Composition comprising Lactobacillus sakei Probio-65 for prevention or treatment of alzheimer's disease

Abstract

The present invention provides a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 as an active ingredient. The composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 according to the present invention has no side effects in the body and has the effect of alleviating Alzheimer's disease through changes in the profile of intestinal microorganisms and the composition of intestinal metabolites, and can be used as a pharmaceutical composition, a food composition, a cosmetic composition, and a feed composition.

Description

Technology Field

[0001] The present invention relates to a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65, and more specifically, to a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 that has no side effects and can alleviate Alzheimer's disease through changes in the profile of intestinal microorganisms and the composition of intestinal metabolites. Background Technology

[0002] Lactobacillus sakei Probio-65 is a strain isolated from kimchi, a fermented food source whose safety has been traditionally ensured through centuries of consumption by Koreans. It exhibits acid resistance, bile resistance, and antibiotic resistance, inhibits the growth of harmful pathogenic microorganisms in the body and intestines of animals, and displays immune-enhancing activity. In particular, it inhibits the growth of Staphylococcus aureus, which is known as a disease-exacerbating factor for atopic dermatitis lesions, making it effective for the treatment or prevention of atopic dermatitis and also effective for the treatment or prevention of allergy-related diseases. Furthermore, it has the efficacy of stabilizing the intestinal microbial community by inhibiting the abnormal proliferation of harmful microorganisms in the intestines. The inventor of the present invention is the inventor of Korean Registered Patent No. 10-0479719, who has proposed the above-mentioned Lactobacillus sakei Probio-65 and its composition.

[0003] While conducting research regarding other efficacy of Lactobacillus sakei Probio-65, the inventors discovered that Lactobacillus sakei Probio-65 can be used as a composition for the prevention or treatment of Alzheimer's disease.

[0004] Alzheimer's disease is the most widespread form of dementia globally, affecting 47 million people and incurring a total cost of $604 billion as of 2018 (Alzheimer's Association, 2018). While Alzheimer's disease shares many pathological aspects with other neurodegenerative diseases, such as neurofibrillary tangles and neuronal loss primarily in the cerebral cortex and hippocampus, the defining characteristic of the disease is the presence of amyloid plaques caused by the deposition of amyloid peptides known as amyloid beta (Aβ). Among all Aβ species formed, Aβ42 is the most toxic and aggressive form that causes neurodegeneration. Inflammation is a central mechanism in Alzheimer's disease that triggers early Aβ pathology, as the disruption of inflammatory homeostasis leads to chronic inflammation.

[0005] Specifically, Aβ, currently known as the most likely cause of Alzheimer's disease, is produced when beta (β) and gamma-secretase (γ-secretase) enzymes abnormally hydrolyze APP (Amyloid Precursor Protein). Aβ is known to contain toxins that destroy neurons in the cerebral cortex and the parts of the brain responsible for producing the neurotransmitter acetylcholine, thereby reducing acetylcholine activity. Furthermore, it is known to induce neuronal death through oxidative damage caused by the generation of Reactive Oxygen Species (ROS).

[0006] Most currently used Alzheimer's disease treatments are Ach-degrading enzyme inhibitors, but they have the problem of causing significant liver toxicity and various other side effects.

[0007] In order to solve the problems of such conventional Alzheimer's disease treatments, the inventors have developed a composition for the prevention or treatment of Alzheimer's disease containing Lactobacillus sakei Probio-65, which has no side effects in the body and can alleviate Alzheimer's disease through changes in the profile of intestinal microorganisms and the composition of intestinal metabolites. The problem to be solved

[0008] The technical problem that the present invention aims to solve is to provide a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65, which has no side effects in the body, can alleviate Alzheimer's disease through changes in the profile of intestinal microorganisms and the composition of intestinal metabolites, and can be used as a pharmaceutical composition, food composition, cosmetic composition, and feed composition based on microorganisms or culture solutions thereof.

[0009] The above and other objectives of the present invention can all be achieved by the present invention described below. means of solving the problem

[0010] A composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 according to one embodiment of the present invention is characterized by comprising Lactobacillus sakei Probio-65 as an active ingredient.

[0011] Here, the above Lactobacillus sakei Probio-65 is characterized as being a Lactobacillus sakei Probio-65 culture medium obtained by liquid culturing the Lactobacillus sakei Probio-65 strain.

[0012] In addition, a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 according to one embodiment of the present invention can be used as a food composition comprising Lactobacillus sakei Probio-65 or a culture medium thereof.

[0013] A composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 according to one embodiment of the present invention can be used as a cosmetic product comprising Lactobacillus sakei Probio-65 or a culture solution thereof.

