CN105311279A - Application of traditional Chinese medicine composition to preparation of medicines for preventing or treating Alzheimer's disease - Google Patents
Application of traditional Chinese medicine composition to preparation of medicines for preventing or treating Alzheimer's disease Download PDFInfo
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Abstract
The invention discloses 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 to preparation of medicines for preventing or treating Alzheimer's disease. The traditional Chinese medicine composition is mainly composed of rhizoma valerianae jatamansi, rhizoma acori tatarinowii, semen pharbitidis, rhizoma nerdostachyos, uncariae ramulus cum uncis, semen euphorbiae, menthol and valeriana officinalis. According to animal experiment results, the traditional Chinese medicine composition has an evident inhibition effect on paralysis phenotype of an animal phthological model of the Alzheimer's disease, thereby having a potential in prevention or treatment of the Alzheimer's disease. The traditional Chinese medicine composition is applicable to preparation of medicines for preventing or treating the Alzheimer's disease.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition, be specifically related to the preparation method of this Chinese medicine composition and the application in preparation prevention or treatment Alzheimer disease drugs thereof, belong to field of traditional Chinese medicine pharmacy.
Background technology
Alzheimer (Alazheimer ' sdisease, AD) alzheimer disease is also called, a kind of based on the nervous system degenerative disease of Progressive symmetric erythrokeratodermia cognitive disorder and memory impairment, in dementia patients, proportion is more than 75%, within more than 85 years old, has the puzzlement that 30% is subject to Alzheimer in crowd at least.Under the background of China's aged tendency of population, effective prevention and therapy Alzheimer becomes the another key subjects of pendulum in face of us.
The key pathological feature of Alzheimer is amyloid-beta in brain (A β) deposition and neurofibrillary tangles.Modern medicine has carried out years of researches to the pathogenesis of Alzheimer, but due to its cause of disease complexity, the pathogenesis of this disease is still indefinite so far, and " A β cascade hypothesis " is current by one of well accepted mechanism of scientific circles.This theory thinks that the A β of abnormal deposition in brain in patients is by a series of cascade reactions such as radical reaction, Mitochondrial oxidative damage and inflammatory reactions; directly or indirect action in neuron and neurogliocyte; finally cause neuronal function abnormal or dead; cause cognitive impairment and amnesia, finally cause dementia.In familial Alzheimer Disease patient, find that amyloid precursor protein (APP) gene and presenilin genes are undergone mutation, cause A β overexpression in the brain, this becomes one of this theory evidence the strongest.Therefore amyloid beta has become generally acknowledged screening anti-Alzheimer disease drug targets.
Alzheimer belongs to the diseases such as Chinese medicine stays disease, literary composition is silly, language is put upside down, forgetful, and the traditional Chinese medical science thinks that marrow of suffering from a deficiency of the kidney loses as this, and expectorant stasis blocking is stagnant is mark, and the five internal organs are lacked of proper care, and brains apraxia is the pathogenesis of this disease.
The Alzheimer cause of disease is complicated, the course of disease is very long and morbidity link is more, need Long-term taking medicine, anti-Alzheimer disease medicine main is at present divided into two classes: acetylcholinesteraseinhibitors inhibitors (as galantamine) and N-methyl-D-aspartate receptor (nmda receptor) antagonist (as memantine), but said medicine is expensive, side effect remarkable (hallucination, misunderstanding, dizziness, headache and tired), and can only control but can not reverse conditions of patients.
Current Western medicine carries out intervening for a certain particular pathologies link of Alzheimer and often occurs drug resistance and serious adverse reaction.And Chinese medicine has the function Characteristics of various active composition, Mutiple Targets, treatment by Chinese herbs Alzheimer is made to have unique curative effect advantage.
Summary of the invention
The object of the invention is to provide a kind of Chinese medicine composition and preparation method thereof, another object of the present invention is to provide the application of a kind of Chinese medicine composition in preparation prevention or treatment Alzheimer disease drugs, and described Chinese medicine composition comprises following crude drug: Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae.
