Agaricus blazei murill extract and application thereof in treating neurodegenerative diseases

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an agaricus blazei murill extract and application thereof in treatment of neurodegenerative diseases, in particular to application of the agaricus blazei murill extract FA-2-b-beta in prevention and / or treatment of the neurodegenerative diseases. Experimental data provided by the invention strongly confirms the potential of FA-2-b-beta as an active ingredient in treatment of neurodegenerative diseases, and FA-2-b-beta can inhibit activation of NF-kappa B, down-regulate expression of NLRP3 protein and inhibit pyroptosis of BV2 cells so as to improve neurological function impairment and has great clinical application value.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to an Agaricus blazei Murrill extract and use thereof in treating neurodegenerative diseases, and in particular to use of the Agaricus blazei Murrill extract FA-2-b-β in the prevention and / or treatment of neurodegenerative diseases. Background Art

[0002] Alzheimer's disease (AD) is a neurodegenerative disease with memory impairment, cognitive impairment and behavioral abnormalities as its main clinical manifestations, which seriously affects the quality of life of the elderly. The pathological characteristics of AD include the accumulation of amyloid beta (Aβ) and the formation of neurofibrillary tangles (NFTs) in the brain, among which the aggregation of Aβ is considered to be one of the core factors in the onset of AD. 1-42 It is a highly neurotoxic Aβ subtype that can trigger pathological processes such as oxidative stress, inflammatory response, and neuronal damage, thereby aggravating the development of AD.

[0003] Neuroinflammation, as one of the important pathological processes of AD, is mainly mediated by the innate immune cells in the brain - microglia. Microglia play a dual role in the Aβ pathogenesis of AD. Under normal circumstances, microglia help eliminate Aβ aggregates through phagocytosis. However, under pathological conditions, excessive production of Aβ leads to overactivation of microglia. Overactivated microglia produce more reactive oxygen species (ROS) and proinflammatory factors, and show degeneration of phagocytic ability, which ultimately leads to aggravated neuronal damage. The cellular contents released by damaged cells further promote microglial activation and enhance the inflammatory response. At this stage, removing the initial inflammatory stimulus, such as Aβ, does not alleviate neuroinflammation, which may be the reason why anti-amyloid protein treatment strategies are ineffective in late-stage AD. Therefore, inhibiting Aβ 1-42 Induced neuroinflammation, especially by modulating pathways of microglial activation, may be an important strategy for preventing and treating AD.

[0004] In the inflammatory signaling pathway, nuclear factor-κB (NF-κB) is a key transcription factor that is involved in regulating the expression of multiple inflammation-related genes. Excessive activation of the NF-κB signaling pathway not only promotes the pro-inflammatory state of microglia, but also exacerbates the neuropathological changes of AD. Therefore, inhibiting the activation of the NF-κB signaling pathway may be an effective anti-inflammatory strategy with potential value in the treatment of AD.

[0005] In recent years, natural products have received extensive attention in the treatment of neurodegenerative diseases. As a selenium-rich Chinese herbal medicine, the active ingredients of Agaricus blazei Murrill have shown significant biological effects in anti-oxidation, anti-inflammation and immunomodulation. In particular, the Agaricus blazei Murrill extract FA-2-b-β was found to have strong anti-inflammatory activity and can play a protective role against a variety of inflammatory diseases by regulating multiple cell signaling pathways. However, whether the Agaricus blazei Murrill extract FA-2-b-β can regulate Aβ by inhibiting the NF-κB signaling pathway 1-42 Mediated neuroinflammation has not been systematically studied. Summary of the invention

[0006] In view of the deficiencies in existing treatments for neuroinflammation, the present invention provides an application of an Agaricus blazei Murrill extract FA-2-b-β in preventing and / or treating neurodegenerative diseases, so as to solve the problems existing in existing treatments.

[0007] In order to achieve the above technical objectives, the present invention hereby proposes the following technical solutions:

[0008] One of the purposes of the present invention is to provide a use of a plant extract in preparing a drug for inhibiting neuroinflammation, wherein the plant extract contains FA-2-b-β.

