CN116173078A - Application of fermented lactobacillus mucilaginosus in preparation of products for preventing or delaying huntington chorea - Google Patents
Application of fermented lactobacillus mucilaginosus in preparation of products for preventing or delaying huntington chorea Download PDFInfo
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- CN116173078A CN116173078A CN202211578249.1A CN202211578249A CN116173078A CN 116173078 A CN116173078 A CN 116173078A CN 202211578249 A CN202211578249 A CN 202211578249A CN 116173078 A CN116173078 A CN 116173078A
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Abstract
The invention discloses an application of lactobacillus fermentum in preparing a product for preventing or delaying huntington's disease, wherein the lactobacillus fermentum is lactobacillus fermentum LFPerfectus001 (Limosilactobacillus fermentum) with the amino acid sequence of SEQ ID NO:1, and experiments show that compared with a blank control group, the neuron survival proportion of the caenorhabditis elegans can be remarkably improved after the lactobacillus mucilaginosus LFP001 is added.
Description
Technical Field
The invention relates to the application field of probiotics, in particular to application of fermented lactobacillus mucilaginosus in preparing a product for preventing or delaying huntington's disease.
Background
Probiotics are active microorganisms beneficial to organisms and have great effects in maintaining microbial balance in intestinal tracts. The fermented lactobacillus mucilaginosus belongs to the genus lactobacillus, is one of common probiotics capable of promoting human health, has a plurality of reports on antioxidation, regulation of intestinal flora balance and the like, but has no report on prevention or delay of huntington disease.
In the related art, huntington's Disease (HD) is an autosomal dominant neurodegenerative Disease, and takes mental, motor and cognitive decline as a main clinical feature, and the main cause is that Huntingtin gene on chromosome four of a patient is mutated, which causes abnormal accumulation of polyQ protein in cells to generate cytotoxicity, break down the intracellular balance and finally cause cell death. Typical patients develop disease in middle-aged years, manifesting as chorea-like movements, gradually losing the ability to speak, act, think and swallow as the condition progresses, with the condition continuing for 10 to 20 years at an approximate date, and ultimately leading to death of the patient. At present, the clinical medicine intervention is mainly carried out on the surface diseases of HD, such as preventing the onset of the disease or delaying the onset of the disease by inhibiting polyQ-mediated protein aggregation and aggregation toxicity, however, the treatment method has high cost, larger side effect and very little effect.
Therefore, there is still a need for a new product that can effectively relieve huntington's disease with little side effects and good safety.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, the invention provides an application of lactobacillus mucilaginosus in preparing products for preventing or treating neurodegenerative diseases.
The invention also provides an application of the culture of the fermented lactobacillus mucilaginosus in preparing products for preventing or treating neurodegenerative diseases.
The invention also provides application of the fermentation lactobacillus mucilaginosus metabolite in preparing products for preventing or treating neurodegenerative diseases.
The invention also provides application of the extract of the fermented lactobacillus mucilaginosus in preparing products for preventing or treating neurodegenerative diseases.
The invention also provides application of the microbial inoculum containing the fermented lactobacillus mucilaginosus in preparing products for preventing or treating neurodegenerative diseases.
In a first aspect, the invention provides the use of a fermented lactobacillus mucilaginosus for the preparation of a product for the prevention or treatment of neurodegenerative diseases, wherein,
the neurodegenerative disease comprises at least one of Huntington chorea, alzheimer disease, parkinson disease, amyotrophic lateral sclerosis, senile dementia, frontotemporal dementia and dementia with lewy bodies.
The application according to the embodiment of the invention has at least the following beneficial effects: the fermented lactobacillus mucilaginosus is applied to preparing products for preventing or treating neurodegenerative diseases, can effectively improve the neuron survival proportion of the caenorhabditis elegans, and compared with a blank control group, the added lactobacillus mucilaginosus LFP001 can obviously improve the neuron survival proportion of the caenorhabditis elegans.
In addition, the invention discovers the application of the fermented lactobacillus mucilaginosus (LFP fectus 001) in the medicine for preventing or delaying the Huntington disease for the first time, and can provide a reference theoretical basis for further development, utilization and popularization of the fermented lactobacillus mucilaginosus.
In a second aspect of the invention, there is provided the use of a culture of fermented lactobacillus mucilaginosus in the manufacture of a product for the prevention or treatment of neurodegenerative diseases.
