Yak yogurt-derived pediococcus acidilactici with neuroprotection and anti-dementia functions as well as metagen and application of yak yogurt-derived pediococcus acidilactici

Abstract

The invention belongs to the technical field of biology, and discloses a yak yogurt-derived pediococcus acidilactici strain with neuroprotection and anti-dementia functions as well as a metagen and application of the yak yogurt-derived pediococcus acidilactici strain, the name of the yak yogurt-derived pediococcus acidilactici strain is MNSR144, the classification name is pediococcus acidilactici, the preservation number is CGMCC NO.37266, the preservation date is January 4, 2026, and the preservation number is CGMCC NO.37266. The strain is preserved in China General Microbiological Culture Collection Center (CGMCC), No. 3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing. The invention finds that pediococcus acidilactici MNSR144 and the metaplast thereof can realize nerve injury protection and prevention and treatment of dementia diseases through anti-inflammatory action, experiments are carried out by using the pediococcus acidilactici MNSR144, and the experiments show that the strain and the metaplast thereof can reduce the content of IL-1beta, IL-6 and TNF-alpha in brain tissues of mice with senile dementia, improve neuroinflammation and prevent and treat dementia diseases, so that the pediococcus acidilactici MNSR144 and the metaplast thereof can be used for preventing and treating dementia diseases. Therefore, the effect of preventing senile dementia is achieved.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to functional food preparation technology, especially a strain of Pediococcus lactis derived from yak yogurt with neuroprotective and anti-dementia functions, its postbiotics and applications. Background Technology

[0002] Lactic acid bacteria have long been considered one of the most abundant microorganisms in the human gastrointestinal tract and are important beneficial bacteria in the human gut.

[0003] Neurological damage to the human nervous system is irreversible and progressive. It not only directly damages neuronal structure and blocks nerve signal transmission, but also triggers degenerative changes in multiple dimensions of function, including cognition, movement, and behavior, devastating patients' health and quality of life. Among various neurological damage-related diseases, dementia, due to its strong correlation with population aging, is becoming a pressing global public health challenge. As the global population continues to age, the risk of neurological damage among the elderly is significantly increasing, and the incidence of dementia is also rising sharply with age. Alzheimer's disease not only leads to memory decline, cognitive loss, and inability to care for oneself, but also places a heavy burden on family care, becoming a major public health challenge for global healthcare systems. Currently, there is no cure for dementia; prevention and intervention have become key strategies for reducing incidence and alleviating the social burden.

[0004] The core pathology of Alzheimer's disease, a representative of senile dementia, is currently believed to mainly involve β-amyloid protein deposition, tau protein hyperphosphorylation, and damage to neurons and synapses in memory-related brain regions. These lesions ultimately lead to memory decline and cognitive impairment. Chronic central nervous system inflammation (release of pro-inflammatory factors such as TNF-α and IL-1β) and oxidative stress are key upstream driving factors, exacerbating neuronal damage and synaptic dysfunction. Furthermore, age-related decline in intestinal function further increases the risk of disease, providing a clear direction for targeted prevention.

[0005] The gut-brain axis is the core connecting bridge between the gut microbiota and the central nervous system. Studies have found that the decline in the abundance of beneficial bacteria in the elderly gut leads to microbiota imbalance, and the metabolites of harmful bacteria trigger systemic inflammation, promoting pathological changes in the brain. Beneficial bacteria and their produced short-chain fatty acids and other active substances can inhibit central inflammation, regulate neurotransmitters, and maintain the function of memory brain regions. Probiotics and postbiotics (inactivated bacteria and active metabolites), as gut microbiota regulators, have great potential to prevent nerve damage and dementia-like diseases.

[0006] The existing technology has the following technical defects: 1. Currently, drug treatment for Alzheimer's disease is not very effective; it can only relieve symptoms and cannot repair damaged nerves or stop the progression of the disease.

[0007] 2. The pathogenesis of Alzheimer's disease is complex. This invention provides a new treatment method targeting the gut-brain axis.

[0008] 3. Existing treatments have significant systemic side effects, while probiotics indirectly regulate the nervous system through the gut-brain axis, resulting in fewer side effects.

[0009] Therefore, this invention is of great significance for developing drugs and functional products for the prevention and treatment of neurological damage and dementia, and for improving human health. Summary of the Invention

[0010] The purpose of this invention is to overcome the shortcomings of the prior art and provide a strain of Pediococcus lactis derived from yak yogurt with neuroprotective and anti-dementia functions, as well as its post-genes and applications.