[0009] The second object of the present invention is to provide a use of a plant extract in preparing a drug for preventing and / or treating a disease caused by Aβ accumulation, wherein the plant extract contains FA-2-b-β.

[0010] In some embodiments, the Aβ is preferably Aβ 1-42 .

[0011] In some embodiments, the disease caused by Aβ accumulation comprises a neurodegenerative disease.

[0012] In some preferred embodiments, the disease caused by Aβ accumulation is selected from at least one of Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS).

[0013] In some more preferred embodiments, the disease caused by Aβ accumulation is Alzheimer's disease.

[0014] In some embodiments, the plant extract is preferably an Agaricus blazei murill extract.

[0015] In some preferred embodiments, the plant extract is Kangzhen selenium-rich Agaricus blazei extract.

[0016] In some embodiments, the total selenium content of the plant extract is greater than 80 mg / kg, greater than 85 mg / kg, greater than 90 mg / kg, or greater than 95 mg / kg.

[0017] In some preferred embodiments, the total selenium content of the plant extract can be 80mg / kg, 81mg / kg, 82mg / kg, 83mg / kg, 84mg / kg, 85mg / kg, 86mg / kg, 87mg / kg, 88mg / kg, 89mg / kg, 90mg / kg, 91mg / kg, 92mg / kg, 93mg / kg, 94mg / kg, 95mg / kg, 96mg / kg, 97mg / kg, 98mg / kg, 99mg / kg, wherein mg is the mass of total selenium and kg is the mass of the fruiting body of the plant.

[0018] In some embodiments, the percentage of organic selenium content of the plant extract to the total selenium content is greater than 80%, greater than 85%, greater than 90%, greater than 95%, or greater than 99%.

[0019] In some preferred embodiments, the percentage of organic selenium content in the plant extract to the total selenium content is greater than 99%.

[0020] In some more preferred embodiments, the percentage of organic selenium content of the plant extract to the total selenium content can be 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98% or 99.99%.

[0021] In some embodiments, the plant extract comprises 90 mg / kg or more, 95 mg / kg or more, 100 mg / kg or more, 110 mg / kg or more, 120 mg / kg or more, 130 mg / kg or more, 140 mg / kg or more, 150 mg / kg or more, 160 mg / kg or more, 170 mg / kg or more, 180 mg / kg or more, 190 mg / kg or more, 200 mg / kg or more, 250 mg / kg or more, or 300 mg / kg or more of FA-2-b-β, wherein mg is the mass of FA-2-b-β and kg is the mass of the fruiting body of the plant.

[0022] The third object of the present invention is to provide a pharmaceutical composition comprising a plant extract as a pharmaceutically active ingredient and a pharmaceutically acceptable excipient, wherein the plant extract comprises FA-2-b-β.

[0023] In some embodiments, the plant extract comprises an Agaricus blazei extract.

[0024] In some preferred embodiments, the plant extract is Kangzhen selenium-rich Agaricus blazei extract.

[0025] In some embodiments, the total selenium content of the plant extract is greater than 80 mg / kg, greater than 85 mg / kg, greater than 90 mg / kg, or greater than 95 mg / kg.

[0026] In some preferred embodiments, the total selenium content of the plant extract can be 80mg / kg, 81mg / kg, 82mg / kg, 83mg / kg, 84mg / kg, 85mg / kg, 86mg / kg, 87mg / kg, 88mg / kg, 89mg / kg, 90mg / kg, 91mg / kg, 92mg / kg, 93mg / kg, 94mg / kg, 95mg / kg, 96mg / kg, 97mg / kg, 98mg / kg, 99mg / kg, wherein mg is the mass of total selenium and kg is the mass of the fruiting body of the plant.

[0027] In some embodiments, the percentage of organic selenium content of the plant extract to the total selenium content is greater than 80%, greater than 85%, greater than 90%, greater than 95%, or greater than 99%.

[0028] In some preferred embodiments, the percentage of organic selenium content in the plant extract to the total selenium content is greater than 99%.

[0029] In some more preferred embodiments, the percentage of organic selenium content of the plant extract to the total selenium content can be 99.0%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98% or 99.99%.

