CN111904994A - Application of traditional Chinese medicine composition in preparation of food, medicine or health-care food for preventing and treating neurodegenerative diseases - Google Patents
Application of traditional Chinese medicine composition in preparation of food, medicine or health-care food for preventing and treating neurodegenerative diseases Download PDFInfo
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- CN111904994A CN111904994A CN201910375482.1A CN201910375482A CN111904994A CN 111904994 A CN111904994 A CN 111904994A CN 201910375482 A CN201910375482 A CN 201910375482A CN 111904994 A CN111904994 A CN 111904994A
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/24—Apocynaceae (Dogbane family), e.g. plumeria or periwinkle
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/287—Chrysanthemum, e.g. daisy
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
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- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- Health & Medical Sciences (AREA)
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- Medicines Containing Plant Substances (AREA)
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to an application of a traditional Chinese medicine composition in preparing food, medicine or health-care food for preventing and treating neurodegenerative diseases, wherein the traditional Chinese medicine composition is prepared from the following raw material components in parts by weight: 1000-3500 parts of mesona chinensis benth, 135-900 parts of frangipani, 45-900 parts of microcos paniculata, 180-900 parts of chrysanthemum, 180-900 parts of honeysuckle, 180-900 parts of selfheal and 45-900 parts of liquorice. Through research, the following results are found: the traditional Chinese medicine composition can obviously relieve caenorhabditis elegans Poly-Q and Abeta1‑42The mediated neurotoxicity can be used for preventing and treating neurodegenerative diseases, and can be used for preparing foods, medicines or health-care foods for preventing and treating the neurodegenerative diseases.
Description
Technical Field
The invention relates to a new application of a traditional Chinese medicine composition, in particular to an application of a traditional Chinese medicine composition in preparing food, medicine or health-care food for preventing and treating neurodegenerative diseases.
Background
As the population aging progresses, Neurodegenerative Diseases (ND) have become a worldwide health problem. Neurodegenerative diseases are a group of irreversible diseases characterized by dysfunction of neurons due to brain, spinal cord neuron damage or myelin loss thereof, and mainly include Alzheimer's Disease (AD), Parkinson's Disease (PD), Amyotrophic Lateral Sclerosis (ALS), Huntington's Disease (HD), Spinocerebellar Ataxia (SA), Pick's small body disease, and the like, and the main pathogenesis thereof involves genetic, oxidative stress, mitochondrial injury, metal ion disorder, energy metabolism disorder, immune abnormality, autophagy, apoptosis, viral infection, and the like. At present, no targeted medicine is available for treating neurodegenerative diseases, and the existing medicine can only temporarily relieve the clinical symptoms of the neurodegenerative diseases, but still cannot prevent the progression of the neurodegenerative diseases.
Although these different neurodegenerative diseases have individual features: different etiology, pathological change parts and gene location, but the clinical manifestations are the decline of cognitive function, so that the common pathological characteristics can be achieved. Numerous studies have shown that the common pathological features of neurodegenerative diseases are all related to intracellular inclusions formed by polymerization and deposition of misfolded proteins, such as the old ones present in the brains of AD patientsAge spots (A beta)1-42) And neuro-tangles; polyglutamine (Poly Q) inclusion bodies in affected neurons in HD and SA patients; lewy bodies remaining in dopaminergic neurons in PD patients; bunina corpuscles in spinal cord and brainstem neurons in ALS patients. Initially, these characteristic pathologically altered neurons were not significantly reduced, while the cellular processes were significantly shortened or eliminated, and the pathologically altered neurons survived for a long period of time, gradually lost their biological function, and finally necrotic and apoptotic. Thus, inhibition of deposition or aggregation of these abnormal proteins (e.g., A.beta.1-42Poly Q, etc.) are expected to be effective intervention strategies against this disease.
Chinese patent CN102793038A discloses a Chinese medicinal composition, which comprises 1000-3500 parts of Mesona chinensis Benth, 135-900 parts of frangipani, 45-900 parts of microcos paniculata, 180-900 parts of chrysanthemum, 180-900 parts of honeysuckle, 180-900 parts of selfheal and 45-900 parts of liquorice. The raw material components are compatible with each other, and the tea has the effects of clearing fire, removing dampness, clearing heat, removing toxicity, promoting fluid production and quenching thirst. At present, no report about the prevention and treatment of neurodegenerative diseases by the traditional Chinese medicine composition exists.
Disclosure of Invention
Therefore, there is a need for an application of a Chinese medicinal composition in preparing foods, medicines or health-care foods for preventing and treating neurodegenerative diseases.
