-drying is -40- -20 ℃; the time of the vacuum freeze-drying is 10-14 h; the probiotic preparation is a solid preparation; and the water content of the probiotic preparation is ≤5%.

[0047] In one embodiment of the present application, the concentration of B. longum subsp. infantis NKU FB3-14 in the bacterial liquid of B. longum subsp. infantis NKU FB3-14 is 2×10 9 -3×10 9 CFU / mL.

[0048] In another embodiment of the present application, the concentration of B. longum subsp. infantis NKU FB3-14 in the bacterial liquid of B. longum subsp. infantis NKU FB3-14 is 1×10 9 -1.5×10 9 CFU / mL.

[0049] In the implementation of the present application, the preparation method of the bacterial liquid of B. longum subsp. infantis NKU FB3-14 includes the following steps: inoculating B. longum subsp. infantis NKU FB3-14 into BS liquid medium, carrying out anaerobic culture in a constant-temperature incubator after inoculation, and when the OD value of the culture liquid is 1.5, the concentration of the bacterial liquid is close to 1×10 10 CFU / mL, and adjusting the concentration of the bacterial liquid to 1×10 8 -3×10 9 CFU / mL using BS liquid medium; the inoculation amount of B. longum subsp. infantis NKU FB3-14 in the BS liquid medium is 5% in volume fraction; the temperature of the anaerobic culture is 37 ℃; and the time of the anaerobic culture is 20-24 h.

[0050] After obtaining the bacterial liquid, the present application carries out centrifugal solid-liquid separation on the bacterial liquid to obtain B. longum subsp. infantis NKU FB3-14 supernatant (live) and B. longum subsp. infantis NKU FB3-14 bacterial bodies (live).

[0051] In one embodiment of the present application, the bacterial liquid of B. longum subsp. infantis NKU FB3-14 is sequentially subjected to heat inactivation and vacuum freeze-drying to obtain heat-inactivated material of the bacterial liquid, i.e., heat-inactivated bacterial bodies + supernatant.

[0052] In another embodiment of the present application, the supernatant in the bacterial solution of Bifidobacterium longum subsp. infantis NKU FB3-14 is sequentially subjected to heat inactivation and vacuum freeze-drying to obtain heat-inactivated supernatant in the bacterial solution, i.e., heat-inactivated supernatant.

[0053] In another embodiment of the present application, the bacterial cells in the bacterial solution of Bifidobacterium longum subsp. infantis NKU FB3-14 are sequentially subjected to heat inactivation and vacuum freeze-drying to obtain heat-inactivated bacterial cells in the bacterial solution, i.e., heat-inactivated bacterial cells.

[0054] In the specific implementation of the present application, the homoeopathy mixture is composed of the following raw materials in parts by mass: 10-20 parts of ginseng, 10-20 parts of longan arillus, 10-20 parts of Poria cocos, 10-20 parts of lotus leaf, 5-15 parts of Chinese yam, 5-15 parts of coix seed, 5-15 parts of polygonatum, 5-15 parts of malt, 5-15 parts of orange peel, 5-15 parts of white lentil, and 4-6 parts of hawthorn.

[0055] As an embodiment, the mass fraction of ginseng in the composition of the present application is 10-20 parts, further 15 parts. As an embodiment, the mass fraction of longan arillus in the composition of the present application is 10-20 parts, further 15 parts. As an embodiment, the mass fraction of Poria cocos in the composition of the present application is 10-20 parts, further 15 parts. As an embodiment, the mass fraction of lotus leaf in the composition of the present application is 10-20 parts, further 15 parts. As an embodiment, the mass fraction of Chinese yam in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of coix seed in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of polygonatum in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of malt in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of orange peel in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of white lentil in the composition of the present application is 5-15 parts, further 10 parts. As an embodiment, the mass fraction of hawthorn in the composition of the present application is 4-6 parts, further 5 parts.

[0056] Compared with the use of postbiotic preparation alone, the addition of the homoeopathy mixture significantly reduces the accumulation of liver triglycerides, liver CYP7A1 enzyme level, and intestinal lipid absorption capacity caused by high-fat high-cholesterol diet, and significantly improves the serum nitric oxide (NO) level of the model mice, indicating that the homoeopathy mixture has outstanding potential in regulating liver lipid metabolism and vascular endothelial function of the body.

[0057] Among them, ginseng can enhance insulin sensitivity, regulate blood sugar, antioxidant, anti-inflammatory, improve immunity, help protect cardiovascular function; longan meat can promote blood circulation, anti-platelet aggregation, reduce blood lipids, has antioxidant and anti-inflammatory effects, protect cardiovascular health; lotus leaf can reduce blood lipids, inhibit fat absorption, promote fat decomposition, prevent obesity and atherosclerosis; Fuling can benefit water and remove dampness, regulate intestinal microenvironment, promote sugar and lipid metabolism, improve insulin resistance, and enhance immunity; yam can enhance insulin sensitivity, regulate intestinal flora, promote gastrointestinal function, and has a regulating effect on sugar and lipid metabolism; Yiyiren can reduce blood sugar, reduce blood lipids, anti-inflammatory, benefit water and remove dampness, improve lipid metabolism and prevent atherosclerosis; Yuzhu nourishes yin and moistens dryness, improves insulin resistance, regulates blood sugar, antioxidant and protects cardiovascular; Malt can promote starch digestion and absorption, regulate gastrointestinal function, and help balance sugar and lipid metabolism; orange peel can promote digestion, regulate fat metabolism, antioxidant, and protect cardiovascular health; white beans can invigorate the spleen and remove dampness, improve insulin resistance, regulate sugar and lipid metabolism, and improve energy utilization; hawthorn can reduce blood lipids, reduce blood sugar, promote gastrointestinal peristalsis, improve blood vessel elasticity, prevent hypertension and arteriosclerosis.

[0058] Regulating sugar and lipid metabolism: ginseng, yam, lotus leaf, hawthorn and white beans together promote insulin sensitivity, inhibit fat synthesis, reduce blood sugar and blood lipid levels; antioxidant and anti-inflammatory: longan meat, Fuling, Yuzhu and orange peel are rich in antioxidant substances, reduce inflammatory response and protect the cardiovascular system; promote digestion and absorption: malt, orange peel, white beans and hawthorn help improve gastrointestinal function and enhance metabolic capacity; enhance immunity and protect blood vessels: ginseng, longan meat, Fuling and Yiyiren maintain cardiovascular health by regulating immune function, anticoagulation and reducing blood lipids. These ingredients are compatible with each other, and through the regulation of intestinal microecology, balance of sugar and lipid metabolism, anti-inflammatory and antioxidant effects, and improvement of vascular function, they jointly play a role in improving metabolic syndrome, preventing diabetes and cardiovascular diseases.