[0030] In some embodiments, the plant extract comprises 90 mg / kg or more, 95 mg / kg or more, 100 mg / kg or more, 110 mg / kg or more, 120 mg / kg or more, 130 mg / kg or more, 140 mg / kg or more, 150 mg / kg or more, 160 mg / kg or more, 170 mg / kg or more, 180 mg / kg or more, 190 mg / kg or more, 200 mg / kg or more, 250 mg / kg or more, or 300 mg / kg or more of FA-2-b-β, wherein mg is the mass of FA-2-b-β and kg is the mass of the fruiting body of the plant.

[0031] In some embodiments, the pharmaceutical composition is used to inhibit neuroinflammation.

[0032] In some embodiments, the pharmaceutical composition is used to prevent and / or treat a disease caused by Aβ accumulation.

[0033] In some preferred embodiments, the pharmaceutical composition is used to prevent and / or treat neurodegenerative diseases.

[0034] In some preferred embodiments, the pharmaceutical composition is used to prevent and / or treat Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis or multiple sclerosis.

[0035] In some preferred embodiments, the pharmaceutical composition is used to prevent and / or treat Alzheimer's disease.

[0036] A fourth object of the present invention is to provide a use of any of the aforementioned pharmaceutical compositions in the preparation of a drug for inhibiting neuroinflammation.

[0037] A fifth object of the present invention is to provide use of any of the aforementioned pharmaceutical compositions in the preparation of a drug for preventing and / or treating diseases caused by Aβ accumulation.

[0038] In some embodiments, the disease caused by Aβ accumulation comprises a neurodegenerative disease.

[0039] In some embodiments, the disease caused by Aβ accumulation comprises Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, or multiple sclerosis.

[0040] In some preferred embodiments, the disease caused by Aβ accumulation is Alzheimer's disease.

[0041] In the present invention, the dosage forms of the drugs or pharmaceutical compositions provided by the present invention include but are not limited to ordinary tablets, double-layer tablets, multi-layer tablets, sustained-release tablets, single-chamber controlled-release tablets, double-chamber controlled-release tablets, microporous controlled-release tablets, sublingual tablets, orally rapidly disintegrating tablets, dispersible tablets, enteric-coated tablets, granules, pills, enteric-coated capsules, delayed-release tablets, timed / positioned release tablets, ordinary capsules, sustained-release capsules, controlled-release capsules, capsules containing micropellets or small tablets, pH-dependent capsules containing micropellets or small tablets, oral liquids, films or patches, etc. It should be particularly noted that the pharmaceutical composition is made into tablets or capsules.

[0042] In the present invention, the term pharmaceutically acceptable carrier or excipient refers to those substances known in the art that can serve as fillers or carrier materials in tablets, pills, capsules, etc. Usually these substances are approved by health administration agencies for this purpose, and they are inactive as pharmaceutical agents. "Pharmaceutical Excipient Handbook" (A.Wade and PJWeller, ed., 2nd edition, American Pharmaceutical Association, Washington and Pharmaceutical Press, London, 1994) compiled pharmaceutically acceptable carriers and excipients. In particular, lactose, starch, cellulose derivatives, etc., and mixtures thereof can be used as carriers of the active components of the present composition.

[0043] In the present invention, pharmaceutically acceptable carriers can be made into common oral preparations, including common tablets, common capsules, granules, etc. When made into tablets, the pharmaceutically acceptable carriers include excipients and adjuvants that help to formulate the active compounds into pharmaceutical preparations, such as starch, microcrystalline cellulose, inorganic salts, sucrose, dextrin, lactose, powdered sugar, glucose, sodium chloride, cysteine, citric acid and sodium sulfite, etc., which are common knowledge in the art.

[0044] The present invention provides an application of an Agaricus blazei extract FA-2-b-β in preventing and / or treating neurodegenerative diseases. Experimental data strongly confirm the potential of FA-2-b-β as an active ingredient in treating neurodegenerative diseases. FA-2-b-β can inhibit the activation of NF-κB, downregulate the expression of NLRP3 protein, and inhibit the pyroptosis of BV2 cells, thereby improving neural function damage, and has great clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 Agaricus blazei extract FA-2-b-β and Aβ 1-42 Effects on the survival rate of BV-2 microglia cells.

