CN109350643A - The medical usage of the anti-ischemia apoplexy of Aglaia odorata extract - Google Patents
The medical usage of the anti-ischemia apoplexy of Aglaia odorata extract Download PDFInfo
- Publication number
- CN109350643A CN109350643A CN201811441732.9A CN201811441732A CN109350643A CN 109350643 A CN109350643 A CN 109350643A CN 201811441732 A CN201811441732 A CN 201811441732A CN 109350643 A CN109350643 A CN 109350643A
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- aglaia odorata
- extract
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- health care
- care product
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/58—Meliaceae (Chinaberry or Mahogany family), e.g. Azadirachta (neem)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The present invention relates to the new medical uses of Aglaia odorata leaf extract, and in particular to Aglaia odorata leaf extract is applied in the drug of preparation treatment ischemia apoplexy.Application the present invention also provides Aglaia odorata leaf extract in preparation prevention and/or treatment in the drug of acute and chronic complication as caused by cerebral ischemia.
Description
Technical field
The present invention relates to the medical usages of Aglaia odorata extract, and in particular to Aglaia odorata extract is preparing neuroprotective drug
Application in object, and in particular to application of the Aglaia odorata in the drug for preparing anti-ischemia apoplexy, and mentioned containing Aglaia odorata
Take the pharmaceuticals of object.
Background technique
Ischemia apoplexy is also known as stroke, is the Acute neuronal necrosis of induction due to the decline of cerebral blood perfusion amount,
Pathogenesis is complicated, can generate irreversible serious damage to brain function.Therefore, ischemia apoplexy has become disables at present
The highest the nervous system disease of rate.How to improve Cranial nerve injury as birth trauma caused by ischemia apoplexy, is always what the world of medicine chased
Target, a variety of neuroprotective agents, including Edaravone, gangliosides, cerebrosid-kinin, butylphenyl phthaleine etc. of clinical use at present,
But their efficacy and saferry needs further to be verified, it is still novel for improving because stroke is led there is an urgent need to research and develop
The neuroprotective agent of the cranial nerve necrosis of cause.
After cerebral ischemia occurs, energy exhaustion is the most critical factor for inducing neurotrosis, and is currently had no for this disease
The specific treatment medicine of reason process.In recent years, there is the original new drug for the treatment of ischemia apoplexy based on Chinese Traditional Medicine exploitation
The concern of people is gradually received, there is very wide development prospect in future.
Aglaia odorata (Aglaia odorata Lour.) is that one kind of Meliaceae (Meliaceae) Aglaia (Aglaia) is normal
Green shrub or dungarunga, alias Zhulan tree, tree orchid and anchovies orchid etc., are distributed mainly on the ground such as India, Malaysia, Australia,
Also often there are cultivation in the provinces such as Guangxi, Guangdong, Yunnan, Guizhou, Fujian and the Sichuan in China.
The effect of Chinese medicine thinks, branch, the leaf of Aglaia odorata can be used as medicine, be mainly used for promoting blood circulation to remove blood stasis and swelling and pain relieving, can use
In treatment rheumatic arthritis, traumatic injury and swollen ulcer drug etc., it is not used for the record of neuroprotection.In modern medicine
Research in, at present about the active report of Aglaia plant pharmacology be concentrated mainly on it is anti-inflammatory with immunoregulation, it is antiviral, anti-
Tumour isoreactivity, the report in terms of there is no for neuroprotection.
Summary of the invention
The present inventor has found that the leaf extract of Aglaia odorata is improving because due to cerebral blood perfusion amount by the research that deepens continuously
The Acute neuronal necrosis aspect for declining and inducing has obviously activity, and it is bad to be suitable for nerve caused by improving apoplexy
Dead neuroprotective agent provides new effective component for acute and chronic complication caused by treatment ischemia apoplexy, cerebral ischemia.
Specifically offer technical solution below of the invention:
Aglaia odorata extract can be applied to nerve protection medicine and/or health care product, and Aglaia odorata extract is in zoopery, thin
In born of the same parents' experiment and molecular biology experiment, the activity of neuronal necrosis caused by improving ischemic is all shown, Aglaia odorata extracts
Object can be used as unique active constituent applied to nerve protection medicine and/or health care product.
