CN107233382B - Application of traditional Chinese medicine composition in preparation of drugs or health-care products for preventing or treating AD - Google Patents
Application of traditional Chinese medicine composition in preparation of drugs or health-care products for preventing or treating AD Download PDFInfo
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Abstract
The invention discloses a new application of a traditional Chinese medicine composition, and particularly relates to an application of the traditional Chinese medicine composition in preparation of a medicine for preventing or treating Alzheimer's disease, wherein the traditional Chinese medicine composition comprises 6 parts by weight of radix astragali preparata and 1 part by weight of radix glycyrrhizae preparata. The traditional Chinese medicine composition acts on a pathologic model of caenorhabditis elegans Alzheimer's disease, and the result shows that the traditional Chinese medicine composition disclosed by the invention can obviously delay the muscle paralysis phenotype of the caenorhabditis elegans of Alzheimer's disease, shows that the traditional Chinese medicine composition has an obvious treatment effect on the caenorhabditis elegans with Alzheimer's disease, and prompts that the traditional Chinese medicine composition provided by the invention has the potential of treating the Alzheimer's disease, and can be applied to preparation of medicines or health products for preventing or treating the Alzheimer's disease.
Description
Technical Field
The invention relates to a new application of a traditional Chinese medicine composition, in particular to an application of the traditional Chinese medicine composition in preparing a medicine or a health-care product for preventing or treating Alzheimer's disease.
Background
Alzheimer's Disease (AD) is a dementia caused by chronic degenerative central nervous system degenerative diseases, the most common cause of dementia and the most common senile dementia. AD is characterized by neuropsychiatric symptoms such as progressive memory impairment, cognitive dysfunction, personality changes, and language disorders.
Alzheimer's disease is the most common and important degenerative disease of the brain, with the aging global population, the incidence of AD is on a markedly rising trend year by year. Most data show that the prevalence is about 5% over 65 years old, 20% over 85 years old, and about 3 times that of men in women. Currently, about 1500 million people worldwide suffer from AD, with the incidence of AD steadily increasing at a rate of 0.5% per year in older people over 65 years of age and 8% per year in older people over 85 years of age. Seriously harms physical and psychological health of old people and influences the quality of life, causes profound pain to patients, brings heavy burden to families and society, becomes a serious social problem, and causes general attention of governments and medical circles.
As the main type of senile dementia, the pathological features of Alzheimer's disease include degeneration of local cerebral neurons, especially hippocampal and cortical neurons, intracellular neurofibrillary tangles and extracellular senile plaque deposits[1,2]Modern medicine has studied the pathogenesis of AD for many years, but the pathogenesis of the AD is still unclear due to its complex etiology, and in the study of the etiology of AD, there is much evidence that a β (β -amyloid, the major component of senile plaque) is the major pathological link that induces AD[3]Second, a β promotes neuronal apoptosis[4]Third, a β can also activate glial cells, release cytokines and inflammatory response mediators, produce inflammatory responses, and a β as a stimulator of cytokine production and immune inflammatory responses of the nervous system by activating peripheral microglia, accelerate neuronal cell death, leading to memory impairment and cognitive impairment[5]Thus, β amyloid has become a recognized target for screening anti-AD drugs.
The AD has complex etiology, long course of disease and more pathogenic links, and needs to be taken for a long time, the current main anti-Alzheimer disease drugs such as acetylcholinesterase inhibitor (such as galantamine) and N-methyl-D-aspartate receptor (NMDA receptor) antagonist (such as memantine) are expensive, the side effects such as hallucination, consciousness chaos, dizziness, headache, tiredness and the like after the drugs are taken are obvious, and the patients' conditions can only be controlled but not reversed.
To date, no specific medicine exists for treating AD, and most of the medicines are still in the stages of basic research and clinical trials. The natural medicine has certain advantages for treating AD proved by long-term application practice. And the pathogenesis of AD is very complex, and the traditional Chinese medicine is multi-target and multi-way to treat AD, thereby arousing multiple aspects of attention.