[0046] Figure 2 The effect of FA-2-b-β on Aβ 1-42 The effect of NF-κB signaling pathway activation and inflammatory level in BV-2 microglia was mediated by TNF-α.

[0047] Figure 3 It was shown that Agaricus blazei extract FA-2-b-β regulates Aβ through NLRP3 pathway 1-42 Mediated inflammatory response.

[0048] Figure 4 and Figure 5 The method for preparing the Agaricus blazei Murrill extract FA-2-b-β is exemplarily described. DETAILED DESCRIPTION

[0049] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly used in the field to which the present invention belongs. For the purpose of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.

[0050] Unless the context clearly dictates otherwise, the expressions "a", "an" and "an" as used herein include plural references. For example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art, and so forth.

[0051] As used herein, the term "about" refers to a range of ±20% of the value that follows. In some embodiments, the term "about" refers to a range of ±10% of the value that follows. In some embodiments, the term "about" refers to a range of ±5% of the value that follows.

[0052] Numerical ranges used herein should be understood to include all numbers within the range. For example, a range of 1 to 20 should be understood to include any number, combination of numbers, or subrange from the following group: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.

[0053] As used herein, the term "comprises" or "comprising" means "including but not limited to". The term is intended to be open-ended to specify the presence of any stated features, elements, integers, steps, or components, but does not exclude the presence or addition of one or more other features, elements, integers, steps, components, or groups thereof. Thus, the term "comprising" encompasses the more restrictive terms "consisting of" and "consisting essentially of". In one embodiment, the term "comprising" as used throughout the application, especially in the claims, may be replaced by the term "consisting of".

[0054] The terms "optional", "either", "any", or "any" as used herein mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. As used herein, "a" and "an" are used herein to refer to one or more than one of the grammatical objects.

[0055] The term "and / or" used herein should be understood to mean any one of the optional items or a combination of any two or more of the optional items.

[0056] As used herein, "FA-2-b-β" or "Agaricus blazei extract RNA-protein complex" are used interchangeably, and both belong to the water-soluble effective component acidic RNA-protein complex of Agaricus blazei polysaccharide. The molecular weight of nucleic acid is 10000, and the base components include A, Ap, Gp, Cp, Up and other modified bases (SUN YQ, GUO TK, XI YM, et al. World journal of gastroenterology, 2007, 13 (31): 4185-4191.). The protein part is composed of polysaccharide: protein = 40: 60 w / w, and its sugar chain contains glucose, xylose, mannose, galactose and fucose (ZHANG J, WANG G, LI H, et al. Bioscience, biotechnology, and biochemistry, 1994, 58 (7): 1195-1201.).

[0057] As used herein, the term "pharmaceutical composition" generally refers to a preparation that is in a form that permits the biological activity of the active ingredient to be effective, and does not contain additional ingredients that are unacceptably toxic to the subject to which the composition is to be administered. The composition is sterile.

[0058] As used herein, the term "pharmaceutically acceptable" means that these structures, materials, compositions and / or dosage forms are suitable for contact with human and animal tissues within the scope of reasonable medical judgment, commensurate with a reasonable benefit / risk ratio, and do not produce excessive toxicity, irritation, allergic response or other problems or complications.

[0059] As used herein, the terms "treatment", "prevention", and the like refer to the administration of an agent or the performance of a procedure in order to obtain an effect. These effects may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of affecting a partial or complete cure of a disease and / or disease symptoms. As used herein, "treatment" may include treating a disease or condition (e.g., cancer) in a mammal, particularly a human, and includes: (a) preventing the occurrence of the disease or disease symptoms in a subject susceptible to the disease but not yet diagnosed as ill (e.g., including a disease that may be associated with or caused by the primary disease); (b) inhibiting the disease, i.e., preventing its development; (c) alleviating the disease, i.e., causing regression of the disease. Treatment may refer to any reference to success in treating or improving or preventing cancer, including any objective or subjective parameters, such as elimination; alleviation; reducing symptoms or making the disease condition more tolerable to the patient; slowing the rate of deterioration or decline; or reducing the debilitation of the endpoint of deterioration. The treatment or improvement of symptoms is based on one or more objective or subjective parameters; including the results of a doctor's examination. Therefore, the term "treatment" includes administering the antibodies or compositions or conjugates disclosed herein to prevent or delay, alleviate, stop or inhibit the development of symptoms or conditions associated with a disease (e.g., cancer). The term "therapeutic effect" refers to the reduction, elimination or prevention of a disease, disease symptom or disease side effect in a subject.