The nerve protection medicine and/or health care product are preferably used to the drug and/or health care product for the treatment of ischemia apoplexy,
Can certainly be as the neuroprotective agent of other purposes, such as can be used for Parkinson's disease, Alzheimer's disease, wound and cause
Cranial nerve injury as birth trauma etc., can be used for the treatment and reparation of other neurotrosises such as vertebra.
Aglaia odorata extract may naturally be used for the drug of preparation prevention and/or the treatment complication as caused by cerebral ischemia
And/or in health care product.Acute and chronic complication caused by neurotrosis caused by cerebral ischemia generally has dementia, blindness, aphasia, brothers
One of inability, hemiplegia, cerebral infarction, cerebral hemorrhage are a variety of, and the pharmaceuticals and health care product for treating these diseases are mainly used to control
The damage of nerve is treated, therefore Aglaia odorata extract can play good therapeutic effect.
Aglaia odorata extract of the invention can be the extract of Aglaia odorata plant parts, wherein preferential Aglaia odorata leaf
Extract.The specific extracting method of extract is not particularly limited, can be water extract, ethanol extract, can also
To be extractive with organic solvent.Consider on both side from dissolvent residual and extraction efficiency, preferred alcohol extract.
No matter the preparation of which kind of drug or health care product is used for, and Aglaia odorata extract of the invention can include a kind of or more
The pharmaceutically acceptable auxiliary material of kind.The pharmaceutically acceptable auxiliary material, is not particularly limited, and generally may include medicine
The solvent (such as water, ethyl alcohol, propylene glycol, oil for injection) of field routine, diluent (such as starch, Icing Sugar, dextrin, lactose, pre-
Gelling starch, microcrystalline cellulose, inorganic calcium salt (such as calcium sulfate, calcium monohydrogen phosphate, medicinal calcium carbonate), mannitol etc., gather vegetable oil
Ethylene glycol etc.), adhesive (such as water, ethyl alcohol, starch slurry, sodium carboxymethylcellulose, hydroxypropyl cellulose, methylcellulose and second
Base cellulose, hydroxypropyl methylcellulose etc.), disintegrating agent (such as dried starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crosslinking
Polyvinylpyrrolidone, croscarmellose sodium etc.), lubricant (such as magnesium stearate, superfine silica gel powder, talcum powder, hydrogenation plant
Object oil, polyethylene glycols, magnesium laurylsulfate etc.), sorbefacient (such as surfactant, Azone (Laurocapram),
EDTA, salicylic acid, amino acid 1-ethanamine derivatives, acetoacetic acid esters, β-dicarboxylic ester, aromatic series acid compound, aliphatic
Acid etc.), preservative (such as benzoic acid, hydroxypropyl butyl ester, hydroxypropyl methyl esters, phenol, metacresol), corrigent (such as sucrose, Steviosin
Deng) etc..
As drug and health care product in use, Aglaia odorata extract can also be with existing for treating in ischemic brain
The active constituent of wind cooperates, as such active constituent, there is no specific limitation, can in include and extracted selected from ginkgo leaf
One of object, aspirin, warfarin, clopidogrel, ticlopidine, Dipyridamole a variety of share.
When being applied to the preparation of drug and health care product, can be used as oral preparation makes Aglaia odorata extract of the invention
With, can also be used as non-oral formulation use, be not particularly limited.Such as can be made as oral preparation tablet,
Capsule, pill, granule, pulvis, pelliculae pro cavo oris or oral solution, can also be made the injection of non-oral formulation, paste,
One of creme and suppository are a variety of.In view of bioavilability and action speed, optimizing injection.From trouble easy to use
From the perspective of person's adaptability is good, preferably all kinds of oral preparations.
When for treating ischemia apoplexy or prevention and/or treatment acute and chronic complication as caused by cerebral ischemia,
The subject of drug of the present invention is behaved or mammal.For this purpose, the matter of the intake of drug of the present invention
The quality measured or given is calculated according to adult 60~70kg of weight, is usually taken in for each person every day in terms of the leaf extract of Aglaia odorata
120~700mg more preferably takes in 300~400mg for each person every day.