[0059] The bile salt binding capacity of the combination of the medicinal and edible mixture after deleting the main components (ginseng, longan meat, Fuling, lotus leaf) and the postbiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 is significantly reduced. The bile salt binding capacity of the medicinal and edible mixture combined with other Bifidobacterium longum subsp. infantis compositions is significantly reduced. It is shown that the postbiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and the medicinal and edible composition are the best lipid-lowering active combination.

[0060] In the specific implementation process of the present application, the preparation method of the medicinal and edible mixture comprises the following steps:

[0061] After mixing the raw materials of the medicinal and edible mixture with water, soaking and decocting are carried out in turn to obtain a medicinal and edible water extract;

[0062] The homoisodiacean extract is freeze-dried and ground into powder to obtain a homoisodiacean mixture.

[0063] The homoisodiacean extract is freeze-dried and ground into powder to obtain a homoisodiacean mixture.

[0064] In the specific implementation of the present application, the mass ratio of the probiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and the homoisodiacean mixture is (2-4):1, and further 2:1, and under this ratio, the combination has the highest binding rate with glycocholate and taurocholate.

[0065] In the specific implementation of the present application, the preparation method of the composition comprises the following steps: mixing the homoisodiacean mixture and the probiotic preparation.

[0066] As an embodiment, the dosage form of the composition of the present application can be, but is not limited to, a liquid agent and / or a powder agent, such as a solid beverage, a liquid beverage, a tablet, a pill or a capsule.

[0067] In the specific implementation of the present application, the composition is applied in food.

[0068] In the specific implementation of the present application, the food includes food for improving glycolipid metabolism; and the food can be health food.

[0069] In the specific implementation of the present application, the glycolipid metabolism includes glycolipid metabolism caused by high-fat and high-cholesterol diet.

[0070] In the specific implementation of the present application, the improvement of glycolipid metabolism includes one or more of the following: reducing body weight, controlling body weight gain, reducing blood sugar content, reducing triglyceride content, reducing cholesterol content, reducing appetite, improving food energy conversion efficiency, reducing organ weight, reducing epididymal fat content, inhibiting intestinal absorption of fat, inhibiting intestinal absorption of cholesterol, reducing fat cell area, increasing leptin level in serum, protecting hepatocytes, regulating bile acid metabolism, inhibiting bile acid reabsorption and improving intestinal flora.

[0071] In the specific implementation of the present application, the organ includes liver and / or epididymis.

[0072] The present application also provides the use of the composition described in the above scheme in the preparation of a medicine for preventing, treating and / or relieving diseases; and the diseases include one or more of the following: obesity, glycolipid metabolism dysfunction, intestinal inflammation, intestinal flora imbalance (intestinal flora structure disorder), hyperlipidemia and cardiovascular and cerebrovascular diseases.

[0073] In the present application, the disease is induced by diet, further induced by high-fat high-cholesterol diet; the cardiovascular and cerebrovascular disease includes cardiovascular and cerebrovascular disease caused by glucose and lipid metabolism dysfunction; the cardiovascular and cerebrovascular disease includes but is not limited to atherosclerosis, coronary heart disease, hypertension, hyperlipidemia and metabolic syndrome.

[0074] In the implementation of the present application, the prevention, treatment and / or alleviation of the cardiovascular and cerebrovascular disease includes one or more of reducing cardiovascular risk factor level, increasing NO level in serum, reducing cardiovascular and cerebrovascular injury, alleviating vascular function impairment, promoting vasodilation and blood flow improvement, inhibiting myocardial cell fibrosis, protecting nervous system, repairing blood-brain barrier and improving brain inflammation.

[0075] In the implementation of the present application, the alleviation of vascular function impairment includes alleviation of vascular endothelial function impairment.

[0076] In the present application, the composition is used for reducing cardiac inflammatory cell infiltration and ventricular injury, improving myocardial cell fibrosis and histological injury, reducing cardiac inflammatory cell infiltration and ventricular injury, improving myocardial cell fibrosis and histological injury, reducing brain inflammation factors, improving blood-brain barrier integrity and neuronal injury.

[0077] In the present application, the intestinal flora imbalance includes high-fat high-cholesterol diet-induced intestinal flora imbalance.

[0078] The technical solutions in the present application will be described clearly and completely in combination with the embodiments in the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the present application.

[0079] The components of the culture medium used in the following embodiments are as follows:

[0080] BS solid medium: 10.0 g / L of proteose peptone, 5.0 g / L of liver extract powder, 3.0 g / L of beef extract powder, 5.0 g / L of yeast extract powder, 8.0 g / L of tryptone, 0.5 g / L of soluble starch, 1 g / L of sodium chloride, 1 g / L of potassium phosphate dibasic, 1 g / L of potassium phosphate monobasic, 10 g / L of glucose, 0.01 g / L of ferric sulfate heptahydrate, 0.005 g / L of manganese sulfate, 0.5 g / L of L-cysteine and 20 g / L of agar.

[0081] BS liquid medium: 10.0 g / L of proteose peptone, 5.0 g / L of liver extract powder, 3.0 g / L of beef extract powder, 5.0 g / L of yeast extract powder, 8.0 g / L of tryptone, 0.5 g / L of soluble starch, 1 g / L of sodium chloride, 1 g / L of potassium phosphate dibasic, 1 g / L of potassium phosphate monobasic, 10 g / L of glucose, 0.01 g / L of ferric sulfate heptahydrate, 0.005 g / L of manganese sulfate, and 0.5 g / L of L-cysteine.

[0082] In the following examples, statistical significance was determined by Student's t-test. The significance level was set as *p (or *p) <0.05, **p (or **p) <0.01, ***p (or ***p) <0.001, ****p (or ****p) <0.0001.

[0083] In the following examples, unless otherwise specified, all are conventional methods.

[0084] In the following examples, unless otherwise specified, the materials, reagents, etc. used can be obtained from commercial channels.

[0085] Test Example 1

[0086] 1. Preparation of probiotic preparations of Bifidobacterium longum subsp. infantis NKUFB 3-14 in different forms

[0087] Bifidobacterium longum subsp. infantis NKUFB 3-14 was inoculated into BS liquid medium at a volume fraction of 5%, and after inoculation, it was cultured anaerobically in a 37°C constant temperature incubator for 20-24 h, until the OD value of the culture solution was 1.5, and the bacterial solution concentration was close to 1×10 10 CFU / mL. The bacterial solution concentration was adjusted to 2-3×10 9 CFU / mL using BS liquid medium. The culture solution (bacterial solution) was centrifuged at 8000 g for 5 min to separate the supernatant and the bacterial pellet.