Wherein the weight portion of the traditional Chinese medicinal composition raw materials of preparation prevention or treatment Alzheimer may be selected to be: Rhizoma valerianae latifoliae 300-600, Rhizoma Acori Graminei 300-600, Semen Pharbitidis 150-300, Radix Et Rhizoma Nardostachyos 150-300, Ramulus Uncariae Cum Uncis 150-300, Semen Euphorbiae 10-35, Mentholum 0.1-0.6, Rhizoma et radix valerianae 0.45-0.75.
The weight portion of the traditional Chinese medicinal composition raw materials of preparation prevention or treatment Alzheimer can be preferably: Rhizoma valerianae latifoliae 500, Rhizoma Acori Graminei 500, Semen Pharbitidis 200, Radix Et Rhizoma Nardostachyos 200, Ramulus Uncariae Cum Uncis 200, Semen Euphorbiae 15, Mentholum 0.3, Rhizoma et radix valerianae 0.62.
Experimental result confirms, Chinese medicine composition disclosed by the invention has significant inhibitory action to the paralysis phenotype that Caenorhabditis elegans Alzheimer animal pathological model A β overexpression causes, experimental result shows, Chinese medicine composition disclosed by the invention has certain effect for the damage for the treatment of A β to body, has the potentiality of prevention or treatment Alzheimer.
This Chinese medicine composition can be made into various pharmaceutical dosage form, Chinese medicine composition of the present invention can according to preparation conventional method in theory, such as, the preparation technology that Fan Biting " pharmacy of Chinese materia medica " (Shanghai Science Press in December, 1997 I version) records, make any dosage form, be preferably decoction, capsule, tablet, granule, powder, oral liquid or pill etc. according to its applicable object and indication; The freeze-dried powder used when intestinal external administration and injection etc.
For enabling above-mentioned dosage form realize, the acceptable adjuvant of pharmacy need be added when preparing these dosage forms, such as: filler, disintegrating agent, lubricant, suspending agent, binding agent, correctives, antiseptic, substrate etc.Filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.; Disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc.; Lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.; Suspending agent comprises: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, agar, hydroxypropyl emthylcellulose etc.; Binding agent comprises: starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc.; Sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.; Correctives comprises: sweeting agent and various essence; Antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters, benzalkonium bromide, acetic acid chloroethene are fixed, Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods; Substrate comprises: PEG6000, PEG4000, insect wax etc.
For realizing making the object of patent medicine, necessary extraction or pulverizing are carried out to the some or all of medical material in this Chinese medicine composition, but do not get rid of and all decoct soup and take.
Chinese medicine composition tablet disclosed by the invention can be prepared as follows:
A, take Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae in proportion;
B, valerian powder are broken into coarse powder, carry out percolation with 60% ethanol as solvent, collect percolate;
3/5 weight portion of c, Rhizoma valerianae latifoliae gross weight, Rhizoma Acori Graminei, Radix Et Rhizoma Nardostachyos extract volatile oil, and medical material residue mixes with Ramulus Uncariae Cum Uncis, decocts with water 1-2 time, are preferably 2 times; Each 1-2 hour, is preferably 1.5 hours, filters, merging filtrate;
D, Semen Pharbitidis, Semen Euphorbiae are ground into coarse powder, carry out percolation with 70% ethanol as solvent, and collect percolate and merge with above-mentioned decocting liquid, be evaporated to the extractum that relative density is 1.26-1.30, preferred relative density is the extractum of 1.28;
2/5 parts by weight of crushed of e, Rhizoma valerianae latifoliae gross weight becomes fine powder to mix homogeneously with above-mentioned extractum, and drying is also ground into fine powder, dry with the alcohol granulation of Rhizoma et radix valerianae percolate and 70%-75%, be preferably the alcohol granulation drying of 70%, add Mentholum and above-mentioned volatile oil, mixing, tabletted.
Transgenic C nematicide strain CL4176 is evaluated the effect of disclosed Chinese medicine composition anti-Alzheimer disease by the present invention as the pathological model screening anti-Alzheimer disease medicine, be based on: the cerebral cortex of patients with Alzheimer disease and Hippocampus occur amyloid-beta (A β) assemble formed senile plaque (SP) be Alzheimer one of major pathologic features the most, amyloid-beta can effectively weaken synaptic structure and function, is the important substance causing Alzheimer.Transgenic C nematicide strain CL4176 expresses mankind A β at 25 DEG C of temperature-induced lower muscle specifics, and A β assembles in muscular tissue, finally causes nematicide to be benumbed.By drug effect in nematicide CL4176, if nematode population paralysis situation improves, be embodied in the remarkable retardation of nematicide paralysis time, then illustrate that medicine has anti-Alzheimer disease effect.