Pharmacological testing shows that the leaf extract of Aglaia odorata can substantially reduce rat cerebral ischemia face caused by ischemia-reperfusion
Product reduces Neurological deficits.Aglaia odorata extract shows caspase apoptotic signal Pathway Activation in pharmacological evaluation
Inhibitory activity, this provides the other mechanism of molecular biology grade for the neuroprotection mechanism of Aglaia odorata extract and explains.
Detailed description of the invention
Fig. 1 shows the brain section photo of each group animal subject in embodiment 1, wherein " sham " group is sham-operation group,
" model " group is model group, and " Aglaia odorata " group is the leaf extract administration group (300mg/kg) of Aglaia odorata.
Fig. 2 shows the opposite cerebral ischemia area percentage values of each group animal subject in embodiment 1, wherein " sham " group is
Sham-operation group, " model " group are model group, and " Aglaia odorata " group is the leaf extract administration group (300mg/kg) of Aglaia odorata.
Fig. 3 shows the neuroethology score of each group animal subject in embodiment 2, wherein " sham " group is sham-operation
Group, " model " group are model group, and " Aglaia odorata " group is the leaf extract administration group (300mg/kg) of Aglaia odorata.
Fig. 4 shows the molecule mechanism of the leaf extract neuroprotection of Aglaia odorata in embodiment 3, wherein 4 (A) are shown
The leaf extract of Aglaia odorata inhibits cells apoptosis caused by induced by oxygen-glucose deprivation/reperfusion, and 4 (B) show the leaf of Aglaia odorata
Extract inhibits cell mitochondrial damaging action caused by induced by oxygen-glucose deprivation/reperfusion, and 4 (C) show the leaf extract of Aglaia odorata
Inhibit the activation of caspase apoptotic signal access caused by induced by oxygen-glucose deprivation/reperfusion.
Specific embodiment
Illustrate the present invention referring to specific drawings and examples.It will be appreciated by those skilled in the art that these are implemented
Example is merely to illustrate the present invention, does not limit the scope of the invention in any way.
Experimental method in following embodiments is unless otherwise specified conventional method.Original as used in the following examples
Material, reagent material etc. are commercially available products unless otherwise specified.Wherein, part material, reagent purchase situation are as follows:
Aglaia odorata medicinal material plants decomposing biological Technology Co., Ltd. purchased from Kunming；
Ethyl alcohol, petroleum ether, ethyl acetate, n-butanol are that analysis is pure, are purchased from Beijing Chemical Plant.
Embodiment 1The preparation of Aglaia odorata leaf extract
Aglaia odorata leaf takes 200g, and pulverizer is ground.It is extracted 3 times with 8 times of 95% EtOH Sonicates of amount, each 1h.Extracting solution subtracts
Pressure recycling design obtains 95% ethanol extract, wait wave most solvent to no alcohol taste, obtains the extract 5g of Aglaia odorata leaf.
Embodiment 2Protective effect of the Aglaia odorata leaf extract to cerebral ischemia/reperfusion injury of rats
2.1 animal subjects: SD rat, 8 weeks, male, Beijing Vital River Experimental Animals Technology Co., Ltd..
2.2 test medicines:
Aglaia odorata leaf extract is dissolved in the drug solution that 5mg/mL is configured in 0.5% CMCNa solution.
2.3 animal packets and processing:
After SD rat 24, adaptable fed 1 week, every group 8, being randomly divided into 3 groups: sham group, (sham-operation group is only cut
Epidermis but without artery ligation), model group (physiological saline 300ml/kg), Aglaia odorata leaf extract group (300mg/kg).Mould
Type group, Aglaia odorata leaf extract group carry out following operations:
SD rat separates right carotid (CCA), internal carotid (ICA) and external carotid artery (ECA), ligatures ECA and cuts
Disconnected, CCA beats slip-knot temporary interruption with silk thread, after closing ICA with artery clamp folder, cuts off notch in ECA, is inserted into nylon wire, causes big
Brain Middle cerebral artery occlusion ischemic.It extracts nylon wire after 1.5h out, decontrols CCA silk thread, realize Reperfu- sion.Meanwhile each group rat oral gavage is given
With corresponding test medicine.Anesthetized animal after 24 hours, broken end takes out brain after euthanasia, is averagely cut into 8 brain pieces, sets TTC
It is protected from light temperature in dye liquor and incubates about 15min for 37 DEG C.Ischemic region is white, and non-ischemic region is in rose.Scan each group cerebral ischemia area
Brain relative ischemia area figures are calculated afterwards, and for statistical analysis.