The application of a traditional Chinese medicine composition in preparing foods, medicines or health-care foods for preventing and treating neurodegenerative diseases is characterized in that the traditional Chinese medicine composition is prepared from the following raw material components in parts by weight:
1000-3500 parts of mesona chinensis benth, 135-900 parts of frangipani, 45-900 parts of microcos paniculata, 180-900 parts of chrysanthemum, 180-900 parts of honeysuckle, 180-900 parts of selfheal and 45-900 parts of liquorice.
In one embodiment, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
1500-2500 parts of mesona chinensis benth, 300-500 parts of frangipani, 300-500 parts of microcos paniculata, 300-500 parts of chrysanthemum, 300-500 parts of honeysuckle, 300-500 parts of selfheal and 300-500 parts of liquorice.
In one embodiment, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
1900-2500 parts of sianchau, 350-450 parts of frangipani, 350-450 parts of microcos paniculata, 350-450 parts of chrysanthemum, 350-450 parts of honeysuckle, 350-450 parts of selfheal and 350-450 parts of liquorice.
In one embodiment, the preparation method of the traditional Chinese medicine composition comprises the following steps:
preparing raw material components according to the formula amount, and adding water for extraction;
concentrating the obtained extractive solution to obtain concentrated solution.
In one embodiment, the water is used in an amount of 6 to 20 times the weight of the feed components.
In one embodiment, the extraction time is 0.5-3 h.
In one embodiment, the extraction is performed after soaking for 10-50 minutes after adding water.
In one embodiment, the concentration of the concentrated solution is 9-14 Be.
In one embodiment, the neurodegenerative disease is a disease of the type in which neurotoxicity is induced by polymerization and deposition of misfolded proteins including Poly-Q and/or a β into neurons1-42。
In one embodiment, the Chinese medicinal composition further comprises auxiliary materials or auxiliary components acceptable for food, medicine and health-care food.
In one embodiment, the food product is herbal tea.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a new application of a traditional Chinese medicine composition with active ingredients mainly comprising mesona chinensis benth, frangipani, microcos paniculata, chrysanthemum, honeysuckle, selfheal and liquorice. Through research, the following results are found: the traditional Chinese medicine composition can obviously relieve caenorhabditis elegans Poly-Q and Abeta1-42Mediated neurotoxicity for preventing and treating neurodegenerative diseases, and can be used for preparing medicine for preventing and treating neurodegenerative diseasesFood, medicine or health food, provides a new application of the traditional Chinese medicine composition, and also provides a new way for preventing and treating neurodegenerative diseases. In addition, the traditional Chinese medicine composition can be drunk as a daily beverage, has good adaptability and no side effect, and meets the requirements of modern people.
Drawings
FIG. 1A is a comparison of the CL4176 paralysis curves of C.elegans in the control and the Chinese medicinal composition group (P10);
FIG. 1B is a comparison of the curves of the control and the Chinese medicinal composition group (P10) for the paralysis of C.elegans AM 140.
Detailed Description
The application of the Chinese medicinal composition in preparing foods, medicines or health-care foods for preventing and treating neurodegenerative diseases is further described in detail with reference to specific examples.
Example 1
The embodiment is an application study of the traditional Chinese medicine composition in preparing food, medicine or health-care food with the effect of preventing and treating neurodegenerative diseases.
1. Materials and methods
1.1 Experimental reagent and apparatus
1.1.1 test reagents
The composition of the caenorhabditis elegans growth solid medium (NGM) comprises: NaCl, peptone, agar, streptomycin, cholesterol, CaCl2，MgSO4Nystatin. The caenorhabditis elegans lysate comprises the following components: NaOH, household bleach. Caenorhabditis elegans is N2Wild type caenorhabditis elegans (C.elegans, the Bristol strain N2) Hermaphrodite. Escherichia coli OP50 (Escherichia coli OP 50; E.coli OP50) is a uracil synthesis-deficient strain, grows slowly on NGM, and does not affect the clarity of observing C.elegans under a microscope.