[0011] The technical solution adopted by this invention to solve its technical problem is: A strain of Pediococcus lactis derived from yak yogurt with neuroprotective and anti-dementia functions ( Pediococcus acidilactici The strain is named MNSR144, classified as Pediococcus lactis, with accession number CGMCCNO.37266, accession date January 4, 2026, and deposited at China General Microbiological Culture Collection Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0012] The post-biotic of *Pediococcus lactis* prepared using *Pediococcus lactis* as described above.

[0013] The preparation method of the postbiotic of *Pediococcus lactis* as described above includes the following steps: Dissolve the bacterial cells in a 0.9% sodium chloride solution and heat in a 100°C metal bath for 10 minutes.

[0014] The use of *Pediococcus lactis* and / or *Pediococcus lactis* postbiotics as described above in the preparation of drugs and / or functional products that protect against nerve damage and / or dementia.

[0015] Furthermore, the functional products include conventional nutritional foods, functional foods, special medical foods, or pharmaceuticals.

[0016] Furthermore, the live bacterial culture and metabiotics of the aforementioned *Pediococcus lactis* can significantly improve the cognitive abilities of individuals with dementia.

[0017] Furthermore, the live bacterial solution of *Pediococcus lactis* MNSR144 and the post-biotic have antioxidant capabilities, which can significantly increase GSH in dementia brain tissue and reduce MDA content in dementia tissue.

[0018] Furthermore, the live bacterial solution of *Pediococcus lactis* MNSR144 and the post-biotic have anti-inflammatory effects and can significantly reduce the content of IL-1β, IL-6 and TNF-α in dementia brain tissue.

[0019] Furthermore, the lysate of *Pediococcus lactis* MNSR144 (post-biotic) has neuroprotective effects and can significantly increase the survival rate of PC12 cells damaged by aluminum chloride.

[0020] Furthermore, the *Pediococcus lactis* MNSR144 is derived from yak yogurt from the Qinghai-Tibet Plateau in my country.

[0021] The advantages and positive effects of this invention are as follows: 1. This invention has discovered that Pediococcus lactis MNSR144 and its post-biotics can be used in the protection of nerve damage and the prevention and treatment of dementia. Experiments using Pediococcus lactis MNSR144 showed that the strain and its post-biotics can reduce the latency in the water maze test of Alzheimer's mice and improve memory impairment.

[0022] 2. This invention has discovered that Pediococcus lactis MNSR144 and its metabolites can achieve the protection against nerve damage and prevention of dementia through antioxidant effects. Experiments using Pediococcus lactis MNSR144 showed that this strain and its metabolites can increase the GSH content in the brain tissue of Alzheimer's mice, reduce the MDA content in the brain tissue, and improve oxidative stress, thereby achieving the effect of preventing Alzheimer's disease.

[0023] 3. This invention has discovered that Pediococcus lactis MNSR144 and its metabolites can achieve the protection against nerve damage and the prevention and treatment of dementia through anti-inflammatory effects. Experiments using Pediococcus lactis MNSR144 showed that this strain and its metabolites can reduce the levels of IL-1β, IL-6, and TNF-α in the brain tissue of Alzheimer's mice and improve neuroinflammation, thereby achieving the effect of preventing Alzheimer's disease.

[0024] 4. The strain of *Pediococcus lactis* MNSR144 used in this invention is a food-grade probiotic and its postbiotic, which is highly safe for research on related conventional foods, functional foods, special medical foods, and drugs.

[0025] 5. This invention provides new theoretical guidance and selection of raw materials for functional products for the prevention and treatment of neurological injury diseases and dementia. 6. This invention relates to *Pediococcus lactis* MNSR144, derived from yak yogurt, which possesses neuroprotective and anti-dementia functions. It overcomes the shortcomings of existing daily intervention technologies for neurodegenerative diseases and dementia, and helps address the practical problems of my country's probiotic industry's excessive dependence on imported strains, the risk of strain bottlenecks, and doubts about the compatibility of imported strains with the intestinal and genetic characteristics of the Chinese population. Experiments show that *Pediococcus lactis* MNSR144 (strain preservation number: CGMCC NO.37266), isolated from yak yogurt of the Qinghai-Tibet Plateau, and its metabiotics can effectively improve memory impairment in aluminum chloride-induced Alzheimer's mice; increase GSH content and decrease MDA content in the brain tissue of Alzheimer's mice to achieve antioxidant effects; reduce IL-1β, IL-6, and TNF-α content in the brain tissue of Alzheimer's mice to improve neuroinflammation; and increase the survival rate of aluminum chloride-induced damaged PC12 cells, thereby improving neurological damage. The *Pediococcus lactis* MNSR144 and its postbiotics of the present invention can be used in the fields of food and medicine, and can be used to develop related food, functional food, special medical food, pharmaceutical and other products. Attached Figure Description