In a third aspect of the invention, the use of a fermented lactobacillus mucilaginosus metabolite in the manufacture of a product for the prevention or treatment of neurodegenerative diseases is presented.
In a fourth aspect of the invention, the use of an extract of lactobacillus mucilaginosus for the preparation of a product for the prevention or treatment of neurodegenerative diseases is proposed.
In a fifth aspect, the invention provides an application of a microbial inoculum containing fermented lactobacillus mucilaginosus in preparing a product for preventing or treating neurodegenerative diseases.
In some embodiments of the invention, the neurodegenerative disease is huntington's disease.
In some embodiments of the invention, the lactobacillus fermentum is lactobacillus fermentum LFPerfectus001 (Limosilactobacillus fermentum);
preferably, the lactobacillus fermentum LFPerfectus001 has the amino acid sequence of SEQ ID NO:1, and a 16S rDNA sequence shown in the formula 1.
In some embodiments of the invention, the products include pharmaceuticals and functional foods.
In some embodiments of the invention, the dosage form of the pharmaceutical product is in solid, semi-solid or liquid form;
preferably, the dosage form of the drug is an aqueous solution, a non-aqueous solution, a suspension or a paste;
more preferably, the medicament is in the form of tablets, capsules, granules, pills, oral liquid, emulsion, dry suspension, dry extract or injection.
In some embodiments of the present invention, the preparation raw materials of the medicine may further include pharmaceutical excipients.
In some preferred embodiments of the present invention, the pharmaceutical excipients are conventional pharmaceutical carriers in the art, and may be any suitable physiologically or pharmaceutically acceptable pharmaceutical excipients;
preferably, the pharmaceutical excipients are selected from at least one of disintegrants, diluents, lubricants, binders, wetting agents, flavoring agents, suspending agents, surfactants, preservatives;
more preferably, the disintegrating agent is at least one selected from corn starch, potato starch, crosslinked polyvinylpyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crosslinked sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, alginic acid;
more preferably, the diluent is selected from at least one of lactose, sucrose, mannitol, corn starch, potato starch, calcium phosphate, calcium citrate, crystalline cellulose;
more preferably, the lubricant is at least one selected from the group consisting of silica gel micropowder, magnesium stearate, calcium stearate, stearic acid, talc, and anhydrous silica gel;
more preferably, the binder is at least one selected from acacia, gelatin, dextrin, hydroxypropyl cellulose, methylcellulose, polyvinylpyrrolidone;
more preferably, the wetting agent is selected from sodium dodecyl sulfate;
more preferably, the flavoring agent is at least one selected from aspartame, stevioside, sucrose, maltitol, citric acid;
more preferably, the suspending agent is at least one selected from acacia, gelatin, methylcellulose, sodium carboxymethylcellulose, hydroxymethyl cellulose, and aluminum stearate gel;
more preferably, the surfactant is at least one selected from lecithin, sorbitan monooleate, and glyceryl monostearate;
more preferably, the preservative is selected from at least one of methylparaben or propylparaben.
In some embodiments of the invention, the functional food comprises at least one of a decoction, a beverage, a candy, an oral liquid, a capsule, a tablet, and a powder.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Drawings
The invention is further described with reference to the accompanying drawings and examples, in which:
FIG. 1 is a neuron fluorescence map of caenorhabditis elegans according to an embodiment of the present invention.
FIG. 2 is a graph showing the neuroprotective efficacy of L.mucilaginosa LFP001 against Huntington's disease according to an embodiment of the present invention.
FIG. 3 is a graph showing the neuroprotective efficacy of different strains against Huntington's disease according to an embodiment of the present invention.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
In the description of the present invention, the descriptions of the terms "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
In an embodiment of the invention, HA759 nematodes are purchased from U.S. CGC (The Caenorhabditis Genetics Center (CGC; http:// www.cbs.umn.edu/CGC).
In an embodiment of the present invention, lactobacillus fermentum LFPerfectus001 (Limosilactobacillus fermentum) (hereinafter, abbreviated as LFP001, patent application No. 201610895895.9) has a preservation number of CGMCC No.10049, a preservation time of 2014, 11 months and 24 days, a preservation unit of China general microbiological culture Collection center, and a preservation address of national institute of microbiological culture Collection, national institute of sciences, national institute of advanced, national center for sciences, 1, 3.
