CN105311279B - Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease - Google Patents
Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease Download PDFInfo
- Publication number
- CN105311279B CN105311279B CN201510354975.9A CN201510354975A CN105311279B CN 105311279 B CN105311279 B CN 105311279B CN 201510354975 A CN201510354975 A CN 201510354975A CN 105311279 B CN105311279 B CN 105311279B
- Authority
- CN
- China
- Prior art keywords
- traditional chinese
- chinese medicine
- medicine composition
- preparation
- parts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a traditional Chinese medicine composition, and particularly relates to a preparation method of the traditional Chinese medicine composition and application of the traditional Chinese medicine composition in preparation of a medicine for preventing or treating Alzheimer disease. The traditional Chinese medicine composition disclosed by the invention mainly comprises the following raw material medicines: valeriana jatamansi jones, rhizoma acori graminei, pharbitidis seed, nard, uncaria, semen euphorbiae, menthol and valerian. Animal experiment results prove that the disclosed traditional Chinese medicine composition has an obvious inhibiting effect on the paralytic phenotype of an animal pathological model with the Alzheimer disease, and the potential of preventing or treating the Alzheimer disease is proved to be realized by the traditional Chinese medicine composition, so that the traditional Chinese medicine composition can be applied to the preparation of medicines for preventing or treating the Alzheimer disease.
Description
Technical Field
The invention relates to a traditional Chinese medicine composition, in particular to a preparation method of the traditional Chinese medicine composition and application of the traditional Chinese medicine composition in preparation of a medicine for preventing or treating Alzheimer disease, and belongs to the field of traditional Chinese medicine pharmacy.
Background
Alzheimer's Disease (AD), also known as senile dementia, is a degenerative disease of the nervous system mainly caused by progressive cognitive impairment and memory impairment, and accounts for over 75% of patients with dementia, and at least 30% of people over 85 years old suffer from Alzheimer's disease. Under the background of aging of the population in China, effective prevention and treatment of Alzheimer disease become another important subject before people.
The main pathological features of alzheimer's disease are β -amyloid (a β) deposition in the brain and neurofibrillary tangles. Modern medicine has studied the pathogenesis of alzheimer's disease for many years, but due to its complex etiology, the pathogenesis of the disease is still unclear, and the "Α β cascade hypothesis" is one of the mechanisms that are currently widely accepted by the scientific community. The theory considers that abnormally deposited A beta 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, inflammatory reaction and the like, finally leads to neuron dysfunction or death, triggers cognitive impairment and memory decline, and finally causes dementia. The discovery of mutations in the Amyloid Precursor Protein (APP) gene and the presenilin gene in patients with familial alzheimer's disease, resulting in overexpression of a β in the brain, is one of the most compelling evidences of this theory. Therefore, the beta amyloid protein becomes a well-known target for screening the anti-Alzheimer disease drugs.
Alzheimer disease belongs to the symptoms of dementia, language reversal, amnesia and the like in traditional Chinese medicine, and the traditional Chinese medicine considers kidney deficiency and marrow depletion as the basis, phlegm stasis as the target, five internal organs are disordered, and brain marrow loss is the pathogenesis of the disease.
Because the alzheimer disease is complex, long in course and many in disease links, the alzheimer disease needs to be taken for a long time, and at present, the main anti-alzheimer disease medicines are divided into two types: acetylcholinesterase inhibitors (e.g., galantamine) and antagonists of the N-methyl-D-aspartate receptor (NMDA receptor) (e.g., memantine), but these drugs are expensive, have significant side effects (hallucinations, disorganized consciousness, dizziness, headache, and tiredness), and can only be controlled but not reversed.
At present, drug resistance and serious adverse reactions often occur when western medicines intervene specific pathological links of the Alzheimer disease. The traditional Chinese medicine has the action characteristics of multiple active ingredients and multiple targets, so that the traditional Chinese medicine has unique curative effect advantage for treating the Alzheimer disease.
Disclosure of Invention
The invention aims to provide a traditional Chinese medicine composition and a preparation method thereof, and the other aim of the invention is to provide an application of the traditional Chinese medicine composition in preparing a medicine for preventing or treating Alzheimer disease, wherein the traditional Chinese medicine composition comprises the following raw material medicines: valeriana jatamansi jones, rhizoma acori graminei, pharbitidis seed, nard, uncaria, semen euphorbiae, menthol and valerian.
The traditional Chinese medicine composition for preventing or treating Alzheimer disease comprises the following raw material medicines in parts by weight: 300-600 of valeriana jatamansi, 300-600 of acorus gramineus, 150-300 of pharbitis seed, 150-300 of nard, 150-300 of uncaria, 10-35 of moleplant seed, 0.1-0.6 of menthol and 0.45-0.75 of valerian.