The invention discloses a traditional Chinese medicine composition, which comprises 6 parts of radix astragali preparata and 1 part of radix glycyrrhizae preparata, and comes from volume five of Taiping Huimin He Ji Ju Fang, which is also named as: astragalus decoction (book Ererjiu from Puji Fang). Prescription: astragalus root (processed with honey) 180g, licorice root (processed) 30 g. Is mainly used for tonifying qi and promoting the production of body fluid. The medicine is mainly used for treating various deficiency of men and women, limb fatigue, vexation and palpitation in chest, frequent thirst, dry lips and mouth, sallow complexion and incapability of eating; it is especially suitable for treating ulcer and furuncle due to thirst, or ulcer and abscess due to thirst. Usually, it can nourish qi and blood, fu organs. Roasted Huang Qi is sweet in taste and warm. It enters lung and spleen meridians. To replenish qi and strengthen the middle-jiao. Can be used for treating qi deficiency, asthenia, anorexia, and loose stool. Prepared licorice root, radix Glycyrrhizae Praeparata, sweet in flavor and neutral in taste. It enters heart, lung, spleen and stomach meridians. Tonify spleen and stomach, replenish qi and recover pulse. Can be used for treating weakness of spleen and stomach, asthenia, palpitation, and intermittent pulse.
Caenorhabditis elegans (Caenorhabditis elegans) is a very useful research platform for multifunctional drug screening and drug action mechanisms. It is cheap and easy to culture; the generation period is short, the number of offspring is large, a large number of individuals with consistent backgrounds can be obtained, the experimental repeatability is ensured, and the experiment is carried out by adopting a large sample amount, so that the influence of individual difference is eliminated; is highly conserved with higher organisms in gene and molecular pathways[6]. It is more and more favored by pharmacologists as a bridge from the primary screening of cell in vitro horizontal drugs to the secondary screening of mouse in vivo horizontal drugs. The American devGen company has been FDA approved for clinical study using humanized nematode screening for antiarrhythmic drugs.
The establishment of the humanized caenorhabditis elegans AD pathological model is to transfer the humanized A β 1-42 gene intoThe expression of the myosin promoter is controlled by temperature, the nematode strain can normally grow at 15 ℃, when the nematode strain is transferred to 25 ℃, A β 1-42 is expressed and accumulated in the muscle of the nematode strain, so that the nematode loses the motor ability and is paralyzed[7]。
The inventor discovers that the traditional Chinese medicine composition disclosed by the invention has an anti-AD effect after screening a large number of traditional Chinese medicine compositions by adopting a caenorhabditis elegans AD pathological model. Therefore, the traditional Chinese medicine composition provided by the invention has the potential of treating the Alzheimer disease, and can be applied to preparation of medicines or health-care products for preventing or treating the Alzheimer disease.
Reference to the literature
[1]Mattson M P.Pathways towards and away from Alzheimer’sdisease.Nature, 2004,430；631-639。
[2]Hardy J,Selkoe D J.The amyloid hypothesis of Alzheimer’s disease:progress and problems on the road to therapeutics.Science,2002,297:353-356。
[3]Fu HJ,Liu B,Frost JL,et al.Amyloid-beta immunotherapy forAlzheimer’s disease[J]. CNS Neurol Disord Drud Targets,2010,9(2):197-206。
[4]NelsonTJ,Alkon DL.Oxidation of cholesterol by amyloid precursorprotein and beta-amyloid peptide[J].J Biol Chem,2005,280(8):7377-7387。
[5] Vimetan, linsuzong, cinnoline, etc. Effect of A β (1-42) -acting microglia on the survival of neural stem cells cultured in vitro [ J ]. Chinese journal of pathophysiology, 2012 (4): 683-688.
[6]Kaletta T,Hengartner MO.Finding function in novel targets:C.elegans as a model organism.Nat Rev Drug Discov 2006,5:387e398。
[7]Link CD.Transgenic invertebrate models of age-associatedneurodegenerative diseases. Mech Ageing Dev 2001,122:1639e1649。
Disclosure of Invention
The invention aims to provide a new application of a traditional Chinese medicine composition. Wherein the application is the application in preparing medicaments or health-care products for preventing or treating Alzheimer's disease.
The traditional Chinese medicine composition comprises 6 parts by weight of radix astragali preparata and 1 part by weight of radix glycyrrhizae preparata.
Preferably, the traditional Chinese medicine composition comprises 15g of radix astragali preparata and 2.5g of radix glycyrrhizae preparata.