[0026] Figure 1 This is a comparison of the latency periods of water maze experiments using Pediococcus lactis MNSR144 and its postbiotics in this invention. Figure 2 This is a comparison of GSH and MDA content in mouse brain tissue after treatment with Pediococcus lactis MNSR144 and its postbiotics in this invention. Figure 3 This is a comparison of the levels of IL-1β, IL-6, and TNF-α in mouse brain tissue after treatment with Pediococcus lactis MNSR144 and its postbiotics in this invention. Figure 4 This is a comparison chart showing the improvement of PC12 cell survival rate after treatment with Pediococcus lactis MNSR144 and subsequent biotics in aluminum chloride-induced damage, as described in this invention. Detailed Implementation

[0027] The present invention will be further described below with reference to the embodiments. The following embodiments are descriptive and not limiting, and should not be used to limit the scope of protection of the present invention.

[0028] The various experimental operations involved in the specific embodiments are all conventional techniques in the art. For parts not specifically annotated herein, those skilled in the art can refer to various commonly used reference books, scientific and technological literature, or related instructions and manuals prior to the filing date of this invention for implementation. The following embodiments are used to illustrate the present invention, but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0029] A strain of Pediococcus lactis derived from yak yogurt with neuroprotective and anti-dementia functions ( Pediococcus acidilactici The strain is named MNSR144, classified as Pediococcus lactis, with accession number CGMCCNO.37266, accession date January 4, 2026, and deposited at China General Microbiological Culture Collection Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0030] The post-biotic of *Pediococcus lactis* prepared using *Pediococcus lactis* as described above.

[0031] The preparation method of the postbiotic of *Pediococcus lactis* as described above includes the following steps: Dissolve the bacterial cells in a 0.9% sodium chloride solution and heat in a 100°C metal bath for 10 minutes.

[0032] The use of *Pediococcus lactis* and / or *Pediococcus lactis* postbiotics as described above in the preparation of drugs and / or functional products that protect against nerve damage and / or dementia.

[0033] Furthermore, the functional products include conventional nutritional foods, functional foods, special medical foods, or pharmaceuticals.

[0034] Furthermore, the live bacterial culture and metabiotics of the aforementioned *Pediococcus lactis* can significantly improve the cognitive abilities of individuals with dementia.

[0035] Furthermore, the live bacterial solution of *Pediococcus lactis* MNSR144 and the post-biotic have antioxidant capabilities, which can significantly increase GSH in dementia brain tissue and reduce MDA content in dementia tissue.

[0036] Furthermore, the live bacterial solution of *Pediococcus lactis* MNSR144 and the post-biotic have anti-inflammatory effects and can significantly reduce the content of IL-1β, IL-6 and TNF-α in dementia brain tissue.

[0037] Furthermore, the lysate of *Pediococcus lactis* MNSR144 (post-biotic) has neuroprotective effects and can significantly increase the survival rate of PC12 cells damaged by aluminum chloride.

[0038] Furthermore, the *Pediococcus lactis* MNSR144 is derived from yak yogurt from the Qinghai-Tibet Plateau in my country.

[0039] Furthermore, the live bacterial culture of *Pediococcus lactis* MNSR144 and the postbiotic significantly improved the latency changes in Alzheimer's mice in the water maze test.

[0040] Furthermore, the live bacterial culture of *Pediococcus lactis* MNSR144 and the postbiotic significantly increased the GSH content and decreased the MDA content in the brain tissue of Alzheimer's mice.

[0041] Furthermore, the live bacterial culture of *Pediococcus lactis* MNSR144 and the post-biotic significantly reduced the levels of IL-1β, IL-6, and TNF-α in the brain tissue of Alzheimer's mice.

[0042] Furthermore, the lysate of *Pediococcus lactis* MNSR144 significantly reduced the damage of aluminum chloride to PC12 cells.