[0060] To make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the examples. If the specific conditions are not specified in the examples, they are carried out according to the normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for all reagents or instruments, they are all conventional products that can be purchased commercially. In order to better illustrate the present invention, numerous specific details are given in the specific embodiments below. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention. Such structures and technologies are also described in many publications, such as "Molecular Cloning Experiment Guide (Fourth Edition)" (Cold Spring Harbor Laboratory Science Press), Ausubel, FM et al., Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley-Interscience.

[0061] The Agaricus blazei extract FA-2-b-β of the present invention is obtained by the following method:

[0062] Provided by Kangzhen Biotechnology (Beijing) Co., Ltd., its composition and quality evaluation were tested by Beijing Qingxi Technology Research Institute. After being identified by Specimen Test, the total selenium content of the extract was 87 mg / kg and the organic selenium content was 86.8 mg / kg. The specific steps are as follows: The obtained Agaricus blazei extracts were dried in a 60°C drying oven for 72 hours and stored. When used, the Kangzhen selenium-rich Agaricus blazei extract was added with 70% methanol solution as an extraction reagent, and the Kangzhen selenium-rich Agaricus blazei extract was obtained by centrifugation as the mother liquor (pH = 7.4, osmotic pressure = 300mmol / L), which was stored in a -20°C refrigerator and diluted with cell culture medium to the corresponding experimental concentration before use. The method for culturing Agaricus blazei can be found in the inventor's prior application CN117581745A; the preparation method of Agaricus blazei extract FA-2-b-β is as follows Figure 4 and Figure 5 As shown (only exemplary description, other conventional extraction methods in the art can also be used in the present invention).

[0063] The purpose of the present invention is achieved through the following research methods:

[0064] Step 1: Cell culture and reagent preparation

[0065] Mouse BV-2 microglial cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin under the following conditions: 5% CO2, 37°C, and saturated humidity. 1-42 Preparation of solution: The lyophilized protein powder was dissolved in hexafluoroisopropanol to a final concentration of 1 mM, and the solution was placed in a fume hood overnight to remove the isopropanol. DMSO was then added to dissolve the Aβ peptide to a concentration of 5 mM, and then diluted to 100 μM with DMEM medium.

[0066] Step 2: Screening of FA-2-b-β and Aβ by CCK8 method 1-42 Optimal dosing concentration

[0067] Investigation of FA-2-b-β and Aβ 1-42 The CCK-8 assay was used to assess the viability of BV-2 cells. 3 cells (5.0×10 4 The cells were inoculated at a density of 100 / mL in a 96-well plate and allowed to grow for 24-36 h. 1-42 After being treated with FA-2-b-β for 24 or 36 h, the cells were incubated with 10 μL CCK-8 solution at 37°C in the dark for 1-2 h. The absorbance at a wavelength of 450 nm was measured using an ELISA reader.

[0068] Step 3: Cell transfection

[0069] BV-2 cells were collected at 1×10 5 Cells were seeded in 6-well plates at a density of 10 cells / well. Cells were transfected with siRNA to knock down NLRP3 using Lipofectamine 2000 according to the manufacturer's instructions. Transfection efficiency was detected by immunoblotting. The sequences of siRNAs used in this study are as follows: siRNA-1: 5'-GGUCUCCACUGAUGCCUCAtt-3' (SEQ ID NO: 1); siRNA-2: 5'-UGGAUGCUUCAAGGUGAAAtt-3 (SEQ ID NO: 2) siRNA-3: 5'-CCUCAAGGGAAGGUGGCAUtt-3' (SEQ ID NO: 3). In subsequent experiments, siRNAs with higher knockdown efficiency were used.