The traditional Chinese medicine composition is prepared by the following method: weighing 15g of radix astragali Preparata and 2.5g of radix Glycyrrhizae Preparata according to a formula, uniformly mixing, adding 150ml of water, soaking for 30min, heating and boiling, keeping for 30min, naturally cooling to room temperature, filtering, separating filtrate and filter residue, soaking the filter residue in 100ml of deionized water for 10min, heating to boil with strong fire, heating for 25min, naturally cooling to room temperature, filtering, discarding the filter residue, combining the two filtrates, fixing the volume to 50ml, filtering, and sterilizing to obtain the traditional Chinese medicine composition.
In addition, the traditional Chinese medicine composition can be prepared into any one of capsules, tablets, granules, powder, oral liquid, pills and other dosage forms by adding appropriate auxiliary materials, which can be understood by researchers in the field.
The experiment is carried out by taking the caenorhabditis elegans as a pathologic model of the Alzheimer's disease, and the result shows that the traditional Chinese medicine composition disclosed by the invention can obviously delay the muscle paralysis phenotype of the caenorhabditis elegans of the Alzheimer's disease, shows that the traditional Chinese medicine composition has an obvious treatment effect on the caenorhabditis elegans with the Alzheimer's disease, and prompts that the traditional Chinese medicine composition provided by the invention has the potential of treating the Alzheimer's disease, and can be applied to the preparation of medicines or health care products for preventing or treating the Alzheimer's disease.
When the radix astragali preparata is used singly, the treatment effect on the AD nematodes is weaker under the concentration adopted by the invention; when the honey-fried licorice root is used alone, the drug has no treatment effect on AD nematodes. When the two are used in combination, the anti-AD effect is remarkable. The invention shows that the radix astragali preparata and the radix glycyrrhizae preparata in the water decoction of the traditional Chinese medicine composition provided by the invention have synergistic effect on resisting AD.
Drawings
FIG. 1 shows the result of the Chinese medicinal composition for delaying the muscular paralysis of AD C. Wherein, the group A is NGM culture medium containing 35mg/ml water decoction of Chinese medicinal composition, the group B is NGM culture medium containing 7mg/ml water decoction of Chinese medicinal composition, the group C is NGM culture medium containing 3.5mg/ml water decoction of Chinese medicinal composition, and the control group is NGM culture medium added with sterile water with the same volume as the water decoction of Chinese medicinal composition.
FIG. 2 shows the result of the synergistic effect of the ingredients in the Chinese medicinal composition for delaying the muscular paralysis of AD C. Wherein the traditional Chinese medicine group is coated with OP50 and contains NGM culture medium of 7mg/ml traditional Chinese medicine composition water decoction, the radix astragali Preparata group is coated with OP50 and contains NGM culture medium of 6mg/ml radix astragali Preparata water decoction, the radix Glycyrrhizae Preparata group is coated with OP50 and contains NGM culture medium of 1mg/ml radix Glycyrrhizae Preparata water decoction, and the control group is coated with OP50 and is added with aseptic water with the same volume as the radix astragali Preparata water decoction.
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 Water decoction of a Chinese medicinal composition significantly delays the muscle paralysis of the AD C
1. Biological material
(1) Caenorhabditis elegans CL4176 was purchased from Caenorhabditis Genetics Center (CGC), is a transgenic line that expresses human A β muscle-specifically under induction of a temperature of 25 ℃1-42，Aβ1-42Accumulation in muscle tissue, ultimately leading to nematode paralysis, the present example used the C.elegans CL4176 strain as a pathological model for screening anti-AD drugs.
(2) Escherichia coli OP50 (uracil leaky mutant), purchased from Caenorhabditis Genetics Center (CGC), was used as feed for C.elegans.
2. Reagent
(1) Solid NGM (Newatode Growth Medium) medium composition and preparation (taking 1 liter as an example):
| composition (I) | Content (wt.) |
| NaCl | 3.00g |
| K2HPO4 | 2.34g |
| KH2PO4 | 17.23g |
| Peptone | 2.50g |
| Agar-agar | 17.00g |
| Supplement H2O to | 1000mL |
After preparing the solid NGM culture medium, sterilizing at 121 deg.C under high pressure and constant temperature for 20min, adding 5mg/mL cholesterol 1mL, 1M MgSO41mL,1M CaCl21mL was shaken well and poured hot into a sterilized 9cm plate, approximately 20 mL/plate. Standing for solidification of the culture medium.
(2) M9 liquid formula
(3) Preparation of lysate: a6.4% NaClO solution and a 1M NaOH solution were mixed in a volume ratio of 1: 1.
