CN-121975664-A - Application of composite probiotic preparation and metagen thereof in reducing neurodegenerative diseases, resisting oxidization and preventing Alzheimer's disease

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a composite probiotic preparation and application of a metagen thereof in reducing neurodegenerative diseases, resisting oxidization and preventing Alzheimer's disease. The invention provides a mixed probiotic preparation which is composed of lactobacillus rhamnosus NKU ML1-2, bifidobacterium breve NKU BB1-2 and bifidobacterium longum subspecies infantis NKU FB 3-14 according to a specific proportion, and further researches the metazoan form of the mixed probiotic preparation. The invention systematically evaluates the efficacy and potential mechanism of the mixed probiotic preparation and its metazoan in preventing AD through in vitro antioxidant screening, in vitro AD cell model verification, metabonomics and molecular docking analysis, and provides a safe, efficient and definite-acting microorganism solution for preventing Alzheimer's disease.

Inventors

• LIU JIANBIN
• SUN YI
• LIU YUAN

Assignees

• 天赋能健康产业集团股份有限公司
• 南开大学

Dates

Publication Date

20260505

Application Date

20260407

Claims (9)

1. 1. A method of preparing a mixed probiotic formulation comprising the steps of: S1, inoculating lactobacillus rhamnosus NKU ML1-2 into an MRS liquid culture medium, inoculating bifidobacterium longum subspecies NKU FB 3-14 into a BS liquid culture medium, inoculating bifidobacterium breve NKU BB1-2 into a BBL liquid culture medium, and culturing at a constant temperature of 37 ℃ under anaerobic conditions for 20-24 h respectively; S2, centrifuging the cultured lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies NKU FB3-14 and bifidobacterium breve NKU BB1-2, discarding the supernatant, mixing and inoculating the mixture in BBL culture medium according to a certain proportion, and adding a growth auxiliary agent to culture the mixture under an anaerobic environment for 24-48 h to obtain a mixed probiotic preparation; the total number of viable bacteria in the mixed probiotic preparation is not less than 1×10 8 CFU/mL.
2. 2. The preparation method according to claim 1, wherein the lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies infantis NKU FB 3-14 and bifidobacterium breve NKU BB1-2 in step S1 are inoculated in the medium in an inoculum size of 5% by volume.
3. 3. The preparation method according to claim 1, characterized in that the mixed inoculation ratio of lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies infancy NKU FB 3-14 and bifidobacterium breve NKU BB1-2 in step S2 is 1:3:1.
4. 4. The preparation method according to claim 1, wherein the concentration of the growth aid in the BBL medium in the step S2 is not less than 1%, and the growth aid is one or more of glucose, fructo-oligosaccharides, galacto-oligosaccharides and inulin.
5. 5. The mixed probiotic metazoan is characterized in that the mixed probiotic metazoan is prepared by culturing, heat-inactivating and drying the mixed probiotic preparation prepared by the preparation method of any one of claims 1-4.
6. 6. Use of a mixed probiotic preparation prepared by the preparation method of any one of claims 1 to 4 or a mixed probiotic metagen according to claim 5 in the preparation of an antioxidant product.
7. 7. Use of a mixed probiotic preparation prepared by a preparation method according to any one of claims 1-4 or a mixed probiotic metagen according to claim 5 for the preparation of a product for inhibiting a neuronal inflammatory response or inhibiting the activity of the key enzyme BACE1 for the production of beta-amyloid.
8. 8. Use of a mixed probiotic preparation prepared by the preparation method of any one of claims 1 to 4 or a mixed probiotic metagen according to claim 5 in the preparation of a product for preventing alzheimer's disease.
9. 9. The use according to claim 8, wherein the mixed probiotic preparation or mixed prebiotic is capable of producing a differential metabolite having high binding affinity to a critical target for alzheimer's disease; the key targets are glycogen synthase kinase-3 beta and beta-secretase; The metabolites include one or more of acetylsparteine, manzamine F, sesbanamide A, taurocholate and tryptamine.