[0070] Step 4: Detection of related inflammatory factor levels by enzyme-linked immunosorbent assay (ELISA)

[0071] After the cells were collected, they were lysed in RIPA lysis buffer and then centrifuged at 13,000 × g for 10 min at 4°C. The supernatant was collected and the protein concentration was determined using the Boster BCA protein concentration assay kit. TNF-α, IL-1β, and IL-18 levels were detected using ELISA kits according to the manufacturer's instructions.

[0072] Step 5: RT-qPCR to detect the mRNA expression level of related proteins

[0073] Total RNA of microglia was extracted using a tissue / cell RNA rapid extraction kit, and then the extracted total RNA was reverse transcribed into cDNA. The PCR reaction conditions were set according to the manufacturer's instructions to detect the mRNA expression level. The test was repeated 3 times, and the RT-qPCR results were analyzed using the ΔΔCT method. The ΔCT value was the difference between the CT value of the target gene and the CT value of the internal reference gene (GAPDH), and the ΔΔCT was the difference between the ΔCT of each experimental group and the ΔCT of the blank control group. The average relative content = 2-ΔΔCT, which is the mRNA level relative to the normal group. The primers were designed by Sangon Biotech (Shanghai) Co., Ltd., and the primer sequences are shown in Table 1.

[0074] Table 1: Primer sequence information

[0075]

[0076] Step 6: Western Blot method to detect the expression of related proteins in each group

[0077] Total protein was extracted from BV-2 cells using RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors. After BCA protein quantification, 20 μg of protein per well was electrophoretically separated on a 10% PAGE gel and transferred to a methanol-pretreated PVDF membrane. After blocking with 5% skim milk solution at room temperature for 2 hours, the membrane was incubated with diluted primary antibodies at 4 ° C overnight. The next day, the PVDF membrane was incubated with secondary antibodies for 2 hours and then exposed.

[0078] Example 1: FA-2-b-β and Aβ 1-42 Effects on the survival rate of BV-2 microglia

[0079] FA-2-b-β was prepared as described above.

[0080] The viability of BV-2 cells was evaluated using the CCK-8 kit (Cell Counting Kit-8). BV-2 cells were plated at 5 × 10 3 cells (5.0×10 4 The cells were inoculated at a density of 100 / mL in a 96-well plate and allowed to grow for 24-36 h. 1-42 After 24 or 36 hours of treatment with FA-2-b-β, the cells were incubated with 10 μL CCK-8 solution at 37°C in the dark for 1-2 hours. The absorbance at a wavelength of 450 nm was detected using an ELISA reader. The results showed that the cell activity was not affected when the concentration of FA-2-b-β was 5 mg / mL. 1-42 When the concentration is 0.5-1μM, the cell activity is not affected ( Figure 1 The optimal concentrations screened were 5 mg / mL FA-2-b-β, 1 μM Aβ 1-42 , all subsequent experiments were conducted using this concentration.

[0081] Example 2: FA-2-b-β vs Aβ 1-42 Effects of NF-κB activation and inflammatory levels in BV-2 microglia mediated by

[0082] A blank control group, FA-2-b-β group, and Aβ 1-42 Group, FA-2-b-β+Aβ 1-42 The expression of NF-κB inflammatory pathway-related proteins (TLR4, p-IκB-α) and pyroptosis key proteins (NLRP3, caspase-1) were detected by WB and q-PCR respectively; the expression of inflammatory factors (TNF-α, IL-1β, IL-18) was analyzed by ELISA. The q-PCR results showed that compared with the control group, the expression of Aβ 1-42After treatment of microglia, the expression of TLR4 and p-IκB-α increased significantly, and NLRP3 and caspase-1 also increased significantly ( Figure 2 A), but both were higher than Aβ after FA-2-b-β treatment. 1-42 The Western Blot also showed the same results ( Figure 2 F). ELISA results showed that 1 μM Aβ 1-42 The release increased significantly after 18h and 2h of stimulation ( Figure 2 C, E). Subsequently, BV2 cells were intervened for 18h and 2h to detect the expression of inflammatory factors (TNF-α, IL-1β). The results showed that Aβ 1-42 After treatment, the addition of FA-2-b-β significantly reduced the secretion of TNF-α and IL-1β ( Figure 2 B and D). These results show that FA-2-b-β reduces Aβ 1-42 Mediated microglial pyroptosis and inflammation levels.