Test sample of the traditional Chinese medicine composition: the feed is prepared from the following raw materials in parts by weight: 2200 parts of mesona chinensis benth, 400 parts of frangipani, 400 parts of microcos paniculata, 400 parts of chrysanthemum, 400 parts of honeysuckle, 400 parts of selfheal and 400 parts of liquorice. The preparation method comprises the following steps:
1) raw material treatment: selecting traditional Chinese medicine raw materials for preparing a traditional Chinese medicine composition sample, cleaning, cutting, and preparing according to a formula amount;
2) water extraction: adding water with the weight 6 times that of the raw materials into the raw materials, soaking for 15 minutes, and extracting for 1 hour to obtain water extract;
3) concentration: and concentrating the water extract to a concentration of 9Be to obtain a traditional Chinese medicine composition sample. The Chinese medicinal composition sample is sterilized and packaged to obtain the herbal tea. Or further adding pharmaceutically, health food, and food acceptable adjuvants to make into other foods besides drugs, health food or herbal tea. Wherein, the food is food in general meaning, and does not emphasize specific functions and specific population; the health food is a food with certain health care effect, has specific functions and is generally suitable for specific people.
1.1.2 devices (Table 1)
TABLE 1 main apparatus of experiment
2.2 preparation of test solutions and media
1M potassium phosphate buffer: KH (Perkin Elmer)2PO4 108.39g，K2HPO435.69g, add water to 1L, adjust pH to 6.0.
(iii) M9 buffer: KH (Perkin Elmer)2PO4 3g，Na2HPO4 6g，NaCl 5g，MgSO40.12g, adding distilled water to 1L, and autoclaving at 121 deg.C for 20 min.
③ C.elegans lysate: NaOH 0.1g, add 4mL pure water and 1mL NaCl, mix and shake. The caenorhabditis elegans lysate is prepared for use.
And fourth, LB liquid culture medium: LB broth 21g, adding water to 1L, sterilizing at 121 deg.C for 15 minutes, preferably using it as ready-to-use.
Solid culture Medium for caenorhabditis elegans Growth (NGM): 3g of sodium chloride, 17g of agar powder and 2.5g of tryptone, 975mL of distilled water is added, the mixture is autoclaved at 121 ℃ for 30min, and then 1mL of 5mg/mL of cholesterol (prepared by absolute ethyl alcohol) is added under aseptic conditions，1M CaCl2、1MMgSO41mL each and 25mL of 1M pH6.0 potassium phosphate buffer. After shaking, the medium was aseptically dispensed into petri dishes (55 mm diameter plates) at 10 mL/plate, left at room temperature for 2 to 3 days before use, and then coated with the broth.
2.3 Experimental methods
2.3.1 preparation of C.elegans
Culturing and subculturing caenorhabditis elegans
According to The literature method (S.Brenner, The Genetics of caenorhabditis elegans, Genetics77:71-94, May 1974), under The aseptic condition, a culture medium containing more caenorhabditis elegans is cut into pieces, and The pieces are transferred to a new NGM culture medium of OP50, and The caenorhabditis elegans can automatically climb to The more-food lawn from The edge of The less-food culture medium under The standard culture condition (20 ℃, The humidity is 40-60%). As the caenorhabditis elegans is easy to penetrate into the agar through the surface gaps of the culture medium, when the caenorhabditis elegans is cultured, an agar plate with smooth surface and no bubbles or scratches is selected as much as possible. Synchronization can be performed by culturing for about 48 hours.
② synchronization of caenorhabditis elegans
Sodium perchlorate bleaching (Brian Onken et al, Methfomin indices a direct reduction-like state and the oxidative stress to extended C.elegans fatty spa AMPK, LKB1, and SKN-1, PLOS one: January 2010, Volume5, Issue1, 8758) was used: under the aseptic condition, using M9 buffer solution sterilized at high temperature to wash caenorhabditis elegans in the oviposition stage into a 2mL aseptic tube, adding 1mL of lysate to crack the caenorhabditis elegans, oscillating for 3-5 min, placing the tube on a low-speed centrifuge for centrifugation at 3000rpm for 1min, discarding supernatant, washing the caenorhabditis elegans for 2 times by using M9, placing the tube on the low-speed centrifuge for centrifugation at 3000rpm for 1min, discarding supernatant, sucking the residual 0.1mL of caenorhabditis elegans at the bottom of the 2mL aseptic tube by using a pipette, dripping the residual caenorhabditis elegans in a sterile area of NGM, culturing under the standard culture condition, basically developing fertilized eggs in the cracked caenorhabditis elegans into L4 stage larvae after about 51h, and completing synchronization.
2.3.2 preparation of NGM without sample containing the Chinese medicinal composition
Culture of E.coli OP50
The strain OP50 was inoculated into 100mL LB liquid medium, and cultured with shaking at 37 deg.C and 170rpm for 12h until OD600 is 0.4, for inoculation of NGM to feed normal group of C.elegans. If the cultured bacteria liquid is not used immediately, the bacteria liquid is subpackaged by 10mL sterile tubes to about 8mL each tube and placed in a medical refrigerator at 4 ℃ for later use.