[0088] The bacterial solution was heated in a 85°C water bath for 15 min and then vacuum freeze-dried to obtain the probiotic preparation of Bifidobacterium longum subsp. infantis NKUFB 3-14, i.e., heat-inactivated bacterial bodies + supernatant (heat-inactivated product of the bacterial solution);

[0089] The supernatant was heated in a 85°C water bath for 15 min and then vacuum freeze-dried to obtain the probiotic preparation of Bifidobacterium longum subsp. infantis NKUFB 3-14, i.e., heat-inactivated supernatant (heat-inactivated product of the supernatant in the bacterial solution);

[0090] The bacterial pellet was heated in a 85°C water bath for 15 min and then vacuum freeze-dried to obtain the probiotic preparation of Bifidobacterium longum subsp. infantis NKUFB 3-14, i.e., heat-inactivated bacterial bodies (heat-inactivated product of the bacterial bodies in the bacterial solution).

[0091] 2. Screening of postbiotic preparation forms for efficiently improving abnormal glycolipid metabolism indicators in mice

[0092] A mouse cardiovascular and cerebrovascular injury model (hereinafter referred to as "model") was induced by a 12-week high-fat high-cholesterol diet (40 kcal% fat, 1.25 gm% cholesterol, 0.5 gm% sodium cholate), and control group (C), model group (H) and intervention group (T) were set up, wherein the intervention group (T) was divided into heat-killed bacteria group (T1), heat-killed supernatant group (T2) and heat-killed bacteria + supernatant group (T3), and the postbiotic preparation form that efficiently relieves glycolipid metabolism disorder in mice was screened through body weight and blood glycolipid indicators.

[0093] The preparation method of each component is step 1.

[0094] After 1 week of adaptive feeding, the intervention period of 12 weeks began. The control group (C) was fed with normal feed (Synergy SPF grade experimental mouse maintenance feed SWS9102, the main ingredients of which are northeast corn, wheat, imported fish meal, chicken meal, soybean meal, soybean oil, amino acids, vitamins and minerals); the model group (H) and the intervention group (T) were fed with high-fat high-cholesterol feed (formula: 40 kcal% fat, 1.25 gm% cholesterol, 0.5 gm% sodium cholate) during the intervention period. All mice were not restricted in eating and drinking during the intervention period; the heat-killed bacteria group (T1) was intragastrically administered with heat-killed bacteria, and the daily intragastric administration amount was 0.2 mL; the heat-killed supernatant group (T2) was intragastrically administered with heat-killed supernatant, and the daily intragastric administration amount was 0.2 mL; the heat-killed supernatant group (T3) was intragastrically administered with heat-killed bacteria + supernatant, and the daily intragastric administration amount was 0.2 mL; the control group (C) and the model group (H) were simultaneously intragastrically administered with an equal volume of BS liquid medium, and the intragastric administration was continuously performed for 12 weeks; the weight was measured once a week, and the intragastric administration dose was adjusted according to the weight change, as shown in Figure 1 .

[0095] On the last day of the 12-week intervention, the mouse fecal samples were collected and stored at -80°C, and then the mice were fasted for 16 h without water. After fasting, the mice were anesthetized with sodium pentobarbital (1% aqueous solution), and blood samples were collected by eyeball blood collection. After the whole blood sample was placed at room temperature for 4 h, the serum sample was collected by centrifugation at 3000 rpm and 4°C for 15 min. After blood collection, the mice were decapitated, and the jejunum, ileum, colon, liver, fat, brain and heart tissues of the mice were collected, quickly frozen in liquid nitrogen and stored at -80°C or placed in a tissue fixative for histopathological analysis. All animal experiment operations comply with the relevant provisions of the Tianjin Experimental Animal Management Regulations.

[0096] From Figure 1It can be seen that the body weight of the model group (H) mice increased significantly after 12 weeks of high-fat high-cholesterol diet induction compared with the control group (C), indicating that the model can significantly increase the degree of obesity in mice. After 4 weeks of intervention, the increase in body weight of the heat-inactivated supernatant group (T2) and the heat-inactivated bacteria group (T1) was significantly lower than that of the control group (C). Notably, the heat-inactivated bacteria group (T1) had the most prominent control over body weight increase compared to other postbiotic formulations, with a body weight increase of only 2.9 g in mice induced by 12 weeks of high-fat high-cholesterol diet. Therefore, the most effective postbiotic formulation for body weight control is heat-inactivated bacteria (T1).

[0097] By measuring the sugar lipid-related indicators in the serum of mice in different groups, it was found that the glucose, total cholesterol and triglyceride levels in the serum of the control group (C) and the intervention group (T) were significantly lower than those in the model group (H) (p<0.01) Figure 2 ). Among them, the serum triglyceride level of the T1 group was significantly lower than that of the T2 group Figure 2 .

[0098] In summary, the most effective postbiotic formulation for body weight control and serum sugar lipid-related indicator regulation in model mice is heat-inactivated bacteria (T1).

[0099] Example 1

[0100] 1. Heat-inactivated bacteria (heat-inactivated material of bacteria in bacterial solution) were prepared by the method of Test Example 1. The concentration of NKUFB 3-14 strain before heat inactivation was 1×10 9 ~ 1.5×10 9 CFU / mL.

[0101] 2. The medicinal and edible mixture is composed of the following raw materials: 15 g of ginseng, 15 g of longan meat, 15 g of lotus leaf, 15 g of hoelen, 10 g of yam, 10 g of coix seed, 10 g of polygonatum, 10 g of wheat germ, 10 g of white lentil, 10 g of orange peel and 5 g of hawthorn.

[0102] Preparation method:

[0103] After mixing the above raw materials of the medicinal and edible mixture with water, soaking and decocting were carried out in sequence to obtain a medicinal and edible water extract; the medicinal and edible water extract was concentrated under reduced pressure to a crude drug amount of 1 g / mL to obtain a medicinal and edible extract; the obtained medicinal and edible water extract was freeze-dried and ground into powder to obtain a medicinal and edible mixture, which was ready for use.

[0104] 3. The heat-inactivated bacteria obtained in step 1 and the medicinal and edible mixture obtained in step 2 were mixed in a mass ratio of 2:1 to obtain a composition.

[0105] Determination of optimal ratio of probiotic preparation of Bifidobacterium longum subsp. infantis NKU FB3-14 and medicinal and edible mixture

[0106] By determining the differences in the binding capacity of different ratios of probiotic preparation (heat-inactivated bacterial body form) and medicinal and edible mixture in the simulated intestinal environment in vitro, the lipid-lowering activity of different ratio combinations is determined, and the composition with the strongest lipid-lowering activity is screened.

[0107] 1. The heat-inactivated bacterial body and the medicinal and edible mixture were prepared according to the method of Example 1, and the heat-inactivated bacterial body and the medicinal and edible mixture were mixed in a mass ratio of 4:1, 2:1, 1:1, 1:2 and 1:4, respectively, to obtain different ratio combinations of probiotic preparation and medicinal and edible mixture.