Environmental change reaction is responsive to external world for Caenorhabditis elegans, and utilize C. Elegans Automatic Screening can carry out accurate evaluation to the toxicity of medicine, body length is one of assessing developmental toxicity index commonly used the most.Drug effect, after nematicide, significantly reduces if nematicide body is long, then illustrate that medicine has significant development toxicity to nematicide.
Compared with prior art, the present invention has following beneficial effect:
The present invention have evaluated the effect of disclosed Chinese medicine composition to Caenorhabditis elegans Alzheimer disease model paralysis phenotype, result confirms that Chinese medicine composition has obvious inhibitory action to Caenorhabditis elegans Alzheimer disease model paralysis phenotype, has the potentiality of prevention or treatment Alzheimer.
Therefore, disclosed Chinese medicine composition can be applied in preparation prevention or treatment Alzheimer disease drugs.
Accompanying drawing explanation
Fig. 1 is the impact of positive drug memantine on Caenorhabditis elegans Alzheimer disease model paralysis phenotype.
Fig. 2 is that the Chinese medicine composition of formula 1 is on the impact of Caenorhabditis elegans Alzheimer disease model paralysis phenotype.
Fig. 3 is that the Chinese medicine composition of formula 2 is on the impact of Caenorhabditis elegans Alzheimer disease model paralysis phenotype.
Fig. 4 is that the Chinese medicine composition of formula 3 is on the impact of Caenorhabditis elegans Alzheimer disease model paralysis phenotype.
Fig. 5 is the evaluating drug effect experimental result of Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis and Chinese medicine composition thereof.
Fig. 6 is the toxicity assessment experimental result of Rhizoma valerianae latifoliae and Chinese medicine composition.
Detailed description of the invention
Below in conjunction with specific embodiment, technical scheme of the present invention is described in further detail, but protection scope of the present invention is not limited to the following stated.
The preparation of embodiment one Chinese medicine composition tablet
Material: Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae, purchased from the Lanzhou chain Co., Ltd of Hui Rentang Pharmaceutical.
Formula 1:
Rhizoma valerianae latifoliae 300, Rhizoma Acori Graminei 300, Semen Pharbitidis 150, Radix Et Rhizoma Nardostachyos 150, Ramulus Uncariae Cum Uncis 150, Semen Euphorbiae 10, Mentholum 0.1, Rhizoma et radix valerianae 0.45.
Formula 2:
Rhizoma valerianae latifoliae 500, Rhizoma Acori Graminei 500, Semen Pharbitidis 200, Radix Et Rhizoma Nardostachyos 200, Ramulus Uncariae Cum Uncis 200, Semen Euphorbiae 15, Mentholum 0.3, Rhizoma et radix valerianae 0.62.
Formula 3:
Rhizoma valerianae latifoliae 600, Rhizoma Acori Graminei 600, Semen Pharbitidis 300, Radix Et Rhizoma Nardostachyos 300, Ramulus Uncariae Cum Uncis 300, Semen Euphorbiae 35, Mentholum 0.6, Rhizoma et radix valerianae 0.75.
Preparation method:
A, take raw material by above-mentioned formula proportion respectively: Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae;
B, valerian powder are broken into coarse powder, carry out percolation with 60% ethanol as solvent, collect percolate;
3/5 weight portion of c, Rhizoma valerianae latifoliae gross weight, Rhizoma Acori Graminei, Radix Et Rhizoma Nardostachyos extract volatile oil, and medical material residue mixes with Ramulus Uncariae Cum Uncis, decocts with water 1-2 time, each 1-2 hour, filter, merging filtrate;
D, Semen Pharbitidis, Semen Euphorbiae are ground into coarse powder, carry out percolation with 70% ethanol as solvent, collect percolate and merge with above-mentioned decocting liquid, being evaporated to the extractum that relative density is 1.26-1.30;
2/5 parts by weight of crushed of e, Rhizoma valerianae latifoliae gross weight becomes fine powder to mix homogeneously with above-mentioned extractum.