When putting to death above-mentioned each group rat, while sham group rat euthanasia is given, according to same operation, obtains brain and cut
Piece, is then scanned and data processing.
2.4 test result
The black-and-white photograph of each group brain section is shown in Fig. 1.The photo of Fig. 1 is shown, and the rat brain slice of Normal group is depth
Color, without ischemic area；Occur the white of large area, display modeling success in the brain sections of model group.Aglaia odorata leaf mentions
The Cerebral Region of object group (300mg/kg) is taken to significantly reduce compared with model group.
The brain relative ischemia area (%) of each group rat, is shown in Table 1 and Fig. 2.
1 each group rat brain relative ischemia area (mean+SD) of table
Note: ## indicates P < 0.01 compared with sham-operation group；* indicates P < 0.01 compared with model group.
The data of table 1 combine Fig. 2 shows: compared with model group, the cerebral ischemia of Aglaia odorata leaf extract group (300mg/kg)
Area reduces (P < 0.01 * *) significantly.
The result of the present embodiment illustrates:
Aglaia odorata leaf extract can efficiently reduce the area of cerebral ischemia re-pouring cerebral Ischemia.Prompt Aglaia odorata leaf mentions
Take object that can improve cerebral injury caused by cerebral ischemia re-pouring well.
Embodiment 3Improvement result of the Aglaia odorata leaf extract to neuroethology caused by cerebral ischemic reperfusion in rats
3.1 animal subjects: SD rat, 8 weeks, male, Beijing Vital River Experimental Animals Technology Co., Ltd..
3.2 test medicines:
Aglaia odorata leaf extract is dissolved in the drug solution that 5mg/mL is configured in 0.5% CMCNa solution.
3.3 animal packets and processing:
After SD rat 24, adaptable fed 1 week, every group 8, being randomly divided into 3 groups: sham group, (sham-operation group is only cut
Epidermis but without artery ligation), model group (physiological saline 300ml/kg), Aglaia odorata leaf extract group (300mg/kg).Mould
Type group, Aglaia odorata leaf extract group animal carry out following operations:
SD rat separates right carotid (CCA), internal carotid (ICA) and external carotid artery (ECA), ligatures ECA and cuts
Disconnected, CCA beats slip-knot temporary interruption with silk thread, after closing ICA with artery clamp folder, cuts off notch in ECA, is inserted into nylon wire, causes big
Brain Middle cerebral artery occlusion ischemic.It extracts nylon wire after 1.5h out, decontrols CCA silk thread, realize Reperfu- sion.Meanwhile each group rat oral gavage is given
With corresponding test medicine.Divide scale statistics each group rat nerve neurological deficit score processed according to longa 5 after 24 hours.
Longa 5 divides standards of grading processed are as follows:
Impassivity injury symptoms, 0 point；
It is unable to fore paw on the left of full extension, 1 point；
It turn-takes to non-ischemic side, 2 points；
It rolls to non-ischemic when walking, 3 points；
It spontaneous cannot walk, consciousness stupor, 4 points.
3.4 test result
The Neurological deficits of each group rat, are shown in Table 2 and Fig. 3.
2 each group rat brain Neurological deficits (mean+SD) of table
Note: ## indicates P < 0.01 compared with sham-operation group；* indicates P < 0.01 compared with model group.
The data and Fig. 3 of table 2 are shown, and the neuroethology of Aglaia odorata leaf extract group (300mg/kg) obtains split pole significantly
Lower than model group (P < 0.01 * *).
The result of the present embodiment illustrates:
Aglaia odorata leaf extract can improve the damage of neuroethology caused by cerebral ischemia re-pouring well.