The raw material medicines of the traditional Chinese medicine composition for preventing or treating the Alzheimer disease in parts by weight are preferably as follows: 500 parts of valeriana jatamansi jones, 500 parts of rhizoma acori graminei, 200 parts of pharbitis seed, 200 parts of nard, 200 parts of uncaria, 15 parts of moleplant seed, 0.3 part of menthol and 0.62 part of valerian.
Experimental results prove that the traditional Chinese medicine composition disclosed by the invention has a remarkable inhibiting effect on a paralytic phenotype caused by excessive expression of A beta in an animal pathological model of caenorhabditis elegans Alzheimer disease, and the experimental results show that the traditional Chinese medicine composition disclosed by the invention has a certain effect on treating the damage of the A beta to an organism and has the potential of preventing or treating Alzheimer disease.
The traditional Chinese medicine composition can be prepared into various pharmaceutical dosage forms, theoretically, the traditional Chinese medicine composition can be prepared into any dosage form according to a preparation general method, for example, a preparation process recorded in Van Biting' traditional Chinese medicine pharmacy (Shanghai science Press 1997 12 month I edition), and decoction, capsules, tablets, granules, powder, oral liquid or pills and the like are preferred according to the application objects and indications of the traditional Chinese medicine composition; lyophilized powder for injection and injection solution for parenteral administration.
In order to make the above dosage forms possible, pharmaceutically acceptable excipients, such as: bulking agent, disintegrating agent, lubricant, suspending agent, binder, correctant, antiseptic, matrix, etc. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crospolyvinylpyrrolidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, etc.; the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like; the adhesive comprises: starch slurry, polyvinylpyrrolidone, hydroxypropylmethylcellulose, and the like; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, and the like; the flavoring agent comprises: sweeteners and various essences; the preservative comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chloroacetidine acetate, eucalyptus oil, etc.; the matrix comprises: PEG6000, PEG4000, insect wax, etc.
In order to realize the purpose of preparing patent medicine, part or all of the medicinal materials in the traditional Chinese medicine composition are extracted or smashed as required, but the whole medicine composition is not taken by decocting.
The Chinese medicinal composition tablet disclosed by the invention can be prepared by the following method:
a. weighing valeriana jatamansi jones, rhizoma acori graminei, pharbitidis seeds, nard, uncaria, semen euphorbiae mollis, menthol and valerian according to a proportion;
b. pulverizing rhizoma et radix Valerianae into coarse powder, percolating with 60% ethanol as solvent, and collecting percolate;
c. 3/5 parts of total weight of radix Pimpinellae Candolleanae, rhizoma Acori Graminei, and radix Et rhizoma Nardostachyos by extracting volatile oil, mixing the residue with ramulus Uncariae cum uncis, decocting in water for 1-2 times, preferably 2 times; filtering for 1-2 hr, preferably 1.5 hr, and mixing filtrates;
d. pulverizing semen Pharbitidis and semen Euphorbiae into coarse powder, percolating with 70% ethanol as solvent, collecting percolate, mixing with the above water decoction, and concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30, preferably 1.28;
e. 2/5 weight parts of radix Pimpinellae Candolleanae total weight is pulverized into fine powder, and mixed with the above extract, dried and pulverized into fine powder, and then granulated and dried with rhizoma et radix Valerianae percolate and 70% -75% ethanol, preferably 70% ethanol, and then added with Mentholum and the above volatile oil, mixed well, and pressed into tablet.
The invention uses the transgenic caenorhabditis elegans strain CL4176 as a pathological model for screening the medicine for resisting Alzheimer disease to evaluate the effect of the disclosed traditional Chinese medicine composition on resisting Alzheimer disease, and is based on that: senile Plaque (SP) formed by beta-amyloid (Abeta) aggregation in cerebral cortex and hippocampus of Alzheimer disease patients is one of the most main pathological features of Alzheimer disease, and beta-amyloid can effectively weaken synaptic structure and function and is an important substance for causing Alzheimer disease. The transgenic caenorhabditis elegans strain CL4176 expresses human A beta specifically through muscles under the induction of the temperature of 25 ℃, and the A beta is gathered in muscle tissues and finally causes nematode paralysis. The drug acts on the nematode CL4176, and if the paralytic state of the nematode is improved, particularly the paralytic time of the nematode is remarkably delayed, the drug has the effect of resisting the Alzheimer disease.