[0043] Specifically, the relevant preparation and testing methods are as follows: This invention relates to a strain of *Pediococcus lactis* derived from yak yogurt that possesses neuroprotective and anti-dementia functions. Pediococcus acidilactici The strain is named MNSR144, classified as *Pediococcus lactis*, with accession number CGMCC NO.37266, accession date January 4, 2026, and depositary institution: China General Microbiological Culture Collection Center, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing. Its 16S rDNA sequence is as follows: Regulatory Effect of Pediococcus acidilactici MNSR144 and Its Postbiotics on Cognitive Function of Alzheimer's Mice (1)Preparation of probiotics and postbiotics: The cultured bacterial liquid in MRS medium was centrifuged to collect the bacterial cells, diluted with normal saline, and inactivated in a water bath at 100 °C for 10 minutes. The remaining drugs for animal experiments were diluted with normal saline. The bacterial lysate was prepared by dissolving the bacterial cells in PBS through the repeated freezing and thawing method (-80 °C freezing + 37 °C water bath thawing; repeated three times).

[0044] The preparation method of Pediococcus acidilactici postbiotics includes the following steps: Dissolve the bacterial cells in a 0.9% sodium chloride solution by mass concentration and heat in a metal bath at 100 °C for 10 minutes.

[0045] (2)Animal grouping and administration of samples: Male Kunming mice, 4 weeks old, weighing 20 - 22 g, were purchased from Beijing Spey Foster Biotechnology Co., Ltd. (License: SCXK(Beijing)2024 - 0001). The animals were randomly divided into a blank group NC (0.1 ml / 10 g normal saline, gavage treatment after 7 days of adaptive feeding), a model group M (200 mg / kg AlCl3, gavage treatment after 7 days of adaptive feeding), a positive drug group PC (3 mg / kg donepezil + 200 mg / kg AlCl3, gavage treatment after 7 days of adaptive feeding), a low-dose live bacteria group L-L (5×10

[0046] CFU / 10 g + 200 mg / kg AlCl3, gavage treatment after 7 days of adaptive feeding), a high-dose live bacteria group H-L (5×10 7 CFU / 10 g + 200 mg / kg AlCl3, gavage treatment after 7 days of adaptive feeding), a low-dose postbiotic group P-L-L (5×10 6 CFU / 10 g + 200 mg / kg AlCl 3 3, gavage treatment after 7 days of adaptive feeding), a high-dose postbiotic group P-H-L (5×10 7 CFU / 10 g + 200 mg / kg AlCl3, gavage treatment after 7 days of adaptive feeding). They were housed in a SPF animal laboratory at 18 °C - 22 °C. Gavage for 30 days.

[0046] (3) Animal behavioral analysis: A water maze experiment was conducted during the last six days. Adaptation period: On day 1, mice were placed in a water maze without a platform and allowed to swim freely for 2 minutes to familiarize themselves with the aquatic environment. Training period: From day 2 to day 5, the platform was placed in the water 1 cm below the surface. Mice were placed in the water in multiple quadrants each day. If the platform was not found within 60 seconds, the mice were herded onto the platform and remained there for 15 seconds; if the platform was found, the mice also remained there for 15 seconds. Testing period: On day 6, the platform was removed, and the animals were placed in the water maze. The time it took to reach the original platform location within 60 seconds was recorded as the latency period. If the animals did not reach the platform, it was recorded as 60 seconds.

[0047] (4) Results: From Figure 1 It can be seen that the latency period of the model group was about 50 seconds, while the latency periods of the live bacteria group (17.6 seconds, 25.2 seconds) and the postbiotic group (21.7 seconds, 15.8 seconds) were all lower than those of the model group (50.2 seconds), which significantly improved the memory impairment of mice.

[0048] Example 2: Detection of levels of oxidative stress-related factors in mouse brain tissue Mice were euthanized by cervical dislocation after 30 days of gavage, and brain tissue was extracted. The levels of GSH and MDA in the brain tissue were determined according to the kit instructions. Figure 2 It can be seen that the GSH content in the brain tissue of the model group was 27 ug / mgprot, while that of the high-dose live bacteria group and the high-dose postbiotic group was 34 ug / mgprot and 36 ug / mgprot, respectively, which were significantly higher than those of the model group. The MDA content in the brain tissue of the high-dose live bacteria group and the high-dose postbiotic group was 1.94 nmol / mgprot and 1.72 nmol / mgprot, respectively, which were significantly lower than those of the model group (2.9 nmol / mgprot), indicating that they have the ability to resist oxidative damage to brain tissue.