The 16S rDNA sequence of the lactobacillus mucilaginosus LFP001 is as follows:
CGAACGCGTTGGCCCAATTGATTGATGGTGCTTGCACCTGATTGATTTTGGTCGCCA
ACGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCTGCCCAGAAGCGGGGGACAACA
TTTGGAAACAGATGCTAATACCGCATAACAACGTTGTTCGCATGAACAACGCTTAAAAG
ATGGCTTCTCGCTATCACTTCTGGATGGACCTGCGGTGCATTAGCTTGTTGGTGGGGTAA
CGGCCTACCAAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACAATGGGAC
TGAGACACGGCCCATACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGGCGC
AAGCCTGATGGAGCAACACCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGT
TGTTAAAGAAGAACACGTATGAGAGTAACTGTTCATACGTTGACGGTATTTAACCAGAA
AGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCG
GATTTATTGGGCGTAAAGAGAGTGCAGGCGGTTTTCTAAGTCTGATGTGAAAGCCTTCG
GCTTAACCGGAGAAGTGCATCGGAAACTGGATAACTTGAGTGCAGAAGAGGGTAGTGG
AACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCG
GCTACCTGGTCTGCAACTGACGCTGAGACTCGAAAGCATGGGTAGCGAACAGGATTAGA
TACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAGGTGTTGGAGGGTTTCCGCCCTTC
AGTGCCGGAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAA
CTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCT
ACGCGAAGAACCTTACCAGGTCTTGACATCTTGCGCCAACCCTAGAGATAGGGCGTTTC
CTTCGGGAACGCAATGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTT
GGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCA
CTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAGATCATCA
TGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTCGCGAAC
TCGCGAGGGCAAGCAAATCTCTTAAAACCGTTCTCAGTTCGGACTGCAGGCTGCAACTC
GCCTGCACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTT
CCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGT
GGGGTAACCTT(SEQ ID NO：1)。
in an embodiment of the invention, caenorhabditis elegans growth solid medium (NGM) is formulated as follows: 3g of NaCl, 2.5g of peptone and 20g of agar are respectively weighed, 975mL of distilled water is added, and after uniform mixing, the mixture is sterilized at the high temperature of 121 ℃ for 30min and then quickly transferred into a 55 ℃ incubator. Taking out the plate before pouring, and adding the following sterilized solutions: 1mol/L CaCl 2 、1mol/L MgSO 4 5mg/L cholesterol solution (ethanol formulation without sterilization) 1mL each and 1mol/L phosphate buffer 25mL; 20mL of each 9cm dish was prepared and sterilized NGM plate was prepared.
S.M buffer was formulated as follows: 2.925g of sodium chloride, 3g of monopotassium phosphate, 0.66g of dipotassium phosphate and 500mL of distilled water are weighed, mixed, sterilized at a high temperature of 121 ℃ for 30min, cooled to room temperature, added with 0.5mL of 5mg/mL cholesterol ethanol solution, 1.5mL of 1mol/L calcium chloride solution, 1.5mL of 1mol/L magnesium sulfate solution, 5mL of 1mol/L potassium citrate solution and 5mL of trace element solution, and shaken well for later use.
Example 1: neuroprotective efficacy test of lactobacillus mucilaginosus LFP001 against huntington's disease
1. Test method
After synchronizing HA759 nematodes at spawning, eggs were collected and incubated in sterile caenorhabditis elegans growth solid medium (NGM), after 24h L1 stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), 100. Mu.L of concentrated NA22 was added to each well (if not noted individually, all bacteria concentrations were at OD after 10-fold dilution) 570 0.5) and 0%, 4%, 8% of lactobacillus mucilaginosus LFP001.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
The number of nematodes with 2 spots in the head is shown as a in fig. 1, indicating ASH neuron survival; the number of nematodes with 1 fluorescent spot in the head is shown as b in fig. 1, indicating ASH neuron death.