Caenorhabditis elegans is sensitive to the change reaction of the external environment, the toxicity of the medicine can be accurately evaluated by using the caenorhabditis elegans, and the body length is one of the most common developmental toxicity evaluation indexes. After the medicament acts on the nematodes, if the body length of the nematodes is remarkably reduced, the medicament has remarkable developmental toxicity to the nematodes.
Compared with the prior art, the invention has the following beneficial effects:
the invention evaluates the effect of the disclosed traditional Chinese medicine composition on the paralytic phenotype of the caenorhabditis elegans Alzheimer disease model, and the result proves that the traditional Chinese medicine composition has an obvious inhibiting effect on the paralytic phenotype of the caenorhabditis elegans Alzheimer disease model and has the potential of preventing or treating Alzheimer disease.
Therefore, the disclosed traditional Chinese medicine composition can be applied to preparation of medicines for preventing or treating Alzheimer disease.
Drawings
FIG. 1 is a graph showing the effect of the positive drug memantine hydrochloride on the paralytic phenotype of the caenorhabditis elegans Alzheimer's disease model.
FIG. 2 is a graph showing the effect of the Chinese medicinal composition of formula 1 on the paralytic phenotype of the caenorhabditis elegans Alzheimer's disease model.
FIG. 3 is a graph showing the effect of the Chinese medicinal composition of formula 2 on the paralytic phenotype of the caenorhabditis elegans Alzheimer's disease model.
FIG. 4 is a graph showing the effect of the Chinese medicinal composition of formula 3 on the paralytic phenotype of the caenorhabditis elegans Alzheimer's disease model.
FIG. 5 shows the results of efficacy evaluation experiments of valeriana jatamansi jones, Acorus tatarinowii Schott, Uncaria rhynchophylla and Chinese medicinal compositions thereof.
FIG. 6 shows the toxicity evaluation test results of Valeriana jatamansi Jones and the Chinese medicinal composition.
Detailed Description
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.
EXAMPLE A preparation of a Chinese medicinal composition tablet
Materials: valeriana jatamansi jones, rhizoma acori graminei, pharbitis seed, nard, uncaria, semen euphorbiae, menthol and valerian are purchased from Hurriertn pharmaceutical chain liability company in Lanzhou.
Formula 1:
300 parts of valeriana jatamansi jones, 300 parts of rhizoma acori graminei, 150 parts of pharbitis seed, 150 parts of nard, 150 parts of uncaria, 10 parts of moleplant seed, 0.1 part of menthol and 0.45 part of valerian.
And (2) formula:
500 parts of valeriana jatamansi jones, 500 parts of rhizoma acori graminei, 200 parts of pharbitis seed, 200 parts of nard, 200 parts of uncaria, 15 parts of moleplant seed, 0.3 part of menthol and 0.62 part of valerian.
And (3) formula:
600 parts of valeriana jatamansi jones, 600 parts of rhizoma acori graminei, 300 parts of pharbitis seed, 300 parts of nard, 300 parts of uncaria, 35 parts of moleplant seed, 0.6 part of menthol and 0.75 part of valerian.
The preparation method comprises the following steps:
a. weighing the following raw materials according to the formula proportion: valeriana jatamansi jones, rhizoma acori graminei, pharbitidis seed, nard, uncaria, semen euphorbiae, menthol and valerian;
b. pulverizing rhizoma et radix Valerianae into coarse powder, percolating with 60% ethanol as solvent, and collecting percolate;
c. 3/5 parts of total weight of valeriana jatamansi, rhizoma acori graminei and rhizoma nardostachyos, extracting volatile oil, mixing the medicinal material residue with uncaria, adding water, decocting for 1-2 times, 1-2 hours each time, filtering, and combining the filtrates;
d. pulverizing semen Pharbitidis and semen Euphorbiae into coarse powder, percolating with 70% ethanol as solvent, collecting percolate, mixing with the above water decoction, and concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30;
e. 2/5 parts by weight of the total weight of the valeriana jatamansi jones are crushed into fine powder and evenly mixed with the extract.
Drying the above extract, pulverizing into fine powder, granulating with rhizoma et radix Valerianae percolate and 70% -75% ethanol, drying, adding Mentholum and the above volatile oil, mixing, and making into tablet.
EXAMPLE Effect of Memantine dihydrochloride on C.elegans CL4176
1. Biological material
(1) Caenorhabditis elegans CL4176 was purchased from Caenorhabditis Genetics Center; for transgenic lines, specifically expressing human a β in muscle at 25 ℃ under induction, a β aggregates in muscle tissue, eventually leading to nematode paralysis, the example used the caenorhabditis elegans line CL4176 as a pathological model for screening anti-alzheimer drugs.
(2) Escherichia coli OP50 (uracil leaky mutant), purchased from Caenorhabditis Genetics Center, as a feed for C.elegans.