[0014] A composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 according to one embodiment of the present invention can be used as a feed composition comprising Lactobacillus sakei Probio-65 or a culture medium thereof. Effects of the invention

[0015] The present invention has the effect of providing a composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65, which has no side effects in the body and can alleviate Alzheimer's disease through changes in the profile of intestinal microorganisms and the composition of intestinal metabolites. The microorganism of the present invention or a culture medium thereof can be used as a pharmaceutical composition, a food composition, a cosmetic composition, and a feed composition. Brief explanation of the drawing

[0016] Figure 1 is a photograph of the eye shape of GMR-OreR supplied with normal food, taken at a magnification of 800x. Figure 2 is a photograph of the eye shape of a transgenic fruit fly (GMR-Aβ42) fed normal food, taken at a magnification of 800x. Figure 3 is a photograph of the eye shape of a transgenic fruit fly (GMR-Aβ42) fed normal food containing Lactobacillus sakei Probio-65 culture medium, taken at a magnification of 800x. Figure 4 is a graph showing the principal coordinate analysis (PcoA) of a microbial community based on the Bray-Curtis distance matrix. Figure 5 is a graph showing the intestinal microbiological characteristics of fruit flies, indicating the relative abundance of the genus observed in different treatment groups. Specific details for implementing the invention

[0017] The above-described objects, features, and advantages of the present invention will become more apparent from the detailed description of preferred embodiments and comparative examples of the present invention. The following detailed description is not intended to be limiting, and the scope of the present invention is limited only by the appended claims, including all equivalents thereof.

[0018] The present invention will be described in more detail below with reference to the attached drawings and embodiments.

[0020] Examples

[0021] Example 1: Preparation of culture solution for Lactobacillus sakei Probio-65

[0022] Lactobacillus sakei Probio-65, isolated from kimchi, a traditional Korean fermented food, was used as a starter culture after superior strains obtained by subculturing 2 to 3 times for 15 hours in an incubator at 37°C in MRS broth (Difco laboratories, USA) to improve the physiological activity of long-term preserved strains and select healthy strains, and then culturing the cells in an incubator at 37°C for 15 hours while maintaining constant temperature in MRS broth. After that, a liquid culture medium comprising 1% by weight of glucose, 1% by weight of yeast extract, 0.25% by weight of soy peptone, 0.1% by weight of polyoxyethylene sorbitan mono-oleate, 0.1% by weight of ammonium sulfate ((NH4)2SO4), 0.01% by weight of magnesium sulfate (MgSO4), 0.1% by weight of potassium dihydrogen phosphate (KH2PO4), and 90-99% by weight of water was sterilized at 121°C for 15 minutes, and then the Lactobacillus sakei Probio-65 was liquid cultured in a 2L flask at 37°C for 24 hours using this medium to prepare a Lactobacillus sakei Probio-65 culture solution.

[0024] Examples 2: Using transgenic fruit flies (GMR-Aβ42) Lactobacillus Sakei Probio Experiment on the preventive or therapeutic effects of -65 on Alzheimer's disease

[0025] Example 2-1 Rearing of Experimental Fruit Fly Group

[0026] For this experiment, the fruit fly population (Drosophila melanogaster) used was the wild Oregon-R, Glass Multiple Reporter (GMR)-GAL4, and UAS-Aβ42. Oregon-R was crossed with GMR-GAL4 to produce GMR-OreR, and UAS-Aβ42 was crossed with GMR-GAL4 to produce transgenic fruit flies (GMR-Aβ42). All fruit fly populations were grown in an environment of 25°C, and the mating conditions were maintained at 30°C.

[0028] Example 2-2 Feeding of the experimental fruit fly group

[0029] Prior to transformation treatment, fruit fly populations for all experiments were cultured at 18°C ​​and fed standard feed. The standard feed was prepared by mixing 4% (w / v) boiled corn starch, 5% (w / v) polenta, 10% (w / v) brown sugar, 0.7% (w / v) agar, 5% (w / v) heat-killed yeast, 3% (w / v) nipagin, and 0.7% (v / v) propionic acid, transferring the mixture into a sterile plastic vial, and then cooling and solidifying it. During the transformation process, the same feed was prepared without nipagin and propionic acid, and just before solidification during the cooling process, Lactobacillus sakei Probio-65 (1×10⁶) 11CFU / mL) was added. Each feed was prepared fresh prior to feeding. Normal and transgenic fruit flies (GMR-Aβ42) were raised on standard feed. On the other hand, transgenic fruit flies (GMR-Aβ42) fed with Lactobacillus sakei Probio-65 were raised on feed containing lactic acid bacteria. Progeny fruit flies were collected from each group after 10 days for analysis.

[0031] Experimental results of Example 2-3

[0032] As fruit flies are similar to mammals but share a simple central nervous system, the use of Drosophila melanogaster among fruit flies in neuroscience research has become a significant breakthrough in this field. Therefore, Drosophila melanogaster (D. melanogaster) was used as the fruit fly group in this experiment. In addition, the culture medium composition containing Lactobacillus sakei Probio-65 according to the present invention was tested on D. melanogaster ectopically expressing human Aβ42 in compound eyes using a pan-retinal GMR-GAL4 driver.