3. Preparing NGM flat plate containing Chinese medicinal composition water extract
Preparing a water decoction of the traditional Chinese medicine composition:
15g of radix astragali preparata and 2.5g of honey-fried licorice root, uniformly mixing, adding 150ml of water, soaking for 30min, heating and boiling for 30min, naturally cooling to room temperature, filtering, separating filtrate from filter residue, soaking the filter residue in 100ml of deionized water for 10min, heating to boil with strong fire, heating for 25min, naturally cooling to room temperature, filtering, discarding the filter residue, combining the two filtrates, fixing the volume to 50ml, filtering, and sterilizing for later use.
Preparing an NGM flat plate containing the medicine water decoction:
adding radix astragali Preparata water decoction, radix Glycyrrhizae Preparata water decoction or Chinese medicinal composition water decoction of different dilutions into NGM culture medium, and pouring.
Finally preparing NGM culture medium containing 30mg/ml water decoction of radix astragali Preparata; NGM culture medium containing 5mg/ml water decoction of radix Glycyrrhizae Preparata; NGM culture medium containing water decoctions of Chinese medicinal composition 35mg/ml, 7mg/ml, and 3.5 mg/ml; standing for culture medium solidification. Coli OP50 was spread evenly on the medium as feed for nematodes.
4. Carrying out the step
(1) And (3) culturing nematodes:
the nematodes were plated on solid NGM plates coated with E.coli OP50 and then incubated in an incubator at 16 ℃ and synchronized when they reached adult growth.
(2) Nematode synchronization:
selecting NGM culture medium containing a large amount of imagoes and part of nematode eggs which have been hatched, flushing the nematodes from the culture medium by using M9 liquid, transferring the nematodes to a centrifuge tube, standing the nematode to enable the nematodes to freely settle to the bottom of the centrifuge tube, and discarding supernatant. And (3) adding the lysate into the centrifuge tube according to the amount of the nematode, oscillating on a vortex stirrer for 5-7min, stopping vortex when the nematode is completely broken, subpackaging the obtained product in a 1.5mL centrifuge tube, and washing the nematode eggs with an M9 solution for three times.
(3) Effect of water decoction of traditional Chinese medicine composition on Alzheimer's disease caenorhabditis elegans CL4176
Transferring the synchronized nematode eggs subpackaged in the centrifuge tubes to a group A of NGM culture medium containing 35mg/ml of Chinese medicinal composition water decoction, a group B of NGM culture medium containing 7mg/ml of Chinese medicinal composition water decoction, a group C of NGM culture medium containing 3.5mg/ml of Chinese medicinal composition water decoction, a control group of NGM culture medium added with sterile water with the same volume as the Chinese medicinal composition water decoction, wherein the culture dishes of each group are coated with OP50 with the same volume as the Chinese medicinal composition water decoction and used as nematode food, 60 nematodes are distributed in each culture dish, the concentration of each nematode is three in parallel, and the nematode is cultured for 3 days at 16 ℃ to the L3 period. In order to express A beta, the L3 phase nematodes were then induced at 25 ℃ and a count of paralyzed nematode sticks was initiated after 35 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 to the body). The results are shown in FIG. 1.
The ordinate of FIG. 1 shows the percentage of C.elegans which had no paralysis, and at the same time point, the larger this value indicates a larger proportion of C.elegans which had no paralysis in the treated group, i.e.a stronger effect of the drug in delaying paralysis.
Experimental results show that the water decoction of the traditional Chinese medicine composition can delay the muscular paralysis of the AD caenorhabditis elegans, and has better effect at medium concentration. The above examples prove that the water decoction of the traditional Chinese medicine composition provided by the invention has a therapeutic effect on AD caenorhabditis elegans, and the traditional Chinese medicine composition provided by the invention has potential of resisting Alzheimer's disease and can be applied to preparation of medicines for preventing and treating Alzheimer's disease.
Example two radix astragali Preparata and radix Glycyrrhizae Preparata exert synergistic effects in delaying muscular paralysis of AD caenorhabditis elegans
(1) Preparing astragalus root decoction:
15g of radix astragali Preparata is added with 150ml of water, soaked for 30min, heated and boiled for 30min, naturally cooled to room temperature and filtered, the filtrate and the filter residue are separated, the filter residue is soaked for 10min with 100ml of deionized water and heated to boiling with strong fire, then heated for 25min, naturally cooled to room temperature and filtered, the filter residue is discarded, the two filtrates are combined, the volume is fixed to 50ml, and the filtrate is filtered and sterilized for later use.