2. Reagent
(1) Memantine hydrochloride (MEM), chemical name: 1-amino-3, 5-dimethyladamantane hydrochloride of formula: c12H21N.hcl, available from carbofuran technologies, CAS: 41100-52-1; memantine hydrochloride is an uncompetitive NMDA receptor antagonist and is used for treating moderate-severe to severe Alzheimer-type dementia, and the literature reports that memantine (2.16mg/ml) can remarkably inhibit the paralytic phenotype of a transgenic nematode strain CL4176, and the embodiment uses the memantine hydrochloride as a positive medicament.
(2) Solid NGM (Newatode Growth Medium) medium composition and preparation (taking 1 liter as an example):
| composition (I) | Content (wt.) |
| NaCl | 3.00g |
| K2HPO4 | 2.34g |
| KH2PO4 | 17.23g |
| Peptone | 2.50g |
| Agar-agar | 17.00g |
| Supplement H2O to | 1000ml |
Preparing 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 CaCl2Shaking at 1m, and pouring into a sterilized 9cm plate at about 20 ml/plate while it is hot. Standing for solidification of the culture medium.
(3) Preparing NGM culture medium containing memantine hydrochloride
Memantine hydrochloride was dissolved in sterile water to prepare a mother solution of 2.16mg/ml, and NGM medium containing memantine hydrochloride at final concentrations of 215.76. mu.g/ml, 107.88. mu.g/ml, 21.58. mu.g/ml, 4.32. mu.g/ml and 0.86. mu.g/ml were prepared, respectively.
Blank control is NGM medium supplemented with an equal volume of sterile water as memantine hydrochloride under the same conditions.
The above medium was coated with E.coli OP50 as nematode food.
(4) M9 liquid formula
| Na2HPO4 | 6.00g |
| KH2PO4 | 3.00g |
| NaCl | 5.00g |
| 1M MgSO4 | 1.00ml |
| Supplement H2O to | 1000ml |
(5) Preparation of lysate: 6.4% NaClO3The solution and 1M NaOH solution were mixed in a volume ratio of 1: 1.
3. 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. The amount of the nematode is measured by adding the nematode lysate into the centrifuge tube, oscillating the centrifuge tube for 5 to 7 minutes on a vortex mixer, stopping vortex when the nematodes are completely broken, subpackaging the centrifuge tube with 1.5mL, and washing nematode eggs with M9 solution for three times.
(3) Action of memantine hydrochloride on caenorhabditis elegans CL4176
After the synchronization of the nematodes, nematode eggs are transferred to NGM culture media containing memantine hydrochloride with different concentrations, 60 nematodes are contained in each culture medium, three culture media are used as each drug concentration in parallel, and the nematodes are cultured for 3 days at 16 ℃ until the L3 stage.
In order to express A beta, the L3 phase nematodes were induced at 25 ℃ and count of paralyzed nematode sticks started after 34 h. Counting every two hours until all nematodes are paralyzed (paralysis means that the nematodes do not move or only the head moves when the nematode is mechanically stimulated in the body), and the results are shown in FIG. 1.
As can be seen from the figure 1, the positive drug memantine hydrochloride can obviously inhibit CL4176 paralytic phenotype at 21.58 mu g/ml, 4.32 mu g/ml and 0.86 mu g/ml, and the memantine hydrochloride at 215.76 mu g/ml and 107.88 mu g/ml strongly inhibits nematode development. The experiment result shows that the experiment system is normal, which indicates that the caenorhabditis elegans strain CL4176 can be used for screening the medicine for resisting the Alzheimer disease.
EXAMPLE III Effect of the Chinese medicinal composition on caenorhabditis elegans CL4176
1. The biomaterial was the same as in example two.
2. Reagent
(1) The traditional Chinese medicine composition tablet is a preparation prepared according to the embodiment method, and specifically comprises the following components: the traditional Chinese medicine composition tablet of the formula 1, the traditional Chinese medicine composition tablet of the formula 2 and the traditional Chinese medicine composition tablet of the formula 3.
(2) Solid NGM (Newatode Growth Medium) medium composition and preparation (taking 1 liter as an example): the same as the second embodiment.
(3) Preparing NGM culture medium containing Chinese medicinal composition
Grinding the Chinese medicinal composition tablet into powder with mortar, dissolving in sterile water to obtain 150mg/ml Chinese medicinal composition water solution, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the aqueous solution of the Chinese medicinal composition into the melted NGM culture medium to obtain NGM culture medium containing the Chinese medicinal composition at final concentration of 1.0mg/ml, 5.0mg/ml, and 15.0 mg/ml.