2. Test results
The neuroprotective effect of the fermented lactobacillus mucilaginosus LFP001 on the Huntington disease is shown in a graph in FIG. 2, compared with a blank group (0), after 4 percent and 8 percent of the fermented lactobacillus mucilaginosus LFP001 are added, the neuron survival proportion is obviously improved by 16.07 percent and 17.20 percent (p is less than 0.0001), and the result shows that the fermented lactobacillus mucilaginosus LFP001 can effectively improve the neuron survival proportion of the Huntington disease model caenorhabditis elegans when the addition amount is 4 percent and 8 percent, and has obvious neuroprotective effect; the effect is better when the addition amount is 8%.
Comparative example 1: neuroprotective efficacy test of Lactobacillus fermentum CECT5716 against Huntington disease
1. Test method
After synchronizing the HA759 nematodes at spawning, eggs were collected and incubated in caenorhabditis elegans growth solid medium (NGM), after 24h L1-stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), and 100. Mu.L of concentrated NA22, 8% Lactobacillus fermentum CECT5716 was added to each well.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
2. Test results
The results of the neuroprotective efficacy of lactobacillus fermentum CECT5716 against huntington's disease are shown in fig. 3, and it is apparent from fig. 3 that lactobacillus fermentum CECT5716 does not have the neuroprotective efficacy against huntington's disease.
Comparative example 2: neuroprotective efficacy test of bifidobacterium animalis Bb-12 against Huntington disease
1. Test method
After synchronizing the HA759 nematodes at spawning, eggs were collected and incubated in caenorhabditis elegans growth solid medium (NGM), after 24h L1-stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), and 100. Mu.L of concentrated NA22, 8% bifidobacterium animalis Bb-12 was added to each well.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
2. Test results
The result of the neuroprotection effect of bifidobacterium animalis Bb-12 on Huntington disease is shown in FIG. 3, and it is apparent from FIG. 3 that bifidobacterium animalis Bb-12 does not have the neuroprotection effect on Huntington disease.
Comparative example 3: neuroprotective efficacy test of lactobacillus rhamnosus HN001 against huntington's disease
1. Test method
After synchronizing the HA759 nematodes at spawning, eggs were collected and incubated in caenorhabditis elegans growth solid medium (NGM), after 24h L1-stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), and 100. Mu.L of concentrated NA22, 8% Lactobacillus rhamnosus HN001 was added to each well.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
2. Test results
The results of the neuroprotective efficacy of lactobacillus rhamnosus HN001 against huntington's disease are shown in fig. 3, and it is apparent from fig. 3 that lactobacillus rhamnosus HN001 does not have the neuroprotective efficacy against huntington's disease.
Comparative example 4: neuroprotective efficacy test of lactobacillus rhamnosus GG against huntington's disease
1. Test method
After synchronizing the HA759 nematodes at spawning, eggs were collected and incubated in caenorhabditis elegans growth solid medium (NGM), after 24h L1-stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), and 100. Mu.L of concentrated NA22, 8% Lactobacillus rhamnosus GG was added to each well.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
2. Test results
The results of the neuroprotective efficacy of lactobacillus rhamnosus GG against huntington's disease are shown in fig. 3, and it is apparent from fig. 3 that lactobacillus rhamnosus GG does not have the neuroprotective efficacy against huntington's disease.
Comparative example 5: neuroprotective efficacy test of Lactobacillus acidophilus NCFM against Huntington disease
1. Test method
After synchronizing the HA759 nematodes at spawning, eggs were collected and incubated in caenorhabditis elegans growth solid medium (NGM), after 24h L1-stage larvae were collected in 50mL Erlenmeyer flasks with S.M buffer, the density of larvae was adjusted to 5/10. Mu.L and transferred to 24 well plates (1000. Mu.L/well), and 100. Mu.L of concentrated NA22, 8% Lactobacillus acidophilus NCFM was added to each well.
Placing the above-mentioned pore plate at 20deg.C and 150rpm for shaking culture for 42 hr (L4 stage), adding 5-FUDR (final concentration of 75 μg/mL) to inhibit nematode oviposition; after 65h of culture (D1 phase), the caenorhabditis elegans of each group are respectively taken out and collected in an EP tube, the buffer solution S.M is used for cleaning to ensure that the supernatant is clear and transparent, then the pure worm solution at the bottom is discarded, 100 mu L of absolute ethyl alcohol is added for immediately and uniformly mixing, about 1.5mL of S.M buffer solution is added after 5min of nematode death to adjust the worm density to 20/80 mu L, the mixture is transferred into 384-well plates, 80 mu L of worm solution is filled into each well, 10 compound wells are filled into each group, and fluorescent photographing counting is carried out to obtain the survival rate of ASH neurons.