Description

Application of composite probiotic preparation and metagen thereof in reducing neurodegenerative diseases, resisting oxidization and preventing Alzheimer's disease Technical Field The invention belongs to the technical field of microorganisms, and particularly relates to a composite probiotic preparation and application of a metagen thereof in reducing neurodegenerative diseases, resisting oxidization and preventing Alzheimer's disease. Background Alzheimer's Disease (AD) is a progressive neurodegenerative disease whose major pathological features include senile plaques formed by extracellular β -amyloid (Abeta) deposition, neurofibrillary tangles formed by intracellular hyperphosphorylation of tau protein, and concomitant synaptic loss, neuronal death and chronic neuroinflammation. Currently, the number of global AD patients continues to grow, placing a heavy burden on society and individuals, however, effective radical drugs are still lacking. In recent years, the intestine-Brain Axis (Gut-Brain Axis) theory provides a new perspective for the prevention and treatment of AD. The intestinal flora communicates bi-directionally with the central nervous system via various pathways such as immunity, neuroendocrine and vagus nerves. It was found that AD patients are often accompanied by a dysregulation of intestinal flora, which may exacerbate neuroinflammation, oxidative stress and aβ deposition. Thus, the regulation of intestinal micro-ecology by probiotics, prebiotics or metazoans has become a potential strategy for the prevention and intervention of AD. Probiotics are living microorganisms that when ingested in sufficient amounts are beneficial to host health. However, viable bacteria formulations have problems with stability, gastric acid bile tolerance, and potential risk of infection (for the particular population). Metazoan (Postbiotics) refers to preparations of inanimate microorganisms and/or their components that are beneficial to host health, including bacterial components (e.g., peptidoglycans, surface proteins) and metabolites (e.g., short chain fatty acids, bacteriocins, vitamins, etc.). The metazoan has the advantages of high safety, good stability, definite action target point and the like, and the problem of bacterial survival is not required to be considered. At present, probiotics or metazoans for AD are concentrated on single strains or simple mixtures, and there is a lack of synergistic strain combination optimization research for multi-target pathological mechanisms of AD, and there is also little systematic difference in neuroprotective efficacy between different metazoan forms (such as supernatant, thallus). Therefore, there is a need to develop a safe, efficient, microbial solution with a defined target for preventing alzheimer's disease that maximizes the antioxidant and beneficial metabolite production capacity of the probiotic system by optimizing the culture conditions and formulation. Disclosure of Invention The invention aims to provide a mixed probiotic preparation and application of a metagen thereof in reducing neurodegenerative disease risk factors or preventing Alzheimer's disease, the mixed probiotic preparation provided by the invention is safe and efficient, has a definite action target point, and can effectively prevent Alzheimer's disease. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides a mixed probiotic preparation, which comprises lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies infancy NKU FB 3-14 and bifidobacterium breve NKU BB1-2; the preservation number of the lactobacillus rhamnosus NKU ML1-2 is GDMCC No.66837, the preservation number of the bifidobacterium breve NKU BB1-2 is GDMCC No.65956, and the preservation number of the bifidobacterium longum subspecies infancy NKU FB3-14 is CGMCC No.25762. Preferably, the total number of viable bacteria in the mixed probiotic preparation is not less than 1×10 8 CFU/mL; Wherein, the mixing ratio of lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies infancy NKU FB 3-14 and bifidobacterium breve NKU BB1-2 in the mixed probiotic preparation is 1:3:1. The invention also provides a preparation method of the mixed probiotic preparation, which comprises the following steps: s1, inoculating lactobacillus rhamnosus NKU ML1-2 into an MRS liquid culture medium, inoculating bifidobacterium longum subspecies NKU FB3-14 into a BS liquid culture medium, inoculating bifidobacterium breve NKU BB1-2 into a BBL liquid culture medium, and culturing for 20-24 hours under the conditions of constant temperature of 37 ℃ and anaerobic condition; S2, centrifuging the cultured lactobacillus rhamnosus NKU ML1-2, bifidobacterium longum subspecies NKU FB3-14 and bifidobacterium breve NKU BB1-2, discarding the supernatant, mixing and inoculating the mixture in BBL culture medium according to a certain proportion, and adding a growth auxili