The blank control is NGM culture medium added with sterile water of the same volume as the Chinese medicinal composition aqueous solution under the same condition.
The above medium was coated with E.coli OP50 as nematode food.
(4) M9 liquid formula: the same as the second embodiment.
(5) Preparation of lysate: the same as the second embodiment.
3. Carrying out the step
(1) And (3) culturing nematodes: the same as the second embodiment.
(2) Nematode synchronization: the same as the second embodiment.
(3) Action of traditional Chinese medicine composition on caenorhabditis elegans CL4176
After the synchronization of the nematodes, transferring nematode eggs to NGM culture medium coated with OP50 and mixed with formula 1 Chinese medicinal composition tablets, formula 2 Chinese medicinal composition tablets and formula 3 Chinese medicinal composition tablets with different concentrations; the blank was NGM medium coated with OP50 and supplemented with equal volume of sterile water, 60 nematodes per medium, three media per drug concentration as a parallel, and cultured at 16 ℃ for 3 days to L3.
In order to express A beta, the L3 phase nematodes were induced at 25 ℃ and count of paralyzed nematode sticks started after 34 h. Counting every two hours until all nematodes are paralyzed (paralysis means that the nematodes cannot move or only the head moves when the nematode is mechanically stimulated in the body), and the results are shown in fig. 2, fig. 3 and fig. 4.
As can be seen from the figures 2, 3 and 4, the formulas 1, 2 and 3 can significantly inhibit the nematode paralysis phenotype caused by the overexpression of A beta (p is less than 0.001), wherein the nematode paralysis phenotype caused by the overexpression of A beta is best inhibited by the formula 2, the nonparalysis proportion of 15mg/ml and 5mg/ml is still kept at about 80% at 44h (figure 3), and the nonparalysis proportion of 5mg/ml is lower than 80% in the formulas 1 and 3 (figures 2 and 4).
In conclusion, the above embodiments show that the traditional Chinese medicine composition has an obvious inhibitory effect on the paralytic phenotype caused by the aggregation of caenorhabditis elegans A beta in an Alzheimer disease model, and the traditional Chinese medicine composition has the potential of preventing or treating Alzheimer disease. Therefore, the traditional Chinese medicine composition disclosed by the invention can be applied to preparation of medicines for preventing or treating Alzheimer's disease.
Example four comparative evaluation experiments on efficacy and toxicity of Chinese medicinal composition
1. The biomaterial is as in example one.
2. Reagent
(1) The traditional Chinese medicine composition tablet is a preparation prepared according to the embodiment method, and specifically comprises the following components: the traditional Chinese medicine composition tablet of the formula 2.
(2) Solid NGM (Newatode Growth Medium) medium composition and preparation (taking 1 liter as an example): the same as the first embodiment.
(3) NGM culture medium for preparing water-soluble Chinese medicinal composition
Grinding the Chinese medicinal composition tablet into powder with mortar, dissolving in sterile water to obtain 150mg/ml Chinese medicinal composition water solution, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the aqueous solution of the Chinese medicinal composition into the melted NGM culture medium to prepare the NGM culture medium containing the Chinese medicinal composition with the final concentration of 15.0 mg/ml.
(4) Preparation of valeriana jatamansi jones extract and NGM culture medium containing valeriana jatamansi jones extract
3/5 parts by weight of the total weight of the valeriana jatamansi jones in the formula 2 are taken to extract volatile oil, and the medicinal material residues are decocted for 1-2 times, preferably 2 times, by adding water; filtering for 1-2 hr, preferably 1.5 hr each time, mixing filtrates, and concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30, preferably 1.28; 2/5 weight parts of the total weight of the valeriana jatamansi jones are crushed into fine powder for standby. Mixing the volatile oil, extract and fine powder, adding sterile water, dissolving to obtain arachnid perfume solution, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the spider perfume solution into the melted NGM culture medium to make the concentration of spider perfume in the culture medium consistent with that of the NGM culture medium containing the traditional Chinese medicine composition in the formula 2.
(5) Preparation of rhizoma acori graminei extract and NGM culture medium containing rhizoma acori graminei extract
Extracting volatile oil from rhizoma Acori Graminei in formula 2, decocting the residue in water for 1-2 times, preferably 2 times; filtering for 1-2 hr, preferably 1.5 hr, and mixing filtrates; concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30, preferably 1.28. Mixing the above volatile oil and extract, dissolving in sterile water, making into rhizoma Acori Graminei water solution, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the rhizoma acori graminei water solution into the melted NGM culture medium to ensure that the concentration of the rhizoma acori graminei in the culture medium is consistent with that of the rhizoma acori graminei in the NGM culture medium containing the traditional Chinese medicine composition in the formula 2.