(2) Preparing roasted liquorice water decoction:
2.5g of honey-fried licorice root, 150ml of water is added, the honey-fried licorice root is soaked for 30min, heated and boiled for 30min, naturally cooled to room temperature and filtered, the filtrate and the filter residue are separated, the filter residue is soaked for 10min by 100ml of deionized water, heated to boiling by strong fire, heated for 25min, naturally cooled to room temperature and filtered, the filter residue is discarded, the two filtrates are combined, the volume is fixed to 50ml, and the filtrate is filtered and sterilized for later use.
The traditional Chinese medicine group is coated with OP50 and contains NGM culture medium of 7mg/ml traditional Chinese medicine composition water decoction, the radix astragali Preparata group is coated with OP50 and contains NGM culture medium of 6mg/ml radix astragali Preparata water decoction, the radix Glycyrrhizae Preparata group is coated with OP50 and contains NGM culture medium of 1mg/ml radix Glycyrrhizae Preparata water decoction, and the control group is coated with OP50 and is added with aseptic water with the same volume as the radix astragali Preparata water decoction. The other operations are the same as those of the first embodiment.
The results of the experiment are shown in FIG. 2, where the ordinate of FIG. 2 indicates the percentage of C.elegans which are not paralyzed, and the larger this value at the same time point indicates that the greater the proportion of C.elegans which are not paralyzed in this treatment group, i.e., the stronger the effect of the drug in delaying paralysis.
Experimental results show that in the embodiment, the effect of delaying the muscle paralysis of the AD nematodes is weaker by using the radix astragali preparata alone, the effect of delaying the muscle paralysis of the AD nematodes by using the radix glycyrrhizae preparata alone is not shown, and the effect of delaying the muscle paralysis of the AD nematodes by using the traditional Chinese medicine composition is obviously stronger than that of using the radix astragali preparata alone or using the radix glycyrrhizae preparata alone. The result shows that the radix astragali preparata and the radix glycyrrhizae preparata can play a synergistic effect in resisting AD when the mass ratio is 6:1, and the traditional Chinese medicine composition can be used for preparing health-care products or medicines for preventing or treating Alzheimer's disease.
Claims (4)
1. The application of the traditional Chinese medicine composition in preparing the medicine for preventing or treating the Alzheimer disease is characterized in that the traditional Chinese medicine composition is composed of 6 parts by weight of radix astragali preparata and 1 part by weight of radix glycyrrhizae preparata.
2. The use of the Chinese medicinal composition of claim 1 in the preparation of a medicament for preventing or treating alzheimer's disease, wherein the Chinese medicinal composition comprises 15g of radix astragali Preparata and 2.5g of radix Glycyrrhizae Preparata.
3. The application of the traditional Chinese medicine composition in preparing the medicine for preventing or treating Alzheimer's disease according to claim 2 is characterized in that the traditional Chinese medicine composition is prepared by weighing radix astragali Preparata and radix Glycyrrhizae Preparata according to a formula, uniformly mixing, adding 150ml of water, soaking for 30min, heating and boiling for 30min, naturally cooling to room temperature, filtering, separating filtrate from filter residue, soaking the filter residue in 100ml of deionized water for 10min, heating to boiling with strong fire, heating for 25min, naturally cooling to room temperature, filtering, removing the filter residue, combining the two filtrates, fixing the volume to 50ml, filtering and sterilizing to obtain the traditional Chinese medicine composition.
4. The use of the Chinese medicinal composition according to any one of claims 1 to 3 in the preparation of a medicament for preventing or treating Alzheimer's disease, wherein the Chinese medicinal composition is prepared into any one dosage form of capsules, tablets, granules, powder, oral liquid and pills.
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| 甘草苷的神经保护及对胆碱酯酶的抑制作用;刘睿婷等;《中国新药杂志》;20081231;第17卷(第7期);第574-577、581页，尤其是第575页左栏第1段，第577页左栏末段 * |
| 黄芪对阿尔茨海默病模型大鼠学习记忆能力的影响;万朋等;《吉林医药学院学报》;20110430;第32卷(第2期);第69-73页，摘要，第70页左栏第1.1节 * |
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