[0108] 2. Standard curve of sodium glycocholate and sodium taurocholate was drawn. Different concentrations of sodium glycocholate standard solution and sodium taurocholate standard solution (0, 0.5, 1, 1.5, 2, 2.5, 3 μg / mL) were prepared in a stoppered test tube, 6 mL of 60% sulfuric acid solution was added, mixed well, heated at 70°C water bath for 20 min, quickly ice bath for 5 min, and the absorbance was measured at 387 nm wavelength by ultraviolet spectrophotometry. The absorbance was taken as the ordinate, and the content of cholic acid salt was taken as the abscissa, and the standard curve of sodium glycocholate and sodium taurocholate was drawn.

[0109] 3. The binding capacity in the simulated intestinal environment in vitro was determined. 3 mL of different ratio combinations of probiotic preparation and medicinal and edible mixture (4:1, 2:1, 1:1, 1:2, 1:4) and 3 mL of 10 mg / mL pepsin solution and 1 mL of 0.01 mol / L HCl solution were digested in a 37°C constant temperature oscillator for 1 h to simulate the gastric digestion process. Then, the pH value of the solution was adjusted to 6.3 with 0.1 mol / L NaOH solution, and then 4 mL of 10 mg / mL trypsin was added, and the mixture was incubated at 37°C to simulate the intestinal environment. 4 mL of 0.4 mmol / L sodium glycocholate or 0.5 mmol / L sodium taurocholate was added to each sample. After constant temperature oscillation at 37°C for 1 h, the mixture was transferred to a centrifuge tube and centrifuged at 4000 r / min for 20 min. The supernatant was determined by colorimetry at 387 nm. Each sample was determined in triplicate, and the residual sodium glycocholate and sodium taurocholate content was calculated according to the standard curve. The difference between the total amount of added sodium glycocholate or sodium taurocholate and the residual amount was calculated, and the ratio was expressed as a percentage. The formula is as follows:

[0110] Cholic acid salt binding rate = (cholic acid salt added - cholic acid salt remaining) / cholic acid salt added × 100%;

[0111] The results show that the higher the binding rate of cholate, the better the effect of reducing cholesterol. From A and B in Figure 3 , it can be known that when the ratio of probiotic preparation to food-medicine mixture is 2:1, the binding rate of probiotic preparation to glycocholate and taurocholate is the highest, which is 78% and 84% respectively. Therefore, the optimal ratio of probiotic preparation to food-medicine mixture is determined to be 2:1.

[0112] Example 3

[0113] The concentration of NKUFB 3-14 strain before heat inactivation is 2-3 x 10 9 CFU / mL, and the rest is the same as in Example 1.

[0114] Example 4 Effect of probiotic preparation and food-medicine mixture composition on improving glucose and lipid metabolism dysfunction

[0115] 1. Effect of probiotic preparation and food-medicine mixture composition of Bifidobacterium longum subsp. infantis NKUFB 3-14 on body weight, appetite, and organ weight of model mice.

[0116] A mouse cardiovascular and cerebrovascular injury model (hereinafter referred to as "model") was induced by a 12-week high-fat high-cholesterol diet (40 kcal% fat, 1.25 gm% cholesterol, 0.5 gm% sodium cholate), and control group (C), model group (H), and intervention group were set up. The intervention group was divided into: heat-inactivated bacterial group (J), food-medicine mixture group (Y), and probiotic preparation and food-medicine mixture composition group (Z).

[0117] The preparation method of each component is the same as in Example 3.

[0118] After 1 week of adaptive feeding, the mice entered the 12-week intervention period. The control group (C) was fed with normal feed; the model group (H), heat-inactivated bacterial group (J), food-medicine mixture group (Y), and probiotic preparation and food-medicine mixture composition group (Z) were fed with high-fat high-cholesterol feed (formula: 40 kcal% fat, 1.25 gm% cholesterol, 0.5 gm% sodium cholate) during the intervention period. All mice were not restricted in food and water intake during the intervention period; the heat-inactivated bacterial group (J) was given heat-inactivated bacteria by gavage, with a daily gavage amount of 0.2 mL; the food-medicine mixture group (Y) was given food-medicine mixture by gavage, with a daily gavage amount of 0.2 mL; the probiotic preparation and food-medicine mixture composition group (Z) was given the composition by gavage, with a daily gavage amount of 0.2 mL; the control group (C) and the model group (H) were simultaneously given an equal volume of BS liquid medium by gavage, and the gavage was continued for 12 weeks. The mice were weighed once a week and the gavage dose was adjusted in time according to the body weight change.

[0119] The fecal samples of mice were collected on the last day of 12-week intervention and stored at -80℃, followed by 16h fasting without water. After fasting, the mice were anesthetized with sodium pentobarbital (1% aqueous solution) and blood samples were collected by eyeball blood collection. After blood collection, the mice were decapitated, and the jejunum, ileum, colon, liver, fat, brain, and heart tissues of the mice were collected, quickly frozen in liquid nitrogen, and stored at -80℃ or placed in a tissue fixative for histopathological analysis. All animal experiments were conducted in accordance with the relevant provisions of the Tianjin Experimental Animal Management Regulations.

[0120] After 12 weeks of high-fat high-cholesterol diet induction, the body weight of the model group (H) mice was significantly higher than that of the control group (C) mice from the 4th week, and the body weight of the heat-killed bacteria group (J), the medicinal food mixture group (Y), and the probiotic preparation combined with medicinal food mixture composition group (Z) mice was significantly lower than that of the model group (H) mice from the 4th week. Figure 4 Notably, the heat-killed bacteria group (J) was superior to the medicinal food mixture group (Y) in controlling the body weight of the model mice, while the probiotic preparation combined with medicinal food mixture composition group (Z) was the most obvious in reducing the body weight of the model mice. Therefore, the probiotic preparation combined with medicinal food mixture composition can effectively alleviate the symptoms of obesity in mice.

[0121] During the 12-week intervention, the food intake of the mice was recorded every week. Food utilization refers to the ratio of the percentage change in body weight to the energy intake after a certain amount of food is ingested. Figure 5 It can be seen that the average energy intake and food utilization of the model group (H) mice were significantly higher than those of the control group (C) mice. The daily food intake and average energy intake of the heat-killed bacteria group (J) and the medicinal food mixture group (Y) had no significant difference with those of the model group (H), while the daily food intake and average energy intake of the probiotic preparation combined with medicinal food mixture composition group (Z) mice were significantly lower than those of the model group (H). In addition, the food utilization of the medicinal food mixture group (Y) mice had no significant difference with that of the model group (H), while the food utilization of the heat-killed bacteria group (J) and the probiotic preparation combined with medicinal food mixture composition group (Z) mice were significantly lower than that of the model group (H). Therefore, the probiotic preparation combined with medicinal food mixture composition can effectively reduce the appetite and food energy conversion efficiency of the model mice.

[0122] After 12 weeks of intervention, the liver tissue and epididymal fat of each sacrificed mouse were collected, and the liver weight and epididymal fat weight were recorded. From Figure 6It can be seen that the liver and epididymal fat weights of the model group (H) mice were significantly higher than those of the control group (C) mice. The liver and epididymal fat weights of the heat-killed cell group (J), the food-medicine mixture group (Y), and the postbiotic preparation and food-medicine mixture composition group (Z) mice were lower than those of the model group (H), and the epididymal fat weight of the postbiotic preparation and food-medicine mixture composition group (Z) mice was the lowest. It can be seen that the postbiotic preparation and food-medicine mixture composition can effectively reduce the organ weights of the model mice.

[0123] 2. Effect of the postbiotic preparation and food-medicine mixture composition of Bifidobacterium longum subsp. infantis NKU FB3-14 on blood glucose and lipid metabolism in model mice.

[0124] After 12 weeks of high-fat high-cholesterol diet induction, the blood glucose level of the model group (H) mice was significantly higher than that of the control group (C) mice ( Figure 7 ). The blood glucose levels of the heat-killed cell group (J), the food-medicine mixture group (Y), and the postbiotic preparation and food-medicine mixture composition group (Z) mice were significantly lower than those of the model group (H), and the blood glucose level of the postbiotic preparation and food-medicine mixture composition group (Z) mice was the lowest.

[0125] Similarly, the total cholesterol and triglyceride levels in the serum of the model group (H) mice were significantly higher than those of the control group (C) mice ( Figure 7 ). The total cholesterol and triglyceride levels in the serum of the heat-killed cell group (J) and the postbiotic preparation and food-medicine mixture composition group (Z) mice were significantly lower than those of the model group (H), and the total cholesterol and triglyceride levels in the serum of the postbiotic preparation and food-medicine mixture composition group (Z) mice were the lowest.

[0126] After 12 weeks of high-fat high-cholesterol diet induction, the HDL level in the serum of the model group (H) mice was significantly lower than that of the control group (C) mice ( Figure 7 ). The HDL levels in the serum of the heat-killed cell group (J) and the food-medicine mixture group (Y) mice were significantly higher than those of the model group (H), and there was no significant difference in the HDL levels in the serum between the postbiotic preparation and food-medicine mixture composition group (Z) and the model group (H) mice. The LDL level in the serum of the model group (H) mice was significantly higher than that of the control group (C) mice ( Figure 7 ). The LDL levels in the serum of the heat-killed cell group (J), the food-medicine mixture group (Y), and the postbiotic preparation and food-medicine mixture composition group (Z) mice were significantly lower than those of the model group (H).

[0127] In summary, the postbiotic preparation and food-medicine mixture composition can effectively reduce the glucose and lipid metabolism disorders in the serum of the model mice.

[0128] 3. Effect of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation and medicinal and edible mixture composition on intestinal fat absorption capacity and fat tissue metabolic capacity of model mice.

[0129] After 12 weeks of high-fat high-cholesterol diet induction, the triglyceride level in the jejunum tissue of the model group (H) mice was significantly higher than that of the control group (C) mice ( Figure 8 ), indicating that the intestinal lipid absorption capacity of the cardiovascular model mice induced by high-fat high-cholesterol diet was significantly enhanced. The triglyceride level in the jejunum tissue of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice was significantly lower than that of the model group (H) mice, and the triglyceride level in the jejunum tissue of the probiotic preparation and medicinal and edible mixture composition group (Z) mice was the lowest, indicating that the probiotic preparation and medicinal and edible mixture composition could effectively inhibit the absorption of fat and cholesterol in the diet by the intestinal tissue of the mice, thereby reducing the accumulation of lipids in the mice.

[0130] The pathological section staining results showed that compared with the control group (C) mice, the fat cell volume of the model group (H) mice increased, showing large vacuoles, and the size of the vacuoles was uneven, and a large number of inflammatory cells in the interstitial fat infiltrated in the form of a corolla structure ( Figure 9 indicated by B black arrows), while the fat cells of the control group (C), the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice were arranged closely, with relatively small volume, clear boundary and uniform morphology, and no inflammatory cell infiltration was observed ( Figure 9 A, C, D, and E in the figure). By statistically analyzing the epididymal fat cell area of mice in different groups, it was found that the fat cell area of the model group (H) mice was significantly higher than that of the control group (C), and the fat cell area of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice was significantly smaller than that of the model group (H) mice ( Figure 9 F in the figure).

[0131] After 12 weeks of high-fat high-cholesterol diet induction, the leptin level in the serum of the model group (H) mice was significantly higher than that of the control group (C) mice ( Figure 10 ), indicating that the increase in fat tissue of the cardiovascular injury mice induced by high-fat high-cholesterol diet led to an increase in the secretion of leptin. The leptin level in the serum of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice was significantly lower than that of the model group (H) mice, and the leptin level in the serum of the probiotic preparation and medicinal and edible mixture composition group (Z) mice was the lowest.

[0132] 4. Effects of Bifidobacterium longum subsp. infantis NKU FB3-14 probiotic preparation and medicinal and edible mixture composition on liver lipid deposition and liver function damage in model mice.

[0133] The pathological section staining results showed that after 12 weeks of high-fat high-cholesterol diet induction, compared with the control group (C) mice, the model group (H) mice showed obvious fatty degeneration in the liver HE results, and the fat vacuoles in the liver cells increased, and the cell edge showed vacuolar changes. The liver tissue structure of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice was regular, the liver cells were arranged in order, the cell nucleus size was uniform, and there was no obvious fatty degeneration or inflammatory reaction. Figure 11

[0134] The pathological section staining results showed that after 12 weeks of high-fat high-cholesterol diet induction, compared with the control group (C) mice, the model group (H) mice showed bright red staining after oil red staining, which reflected that there was a large amount of fat accumulation in the liver, especially the increase of fat vacuoles in the liver cells. The liver sections of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the probiotic preparation and medicinal and edible mixture composition group (Z) mice had no obvious fat staining, indicating that there was almost no neutral fat deposition in the liver cells. Figure 12

[0135] After 12 weeks of high-fat high-cholesterol diet induction, the ALT and AST levels in the liver of the model group (H) mice were significantly higher than those in the control group (C) mice Figure 13 ), indicating that the high-fat high-cholesterol diet induced liver cell damage. Among them, the ALT level in the liver of the probiotic preparation and medicinal and edible mixture composition group (Z) mice was significantly lower than that in the model group (H) mice, while the other intervention groups had no significant difference with the model group (H) mice. The AST level in the liver of the heat-killed bacteria group (J) and the probiotic preparation and medicinal and edible mixture composition group (Z) mice was significantly lower than that in the model group (H) mice, while the medicinal and edible mixture group (Y) mice had no significant difference with the model group (H) mice. Therefore, compared with other intervention groups, the probiotic preparation and medicinal and edible mixture composition group can effectively protect the integrity of liver cells and prevent liver cell damage induced by high-fat high-cholesterol diet.

[0136] 5. Effects of probiotic preparation and medicinal and edible mixture composition on cardiovascular risk factors and vascular function damage.

[0137] ​​Cardiovascular risk factors (TG / HDL, HDL / LDL, TC / HDL ratio) are a common indicator for assessing metabolic syndrome, diabetes, and cardiovascular disease. High TG / HDL ratio is considered an independent risk factor for atherosclerosis and coronary heart disease, and a higher TG / HDL ratio is usually associated with an increased cardiovascular risk.

[0138] The results show that the TG / HDL, HDL / LDL, and TC / HDL ratios of mice in the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the composition of the postbiotic preparation combined with the medicinal and edible mixture group (Z) are significantly lower than those in the model group (H), and the TG / HDL ratio of mice in the heat-killed bacteria group (J) is the lowest ( Figure 14 ), indicating that the postbiotic preparation, the medicinal and edible mixture, and the composition of the two can significantly reduce the cardiovascular risk factors of model mice.

[0139] Nitric oxide (NO) is an important signaling molecule, especially in vascular endothelial function and metabolic regulation. The results show that the serum NO level of mice in the high-fat high-cholesterol diet-induced cardiovascular and cerebrovascular injury model group (H) is significantly lower than that in the control group (C). High-fat high-cholesterol diet can cause endothelial cell dysfunction, leading to reduced activity of endothelial nitric oxide synthase (eNOS) and thus reduced NO production, impaired vascular endothelial function, and homeostasis imbalance. Compared with the model group, the serum NO level of mice in the medicinal and edible mixture group (Y) and the composition of the postbiotic preparation combined with the medicinal and edible mixture group (Z) is increased, and is extremely significantly higher than that in the model group, and the serum NO level of mice in the composition of the postbiotic preparation combined with the medicinal and edible mixture group is the highest ( Figure 15 ), indicating that the combination of postbiotics and medicinal and edible mixture helps to alleviate the impaired vascular function induced by high-fat high-cholesterol diet, promote vasodilation and blood flow improvement, and has great potential for protecting cardiovascular health.

[0140] 6. Effect of the composition of the postbiotic preparation combined with the medicinal and edible mixture on myocardial tissue damage and fibrosis.

[0141] The results of pathological section staining show that after 12 weeks of high-fat high-cholesterol diet induction, the myocardial cell nuclei of mice in the control group (C) are blue, the myocardial cells are arranged in order, the muscle bundles are full, and the cytoplasm is bright red, while the myocardium of mice in the model group (H) is disordered, with inflammatory cell infiltration and ventricular damage. Compared with the model group, the myocardial cells of mice in the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the composition of the postbiotic preparation combined with the medicinal and edible mixture group (Z) appear normal, the inflammatory cell infiltration is significantly reduced, and the myocardial cells are arranged in order ( Figure 16 ).

[0142] Myocardial fibrosis is usually manifested as blue collagen fiber proliferation, mainly in the cardiac interstitium or subendocardial layer. The pathological section staining results show that the heart of the model group (H) mice induced by high-fat high-cholesterol diet shows more obvious collagen fiber deposition under Masson staining. Compared with the control group (C) mice, the heart of the model group (H) mice has a more obvious degree of fibrosis, which may further affect the systolic function of the heart. Compared with the model group, the degree of myocardial cell blue fibrosis of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the composition of the probiotic preparation and the medicinal and edible mixture group (Z) mice is reduced Figure 17

[0143] 7. The effect of the composition of the probiotic preparation and the medicinal and edible mixture on the blood-brain barrier and neuronal damage.

[0144] Nissl staining is mainly used to observe the cell body of nerve cells, especially the distribution of rough endoplasmic reticulum (i.e. Nissl body), which can reveal the degree of neuronal damage in model mice and evaluate the specific effects of high-fat high-cholesterol diet on the health of the nervous system. After 12 weeks of high-fat high-cholesterol diet induction, the Nissl body of the model group (H) mice significantly decreased and the morphology changed, showing swelling or irregular structure of the rough endoplasmic reticulum Figure 18 , indicating that high-fat high-cholesterol diet induced neuronal damage and degeneration in the brain of mice. Notably, the degree of damage to the Nissl body of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the composition of the probiotic preparation and the medicinal and edible mixture group (Z) mice was improved, and the number of Nissl bodies of the mice in the composition of the probiotic preparation and the medicinal and edible mixture group (Z) significantly increased, and the morphology returned to normal Figure 18 , indicating that the probiotic and the composition of the medicinal and edible mixture have an important positive effect on the health of the nervous system of model mice.

[0145] After 12 weeks of high-fat high-cholesterol diet induction, the IL-1β and TNF-α levels in the brain of the model group (H) mice were significantly higher than those in the control group (C) mice, indicating that high-fat high-cholesterol diet induced systemic inflammatory response in mice, and this inflammatory response may affect the central nervous system. Long-term high-fat high-cholesterol diet may also lead to a decrease in the function of the blood-brain barrier, which makes it easier for peripheral immune factors (such as IL-1β and TNF-α) to enter the brain, thereby exacerbating local inflammation.

[0146] Notably, the IL-1β and TNF-α levels in the brain of the heat-killed bacteria group (J), the medicinal and edible mixture group (Y), and the composition of the probiotic preparation and the medicinal and edible mixture group (Z) mice were significantly lower than those in the model group (H) mice Figure 19 ​), indicating that the probiotics and the mixture of food-herbs combination have a significant effect on repairing the blood-brain barrier and reducing the inflammation level of the model mice.

[0147] 8. The effect of probiotics and the mixture of food-herbs combination on the intestinal flora disorder of the model mice.