(6) Preparing ramulus Uncariae cum uncis extract and NGM culture medium containing the same
Decocting ramulus Uncariae cum uncis in formula 2 with water for 1-2 times, preferably 2 times; filtering for 1-2 hr, preferably 1.5 hr, and mixing filtrates; concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30, preferably 1.28. Dissolving the extract in sterile water, preparing into ramulus Uncariae cum uncis aqueous solution, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the uncaria aqueous solution into the melted NGM culture medium to ensure that the concentration of the uncaria in the culture medium is consistent with that of the uncaria in the NGM culture medium containing the traditional Chinese medicine composition in the formula 2.
(7) Preparing extracts of radix Pimpinellae Candolleanae, rhizoma Acori Graminei and ramulus Uncariae cum uncis and NGM culture medium containing the extracts
3/5 parts by weight of the total weight of valeriana jatamansi jones and rhizoma acori graminei in the formula 2 are taken to respectively extract volatile oil, the obtained residue is combined with uncaria, and water is added for decoction for 1-2 times, preferably 2 times; filtering for 1-2 hr, preferably 1.5 hr, and mixing filtrates; concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30, preferably 1.28. 2/5 weight parts of the total weight of the valeriana jatamansi jones are crushed into fine powder for standby. Mixing the above volatile oil, extract and fine powder, adding sterile water for dissolving, preparing into aqueous solution of extract of Valeriana jatamansi Jones, rhizoma Acori Graminei and ramulus Uncariae cum uncis, performing ultrasonic treatment at room temperature for 30min, centrifuging at 10000rpm for 10min, collecting supernatant, and storing at 4 deg.C. Adding the aqueous solution of the extracts of the spider aroma, the grassleaf sweelflag rhizome and the gambir plant into the melted NGM culture medium to ensure that the concentration of the spider aroma, the grassleaf sweelflag rhizome and the gambir plant in the culture medium is consistent with that of the spider aroma, the grassleaf sweelflag rhizome and the gambir plant in the NGM culture medium containing the traditional Chinese medicine composition in the formula 2.
The blank control is NGM culture medium added with sterile water of the same volume as the Chinese medicinal composition aqueous solution under the same condition.
The above medium was coated with E.coli OP50 as nematode food.
(8) M9 liquid formula: the same as the first embodiment.
(9) Preparation of lysate: the same as the first embodiment.
3. Carrying out the step
(1) And (3) culturing nematodes: the same as the first embodiment.
(2) Nematode synchronization: the same as the first embodiment.
(3) Comparative evaluation of drug effect of traditional Chinese medicine composition
After the synchronization of the nematodes, transferring nematode eggs to an NGM culture medium coated with OP50 and mixed with a traditional Chinese medicine composition, an NGM culture medium of an valeriana jatamansi jones extract, an NGM culture medium of a grassleaf sweelflag rhizome extract, an NGM culture medium of an uncaria rhynchophylla extract, and an NGM culture medium of a valeriana jatamansi jones, a grassleaf sweelflag rhizome and uncaria rhynchophylla formula extract; blank control was NGM medium coated with OP50 and added with sterile water of equal volume to the aqueous solution of the Chinese medicinal composition. 60 nematodes were cultured in each medium at 16 ℃ for 3 days to L3 stage.
In order to express A beta, the L3 phase nematodes were induced at 25 ℃ and count of paralyzed nematode sticks started after 34 h. Counting was performed every two hours until all nematodes were paralyzed (paralysis means that the nematodes could not move or only the head moved when the nematode was mechanically stimulated in the body), and the results are shown in FIG. 5.
The experimental results show that the valeriana jatamansi jones, the uncaria rhynchophylla, the rhizoma acori graminei and the traditional Chinese medicine composition can obviously inhibit the nematode paralysis phenotype (p is less than 0.001) caused by the overexpression of the A beta, wherein the effect of the valeriana jatamansi jones on inhibiting the nematode paralysis phenotype caused by the overexpression of the A beta is the best, and is almost equivalent to that of the traditional Chinese medicine composition; ramulus Uncariae cum uncis and rhizoma Acori Graminei have inferior inhibitory effect on nematode paralytic phenotype to traditional Chinese medicinal composition (P is less than 0.001). And the combination of valeriana jatamansi jones, uncaria and acorus gramineus soland seriously affects the growth of nematodes, and the nematodes cannot hatch, which indicates that the toxicity of the combination of the valeriana jatamansi jones, the uncaria and the acorus gramineus is increased.