4 meters of embodimentYoung orchid leaf extract realizes the mechanism of neuroprotection
Inventor has carried out various molecular biology experiments, in the exploration of numerous active mechanisms, invention of the invention
People has found that Aglaia odorata extract shows apparent activity in terms of the regulation of mitochondrial apoptosis signal path.Related experiment process
It is as follows:
PC12 cell line be purchased from Chinese Academy of Medical Sciences's cell centre, with DMEM in high glucose culture medium (contain 10% fetal calf serum,
100U·L-1Penicillin, 100 μ gL-1Streptomysin) secondary culture is carried out, it passes on 1 time within every 2 days.The dissolution of Aglaia odorata leaf extract
In DMSO, it is configured to stock solution.
It is carried out again after handling PC12 cell 2h with Aglaia odorata leaf extract (5,10,50ng/mL) under the conditions of induced by oxygen-glucose deprivation
Perfusion injury processing.Then cell continues culture for 24 hours under normal operation, carries out cell fluorescence dyeing or passes through after collecting cell
Western blot method analyzes protein expression and carries out Study on mechanism.
Hoechst33258 cell fluorescence Coloration experiment condition is as follows: to be inoculated in 24 orifice plates, model group lures PC12 cell
Induced by oxygen-glucose deprivation/reperfusion damage is led, Aglaia odorata leaf extract processing group handles cell according to above-mentioned concentration.Reoxygenation is for 24 hours after anoxic.
The fixed 30min of 4% paraformaldehyde is added；Paraformaldehyde is siphoned away, PBS cleaning is added, cleans 3 times；PBS is discarded, is added
33258 dye liquor of Hoechst dyes 30min；Dye liquor is siphoned away, PBS cleaning is added, each 5min is cleaned 3 times.It is aobvious with fluorescence
Micro mirror (excitation wavelength 352nm, launch wavelength 461nm) observes (× 200) karyomorphism.Finally obtain 4 (A) pictures.
JC-1 cell mitochondrial fluorescent staining experiment condition is as follows: PC12 cell inoculation is lacked in 24 orifice plates, model group induction
Oxygen/lack sugared reperfusion injury, Aglaia odorata leaf extract processing group handles cell according to above-mentioned concentration.Reoxygenation for 24 hours, presses examination after anoxic
Agent box requires for mother liquor to be configured to working solution, and working solution is added, is protected from light, and dyes 30min；Working solution is siphoned away, it is clear that PBS is added
It washes, cleans 3 times, observe (× 200) cell mitochondrial shape with fluorescence microscope (excitation wavelength 514nm, launch wavelength 529nm)
State.Finally obtain 4 (B) pictures.
Western blot experiment condition is as follows: collecting group of cells and with RIPA lysate (inner protein enzyme inhibitor)
Lytic cell obtains total protein.Then 6%-15%SDS-PAGE protein isolate is used, semidry method is by protein delivery to pvdf membrane
On, 5% skimmed milk power closing 20min is added primary antibody (1:1000), is incubated at room temperature 2h, sufficiently washs, addition secondary antibody (1:
1000) 1h, is incubated at room temperature to add ECL reagent colour development sufficiently after washing, take pictures on gel imaging system, finally obtain 4 (C) figures
Piece.With Gel-Pro software 5 (C) pictures are carried out with the absorbance integral analysis of band, draws column diagram.All data are with averagely
Value ± standard deviation indicates.T inspection and one-way analysis of variance, P < 0.05 are carried out using GraphPad Prism5 statistics software
Think statistically significant.
The experimental result dyed from Hoechst33258 cell fluorescence can be seen that (shown in such as Fig. 4 (A)), with Aglaia odorata
The cells show of the addition of leaf extract, induced by oxygen-glucose deprivation/reperfusion damage group goes out apparent Chromatin condensation and karyopyknosis
Equal apoptosis feature, and after the processing of Aglaia odorata leaf extract is added, the appearance of above-mentioned apoptosis feature is significantly suppressed, Aglaia odorata is prompted
Leaf extract has the function of preferable anti anoxia/scarce sugared reperfusion injury.In addition, from JC-1 cell mitochondrial fluorescent staining
Experimental result can be seen that the cells show of induced by oxygen-glucose deprivation/reperfusion damage group goes out apparent mitochondria (shown in such as Fig. 4 (B))
Membrane depolarization equivalent damage feature (green fluorescence), and after the processing of Aglaia odorata leaf extract is added, significantly suppress above-mentioned mitochondria
The appearance of damage characteristic prompts Aglaia odorata leaf extract to have injury of mitochondria caused by preferable anti anoxia/scarce sugared Reperfu- sion
Effect.Fig. 4 (C) shows Aglaia odorata leaf extract can promote caspase-9 under induced by oxygen-glucose deprivation/reperfusion damaging condition,
The activation of caspase-3 and PARP albumen illustrates that Aglaia odorata leaf extract may be by inhibiting mitochondria to mediate
The activation of caspase-9/3/PARP apoptotic signal access, and then play anti anoxia/scarce sugared reperfusion injury effect.Therefore it pushes away
It surveys, the neurocyte protection effect that Aglaia odorata leaf extract mediates may be by inhibiting mitochondrial membrane depolarising, and then inhibit
Mitochondria rely on caspase apoptotic signal access and realize.The mechanism of action had not been reported in the past.
The present invention provides the novel medical uses of Aglaia odorata leaf extract, thus for clinical treatment ischemia apoplexy, with
And prevention and/or treatment the acute and chronic complication as caused by cerebral ischemia provide new selection.
Claims (10)
1. Aglaia odorata extract is preparing the application in nerve protection medicine and/or health care product.
2. application according to claim 1, wherein the nerve protection medicine and/or health care product are treatment ischemic brains
The drug and/or health care product of apoplexy.
3. application according to claim 1, wherein the Aglaia odorata extract is the extract of Aglaia odorata leaf.
4. application according to claim 1, wherein the Aglaia odorata extract is the ethanol extract of Aglaia odorata leaf.
5. application according to claim 1, which is characterized in that described to prepare nerve protection medicine and/or health care product includes
One or more pharmaceutically acceptable auxiliary materials.
6. application according to claim 1, which is characterized in that the nerve protection medicine and/or health care product are oral systems
Agent.
7. the drug and/or health care product of Aglaia odorata leaf extract complication as caused by cerebral ischemia in preparation prevention and/or treatment
In application.
8. application according to claim 7, which is characterized in that the complication is selected from dementia, blindness, aphasia, Shou Zuwu
One of power, hemiplegia, cerebral infarction, cerebral hemorrhage are a variety of.
9. the application according to any one of claim 7, which is characterized in that the prevention and/or treatment are led by cerebral ischemia
The drug and/or health care product of the acute and chronic complication of cause can also include one or more pharmaceutically acceptable auxiliary materials.
10. the application according to any one of claim 7, which is characterized in that the prevention and/or treatment are led by cerebral ischemia
The drug and/or health care product of the acute and chronic complication of cause are oral preparations.
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| CN201811441732.9A CN109350643B (en) | 2018-11-29 | 2018-11-29 | Medical application of aglaia odorata extract in resisting ischemic cerebral apoplexy |
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Citations (3)
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| CN105126002A (en) * | 2015-09-24 | 2015-12-09 | 杨立珍 | Traditional Chinese medicine preparation for preventing and curing epilepsy and preparation method |
| CN105287910A (en) * | 2015-11-10 | 2016-02-03 | 王冬梅 | Anti-atherosclerosis preparation and preparation method |
| CN105287957A (en) * | 2015-11-10 | 2016-02-03 | 王冬梅 | Traditional Chinese medicine composition for preventing and treating arteriosclerosis and preparing method |
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Patent Citations (3)
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| CN105126002A (en) * | 2015-09-24 | 2015-12-09 | 杨立珍 | Traditional Chinese medicine preparation for preventing and curing epilepsy and preparation method |
| CN105287910A (en) * | 2015-11-10 | 2016-02-03 | 王冬梅 | Anti-atherosclerosis preparation and preparation method |
| CN105287957A (en) * | 2015-11-10 | 2016-02-03 | 王冬梅 | Traditional Chinese medicine composition for preventing and treating arteriosclerosis and preparing method |
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