Wherein the survival rate of ASH neurons is calculated as follows:
neuronal survival = number of nematodes with 2 spots in the head/(number of nematodes with 2 spots in the head + number of nematodes with 1 spot in the head).
2. Test results
The neuroprotective effect of lactobacillus acidophilus NCFM against huntington's disease results are shown in fig. 3, and it is apparent from fig. 3 that lactobacillus acidophilus NCFM does not have the neuroprotective effect of huntington's disease.
From this, it was found that the neuronal survival rate of caenorhabditis elegans was significantly increased by 20.19% (p < 0.01) after the addition of lactobacillus mucilaginosus LFP001 compared to the blank group (i.e. no 8% of the test strain was added, and the other was the same as in example 1) under the same experimental conditions at the same addition level (8%), whereas the neuronal survival rate of caenorhabditis elegans was not significantly increased by the addition of CECT5716 belonging to lactobacillus mucilaginosus; the neuron survival proportion of the caenorhabditis elegans is not obviously improved by adding other strains of bifidobacterium animalis Bb-12, lactobacillus rhamnosus HN001, lactobacillus rhamnosus GG and lactobacillus acidophilus NCFMLPF 001.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.
Claims (10)
1. Use of fermented lactobacillus mucilaginosus in the preparation of a product for preventing or treating neurodegenerative diseases, wherein,
the neurodegenerative disease comprises at least one of Huntington chorea, alzheimer disease, parkinson disease, amyotrophic lateral sclerosis, senile dementia, frontotemporal dementia and dementia with lewy bodies.
2. Use of a culture of fermented lactobacillus mucilaginosus for the preparation of a product for the prevention or treatment of neurodegenerative diseases.
3. Use of a fermented lactobacillus mucilaginosus metabolite in the preparation of a product for preventing or treating neurodegenerative diseases.
4. Use of an extract of lactobacillus mucilaginosus for the preparation of a product for the prevention or treatment of neurodegenerative diseases.
5. Use of a microbial inoculum containing fermented lactobacillus mucilaginosus in the preparation of a product for preventing or treating neurodegenerative diseases.
6. The use according to any one of claims 1 to 5, wherein the neurodegenerative disease is huntington's disease.
7. The use according to claim 6, wherein the lactobacillus fermentum is lactobacillus fermentum LFPerfectus001 (Limosilactobacillus fermentum);
preferably, the lactobacillus fermentum LFPerfectus001 has the amino acid sequence of SEQ ID NO:1, and a 16S rDNA sequence shown in the formula 1.
8. The use according to claim 7, wherein the products include pharmaceuticals and functional foods.
9. The use according to claim 8, wherein the pharmaceutical product is in the form of a solid, semi-solid or liquid.
10. The use according to claim 8, wherein the functional food is in the form of at least one of a decoction, a beverage, a candy, an oral liquid, a capsule, a tablet, a powder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211578249.1A CN116173078A (en) | 2022-12-06 | 2022-12-06 | Application of fermented lactobacillus mucilaginosus in preparation of products for preventing or delaying huntington chorea |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211578249.1A CN116173078A (en) | 2022-12-06 | 2022-12-06 | Application of fermented lactobacillus mucilaginosus in preparation of products for preventing or delaying huntington chorea |
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| Publication Number | Publication Date |
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| CN116173078A true CN116173078A (en) | 2023-05-30 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202211578249.1A Pending CN116173078A (en) | 2022-12-06 | 2022-12-06 | Application of fermented lactobacillus mucilaginosus in preparation of products for preventing or delaying huntington chorea |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117721033A (en) * | 2023-10-13 | 2024-03-19 | 广东悦创生物科技有限公司 | Lactobacillus mucilaginosus KS6 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines |
-
2022
- 2022-12-06 CN CN202211578249.1A patent/CN116173078A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117721033A (en) * | 2023-10-13 | 2024-03-19 | 广东悦创生物科技有限公司 | Lactobacillus mucilaginosus KS6 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines |
| CN117721033B (en) * | 2023-10-13 | 2024-05-17 | 广东悦创生物科技有限公司 | Lactobacillus mucilaginosus KS6 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines |
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