[0148] 16S rRNA gene microbial sequencing: The total DNA in the feces was extracted using QIAamp DNA Stool minikit kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions. The total DNA content was normalized to 1 ng / μL. The V3-V4 of 16S rRNA was amplified, and the amplification primer was a universal primer. The upstream primer was 341F (5'-CCTACGGGNGGCWGCAG-3', SEQ ID NO. 1), and the downstream primer was 802R (5'-TACNVGGGTATCTAATCC-3', SEQ ID NO. 2), wherein N, W, and V used IUPAC (International Union of Pure and Applied Chemistry) nucleotide code to represent the polymorphism of the base; N represents any base (A / T / C / G), which is suitable for the case where there may be a variation at this site; W represents A or T, which ensures the adaptation of the 16S rRNA sequence of different bacteria; V represents A, C, or G; the PCR reaction system is as follows: 10 ng of pure DNA, 15 μL of high-fidelity DNA polymerase and PCR premix, 200 nmol / L of upstream and downstream primers, nuclease-free water, and the final volume is 30 μL. The PCR cycle conditions include initial denaturation at 98°C for 1 min, 30 cycles of 98°C for 10 s, 50°C for 30 s, and 72°C for 5 min. The PCR product was quantified, mixed in equimolar proportion, then purified and sequenced using the Illumina MeSeq platform, and the sequencing depth was at least 20,000 sequences per sample

[0149] QIIME analysis software used FLASH procedure to merge sequences. Paired-end added sequences were merged into one operational taxonomic unit (OTUs) at 97% sequence similarity using GreenGenes dataset and UPARSE algorithm. Representative sequences of each OTU were aligned, and then RDP classifier was used to annotate taxonomic information of each representative sequence. Analysis included each taxonomic level (e.g. phylum, class, order, family, genus, etc.). In-house Perl scripts were used to analyze intra-sample alpha diversity and inter-sample beta diversity. Principal coordinate analysis (PCoA) was used to assess differences between experimental samples using Unweighted unifrac method. All samples were used to calculate alpha diversity including richness and diversity. Taxonomic information of 16S rRNA gene sequence information was analyzed using UCLUST version 1.2.22 against Silva119 16S rRNA dataset using 90% confidence interval.

[0150] By analyzing the microbial community structure of the intestines of different groups of mice, as shown in Figure 20 Figure 2, after 12 weeks of high-fat high-cholesterol diet induction, the relative abundance of Escherichia-Shigella and Parasutterella in the intestines of model group (H) mice increased significantly Figure 20 The increase of Escherichia-Shigella bacteria, especially in the case of glycolipid metabolism disorder, is related to the inflammatory response of the intestinal tract, can trigger the inflammatory response of the intestinal mucosa, and will further affect the intestinal barrier function and metabolic health of the host. The increase of Parasutterella abundance is related to the metabolic health of the host, and excessive consumption of high-fat, ultra-processed food will lead to an increase in the abundance of Parasutterella. The relative abundance of Escherichia-Shigella and Parasutterella in the intestines of heat-killed bacteria group (J), medicinal and edible mixture group (Y) and probiotics preparation combined with medicinal and edible mixture composition group (Z) mice decreased significantly, indicating that probiotics and medicinal and edible mixture have potential relieving effect on intestinal inflammation and dysbiosis induced by high-fat high-cholesterol diet.

[0151] Notably, the relative abundance of Clostridium sensu stricto 1, a bile acid-regulated anaerobic bacteria present in the intestine, which can affect the host's bile acid levels, intestinal health and metabolic function by changing the composition and metabolic pathways of bile acids in the intestine, was significantly lower in the model group (H) mice than in the control group (C) mice. The relative abundance of Clostridium sensu stricto 1 in the intestines of mice in the heat-killed bacteria group (J), the mixture of food and medicine group (Y), and the probiotic preparation combined with the mixture of food and medicine group (Z) was significantly increased, suggesting that the probiotic preparation and the mixture of food and medicine may regulate the glucose and lipid metabolism of model mice through bile acid metabolism.

[0152] PICRUSt analysis was used to predict microbial functions, and the abundance of annotated OTUs was used to identify high-representative flora in different treatment groups using the LEfSe online tool. Further study of the functional clustering differences of intestinal flora in different groups of mice showed that Figure 21 As shown in the figure, after 12 weeks of high-fat high-cholesterol diet induction, the organization signal transduction pathway, OmpR family response regulator, XerD-specific recombinase, bacterial RNA polymerase, glycosyltransferase, sodium ion-driven multi-drug efflux pump, ABC-type multi-drug transport system, AraC-type DNA binding domain, and AcrR family DNA binding transcriptional regulator pathway of the intestinal flora of the model group (H) mice were significantly down-regulated. These functional prediction results show that certain pathways in the intestinal flora have changed under the influence of high-fat high-cholesterol diet, especially the pathways related to anti-pathogenic bacterial infection, stress response, cell wall synthesis, signal transduction, and transcriptional regulation.

[0153] Notably, the expression of these pathways in the intestines of mice in the heat-killed bacteria group (J) and the probiotic preparation combined with the mixture of food and medicine group (Z) increased, indicating that the probiotic preparation and the probiotic combined with the mixture of food and medicine can enhance the adaptability of intestinal flora in glucose and lipid metabolism disorders, chronic inflammation, or in response to drug stress, further suggesting that the probiotic preparation and the probiotic combined with the mixture of food and medicine may play an important role in anti-infection, environmental adaptation, and interaction with the host.

[0154] 9. The probiotic preparation combined with the mixture of food and medicine regulates the glucose and lipid metabolism of model mice by inhibiting bile acid reabsorption.

[0155] Analysis of gut microbiota sequencing results revealed a significant increase in the abundance of bile acid-associated bacteria. Therefore, this study further validated whether the combination of postbiotics and a food-medicine homologous mixture plays a role in improving hyperlipidemia and its induced cognitive impairment in mice with metabolic dysfunction by regulating this key pathway of bile acid metabolism, through measuring bile acid levels in the liver, ileum, and serum of different groups of mice.

[0156] After 12 weeks of induction with a high-fat, high-cholesterol diet, the total bile acid level in the liver of the model group (H) mice was significantly higher than that of the control group (C) mice, indicating that the high-fat, high-cholesterol diet induced increased bile acid synthesis in the liver of mice. The total bile acid levels in the liver tissue of mice in the heat-inactivated bacterial culture group (J), the food-medicine homology mixture group (Y), and the combination group of post-biotic preparation and food-medicine homology mixture (Z) were all significantly lower than those in the model group (H) mice. Among them, the total bile acid level in the liver of mice in the combination group of post-biotic preparation and food-medicine homology mixture (Z) was the lowest.

[0157] The CYP7A1 enzyme level in the liver of model group (H) mice was significantly higher than that in control group (C) mice, indicating that a high-fat, high-cholesterol diet induced increased bile acid synthesis in the liver of mice. There were no significant differences between the heat-inactivated bacterial group (J), the food-medicine mixture group (Y), and the model group (H) mice. However, the CYP7A1 enzyme level in the liver of mice in the combination of postbiotic preparation and food-medicine mixture (Z) mice was significantly lower than that in the model group (H) mice. Figure 22 ).

[0158] The results showed that the total bile acid level in the ileum tissue of the model group (H) mice was significantly higher than that of the control group (C) mice, indicating that a high-fat, high-cholesterol diet induced an enhanced reabsorption capacity of bile acids from the blood in the ileum of mice. The total bile acid levels in the ileum tissue of mice in the heat-inactivated bacterial culture group (J), the food-medicine mixture group (Y), and the combination group of post-biotic preparation and food-medicine mixture (Z) mice were all significantly lower than those in the model group (H) mice. Among them, the total bile acid level in the ileum tissue of mice in the combination group of post-biotic preparation and food-medicine mixture (Z) mice was the lowest. Figure 23 ).

[0159] The serum total bile acid level in the model group (H) mice was significantly higher than that in the control group (C) mice, indicating that a high-fat, high-cholesterol diet induces increased bile acid synthesis in the liver of mice, which then enters the bloodstream. There was no significant difference between the food-medicine mixture group (Y) and the model group (H) mice, while the serum total bile acid level in the heat-inactivated bacterial culture group (J) and the combination group of post-biotic preparation and food-medicine mixture (Z) mice was significantly lower than that in the model group (H) mice.

[0160] In summary, the combination of postbiotic preparations and food-medicine homologous mixtures regulates glucose and lipid metabolism in model mice by inhibiting bile acid reabsorption.

[0161] The main components in the raw materials of the medicinal and edible mixture were deleted in Comparative Example 1, which had a significant influence on the lipid-lowering activity

[0162] The influence of the main components on the lipid-lowering activity was determined by measuring the difference in the binding capacity to cholate in the simulated intestinal environment in vitro of the probiotic preparation (in the form of heat-killed bacteria) and the medicinal and edible mixture before and after the deletion of the main components (ginseng, longan arillus, lotus leaf, and Poria cocos).

[0163] 1. The heat-killed bacteria, the medicinal and edible mixture, and the medicinal and edible mixture after the deletion of the main components were prepared according to the method of Reference Example 1, and the heat-killed bacteria and the two medicinal and edible mixtures were mixed in a mass ratio of 2:1 to obtain different probiotic preparations and medicinal and edible mixture compositions.

[0164] 2. The standard curve of sodium glycocholate and sodium taurocholate was drawn, and the binding capacity to cholate in the simulated intestinal environment in vitro was determined according to Reference Example 2.

[0165] The results showed that: Figure 24 As shown in A and B in Table 1, after the deletion of the main components, the binding capacity to cholate of the probiotic preparation and the medicinal and edible mixture composition decreased significantly, indicating that the main components played an important role in the lipid-lowering activity.

[0166] In Comparative Example 2, other probiotic preparations of Bifidobacterium longum subsp. infantis were selected, and the combination of the probiotic preparations and the above-mentioned medicinal and edible raw material components had a significant influence on the lipid-lowering activity

[0167] The influence on the lipid-lowering activity was determined by measuring the difference in the binding capacity to cholate in the simulated intestinal environment in vitro of the probiotic preparations (in the form of heat-killed bacteria) of other Bifidobacterium longum subsp. infantis (ATCC15697, BI1-4, BI1-9, and BI3-5) and the medicinal and edible mixture. The ATCC15697 is a standard strain, and the other three strains of Bifidobacterium longum subsp. infantis are laboratory strains isolated and preserved by the applicant.

[0168] 1. The heat-killed bacteria and the medicinal and edible mixture were prepared according to the method of Reference Example 1, and the heat-killed bacteria and the two medicinal and edible mixtures were mixed in a mass ratio of 2:1 to obtain different probiotic preparations and medicinal and edible mixture compositions.

[0169] 2. The standard curve of sodium glycocholate and sodium taurocholate was drawn, and the binding capacity to cholate in the simulated intestinal environment in vitro was determined according to Reference Example 2.

[0170] The results showed that: Figure 25A and B in the results showed that the bile salt binding capacity of different B. longum subsp. infantis combined with the medicinal and edible components had significant differences, and only NKU FB3-14 combined with the medicinal and edible components had the strongest lipid-lowering activity.

[0171] Although the above embodiments have made a detailed description of the present application, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiments without creativity, which all belong to the protection scope of the present application.

Claims

1. A composition of a metabiotic preparation of *Bifidobacterium longum* subsp. infantis and a mixture of food and medicine homologous substances, characterized in that, Including Bifidobacterium longum infantis subspecies ( Bifidobacterium longum subsp. Infantis The metabiotic preparation and food-medicine homologous mixture of Bifidobacterium longum subsp. infantis NKU FB3-14; the mass ratio of the metabiotic preparation and food-medicine homologous mixture of Bifidobacterium longum subsp. infantis NKU FB3-14 is (2~4):1; The preservation number of *Bifidobacterium longum* subsp. infantis NKU FB3-14 is CGMCC No. 25762; the postbiotic preparation of *Bifidobacterium longum* subsp. infantis NKU FB3-14 is a heat-inactivated product of the bacterial culture of *Bifidobacterium longum* subsp. infantis NKU FB3-14; the heat-inactivated product is further subjected to vacuum freeze-drying; the concentration of *Bifidobacterium longum* subsp. infantis NKU FB3-14 in the bacterial culture is 1×10⁻⁶. 8 ~ 3×10 9 CFU / mL; According to the mass fraction, the medicinal and edible homologous mixture is composed of the following raw materials: 10-20 parts ginseng, 10-20 parts longan pulp, 10-20 parts poria cocos, 10-20 parts lotus leaf, 5-15 parts yam, 5-15 parts coix seed, 5-15 parts polygonatum odoratum, 5-15 parts malt, 5-15 parts tangerine peel, 5-15 parts white hyacinth bean, and 4-6 parts hawthorn.

2. The composition according to claim 1, characterized in that, The concentration of *Bifidobacterium longum* subsp. infantis NKU FB3-14 in the bacterial culture was 2 × 10⁻⁶. 9 ~ 3×10 9 CFU / mL.

3. The use of the composition according to claim 1 or 2 in the preparation of health foods that help control body fat, help maintain healthy blood sugar levels, help maintain healthy triglyceride levels, help maintain healthy cholesterol levels, and help regulate gut microbiota.

4. The use of the composition according to claim 1 or 2 in the preparation of a medicament for the prevention, treatment and / or relief of a disease; wherein the disease is one or more of obesity, glucose and lipid metabolism disorders, intestinal inflammation, intestinal flora imbalance and cardiovascular and cerebrovascular diseases; wherein the intestinal inflammation, intestinal flora imbalance and cardiovascular and cerebrovascular diseases are intestinal inflammation, intestinal flora imbalance and cardiovascular and cerebrovascular diseases induced by a high-fat and high-cholesterol diet.

5. The application according to claim 4, characterized in that, Prevention, treatment, and / or relief of cardiovascular and cerebrovascular diseases include one or more of the following: reducing cardiovascular risk factor levels, increasing serum NO levels, reducing cardiovascular and cerebrovascular damage, alleviating impaired vascular function, promoting vasodilation and improving blood flow, inhibiting myocardial cell fibrosis, protecting the nervous system, repairing the blood-brain barrier, and improving brain inflammation.

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