Above-mentioned extract dry is ground into fine powder, dry with the alcohol granulation of Rhizoma et radix valerianae percolate and 70%-75%, add Mentholum and above-mentioned volatile oil, mixing, tabletted.
Embodiment two memantine is to the effect of Caenorhabditis elegans CL4176
1. biomaterial
(1) Caenorhabditis elegans CL4176 is purchased from CaenorhabditisGeneticsCenter; For transgenic strain, express mankind A β at 25 DEG C of temperature-induced lower muscle specifics, A β assembles in muscular tissue, finally causes nematicide to be benumbed, and the present embodiment adopts C. Elegans Automatic Screening strain CL4176 as the pathological model of screening anti-Alzheimer disease medicine.
(2) escherichia coli OP50 (uracil leaky mutant), purchased from CaenorhabditisGeneticsCenter, as the food of Caenorhabditis elegans.
2. reagent
(1) memantine (MEM), chemical name: MEM amine hydrochlorate, molecular formula: C
12h
21nHCl, purchased from lark prestige science and technology, CAS:41100-52-1; Memantine is uncompetitive nmda receptor antagonist, be used for the treatment of middle severe to severe dementia of the Alzheimer type, bibliographical information memantine (2.16mg/ml) can significantly suppress transgenic nematode strain CL4176 to benumb phenotype, and the present embodiment it can be used as positive drug.
(2) solid NGM (NematodeGrowthMedium) medium component and making (being upgraded to example with 1):
| Composition | Content |
| NaCl | 3.00g |
| K 2HPO 4 | 2.34g |
| KH 2PO 4 | 17.23g |
| Peptone | 2.50g |
| Agar | 17.00g |
| Supplement H 2O extremely | 1000ml |
After solid NGM culture medium prepares, high-pressure constant temp sterilizing 20min at 121 DEG C, adds 5mg/ml cholesterol 1ml, 1MMgSO under aseptic operating platform
41ml, 1MCaCl
21m shakes up, and pours sterilized 9cm culture plate while hot into, about 20ml/ plate.Leave standstill and wait for culture medium solidifying, for subsequent use.
(3) the NGM culture medium of hydrochloric memantine is prepared
Memantine is dissolved in sterilized water, and preparation becomes the mother solution of 2.16mg/ml, prepares the NGM culture medium that hydrochloric memantine final concentration is 215.76 μ g/ml, 107.88 μ g/ml, 21.58 μ g/ml, 4.32 μ g/ml, 0.86 μ g/ml respectively.
Blank is add the NGM culture medium with memantine equal-volume sterilized water under similarity condition.
Above-mentioned culture medium is all coated with the food of escherichia coli OP50 as nematicide.
(4) M9 formula of liquid
| Na 2HPO 4 | 6.00g |
| KH 2PO 4 | 3.00g |
| NaCl | 5.00g |
| 1M MgSO 4 | 1.00ml |
| Supplement H 2O extremely | 1000ml |
(5) preparation of lysate: 6.4%NaClO
3the 1:1 mixing by volume of solution and 1MNaOH solution.
3. implementation step
(1) cultivation of nematicide:
Be connected on by nematicide and scribble on the solid NGM plate of escherichia coli OP50, the incubator being then placed in 16 DEG C is cultivated, and carries out synchronization process when nematicide grows to adult.
(2) nematicide synchronization:
Select containing a large amount of adult and the NGM culture medium having partial line worm's ovum to hatch, with M9 liquid, nematicide is swept away from culture medium, transfer in centrifuge tube, leave standstill and make, at the bottom of nematicide free settling to pipe, to abandon supernatant.How many sight line worm amounts adds nematicide lysate in centrifuge tube, whirlpool misfortune agitator vibrates 5-7 minute until nematicide complete rupture time stop vortex, and be sub-packed in 1.5mL centrifuge tube, wash line eggs three times with M9 solution.
(3) memantine is to the effect of Caenorhabditis elegans CL4176
After nematicide synchronization, line eggs is transferred to the NGM culture medium containing variable concentrations memantine, each culture medium 60 nematicides, each drug level, using three culture medium as parallel, cultivates 3 days to the L3 phase for 16 DEG C.