② preparation of NGM sample without the traditional Chinese medicine composition (contrast group)
An appropriate amount of OP50 bacterial liquid (generally a plate with a diameter of 55mm and 100 mu L) is added on each NGM plate, 100 mu L of OP50 bacterial liquid is taken by a pipette and evenly coated on the center of the NGM plate, and the edge of the bacterial liquid is as far as about 0.5cm away from the edge of the plate. The bacterial coated NGM flat plate can be used after being used at room temperature (21-25 ℃) overnight, and if the bacterial coated NGM flat plate is not used immediately, the sealed membrane is placed in a 4 ℃ medical refrigerator for later use.
③ preparation of the sample NGM containing the traditional Chinese medicine composition (traditional Chinese medicine composition group)
100mg of the traditional Chinese medicine composition samples are respectively weighed by a 10mL volumetric flask under the aseptic environment, the volume is adjusted to 10mL by OP50 bacterial liquid, and a traditional Chinese medicine composition sample with the concentration of 10mg/mL is prepared (P10). The coating method is the same as the preparation of the sample NGM without the traditional Chinese medicine composition. The plate was placed in a freezer at 4 ℃ for 2 weeks and used up as soon as possible.
2.3.3 the Chinese medicinal composition is used for treating A beta in caenorhabditis elegans CL41761-42Aggregation-induced neurotoxic effects
Caenorhabditis elegans transgene CL4176dvIs2[ pCL12(unc-54/human Abeta 1-42minigene) lpRF4]And (4) observation: reference is made to the literature methods (Martorell P, Bataller E, Llopis S, et al, A cocoa peptide precursors from oxidative stress and β -amyloid peptide toxicity [ J]PloS one, 2013, 8 (5): e63283) The mutant drives A beta micro gene to express human A beta through a body wall muscle cell myosin promoter1-42. In constructing the CL4176 transgenic strain, the smg-1mRNA monitoring system was used as a temperature induction system. The smg-1 system was active when the mutant was cultured at 15 ℃ for optimal growth temperature, and was able to recognize and degrade the A.beta.transgene with the wrong 3' noncoding region. E.g. increasing the temperature at larval stage 3 (L3)When the temperature is 25 ℃ which is not suitable for growth, smg-1 system is inactivated, and the transgenic fragment is human Abeta1-42mRNA can be expressed in the somatic cells of the mutant to form Abeta oligomers, and the caenorhabditis elegans gradually loses the motor ability within 24-36 hours. The temperature was raised to 25 ℃ to induce human Α β 1-42 oligomer expression when the contemporaneous worm eggs were cultured at 15 ℃ to L3. The traditional Chinese medicine composition treatment starts from eggs, when the caenorhabditis elegans enters the L3 stage, the temperature is increased to 25 ℃, and the movement of the caenorhabditis elegans is observed every 2 hours. When the head of the caenorhabditis elegans only does a micro-swing motion, the body does not do a rolling circle-drawing motion any more, and the body can not be induced to move under the touch stimulation, paralysis is defined. Because the paralyzed caenorhabditis elegans only feed on the slightly swayed head, a fan-shaped bacterial blank area (halos) is usually formed around the head, and the caenorhabditis elegans with halos can also be defined as paralyzed.
2.3.4 Effect of the Chinese medicinal composition on poly-Q aggregation-induced neurotoxicity in caenorhabditis elegans AM140
Caenorhabditis elegans mutant AM140(rmIsl32[ P (unc-54) Q35:: YFP ]): YFP is expressed in somatic muscle cells, poly-Q repeated extension mutation fused with the YFP is expressed, poly-Q progressive coalescence is insoluble aggregate along with the increase of age, and the paralysis of the caenorhabditis elegans is induced, and the paralysis of the caenorhabditis elegans is defined when the tail of the leptospira and the caenorhabditis elegans do not move forwards. The number of paralyzed caenorhabditis elegans was recorded daily until all caenorhabditis elegans were paralyzed.
2.3.5 data statistics
The survival and death numbers of the caenorhabditis elegans were recorded every 48h during their life cycle, and statistical analysis was performed on the data using Graph Pad Prism 5 software, SPSS Statistics 19.0 software, and Excel tables.
3. Results and analysis
3.1 the Chinese medicinal composition is used for treating A beta in caenorhabditis elegans CL41761-42Aggregation-induced neurotoxic effects
TABLE 2 Chinese medicinal composition for treating A beta in caenorhabditis elegans CL41761-42Aggregation-induced neurotoxic effects
Note: different lower case letters in the same column indicate significant differences.