[0034] Figure 1 is a photograph of the eye shape of GMR-OreR supplied with normal food at a magnification of 800x, Figure 2 is a photograph of the eye shape of a transgenic fruit fly (GMR-Aβ42) supplied with normal food at a magnification of 800x, and Figure 3 is a photograph of the eye shape of a transgenic fruit fly (GMR-Aβ42) supplied with normal food containing Lactobacillus sakei Probio-65 culture at a magnification of 800x.

[0036] As can be seen in Figures 1 to 3, the eye morphology of GMR-OreR (control group) fed a normal diet consists of regularly arranged small hexagonal shapes with bristles formed between them. However, in transgenic fruit flies (GMR-Aβ42) (negative control) fed a normal diet, a severe neurodegenerative phenotype characterized by a coarse eye phenotype (REP) was observed. Compared to the negative control group, Lactobacillus sakei Probio-65 (1×10⁶ 11 Transgenic fruit flies (GMR-Aβ42) fed a normal diet containing CFU / mL showed that omatidia with small hexagonal shapes and less visible pores were observed, the number of bristles increased significantly, and the angles of the bristles were rectified.

[0038] Degree of structural effect on the rough eye phenotype strain Degree of structural effect Hexagonal compound eyes Kangmo Su Presence or absence of a hole GMR-Aβ42 without treatment - - - GMR-Aβ42 treated with L. sakei ProBio65 ++ + ++ Control +++ +++ +++

[0040] Table 1 above shows the degree of structural effect on the coarse eye phenotype in relation to Figures 1 to 3. As can be seen in Table 1, the supply of Lactobacillus sakei Probio-65 showed that the neurotoxic effect was reduced as Aβ42 decreased. The degree of reduction was very pronounced in the hexagonal compound eyes and the pores of the omatidia.

[0042] Figure 4 is a graph showing the Principal Coordinate Analysis (PcoA) of the microbial community based on the Bray-Curtis distance matrix, and Figure 5 is a graph showing the intestinal microbiological characteristics of Drosophila indicating the relative abundance within the genera observed in different treatment groups. In Figures 4 and 5, Nf represents the transgenic Drosophila (GMR-Aβ42) fed a normal diet (negative control), and Probio65 represents Lactobacillus sakei Probio-65 (1×10⁻⁶). 11The transgenic fruit flies (GMR-Aβ42) (Probio65 treatment group) fed a normal diet containing CFU / mL) are shown, and the Control represents GMR-OreR (control group) fed a normal diet.

[0044] As can be seen in Figure 4, the analysis of the gut microbiome showed that the Probio65-treated group was more similar to the control group than the negative control group. Additionally, as can be seen in Figure 5, after treating transgenic fruit flies with Lactobacillus sakei Probio-65, the relative abundance of Wolbachia, a type of parasite, decreased by 37.9% in the Probio65-treated group, whereas Corynebacterium_1 showed high levels only in the Probio65-treated group. Wolbachia is reported as a potential biomarker for Alzheimer's disease because it is a bacterium that initiates or dominates the pathogenesis of Alzheimer's disease in D. melanogaster. Furthermore, Wolbachia is largely associated with neurodegenerative diseases, particularly Parkinson's disease, Huntington's disease, and Alzheimer's disease. Corynebacterium is associated with the production of glutamate. Based on preliminary studies of patients with Alzheimer's disease, it has been reported that these patients have low plasma D-glutamate levels. Furthermore, reduced D-glutamate levels in the brain are associated with cognitive impairment in Alzheimer's disease. These data highlight the importance of D-glutamate in the mental health of patients with Alzheimer's disease. Additionally, Lactobacillus sakei Probio-65 enhances the proliferation of Corynebacterium present in the gut of transgenic fruit flies, thereby potentially promoting the production of glutamate. Consequently, the transgenic fruit flies demonstrated significant Alzheimer's disease alleviation effects. From the above experimental results, it was confirmed that Lactobacillus sakei Probio-65 has the effect of improving neurodegenerative phenomena manifested in the eye defects of fruit flies through changes in the gut microbial profile and gut metabolite composition.

[0046] Although preferred embodiments of the present invention have been described above, the present invention is not limited to the specific embodiments described above. That is, those skilled in the art to which the present invention pertains can make numerous changes and modifications to the present invention without departing from the spirit and scope of the appended claims, and all such appropriate changes and modifications and equivalents should be deemed to fall within the scope of the present invention.

Claims

Claim 1 A composition for the prevention or treatment of Alzheimer's disease comprising Lactobacillus sakei Probio-65 as an active ingredient. Claim 2 A composition for the prevention or treatment of Alzheimer's disease according to claim 1, characterized in that the Lactobacillus sakei Probio-65 is a Lactobacillus sakei Probio-65 culture solution obtained by liquid culturing the Lactobacillus sakei Probio-65 strain. Claim 3 delete Claim 4 delete Claim 5 delete

Images