In order to make nematicide express A β, L3 phase nematicide being turned and induces at 25 DEG C, start after 34h to count nematicide paralysis number.Every two hours count once, until all nematicides are all benumbed (when paralysis refers to mechanical stimulus nematode body, nematicide can not move or only have head movement), the results are shown in Figure 1.
As seen from Figure 1, positive drug memantine can significantly suppress CL4176 to benumb phenotype, the memantine strong inhibition elegans development of 215.76 μ g/ml and 107.88 μ g/ml when 21.58 μ g/ml, 4.32 μ g/ml, 0.86 μ g/ml.Experimental result shows that this experimental system is normal, illustrates that the present invention can adopt C. Elegans Automatic Screening strain CL4176 as screening anti-Alzheimer disease medicine.
Embodiment three Chinese medicine composition is to the effect of Caenorhabditis elegans CL4176
1. biomaterial is with embodiment two.
2. reagent
(1) Chinese medicine composition sheet is for according to the preparation prepared by embodiment one method, is specially: the Chinese medicine composition sheet of the Chinese medicine composition sheet of formula 1, the Chinese medicine composition sheet of formula 2, formula 3.
(2) solid NGM (NematodeGrowthMedium) medium component and making (being upgraded to example with 1): with embodiment two.
(3) preparation is containing the NGM culture medium of Chinese medicine composition
By Chinese medicine composition sheet mortar pulverize, be dissolved in sterilized water, and preparation become 150mg/ml Chinese medicine composition aqueous solution, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.Added by Chinese medicine composition aqueous solution in the NGM culture medium of thawing, it is 1.0mg/ml, 5.0mg/ml, 15.0mg/mlNGM culture medium that preparation becomes containing Chinese medicine composition final concentration.
Blank is add the NGM culture medium with Chinese medicine composition aqueous solution equal-volume sterilized water under similarity condition.
Above-mentioned culture medium is all coated with the food of escherichia coli OP50 as nematicide.
(4) M9 formula of liquid: with embodiment two.
(5) preparation of lysate: with embodiment two.
3. implementation step
(1) cultivation of nematicide: with embodiment two.
(2) nematicide synchronization: with embodiment two.
(3) Chinese medicine composition is to the effect of Caenorhabditis elegans CL4176
After nematicide synchronization, line eggs is transferred to and be coated with OP50 and mix in the NGM culture medium of variable concentrations formula 1 Chinese medicine composition sheet, formula 2 Chinese medicine composition sheet, formula 3 Chinese medicine composition sheet; Blank is the NGM culture medium being coated with OP50 and being added with equal-volume sterilized water, each culture medium 60 nematicides, and each drug level, using three culture medium as parallel, cultivates 3 days to the L3 phase for 16 DEG C.
In order to make nematicide express A β, L3 phase nematicide being turned and induces at 25 DEG C, start after 34h to count nematicide paralysis number.Every two hours count once, until all nematicides are all benumbed (when paralysis refers to mechanical stimulus nematode body, nematicide can not move or only have head movement), the results are shown in Figure 2, Fig. 3, Fig. 4.
As can be seen from Fig. 2, Fig. 3, Fig. 4, nematicide paralysis phenotype (p < 0.001) that formula 1, formula 2, formula 3 all can significantly suppress A β overexpression to cause, wherein, the nematicide paralysis phenotypic effect that formula 2 suppresses A β overexpression to cause is best, during 44h, the ratio of not benumbing of 15mg/ml, 5mg/ml also remains on about 80% (Fig. 3), and in formula 1 and formula 3, do not benumb ratio during 5mg/ml all lower than 80% (Fig. 2, Fig. 4).
In a word, as can be seen from the above-described embodiment, this Chinese medicine composition has obvious inhibitory action to the paralysis phenotype that Alzheimer disease model Caenorhabditis elegans A beta peptide aggregation causes, and illustrates that this Chinese medicine composition has the potentiality of prevention or treatment Alzheimer.Therefore Chinese medicine composition disclosed in this invention can prevent or treat in the medicine of Alzheimer to apply in preparation.