[0083] Example 3: FA-2-b-β regulates Aβ via the NLRP3 pathway 1-42 NF-κB activation and inflammatory response mediated by

[0084] According to the above experimental results, Aβ 1-42 Treatment of BV2 microglia can trigger its inflammatory response, and FA-2-b-β can inhibit this process. Our previous studies have found that NLRP3 plays a key role in this process. Virus transfection was used to construct a cell model with NLRP3 knockdown, and the transfection efficiency was detected by immunoblotting ( Figure 3 A).

[0085] Experimental group: Aβ 1-42 Group (control), Aβ 1-42 +siNLRP3 group, Aβ1-42+FA-2-b-β group. The results showed that compared with the simple Aβ 1-42 Compared with the intervention, knocking down NLRP3 and adding FA-2-b-β showed the same decrease in the levels of inflammatory substances such as NLRP3, Casp1 and ASC ( Figure 3 B and D), knocking down NLRP3 and adding FA-2-b-β also showed the same effect of reducing inflammatory response for cytokines such as (TNF-α, IL-1β) ( Figure 3 C).

[0086] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. Use of a plant extract in the preparation of a drug for inhibiting neuroinflammation, characterized in that: The plant extract comprises FA-2-b-β, wherein the plant extract is preferably an Agaricus blazei Murrill extract.

2. Use of a plant extract in the preparation of a medicament for preventing and / or treating a disease caused by Aβ accumulation, characterized in that: The plant extract comprises FA-2-b-β, wherein the plant extract is preferably an Agaricus blazei Murrill extract; Preferably, the disease caused by Aβ accumulation comprises a neurodegenerative disease, selected from at least one of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis and multiple sclerosis.

3. The use according to claim 1 or 2, characterized in that The total selenium content of the plant extract is above 80 mg / kg, above 85 mg / kg, above 90 mg / kg or above 95 mg / kg.

4. The use according to claim 3, characterized in that The percentage of organic selenium content in the plant extract to total selenium content is more than 80%, more than 85%, more than 90%, more than 95% or more than 99%.

5. The use according to claim 3, characterized in that: The plant extract contains 90 mg / kg or more, 95 mg / kg or more, 100 mg / kg or more, 110 mg / kg or more, 120 mg / kg or more, 130 mg / kg or more, 140 mg / kg or more, 150 mg / kg or more, 160 mg / kg or more, 170 mg / kg or more, 180 mg / kg or more, 190 mg / kg or more, 200 mg / kg or more, 250 mg / kg or more, or 300 mg / kg or more of FA-2-b-β.

6. A pharmaceutical composition, characterized in that The invention comprises a plant extract as a pharmaceutically active ingredient and a pharmaceutically acceptable excipient, wherein the plant extract comprises FA-2-b-β, and wherein the plant extract is preferably an Agaricus blazei Murrill extract.

7. The pharmaceutical composition according to claim 6, characterized in that The total selenium content of the plant extract is above 80 mg / kg, above 85 mg / kg, above 90 mg / kg or above 95 mg / kg.

8. The pharmaceutical composition according to claim 6, characterized in that The percentage of organic selenium content in the plant extract to total selenium content is more than 80%, more than 85%, more than 90%, more than 95% or more than 99%.

9. The pharmaceutical composition according to claim 6, characterized in that The plant extract contains 90 mg / kg or more, 95 mg / kg or more, 100 mg / kg or more, 110 mg / kg or more, 120 mg / kg or more, 130 mg / kg or more, 140 mg / kg or more, 150 mg / kg or more, 160 mg / kg or more, 170 mg / kg or more, 180 mg / kg or more, 190 mg / kg or more, 200 mg / kg or more, 250 mg / kg or more, or 300 mg / kg or more of FA-2-b-β.

10. Use of the pharmaceutical composition according to any one of claims 6 to 9 in the preparation of a drug for a disease caused by Aβ accumulation; Preferably, the disease caused by Aβ accumulation comprises a neurodegenerative disease, selected from at least one of Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis and multiple sclerosis.

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