[0049] Example 3: Detection of inflammation-related factor levels in mouse brain tissue Mice were euthanized by cervical dislocation, and brain tissue was extracted. The levels of IL-1β, IL-6, and TNF-α in the brain tissue were determined using ELISA according to the kit instructions. Figure 3 It can be seen that the IL-1β content in the brain tissue of the high-dose live bacteria group and the high-dose post-biotic group were 34.25 pg / ml and 34.28 pg / ml, respectively, which were significantly lower than those of the model group (47.51 pg / ml); the IL-6 content in the brain tissue of the high-dose live bacteria group and the high-dose post-biotic group were 52.57 pg / ml and 61.37 pg / ml, respectively, which were significantly lower than those of the model group (151.9 pg / ml); the IL-6 content in the brain tissue of the high-dose live bacteria group and the high-dose post-biotic group were 29.66 pg / ml and 26.41 pg / ml, respectively, which were significantly lower than those of the model group (38.65 pg / ml); indicating that they have anti-neuroinflammatory capabilities.

[0050] Example 4: Detection of PC12 cell viability after aluminum chloride-induced damage PC12 cells were seeded at 5000 cells / well in 96-well plates and treated with the bacterial lysis buffer prepared in Example 1 and 8 mmol / L AlCl3 for 24 h. Cell viability was detected using the CCK-8 assay. Cell experimental groups were: control group (no treatment); model group M (8 mmol / L AlCl3); low-dose group L-14-4 (8 mmol / L AlCl3 + 1 × 10⁻⁶ cells / well). 9 CFU / mlMNSR144); Medium-dose group M-14-4 (8mmol / L AlCl3+2×10) 9 CFU / ml MNSR144); High-dose group H-14-4 (8mmol / L AlCl3+4×10) 9 CFU / ml MNSR144). From Figure 4 It can be seen that all three dose groups of MNSR144 (low, medium, and high) can improve the survival rate of PC12 cells damaged by aluminum chloride, with the high dose group showing a significant difference.

[0051] Although embodiments of the invention have been disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, variations, and modifications are possible without departing from the spirit and scope of the invention and the appended claims. Therefore, the scope of the invention is not limited to the contents disclosed in the embodiments.

Claims

1. A strain of Pediococcus lactis derived from yak yogurt with neuroprotective and anti-dementia functions ( Pediococcus acidilactici ), characterized by: The strain is named MNSR144, classified as Pediococcus lactis, with accession number CGMCC NO.37266, accession date January 4, 2026, and deposited at the China General Microbiological Culture Collection Center, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

2. A postbiotic prepared using *Pediococcus lactis* as described in claim 1.

3. The method for preparing postbiotics from *Pediococcus lactis* as described in claim 2, characterized in that: Includes the following steps: Dissolve the bacterial cells in a 0.9% sodium chloride solution and heat in a 100°C metal bath for 10 minutes.

4. The use of Pediococcus lactis and / or Pediococcus lactis postbiotics as described in claim 1 or 2 in the preparation of drugs and / or functional products that protect against nerve damage and / or dementia.

5. The application according to claim 4, characterized in that: The functional products include conventional nutritional foods, functional foods, special medical foods, or medicines.

6. The application according to claim 4, characterized in that: The live bacterial culture and postbiotics of the aforementioned *Pediococcus lactis* can significantly improve cognitive abilities in individuals with dementia.

7. The application according to claim 4, characterized in that: The live bacterial solution of *Pediococcus lactis* MNSR144 and the post-biotic have antioxidant capabilities, which can significantly increase GSH in dementia brain tissue and reduce MDA content in dementia tissue.

8. The application according to claim 4, characterized in that: The live bacterial solution of *Pediococcus lactis* MNSR144 and post-biotic have anti-inflammatory effects and can significantly reduce the content of IL-1β, IL-6 and TNF-α in dementia brain tissue.

9. The application according to claim 4, characterized in that: The lysate of *Pediococcus lactis* MNSR144 (post-biotic) has neuroprotective effects and can significantly increase the survival rate of PC12 cells damaged by aluminum chloride.

10. The application according to claim 4, characterized in that: The lactic acid cocci MNSR144 is derived from yak yogurt from the Qinghai-Tibet Plateau in my country.

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