The drug effect of embodiment four Chinese medicine composition and toxicity comparative evaluation experiment
1. biomaterial is with embodiment one.
2. reagent
(1) Chinese medicine composition sheet is for according to the preparation prepared by embodiment one method, is specially: the Chinese medicine composition sheet of formula 2.
(2) solid NGM (NematodeGrowthMedium) medium component and making (being upgraded to example with 1): with embodiment one.
(3) preparation is containing the NGM culture medium of water-soluble Chinese medicine composition
By Chinese medicine composition sheet mortar pulverize, be dissolved in sterilized water, and preparation become 150mg/ml Chinese medicine composition aqueous solution, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.Added by Chinese medicine composition aqueous solution in the NGM culture medium of thawing, it is the NGM culture medium of 15.0mg/ml that preparation becomes containing Chinese medicine composition final concentration.
(4) Rhizoma valerianae latifoliae extract and the NGM culture medium containing Rhizoma valerianae latifoliae extract thereof is prepared
3/5 weight portion getting Rhizoma valerianae latifoliae gross weight in formula 2 extracts volatile oil, and medical material residue decocts with water 1-2 time, is preferably 2 times; Each 1-2 hour, is preferably 1.5 hours, and filter, merging filtrate, is evaporated to the extractum that relative density is 1.26-1.30, and preferred relative density is the extractum of 1.28; 2/5 parts by weight of crushed of Rhizoma valerianae latifoliae gross weight becomes fine powder for subsequent use.By above volatile oil, extractum and fine powder mix, add sterilized water dissolve, be formulated as Rhizoma valerianae latifoliae aqueous solution, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.Rhizoma valerianae latifoliae aqueous solution is added in the NGM culture medium of thawing, makes the Rhizoma valerianae latifoliae concentration in culture medium consistent with containing the Rhizoma valerianae latifoliae concentration in the NGM culture medium of formula 2 Chinese medicine composition.
(5) Rhizoma Acori Graminei extract and the NGM culture medium containing Rhizoma Acori Graminei extract thereof is prepared
Get Rhizoma Acori Gramineii extract volatile oil in formula 2, medical material residue decocts with water 1-2 time, is preferably 2 times; Each 1-2 hour, is preferably 1.5 hours, filters, merging filtrate; Be evaporated to the extractum that relative density is 1.26-1.30, preferred relative density is the extractum of 1.28.By above volatile oil and extractum mixing, add sterilized water dissolve, be formulated as Rhizoma Acori Graminei aqueous solution, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.Rhizoma Acori Graminei aqueous solution is added in the NGM culture medium of thawing, makes the Rhizoma Acori Graminei concentration in culture medium consistent with containing the Rhizoma Acori Graminei concentration in the NGM culture medium of formula 2 Chinese medicine composition.
(6) prepare Ramulus Uncariae Cum Uncis extract and contain Ramulus Uncariae Cum Uncis extract NGM culture medium
Get Ramulus Uncariae Cum Uncis in formula 2 to decoct with water 1-2 time, be preferably 2 times; Each 1-2 hour, is preferably 1.5 hours, filters, merging filtrate; Be evaporated to the extractum that relative density is 1.26-1.30, preferred relative density is the extractum of 1.28.Add in extractum sterilized water dissolve, be formulated as Ramulus Uncariae Cum Uncis aqueous solution, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.Ramulus Uncariae Cum Uncis aqueous solution is added in the NGM culture medium of thawing, makes the Ramulus Uncariae Cum Uncis concentration in culture medium consistent with containing the Ramulus Uncariae Cum Uncis concentration in the NGM culture medium of formula 2 Chinese medicine composition.
(7) prepare Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis prescription extract and contain prescription NGM culture medium
3/5 weight portion and the Rhizoma Acori Graminei of getting Rhizoma valerianae latifoliae gross weight in formula 2 extract volatile oil respectively, merge, decoct with water residue obtained 1-2 time with Ramulus Uncariae Cum Uncis, are preferably 2 times; Each 1-2 hour, is preferably 1.5 hours, filters, merging filtrate; Be evaporated to the extractum that relative density is 1.26-1.30, preferred relative density is the extractum of 1.28.2/5 parts by weight of crushed of Rhizoma valerianae latifoliae gross weight becomes fine powder for subsequent use.By above volatile oil, extractum and fine powder mix, add sterilized water dissolve, be formulated as the aqueous solution of Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis prescription extract, ultrasonic 30min under room temperature, and under 10000rpm centrifugal 10min, get supernatant, in 4 DEG C of preservations.The aqueous solution of Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis prescription extract is added in the NGM culture medium of thawing, makes the Rhizoma valerianae latifoliae in culture medium, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis concentration consistent with the concentration containing the Rhizoma valerianae latifoliae in the NGM culture medium of formula 2 Chinese medicine composition, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis.
