CN106563061B - Yuye decoction composition for treating Alzheimer disease - Google Patents
Yuye decoction composition for treating Alzheimer disease Download PDFInfo
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Abstract
The invention belongs to the technical field of traditional Chinese medicines, provides a new application of Yuye decoction, and particularly relates to an application of Yuye decoction in preparation of a medicine for treating Alzheimer's disease. Experiments are carried out by using a caenorhabditis elegans pathological model, and the results show that: the cajanus cajan decoction has a remarkable treatment effect on caenorhabditis elegans suffering from Alzheimer's disease, shows that the cajanus cajan decoction has a remarkable effect of delaying the paralytic phenotype of the caenorhabditis elegans suffering from Alzheimer's disease, and suggests that the cajan decoction has the potential of treating Alzheimer's disease, and can be applied to preparation of medicaments for treating Alzheimer's disease.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, provides a new application of Yuye decoction, and particularly relates to an application of Yuye decoction in preparation of a medicine for treating Alzheimer's disease.
Background
Alzheimer's Disease (AD), a degenerative disease of the nervous system that predominates in progressive cognitive impairment and memory impairment, is one of the most common forms of senile dementia. Clinically, it is characterized by generalized dementia such as memory impairment, aphasia, agnosia, impairment of visual spatial skills, impairment of executive function, and personality and behavioral changes. The proportion of the medicine in the patients with dementia is more than 75%. With the increasing aging degree of China, AD can become one of the main diseases affecting the health of the old, and the effective prevention and treatment of AD becomes another important subject before people.
Modern medicine has studied the pathogenesis of AD for many years, but due to its complex etiology, the pathogenesis of the AD is still unclear, and the "a β cascade hypothesis" is one of the mechanisms widely accepted by the scientific community at present, the hypothesis is that a β abnormally deposited in the brain of a patient directly or indirectly acts on neurons and glial cells through a series of cascade reactions such as free radical reaction, mitochondrial oxidative damage and inflammatory reaction, finally leads to neuron dysfunction or death, causes cognitive impairment and memory decline, and finally causes dementia, and the Amyloid Precursor Protein (APP) gene and presenilin gene are found to be mutated in patients with familial alzheimer disease, so that the overexpression of a β in the brain becomes the most powerful evidence of the hypothesis, so β amyloid has become a well-known anti-AD drug target.
The AD has complex etiology, long course of disease and more pathogenic links, and needs to be taken for a long time, the current main anti-Alzheimer's disease drugs such as acetylcholinesterase inhibitor (such as galantamine) and N-methyl-D-aspartate receptor (NMDA receptor) antagonist (such as memantine) are expensive, the side effects such as hallucination, consciousness chaos, dizziness, headache and tiredness after taking the drug are obvious, and the disease condition of a patient can only be controlled but can not be reversed.
At present, the intervention of western medicines aiming at a certain specific pathological link of AD often causes drug resistance and serious adverse reaction. The traditional Chinese medicine composition has the characteristics of multiple active components and multiple targets, so that the prospect of screening drug candidates for treating AD from the traditional Chinese medicine composition treasury is wide.
Yuye Tang is prepared from astragalus root, Chinese yam, anemarrhena rhizome, kudzu vine root, schisandra fruit, trichosanthes root and chicken's gizzard-membrane (Zhang Xi, records of Zhongzhao Zhong xi, 2009), and has the functions of invigorating spleen, reinforcing kidney, replenishing qi, nourishing yin, promoting the production of body fluid to quench thirst, and can be used for treating diabetes caused by deficiency of both spleen and kidney, deficiency of vital energy and deficiency of true yin. In the prior art, Yuye decoction is used for preparing a medicament for treating diabetes.
Caenorhabditis elegans (C.)Caenorhabditis elegans) Is a very useful research platform for multifunctional drug screening and drug action mechanism. It is cheap and easy to culture; the generation period is short, the number of offspring is large, a large number of individuals with consistent backgrounds can be obtained, the experimental repeatability is ensured, and the experiment is carried out by adopting a large sample amount, so that the influence of individual difference is eliminated; is highly conserved in genes and molecular pathways with higher organisms (Kaletta and Hengartner, 2006). It is more and more favored by pharmacologists as a bridge from the primary screening of cell in vitro horizontal drugs to the secondary screening of mouse in vivo horizontal drugs. The American devGen company has been FDA approved for clinical study using humanized nematode screening for antiarrhythmic drugs.