(4) Comparative evaluation of toxicity of traditional Chinese medicine composition
Immediately after the paralysis experiment, the nematodes were treated at 50 ℃ for 15min to straighten the body of the nematodes for easy measurement. Nematode body length was observed under a microscope and photographed and measured using the MvImage image processing system. The results are shown in FIG. 6.
The experimental result shows that compared with a blank control and the traditional Chinese medicine composition thereof, the aragonite has the advantages that the aragonite has the obvious effect of reducing the body length of the aragonite, and the aragonite has the obvious effect of inhibiting the growth and the development of the aragonite and has higher toxicity than the traditional Chinese medicine composition.
The effect evaluation experiment of the embodiment shows that the valeriana jatamansi jones, the uncaria, the rhizoma acori graminei and the traditional Chinese medicine composition thereof can obviously inhibit nematode paralysis phenotype caused by A beta aggregation, wherein the effects of the valeriana jatamansi jones and the traditional Chinese medicine composition are optimal; as can be seen from the toxicity evaluation experiment of the embodiment, compared with the blank control and the traditional Chinese medicine composition thereof, the valeriana jatamansi jones can obviously inhibit the growth and development of nematodes, the toxicity is obviously higher than that of the traditional Chinese medicine composition, and the traditional Chinese medicine composition hardly influences the growth of nematodes, which indicates that the toxicity of the traditional Chinese medicine composition is obviously lower than that of the valeriana jatamansi jones.
In a word, the results of the embodiment show that the traditional Chinese medicine composition has the characteristics of remarkable effect and low toxicity in the aspect of preventing or treating the alzheimer disease, and the traditional Chinese medicine composition has the potential of preventing or treating the alzheimer disease and can be applied to the preparation of the medicines for preventing or treating the alzheimer disease.
Claims (4)
1. An application of a Chinese medicinal composition in preparing a medicament for preventing or treating Alzheimer disease is characterized in that
The Chinese medicinal composition comprises the raw material medicaments of 300-
150-
0.15-0.75.
2. The use of the Chinese medicinal composition of claim 1 in the preparation of a medicament for the prevention or treatment of alzheimer's disease
The application of the traditional Chinese medicine composition is characterized in that the traditional Chinese medicine composition comprises the following raw material medicines in parts by weight: 500 g of valeriana root and rhizome of grassleaved sweetflag
500. 200 parts of pharbitis seed, 200 parts of nard, 200 parts of uncaria, 15 parts of moleplant seed, 0.3 part of menthol and 0.62 part of valerian.
3. Use of a Chinese medicinal composition according to any one of claims 1-2 in the preparation of a medicament for the prevention or treatment of alzheimer's disease
The application of the medicine is characterized in that the medicine preparation of the traditional Chinese medicine composition as an active ingredient is a capsule
Preparation, tablet, granule, powder, oral liquid or pill.
4. The use of the Chinese medicinal composition according to claim 3 in the preparation of a medicament for the prevention or treatment of Alzheimer's disease
Is characterized in that the preparation method of the traditional Chinese medicine composition tablet comprises the following steps:
a. weighing Valeriana jatamansi jones, rhizoma acori graminei, semen pharbitidis, rhizoma nardostachyos, uncaria, semen euphorbiae mollis, menthol,
Valerian root;
b. pulverizing rhizoma et radix Valerianae into coarse powder, percolating with 60% ethanol as solvent, and collecting percolate;
c. 3/5 parts of total weight of valeriana jatamansi, rhizoma acori graminei and rhizoma nardostachyos, extracting volatile oil, and mixing medicinal material residues with rhizoma acori graminei
Mixing the above medicinal materials, decocting in water for 1-2 times (each for 1-2 hr), filtering, and mixing filtrates;
d. pulverizing semen Pharbitidis and semen Euphorbiae, percolating with 70% ethanol, and collecting percolate
Mixing with the above water decoction, and concentrating under reduced pressure to obtain extract with relative density of 1.26-1.30;
e. 2/5 weight parts of radix Pimpinellae Candolleanae total weight are pulverized into fine powder, and the fine powder and the extract are mixed uniformly, dried and powdered
Pulverizing into fine powder, granulating with percolate of rhizoma et radix Valerianae and 70% -75% ethanol, drying, adding Mentholum and above volatile components
Mixing with oil, and making into tablet.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510354975.9A CN105311279B (en) | 2014-06-26 | 2015-06-25 | Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2014/080790 WO2015196402A1 (en) | 2014-06-26 | 2014-06-26 | Use of traditional chinese medicine composition in preparing medicines for prevention or treatment of alzheimer's disease |