Blank is add the NGM culture medium with Chinese medicine composition aqueous solution equal-volume sterilized water under similarity condition.
Above-mentioned culture medium is all coated with the food of escherichia coli OP50 as nematicide.
(8) M9 formula of liquid: with embodiment one.
(9) preparation of lysate: with embodiment one.
3. implementation step
(1) cultivation of nematicide: with embodiment one.
(2) nematicide synchronization: with embodiment one.
(3) to the drug effect comparative evaluation of Chinese medicine composition
After nematicide synchronization, line eggs is transferred to and be coated with OP50 and the NGM culture medium mixing Chinese medicine composition, the NGM culture medium of Rhizoma valerianae latifoliae extract, the NGM culture medium of Rhizoma Acori Graminei extract, the NGM culture medium of Ramulus Uncariae Cum Uncis extract, the NGM culture medium of Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Ramulus Uncariae Cum Uncis prescription extract; Blank is be coated with OP50 and be added with the NGM culture medium with Chinese medicine composition aqueous solution equal-volume sterilized water.Each culture medium 60 nematicides, cultivate 3 days to the L3 phase for 16 DEG C.
In order to make nematicide express A β, L3 phase nematicide being turned and induces at 25 DEG C, start after 34h to count nematicide paralysis number.Every two hours count once, until all nematicides are all benumbed (when paralysis refers to mechanical stimulus nematode body, nematicide can not move or only have head movement), the results are shown in Figure 5.
As can be seen from experimental result, nematicide paralysis phenotype (p < 0.001) that Rhizoma valerianae latifoliae, Ramulus Uncariae Cum Uncis, Rhizoma Acori Graminei and Chinese medicine composition can both significantly suppress A β overexpression to cause, wherein, the nematicide paralysis phenotypic effect that Rhizoma valerianae latifoliae suppresses A β overexpression to cause is best, almost suitable with Chinese medicine composition effect; Ramulus Uncariae Cum Uncis and Rhizoma Acori Graminei benumb phenotype inhibition not as good as Chinese medicine composition (P < 0.001) to nematicide.And after the combination of Rhizoma valerianae latifoliae, Ramulus Uncariae Cum Uncis and Rhizoma Acori Graminei thereof, have a strong impact on nematode growth, nematicide can not hatch, illustrate that three combines rear toxicity and increases.
(4) to the toxicity comparative evaluation of Chinese medicine composition
Immediately nematicide is placed in 50 DEG C of process 15min after paralysis experiment terminates, stretches to make nematode body and be convenient to measure.Examine under a microscope and use MvImage image processing system to take pictures and slotted line polypide length.The results are shown in Figure 6.
As can be seen from experimental result, compared with blank and Chinese medicine composition thereof, Rhizoma valerianae latifoliae group nematicide body is long significantly to be reduced, and show that Rhizoma valerianae latifoliae has inhibitory action significantly to nematode growth growth, toxicity is large compared with Chinese medicine composition.
Tested as can be seen from the effect assessment of the present embodiment, the nematicide paralysis phenotype that Rhizoma valerianae latifoliae, Ramulus Uncariae Cum Uncis and Rhizoma Acori Graminei and Chinese medicine composition thereof all can significantly suppress A beta peptide aggregation to cause, wherein Rhizoma valerianae latifoliae and Chinese medicine composition effect optimum; Tested as can be seen from the toxicity assessment of the present embodiment, compared with blank and Chinese medicine composition thereof, Rhizoma valerianae latifoliae significantly suppresses nematode growth to be grown, toxicity is significantly higher than Chinese medicine composition, and Chinese medicine composition does not almost affect nematicide body is long, show that Chinese medicine composition toxicity is significantly lower than Rhizoma valerianae latifoliae.
In a word, the present embodiment result shows that Chinese medicine composition has Be very effective, feature that toxicity is little in prevention or treatment Alzheimer, illustrate that Chinese medicine composition has the potentiality of prevention or treatment Alzheimer, can apply in preparation prevention or treatment Alzheimer disease drugs.
Claims (5)
1. the application of Chinese medicine composition in preparation prevention or treatment Alzheimer disease drugs, it is characterized in that, this Chinese medicine composition comprises following crude drug: Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae.
2. the application of a kind of Chinese medicine composition as claimed in claim 1 in preparation prevention or treatment Alzheimer disease drugs, it is characterized in that, the weight portion of this traditional Chinese medicinal composition raw materials is as follows: Rhizoma valerianae latifoliae 300-600, Rhizoma Acori Graminei 300-600, Semen Pharbitidis 150-300, Radix Et Rhizoma Nardostachyos 150-300, Ramulus Uncariae Cum Uncis 150-300, Semen Euphorbiae 10-35, Mentholum 0.1-0.6, Rhizoma et radix valerianae 0.45-0.75.
3. the application of a kind of Chinese medicine composition as claimed in claim 2 in preparation prevention or treatment Alzheimer disease drugs, it is characterized in that, the weight portion of this traditional Chinese medicinal composition raw materials is as follows: Rhizoma valerianae latifoliae 500, Rhizoma Acori Graminei 500, Semen Pharbitidis 200, Radix Et Rhizoma Nardostachyos 200, Ramulus Uncariae Cum Uncis 200, Semen Euphorbiae 15, Mentholum 0.3, Rhizoma et radix valerianae 0.62.
4. the application of a kind of Chinese medicine composition as described in any one of claim 1-3 in preparation prevention or treatment Alzheimer disease drugs, it is characterized in that, this Chinese medicine composition is capsule, tablet, granule, powder, oral liquid or pill as the pharmaceutical preparation of active component.
5. the application of a kind of Chinese medicine composition as claimed in claim 4 in preparation prevention or treatment Alzheimer disease drugs, is characterized in that the preparation method of described Chinese medicine composition tablet comprises following steps:
A, take Rhizoma valerianae latifoliae, Rhizoma Acori Graminei, Semen Pharbitidis, Radix Et Rhizoma Nardostachyos, Ramulus Uncariae Cum Uncis, Semen Euphorbiae, Mentholum, Rhizoma et radix valerianae in proportion;
B, valerian powder are broken into coarse powder, carry out percolation with 60% ethanol as solvent, collect percolate;
3/5 weight portion of c, Rhizoma valerianae latifoliae gross weight, Rhizoma Acori Graminei, Radix Et Rhizoma Nardostachyos extract volatile oil, and medical material residue mixes with Ramulus Uncariae Cum Uncis, decocts with water 1-2 time, each 1-2 hour, filter, merging filtrate;
D, Semen Pharbitidis, Semen Euphorbiae are ground into coarse powder, carry out percolation with 70% ethanol as solvent, collect percolate and merge with above-mentioned decocting liquid, being evaporated to the extractum that relative density is 1.26-1.30;
2/5 parts by weight of crushed of e, Rhizoma valerianae latifoliae gross weight becomes fine powder to mix homogeneously with above-mentioned extractum, and drying is also ground into fine powder, adds Mentholum and above-mentioned volatile oil with the alcohol granulation drying of Rhizoma et radix valerianae percolate and 70%-75%, mixing, tabletted.
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Cited By (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 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732515A (en) * | 2010-01-26 | 2010-06-16 | 昆明中药厂有限公司 | Quality test method of Dianxianning tablets |
| CN102058630A (en) * | 2010-12-16 | 2011-05-18 | 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
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101732515A (en) * | 2010-01-26 | 2010-06-16 | 昆明中药厂有限公司 | Quality test method of Dianxianning tablets |
| CN102058630A (en) * | 2010-12-16 | 2011-05-18 | 苏州大学 | Valerian preparation for tranquilizing and allaying excitement and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张暋: "《实用中医老年病学》", 30 April 2000, 人民军医出版社 * |
Cited By (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 |
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