COMPOSITION FOR PREVENTING, ALLEVIATING OR TREATING NEURODEGENERATIVE DISEASES, COMPRISING PEDIOCOCCUS INOPINATUS

Abstract

The present disclosure relates to a composition for preventing, alleviating or treating a neurodegenerative disease, which contains Pediococcus inopinatus as an active ingredient. The Pediococcus inopinatus strain according to the present disclosure can suppress neuroinflammation and can also alleviate the symptoms of ataxia in an animal model of Parkinson's disease and, thus, can be utilized for the prevention, alleviation and treatment of various neurodegenerative diseases including Parkinson's disease.

Claims

1-12. (canceled)

13. A method for preventing, alleviating or treating a neurodegenerative disease, comprising administering a composition comprising a therapeutically effective amount of Pediococcus inopinatus, a culture thereof, a lysate thereof, a pulverization product thereof, a fermentation product thereof or an extract thereof to a subject in need thereof.

14. The method according to claim 13, wherein the composition is a pharmaceutical composition, a food composition or a feed additive composition.

15. The method according to claim 13, wherein the neurodegenerative disease is one or more selected from a group consisting of Parkinson's disease, Alzheimer's disease, Huntington's disease, Lou Gehrig's disease (amyotrophic lateral sclerosis), Creutzfeldt-Jakob disease, stroke, multiple sclerosis, neuroinflammation, learning disorder, cognitive impairment and memory impairment.

16. The method according to claim 14, wherein the pharmaceutical composition is administered via an administration route selected from a group consisting of oral administration, intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration, intradermal administration, topical administration, intranasal administration, intrapulmonary administration and intrarectal administration.

17. The method according to claim 14, wherein the pharmaceutical composition comprises 1×10.sup.6 CFU/mL to 1×10.sup.12 CFU/mL of Pediococcus inopinatus.

18. The method according to claim 14, wherein the pharmaceutical composition prevents or treats the symptoms of ataxia in a subject having neurodegenerative disease.

19. The method according to claim 13, wherein the Pediococcus inopinatus is Pediococcus inopinatus WIKIM27 deposited with an accession number KCCM12653P.

20. The method according to claim 14, wherein the food is a functional health food.

21. The method according to claim 14, wherein the food is a beverage, a bar or a fermented milk.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0068] FIG. 1 schematically shows an experimental procedure for investigating the neuroinflammation-suppressing effect of a Pediococcus inopinatus WIKIM27 strain of the present disclosure.

[0069] FIG. 2 shows a result of observing the result of treating neuroglial cells in which inflammation is induced by LPS with a Pediococcus inopinatus WIKIM27 strain of the present disclosure through immunofluorescence staining.

[0070] FIG. 3 shows a result of treating neuroglial cells in which inflammation is induced by LPS with a Pediococcus inopinatus WIKIM27 strain of the present disclosure.

[0071] FIG. 4 shows a result of orally administering a Pediococcus inopinatus WIKIM27 strain of the present disclosure to a mouse model of Parkinson's disease and then evaluating the motor ability of the mouse using a rotarod.

[0072] FIG. 5 shows a result of orally administering a Pediococcus inopinatus WIKIM27 strain of the present disclosure to a mouse model of Parkinson's disease and then evaluating the grip strength of the mouse.

BEST MODE

[0073] Hereinafter, the present disclosure is described in detail through examples. The following examples merely illustrate the present disclosure, and the scope of the present disclosure is not limited by the examples.

EXAMPLES

Example 1: Preparation of Pediococcus inopinatus Strain

[0074] A Pediococcus inopinatus WIKIM27 strain, which is derived from kimchi and is deposited in the Korea Culture Center of Microorganisms with the accession number KCCM12653P, was used for experiment. After culturing the Pediococcus inopinatus WIKIM27 strain in an MRS medium at 30° C. for 24 hours, the cells were centrifuged at 8,000 rpm for 5 minutes and the remaining medium was removed by rinsing with PBS. After inoculating the cells into DMEM (Dulbecco's modified Eagle's medium, HyClone, USA) medium at a density of 1×10.sup.9 CFU/mL and culturing at 30° C. for 24 hours, the cells were removed by centrifuging at 8,000 rpm for 5 minutes. After adjusting the pH of the supernatant to 7.2, it was filtered through a syringe filter (pore size: 0.22 μm).

Example 2: Primary Culture of Neuroglial Cells

[0075] After disinfecting the scalp of a 2-day-old Sprague-Dawley rat with alcohol, the cerebrum was taken out immediately without damaging. The extracted cerebrum was immersed in cold HBSS (Hank's balanced salt solution) and only the cerebral cortex was separated under a dissecting microscope. After preparing a single cell suspension using a Pasteur pipette, the cells were cultured for 10 days. Neuroglial cells were cultured in a 5% CO.sub.2 incubator at 37° C. using DMEM containing 10% fetal bovine serum (HyClone, USA), 10% horse serum (HyClone, USA) and 1% penicillin/streptomycin (GIBCO, USA).

Example 3: Investigation of Neuroinflammation-Suppressing Effect of Pediococcus inopinatus Strain

[0076] In order to investigate the neuroinflammation-suppressing effect of the Pediococcus inopinatus WIKIM27 strain prepared in Example 1, inflammation was induced in neuroglial cells with LPS (lipopolysaccharide) and the cells were treated with the culture of WIKIM27. After seeding the neuroglial cells onto a 24-well plate at a concentration of 1.0×10.sup.5 cells/mL and replacing with a serum-free medium 24 hours later, the cells were treated with 100 ng/mL LPS 16 hours later. Two hours after the treatment with LPS, the cells were treated with the culture of WIKIM27 with a volume of ⅕ of that of the neuroglial cell medium. The experimental procedure is schematically shown in FIG. 1. A Weissella cibaria WIKIM28 (accession number KFCC11625P) strain and a Lactobacillus sakei WIKIM30 (accession number KFCC11618P) strain were used as control groups.

[0077] As seen from FIGS. 2 and 3, it was confirmed through immunohistochemical analysis that the WIKIM27 treatment group (W27_CM) showed significantly decreased activity of neuroglial cells (microglia and astrocytes) despite the induction of neuroinflammation with LPS. As selective markers for microglia and astrocytes, Iba-1 (ionized calcium-binding adapter molecule 1) and GFAP (glial fibrillary acidic protein) were used, respectively. It was confirmed that the culture of the WIKIM27 strain alleviates neuroinflammation.

Example 4: Preparation and Administration of Pediococcus inopinatus Strain

[0078] After culturing a Pediococcus inopinatus WIKIM27 strain in MRS medium at 30° C. for 24 hours, the cells were centrifuged at 8,000 rpm for 5 minutes and the remaining medium was removed by rinsing with PBS. 1×10.sup.10 CFU/mL of cells were quantitated using PBS. 0.2 mL (1×10.sup.9 CFU) was orally administered to an experimental animal 5 times a week. Sterilized PBS was administered to negative and positive control groups.

Example 5: Investigation of Treatment of Animal Model with Pediococcus inopinatus Strain on Prevention and Treatment of Parkinson's Disease

[0079] In order to investigate the therapeutic effect of the Pediococcus inopinatus WIKIM27 strain prepared in Example 1 on Parkinson's disease, rotarod test and grip strength test were conducted for a mouse model of Parkinson's disease. The mouse model of Parkinson's disease was established by accustoming 8-week-old male mice (C57BL/6) in a laboratory room for a week, orally administering the Pediococcus inopinatus WIKIM27 strain for 30 days and then inducing Parkinson's disease by intraperitoneally injecting MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, 30 mg/kg, once a day for 5 days). Naive stands for a non-treatment group, MPTP a negative control group, Selegiline a positive control group. Selegiline is an inhibitor of MAO-B, which is a neuroprotective material, and was orally administered (3 mg/kg) 3 days before the MPTP injection.

[0080] 1) Rotarod Test

[0081] Rotarod test was performed by measuring riding time on a rotarod consisting of a base platform and a rotating rod with a non-slippery surface. Evaluation was conducted at 30 minutes after training the mouse at a speed of 4 rpm for 2 minutes for accustomation. The rotation speed was increased from 4 rpm up to 40 rpm, with an acceleration paradigm of 1 rpm per 8 seconds, and the maximum rotation speed was maintained at 40 rpm for 300 seconds.

[0082] As a result (FIG. 4), the negative control group treated with PBS maintained balance at a speed of 7.7 rpm on average, whereas the test group to which the Pediococcus inopinatus WIKIM27 strain was administered orally could maintain balance at a speed of 10.6 rpm on average.

[0083] 2) Grip Strength Test

[0084] Grip strength test was performed by measuring the grip strength of the paw of the mice with a grip strength meter. The grip strength of the paw of the mice was measured 5 times and the highest 3 values were averaged.

[0085] As a result (FIG. 5), the negative control group showed a paw grip strength of 77.78 gram, whereas the test group to which the Pediococcus inopinatus WIKIM27 strain was administered orally showed a very high grip strength of 85.79 gram as compared to the negative control group.

[0086] That is to say, since the ingestion of the Pediococcus inopinatus WIKIM27 strain resulted in better results of rotarod and grip strength tests in the mouse model of Parkinson's disease as compared to the negative control group, it was confirmed that the symptoms of ataxia in the mouse model of Parkinson's disease can be improved significantly (p<0.05).