As shown in Table 2, the mean paralysis time of the Chinese medicinal composition group is 11.67 + -0.34 h, the maximum paralysis time is 15.33 + -0.58 h, and compared with the mean paralysis time of the control group of 10.02 + -0.69 h and the maximum paralysis time of 14.33 + -0.58 h, the mean paralysis time is prolonged by 30.24%, and the maximum paralysis time is prolonged by 6.9%, so that the paralysis time (P) of the caenorhabditis elegans can be remarkably prolonged<0.05). As shown in FIG. 1A, the Chinese medicinal composition can remarkably inhibit A beta1-42The abnormal deposition induced neurotoxicity delays the occurrence and development of the paralytic phenotype of the caenorhabditis elegans mutant strain CL4176, so that the traditional Chinese medicine composition can remarkably prolong the paralytic time (P) of the caenorhabditis elegans<0.001)。
3.2 the influence of the Chinese medicinal composition on Poly-Q aggregation-induced neurotoxicity in caenorhabditis elegans AM140
TABLE 3 Effect of Chinese medicinal compositions on poly-Q aggregation-induced neurotoxicity in caenorhabditis elegans AM140
Note: different lower case letters in the same column indicate significant differences.
As shown in Table 3, the mean paralysis time of the Chinese medicinal composition group was 13.08 d; compared with the control group, the paralysis time of the caenorhabditis elegans can be obviously improved (P < 0.05). As shown in FIG. 1B, the paralytic curve of C.elegans in the Chinese medicinal composition group shifted to the right. Therefore, the traditional Chinese medicine composition can obviously inhibit the neurotoxicity induced by abnormal Poly-Q deposition, delay the generation and development of the paralytic phenotype of the caenorhabditis elegans mutant strain AM140 and obviously cause the paralytic curve of the caenorhabditis elegans to move to the right.
In conclusion, the traditional Chinese medicine composition with the active ingredients of mesona chinensis benth, frangipani, microcos paniculata, chrysanthemum, honeysuckle, selfheal and liquorice can obviously reduce nematodes Poly-Q and Abeta1-42Induced neurotoxicity, so that the medicine can be used for preventing and treating neurodegenerative diseases and can also be used for preparing foods, medicines or health-care foods with the effect of preventing and treating the neurodegenerative diseases.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. The application of a traditional Chinese medicine composition in preparing foods, medicines or health-care foods for preventing and treating neurodegenerative diseases is characterized in that the traditional Chinese medicine composition is prepared from the following raw material components in parts by weight:
1000-3500 parts of mesona chinensis benth, 135-900 parts of frangipani, 45-900 parts of microcos paniculata, 180-900 parts of chrysanthemum, 180-900 parts of honeysuckle, 180-900 parts of selfheal and 45-900 parts of liquorice.
2. The application of claim 1, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
1500-2500 parts of mesona chinensis benth, 300-500 parts of frangipani, 300-500 parts of microcos paniculata, 300-500 parts of chrysanthemum, 300-500 parts of honeysuckle, 300-500 parts of selfheal and 300-500 parts of liquorice.
3. The application of claim 2, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
1900-2500 parts of sianchau, 350-450 parts of frangipani, 350-450 parts of microcos paniculata, 350-450 parts of chrysanthemum, 350-450 parts of honeysuckle, 350-450 parts of selfheal and 350-450 parts of liquorice.
4. The use of claim 1, wherein the preparation method of the Chinese medicinal composition comprises the following steps:
preparing raw material components according to the formula amount, and adding water for extraction;
concentrating the obtained extractive solution to obtain concentrated solution.
5. The use according to claim 4, wherein the amount of water is 6-20 times the weight of the feed components; and/or the extraction time is 0.5-3 h.
6. The use as claimed in claim 4, wherein the extraction is carried out after the addition of water and the soaking for 10-50 minutes.
7. Use according to claim 4, wherein the concentrate has a concentration of 9-14 Be.
8. The use of any one of claims 1 to 7, wherein the neurodegenerative disease is a disease of the type resulting from neurotoxicity induced by polymerization and deposition of misfolded proteins comprising Poly-Q and/or A β into neurons, wherein the misfolded proteins comprise1-42。
9. The use of any one of claims 1 to 7, wherein the composition further comprises an adjuvant or auxiliary ingredient acceptable for food, pharmaceutical, health food.
10. The use according to claim 9, wherein the food product is herbal tea.
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