A humanized caenorhabditis elegans AD pathological model is established by transferring humanized A β 1-42 gene to the downstream of a myosin promoter and expressing the gene under the control of temperature, wherein the nematode strain can normally grow at 15 ℃, and when the nematode strain is transferred to the condition of 25 ℃, A β 1-42 gene is expressed and accumulated in the muscle of the nematode, so that the nematode loses the motor ability and is paralyzed.
Unexpectedly, the caenorhabditis elegans AD pathological model is adopted by the inventor to screen a large amount of traditional Chinese medicine compositions, so that the obtained caenorhabditis elegans decoction has a remarkable AD resisting effect, and all components play a synergistic effect.
Therefore, the invention provides a new application of Yuye decoction, and particularly relates to an application of Yuye decoction in preparation of a medicament for treating Alzheimer disease.
Reference documents:
Kaletta T, Hengartner MO. Finding function in novel targets:C. elegansas a model organism. Nat Rev Drug Discov 2006, 5: 387e398.
Link CD. Expression of human beta-amyloid peptide in transgenicCaenorhabditis elegans. Proc Natl Acad Sci USA 1995, 92: 9368e9372.
Link CD. Transgenic invertebrate models of age-associatedneurodegenerative diseases. Mech Ageing Dev 2001, 122: 1639e1649.
disclosure of Invention
The invention aims to provide a new application of Yuye decoction, namely a new application in pharmacy.
In particular to application of Yuye decoction in preparing a medicament for treating Alzheimer disease.
The jade liquid soup provided by the invention contains the following raw material medicines in parts by weight: 1.9-15 parts of astragalus root, 3.7-30 parts of Chinese yam, 2.2-18 parts of anemarrhena, 0.6-5 parts of kudzu root, 1.1-9 parts of schisandra fruit, 1.1-9 parts of trichosanthes root and 0.8-6 parts of chicken's gizzard-membrane.
The Yuye decoction as an active ingredient can be prepared into capsules, tablets, granules, powder, oral liquid and pills.
The invention has the beneficial effects that:
the caenorhabditis elegans is used as a model organism for carrying out experiments, and the result shows that the caenorhabditis elegans decoction has a remarkable treatment effect on caenorhabditis elegans suffering from Alzheimer's disease, has a remarkable inhibition effect on a paralytic phenotype caused by the overexpression of A β in the muscle of AD caenorhabditis elegans, can remarkably delay the paralytic phenotype of the caenorhabditis elegans suffering from Alzheimer's disease, and prompts that the caenorhabditis elegans decoction has the potential of treating Alzheimer's disease, and can be applied to the preparation of medicaments for treating Alzheimer's disease.
The technical solution of the present invention is further described in detail with reference to the following specific examples, but the scope of the present invention is not limited to the following.
Drawings
FIG. 1 shows the effect of Yuye decoction on delaying paralysis of AD caenorhabditis elegans
C-blank control group, L-low-concentration Yuye decoction group, M-medium-concentration Yuye decoction group, and H-high-concentration Yuye decoction group
FIG. 2 Yuye decoction for delaying half of paralysis time of AD caenorhabditis elegans
FIG. 3 the effect of Yuye decoction components on delaying paralysis of AD caenorhabditis elegans
Control-blank Control group, YY-Yuye decoction group, J-J drug group, J + C-J drug group + C drug group, J + C + Z-J drug group + C drug group + Z drug group.
Detailed Description
Example A therapeutic Effect of Yuye decoction on AD caenorhabditis elegans
1. Biological material
(1) Caenorhabditis elegans CL4176 was purchased from Caenorhabditis Genetics Center (CGC), is a transgenic line that expresses human A β muscle-specifically under induction of a temperature of 25 ℃1-42，Aβ1-42Accumulation in muscle tissue, ultimately leading to nematode paralysis, the present example used the C.elegans CL4176 strain as a pathological model for screening anti-AD drugs.
(2) Escherichia coli OP50 (uracil leaky mutant), purchased from Caenorhabditis Genetics Center (CGC), was used as feed for C.elegans.
2. Reagent
(1) Solid NGM (Newatode Growth Medium) medium composition and preparation (taking 1 liter as an example):
after preparing the solid NGM culture medium, sterilizing at 121 deg.C under high pressure and constant temperature for 20min, adding 5mg/mL cholesterol 1mL, 1M MgSO41mL,1M CaCl21mL was shaken well and poured hot into a sterilized 9cm plate, approximately 20 mL/plate. Standing for solidification of the culture medium.
(2) M9 liquid formula
(3) Preparation of lysate: a6.4% NaClO solution and a 1M NaOH solution were mixed in a volume ratio of 1: 1.
3. Preparation of decoction of Yuye decoction
Blank control group: sterile water.
High-concentration Yuye decoction group: 60g of astragalus, 120g of Chinese yam, 72g of rhizoma anemarrhenae, 20g of radix puerariae, 36g of schisandra chinensis, 36g of radix trichosanthis, 24g of endothelium corneum gigeriae galli and 3000mL of water are heated to boil and kept for 30 minutes, the volume is kept to 1000mL, and the filtrate is filtered and sterilized for later use.
Medium concentration yuye decoction group: 30g of astragalus, 60g of Chinese yam, 36g of rhizoma anemarrhenae, 10g of radix puerariae, 18g of schisandra chinensis, 18g of radix trichosanthis, 12g of endothelium corneum gigeriae galli and 3000mL of water are heated to boil and kept for 30 minutes, the volume is kept to 1000mL, and the filtrate is filtered and sterilized for later use.
Low-concentration Yuye decoction group: 15g of astragalus, 30g of Chinese yam, 18g of rhizoma anemarrhenae, 5g of radix puerariae, 9g of schisandra chinensis, 9g of radix trichosanthis, 6g of endothelium corneum gigeriae galli and 3000mL of water are heated to boil and kept for 30 minutes, the volume is kept to 1000mL, and the filtrate is filtered and sterilized for later use.
4. Carrying out the step
(1) And (3) culturing nematodes:
the nematodes were plated on solid NGM plates coated with E.coli OP50 and then incubated in an incubator at 16 ℃ and synchronized when they reached adult growth.
(2) Nematode synchronization:
selecting NGM culture medium containing a large amount of imagoes and part of nematode eggs which have been hatched, flushing the nematodes from the culture medium by using M9 liquid, transferring the nematodes to a centrifuge tube, standing the nematode to enable the nematodes to freely settle to the bottom of the centrifuge tube, and discarding supernatant. And adding the lysate into the centrifuge tube according to the quantity of the nematode, oscillating the centrifuge tube on a vortex mixer for 5 to 7 minutes, stopping vortex when the nematode is completely broken, subpackaging the centrifuge tube with 1.5mL, and washing the nematode eggs with M9 solution for three times.
(3) Action of Yuye decoction on caenorhabditis elegans CL4176
Adding sterile water 180 μ l into 20 μ l of the above jade liquid soup with different concentrations to obtain diluted jade liquid soup, mixing the diluted jade liquid soup with 1.8 mL of NGM culture medium, pouring one NGM plate (diameter of 3.5 cm), and replacing jade liquid soup with sterile water in blank group. Coating OP50 on the culture medium, transferring the synchronized nematode eggs to the culture medium, setting 60 nematodes in each culture dish at three concentrations, and culturing at 16 deg.C for 3 days to L3.
In order to express A β, the nematodes in the L3 phase are transferred to 25 ℃ for constant temperature induction, the number of paralyzed nematodes is counted after 34h, the nematode is counted every two hours until all the nematodes are paralyzed (paralysis means that the nematodes can not move or only the head moves when the nematode is mechanically stimulated).
The ordinate of FIG. 1 shows the percentage of C.elegans which had no paralysis, and at the same time point, the larger this value indicates a larger proportion of C.elegans which had no paralysis in the treated group, i.e.a stronger effect of the drug in delaying paralysis.
The experimental result shows that the Yuye decoction has the treatment effect on the AD caenorhabditis elegans depending on the dosage.
The ordinate of figure 2 is the paralysis time of half of the nematodes, and a larger value indicates a stronger effect of the drug in delaying paralysis.
Since the number of paralyzed C.elegans in the high-concentration Yuye decoction group was still not more than half at the end of the experiment, the half paralysis time (PT 50) of this dose was recorded as the length of time taken for the end of the experiment.
Experimental results show that the dosage of the jade liquid decoction increases half of the paralysis time of the AD caenorhabditis elegans dependently.
Example therapeutic Effect of synergistic Effect of the Components of Eryuye decoction on AD caenorhabditis elegans
The prescription of the Yuye decoction is removed, and the synergistic effect of the components is verified on the treatment effect of the AD caenorhabditis elegans:
drug group J (J): radix astragali and Chinese yam
Drug group C (C): rhizoma anemarrhenae and radix Trichosanthis
Z drug group (Z): kudzu root
1. Experiment group setting
Blank Control (Control): sterile water.
Yuye decoction (YY) comprising radix astragali 30g, rhizoma Dioscoreae 60g, rhizoma anemarrhenae 36g, radix Puerariae 10g, fructus Schisandrae 18g, Trichosanthis radix 18g, endothelium corneum Gigeriae Galli 12g, and water 3000mL, heating to boil and keeping for 30 min, diluting to 1000mL, filtering, sterilizing, and collecting filtrate.
Drug group J: 30g of astragalus and 60g of Chinese yam, adding 3000mL of water, heating to boil and keeping for 30 minutes, metering volume to 1000mL, and filtering and sterilizing for later use.
Drug group J + drug group C (J + C): 30g of astragalus, 60g of Chinese yam, 36g of rhizoma anemarrhenae and 18g of trichosanthes root, adding 3000mL of water, heating to boil and keeping for 30 minutes, fixing the volume to 1000mL, and filtering and sterilizing for later use.
Drug group J + drug group C + drug group Z (J + C + Z): adding 3000mL of water into 30g of astragalus, 60g of Chinese yam, 36g of rhizoma anemarrhenae, 18g of trichosanthes root and 10g of kudzu root, heating to boil and keeping for 30 minutes, metering volume to 1000mL, and filtering and sterilizing for later use.
2. Experimental procedure
The specific implementation steps are the same as those in the first embodiment.
3. Results of the experiment
The ordinate of FIG. 3 shows the percentage of C.elegans which had no paralysis, and at the same time point, the larger this value indicates a larger proportion of C.elegans which had no paralysis in this treatment group, i.e.a stronger effect of the drug in delaying paralysis.
Experimental results show that the effect of the jadeite soup on delaying the paralysis of the AD caenorhabditis elegans is obviously higher than that of the (astragalus mongholicus + yam) group, the (astragalus mongholicus + yam group + anemarrhena asphodeloides + trichosanthes root) group and the (astragalus mongholicus + yam group + anemarrhena asphodeloides + trichosanthes root) group, so that the components in the jadeite soup play a synergistic role in treating the AD caenorhabditis elegans.
The above examples prove that the Yuye decoction provided by the invention has a remarkable treatment effect on AD caenorhabditis elegans, and the Yuye decoction provided by the invention has a potential of treating Alzheimer's disease and can be applied to preparation of a medicine for treating Alzheimer's disease.
Claims (2)
1. The application of Yuye decoction in preparing the medicine for treating Alzheimer disease is characterized in that the Yuye decoction contains the following raw material medicines in parts by weight: 15 parts of astragalus, 30 parts of Chinese yam, 18 parts of rhizoma anemarrhenae, 5 parts of kudzu root, 9 parts of schisandra chinensis, 9 parts of trichosanthes root and 6 parts of endothelium corneum gigeriae galli.
2. The use of Yuye decoction in the preparation of a medicament for treating Alzheimer's disease according to claim 1, wherein the medicinal preparation of Yuye decoction as the active ingredient is capsule, tablet, granule, powder, oral liquid or pill.
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| 阿尔茨海默病发生机制的microＲNA水平研究进展;吴越;《中国老年学杂志》;20140630;第34卷(第12期);3506-3508 * |
| 阿尔茨海默病的秀丽隐杆线虫模型及其应用;杨平;《中国细胞生物学学报》;20101231;第32卷(第1期);37-42 * |
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