| CNPCT/CN2014/080790 | 2014-06-26 | ||
| CN201510354975.9A CN105311279B (en) | 2014-06-26 | 2015-06-25 | Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105311279A CN105311279A (en) | 2016-02-10 |
| CN105311279B true CN105311279B (en) | 2021-02-19 |
Family
ID=55240236
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510354975.9A Active CN105311279B (en) | 2014-06-26 | 2015-06-25 | Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105311279B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107519323A (en) * | 2017-04-25 | 2017-12-29 | 兰州大学 | Rhizoma valerianae latifoliae extract is preparing the application in activating HSP70 medicines |
| CN109223924A (en) * | 2018-11-10 | 2019-01-18 | 兰州大学 | Epilepsy medicine (Dianxianning) adjusts the application in beta-amyloid protein drug in preparation |
| CN109293621A (en) * | 2018-11-21 | 2019-02-01 | 兰州大学 | A kind of Rhizoma valerianae latifoliae extract monomeric compound and its application in treatment Alzheimer disease drug |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732515B (en) * | 2010-01-26 | 2011-06-01 | 昆明中药厂有限公司 | Quality test method of Dianxianning tablets |
| CN102058630B (en) * | 2010-12-16 | 2014-03-19 | 苏州大学 | Valerian preparation for tranquilizing and allaying excitement and preparation method thereof |
-
2015
- 2015-06-25 CN CN201510354975.9A patent/CN105311279B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105311279A (en) | 2016-02-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101126117B1 (en) | A chinese medicine composition and preparation method and use thereof | |
| CA2960075C (en) | A composition comprising extracts of green tea and turmeric and use thereof for reducing body weight or body fat | |
| CN105311279B (en) | Application of traditional Chinese medicine composition in preparation of medicine for preventing or treating Alzheimer disease | |
| CN103393736B (en) | A kind of for pharmaceutical composition treating Alzheimer and its production and use | |
| CN109674958A (en) | A kind of Chinese medicine composition and its preparation method and application with anti-trioxypurine | |
| WO2013176567A1 (en) | Pharmaceutical composition for the prophylaxis and treatment of psychological, behavioral and cognitive disorders | |
| US10226500B2 (en) | Use of the Chinese medicine composition and Dianxianning in the preparation of a medicament for preventing or treating Alzheimer's diseases | |
| CN112656877A (en) | Pharmaceutical composition for treating cervical disc herniation and preparation method and application thereof | |
| DE112006002340T5 (en) | Medicines used to treat prostate cancer | |
| CN103735761B (en) | A kind of pharmaceutical composition preventing and treating Alzheimer disease and preparation method thereof | |
| WO2015196402A1 (en) | Use of traditional chinese medicine composition in preparing medicines for prevention or treatment of alzheimer's disease | |
| CN109289009A (en) | A kind of Chinese medicine composition and its preparation method and application | |
| CN112618591B (en) | A pharmaceutical composition containing total flavonoids of herba Blumeae Balsamiferae | |
| JP2017515902A (en) | Drug composition for controlling lipemia and body weight and its application method | |
| WO2018042384A1 (en) | Compositions of medicinal plants for reducing the effects of aging, prevention and treatment of age-related neurodegenerative diseases, and treatment of anxiety and sleep disorders | |
| CN101224247B (en) | Chinese traditional medicine compounds for treating Alzheimer's disease and preparing method thereof | |
| CN106563061B (en) | Yuye decoction composition for treating Alzheimer disease | |
| CN109453159B (en) | Application of valeriana jatamansi jones extract monomer compound in preparation of anti-AD drugs | |
| CN102727484A (en) | Use of silymarin in preparations of drugs for treatment of Alzheimer's disease | |
| CN107233454B (en) | Traditional Chinese medicine and Tibetan medicine composition for preventing and treating livestock diarrhea | |
| CN107823239B (en) | Preparation method of carmine radish extract and application of carmine radish extract in preparation of medicine for preventing and treating Alzheimer's disease | |
| CN114272354B (en) | Traditional Chinese medicine composition for preventing and improving senile dementia, preparation method and application thereof | |
| CN113116934A (en) | Application of maggot for treating senile dementia | |
| JP2022511544A (en) | Chinese herbal medicine composition for bowel movement, its preparation method and its use | |
| CN104610025A (en) | Lignans compounds separated from rhizoma acori graminei as well as preparation and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20160728 Address after: Chengguan District of Gansu city of Lanzhou province Donggang West Road 730000 No. 249 Lanzhou University Science and technology garden room 119-5 Applicant after: Li Zhongqing Address before: 119-5 room 730000, Lan Da Science and Technology Park, Lanzhou, Gansu Applicant before: Lanzhou Xiaolianghe Biological Technology Co., Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |