CN114306543B - Traditional Chinese medicine composition for treating Alzheimer disease and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition for treating Alzheimer disease and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a traditional Chinese medicine composition for treating Alzheimer disease, and a preparation method and application thereof. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10-15 parts of eclipta, 3-8 parts of diverse wormwood herb, 2-6 parts of semen katsumadai, 5-15 parts of malva seed, 2-6 parts of cortex caryophylli and 10-15 parts of fiveleaf akebia fruit. The Chinese medicinal composition has the basic effects of invigorating kidney, removing blood stasis, invigorating stomach, moisturizing intestine, eliminating dampness, and regulating qi-flowing. Animal experiment results show that the high dosage (12.74 g/kg) of the traditional Chinese medicine composition and the dosage (6.37 g/kg) of the traditional Chinese medicine composition can improve the learning ability of the Kunming mice with the Alzheimer disease induced by beta-amyloid protein, the expression level of the precursor protein of the beta-amyloid protein, the expression level of interleukin-1 beta and the expression level of the acetylcholinergic enzyme, and have no influence on the main organ indexes and the body weight of the mice, so that the traditional Chinese medicine composition is effective in treating the Alzheimer disease and has no side effect.
Description
Technical Field
The invention relates to a traditional Chinese medicine composition, a preparation method and application thereof, in particular to a traditional Chinese medicine composition for treating Alzheimer disease, a preparation method and application thereof. The invention belongs to the technical field of traditional Chinese medicine research.
Background
Alzheimer's Disease is a common senile Disease (Hampel H, vassar R, de Strooper B, et al, beta-secret BACE1 in Alzheimer's Disease [ J ]. Biol Psychiary, 2021, 89 (8): 745-756.), which is often associated with increased beta-amyloid levels in different areas of brain tissue (Busche MA, hyman BT. Synergy beta-amylase amyoid-beta and tau in Alzheimer's Disease [ J ]. Nat Neurosci,2020, 23 (10): 1183-1193.). Beta-amyloid is a very powerful stimulator of Alzheimer's Disease (Chen WT, lu A, craassaerts K, et al spatial Transcriptomics and In Situ Sequencing to Study Alzheimer's Disease [ J ]. Cell,2020, 182 (4): 976-991.) which promotes the formation of positive feedback loops in the brain tissue of the body that are closely related to the inflammatory response, and the beta-amyloid content continues to increase (Breijyeh Z, karaman R. Compilense Review on Alzheimer's Disease Causes and Treatment [ J ]. Molecular, 2020, 25 (24): 5789.) which continuously aggravates the sterile inflammatory response of the brain tissue, thereby causing the corresponding symptoms and signs.
Research data shows that Alzheimer's disease is mostly accompanied by reduced learning ability (Kent SA, spires-Jones TL, durrant CS.S. physiological roles of tau and Abeta: implications for Alzheimer's disease pathology and therapeutics [ J ]. Acta neuroskill, 2020, 140 (4): 417-447.), recent events are extremely forgotten, which have a relationship with increased inflammatory mediator content in brain tissue and damage to hippocampal regions, and also with increased β -amyloid precursor protein content in brain tissue, whereas abnormal increases in inflammatory mediators in brain tissue and β -amyloid precursor protein occur, and positive feedback increases in β -amyloid content in different regions of brain tissue, resulting in reduced learning ability (Roussarie JP, yao V, rodriguez-Rodriguez P, et al selective Neuronal Vulnerability in Alzheimer's Disease A Network-base Analysis [ J ]. Neuron 2020, 107 (5): 821-835).
As the body ages, the amount of free radicals produced in brain tissue increases gradually, and the astrocyte structural abnormalities cause abnormal blood brain barrier structures (Serrano-Pozo A, das S, hyman BT. APOE and Alzheimer 'S disease advances in genetics pathophysiology and therapeutic approaches [ J ]. Lancet Neurol,2021, 20 (1): 68-80.), with imbalance in the choline system of brain tissue, abnormal levels of acetylcholine metabolizing enzymes (Ren JM, zhang SL, wang XL, et al expression levels of the alpha nicotinic acetylcholine receptor in the brains of patients with Alzheimer' S disease and their effect on synaptic proteins in SH-SY5Y cells [ J ]. Mol Med Rep,2020, 22 (3): 2063-2075.), causing a further increase in the free radical levels (Uversky VN, elrashdy F, aljadawi A, et al SEver acute respiratory syndrome coronavirus 2infection reaches the human nervous system How[J ]. J Neurosci Res, 1, 99 (3): 750-777.), and further promote a waterfall inflammatory reaction of the free radicals in brain tissue.
In fact, the elderly and the weak are mostly accompanied by emotional disorders, and the pathological product accumulation is a relatively normal physiological phenomenon, and the pathological product accumulation is related to microglia modulate neurodegeneration in Alzheimer ' S and Parkinson ' S diseases (J. Scientific, 2020, 370 (6512): 66-69.), the microglia activation can promote the increase of interleukin-1 beta content (Rahman MA, islam K, rahman S, et al, biological Cross-talk Between COVID-19and Alzheimer's Disease[J ]. Mol neuromol, 2021, 58 (3): 1017-1023), the inflammatory reaction of brain tissue is promoted, the gradual decomposition of beta-amyloid precursor protein into beta-amyloid protein (Hur JY GR, wu X, et al, innate immunity protein IFITM modates gamma-secretase in Alzheimer ' S diseases (J. 35), the increase of the cholinergic metabolism of the brain tissue is promoted (J. 35) by the use of the human being promoted by the human body, the increase of the cholesterol metabolism of the human being promoted by the human body (J. Biological Cross-talk Between COVID-19and Alzheimer's Disease[J), the human brain tissue is promoted by the human being 2021, the human body (3) [ J. Bioperipheral 1-35 ], and the physiological stimulus content of the human organism is promoted by the human body (J. Biopsi, the human organism) is promoted by the human organism, and the human brain tissue inflammatory reaction is promoted by the human brain tissue.
Lesions of brain tissue with Alzheimer's disease have been shown to focus primarily on the hippocampal region (Richetin K, steullet P, pachoud M, et al, tau accumulation in astrocytes of the dentate gyrus induces neuronal dysfunction and memory deficits in Alzheimer's disease [ J ]. Nat. Neurosci,2020, 23 (12): 1567-1579.). The hippocampal region can be generally divided into multiple nerve segments, with hippocampal 1 and hippocampal 3 regions being extremely closely related to Alzheimer's disease (Lysenko NN, pratico D. ABCA7 and the altered lipidostasis hypothesis of Alzheimer's disease [ J ]. Alzheimers Dement,2021, 17 (2): 164-174.). Increased free radical production in the hippocampal region, increased beta-amyloid content (Ruan Z, pathak D, venkatesan Kalavai S, et al alzheimer's disease Brain-derived extracellular vesicles spread tau pathology in interneurons J. Brain,2021, 144 (1): 288-309.), increased interleukin-1 beta content, etc., can rapidly induce the body to develop very pronounced symptoms and signs thereof, and many researchers have established a conventional alzheimer's disease model by injecting large amounts of beta-amyloid in the hippocampal 1 and 3 regions.
It is well known that many drugs have good effects on beta-amyloid-induced Alzheimer's disease, including anti-beta-amyloid drugs (Lozupon M, solfrizzi V, DUrsoF, et al anti-amyoid-beta protein agents for the treatment of Alzheimer's disease: an update on emerging drugs [ J ]. Expert Opin Emerg Drugs,2020, 25 (3): 319-335.), estrogens, vitamin B6, vitamin C (Yu JT, xu W, tan CC, et al [ Evence-based prevention of Alzheimer's disease systematic review and meta-analysis 243observational prospective studies and 153randomised controlled trials[J ]. J Neurol Neurosurg Psychiatry,2020, 91 (11): 1201-1209.), antioxidants, resveratrol (Han Y, chu X, cui L, et al Neuronal Michondria-targeted therapy for Alzheimer's disease by systemic delivery of resveratrol using dual-modified novel biomimetic nanosystems [ J ]. Drug Deliv,2020, 27 (1): 502-518 ]), novel nano-selenium, etc., but many drugs have certain side effects. In recent years, in order to overcome these side effects, many researchers have developed researches on the prevention and treatment of beta-amyloid-induced Alzheimer's disease by selecting drugs with small side effects, overall regulatory function and relatively many targets.
The theory of traditional Chinese medicine holds that Alzheimer's disease is mostly the principal deficiency and secondary excess, in particular deficiency of kidney-qi (Feng Yijun, zheng Wei, wang Xingjia, etc. the research progress of the kidney-tonifying method for treating Alzheimer's dementia [ J ]. Sichuan traditional Chinese medicine, 2021, 39 (2): 214-217.) has a certain relation with the older age. When the kidney qi is abundant, the kidney yang is sufficient, the learning ability is strong, the deficiency of marrow and sea is avoided, and the decline of the learning ability of the old is relieved, so that the medicine with the function of tonifying the kidney is needed to be selected for treating the Alzheimer disease. The elderly often have weak dietary digestion, which results in nutrient Wei Shihe, kidney essence deficiency and loss of life (Chen Huize, talking about the beginning, meng Shengxi. The research progress of combined Chinese and Western medicine for preventing and treating Alzheimer's disease [ J ]. The combined Chinese and Western medicine combines J. Cardiovascular and cerebrovascular diseases, 2021, 19 (12): 2025-2029.) beta-amyloid is gradually deposited in brain tissues, aggravates the symptoms and signs of the organism, so the treatment of Alzheimer's disease requires the selection of medicines with the functions of invigorating stomach and moistening intestines. The senile kidney qi deficiency is also gradually accumulated (Zhang Yige, liang Yuqing, li Yali, etc.) from the evolution of traditional Chinese medicine pathogenesis of kidney deficiency and phlegm stasis Aer Alzheimer disease [ J ]. World science and technology-modernization of traditional Chinese medicine, 2021, 23 (1): 159-164), which has a certain relation with qi movement disorder and yin-yang imbalance in the elderly (Zhao Ming, li Hao, liu Nayang, etc. the theory of preventing and treating senile dementia by the light-resolution ascending and descending turbidity method and the application of [ J ]. Circumferential traditional Chinese medicine, 2021, 14 (2): 278-281.) are further promoted, thereby increasing the free radical production amount in the hippocampal region, increasing the beta-amyloid content, increasing the interleukin-1 beta content, etc. to induce Alzheimer disease. Therefore, the treatment of Alzheimer disease requires the removal of blood stasis and damp. Comprehensive analysis shows that the treatment of beta-amyloid induced Alzheimer disease requires the functions of tonifying the kidney, removing blood stasis, invigorating the stomach, moistening the intestines, eliminating dampness and regulating qi.
The invention provides a traditional Chinese medicine composition comprising 6 traditional Chinese medicinal materials, which comprises 10-15 parts by weight of eclipta, 3-8 parts by weight of diverse wormwood herb, 2-6 parts by weight of semen katsumadai, 5-15 parts by weight of semen Abutili, 2-6 parts by weight of cortex Periplocae Radicis and 10-15 parts by weight of akebia fruit. Animal experiment results show that the high dosage (12.74 g/kg) and the dosage (6.37 g/kg) of the traditional Chinese medicine composition can improve the learning ability, the beta-amyloid precursor protein expression level, the interleukin-1 beta expression level and the acetylcholine metabolizing enzyme expression level of the Kunming mice with the beta-amyloid induced Alzheimer disease, and meanwhile, the traditional Chinese medicine composition has no influence on the main organ index and the body weight of the Kunming mice with the beta-amyloid induced Alzheimer disease, so that the traditional Chinese medicine composition can be used for treating the Alzheimer disease and has no side effect.
Disclosure of Invention
The invention aims to solve the technical problem of providing a traditional Chinese medicine composition for preventing and treating Alzheimer's disease, and a preparation method and application thereof.
In order to achieve the above purpose, the invention adopts the following technical means:
the invention relates to a traditional Chinese medicine composition for treating Alzheimer disease, which is characterized by comprising the following raw materials in parts by weight: 10-15 parts of eclipta, 3-8 parts of diverse wormwood herb, 2-6 parts of semen katsumadai, 5-15 parts of malva seed, 2-6 parts of cortex caryophylli and 10-15 parts of fiveleaf akebia fruit.
Wherein, the eclipta alba, has cold nature, sour taste and sweet taste, belongs to kidney channel and liver channel, and has the basic effects of tonifying kidney, cooling blood, nourishing liver and stopping bleeding; herba Artemisiae Anomalae, has slightly warm nature, bitter and pungent taste, belongs to spleen meridian, heart meridian and liver meridian, and has the basic effects of removing blood stasis, dredging meridian, resolving food stagnation, relieving pain, breaking blood and promoting digestion; semen Alpiniae, is aliased with semen Alpiniae, has warm nature and pungent taste, belongs to stomach channel and spleen channel, and has the basic effects of invigorating stomach, relieving distention, eliminating dampness and warming middle warmer; the malva seed is cold in nature and sweet in taste, belongs to the small intestine channel, the large intestine channel and the bladder channel, and has the basic effects of moistening the intestines, treating stranguria, promoting diuresis and promoting lactation; the cortex Periplocae Radicis, named with periploca mongolica bark, has slightly warm nature, bitter and pungent taste, belongs to heart meridian, kidney meridian and liver meridian, and has the basic effects of eliminating dampness, strengthening tendons, promoting diuresis, dispelling pathogenic wind, detumescence and strengthening bones; akebiae fructus, it is cold in nature, sweet in taste and bitter in flavor, belongs to kidney meridian, spleen meridian and liver meridian, and has the basic effects of regulating qi, relieving pain, soothing liver and activating blood. In the traditional Chinese medicine composition, the monarch drugs of the jean lotus and the diverse wormwood herb have the basic effects of tonifying kidney and removing blood stasis; the ministerial drugs semen Alpiniae and semen Abutili have the basic effects of invigorating stomach and moistening intestines; the adjuvant drug of cortex Acanthopancis has the basic effect of eliminating dampness; the fiveleaf akebia fruit has the basic effect of regulating qi. The compatibility of the medicines can produce the effects of tonifying kidney, removing blood stasis, invigorating stomach, moistening intestines, eliminating dampness and regulating qi.
Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 12.5 parts of eclipta, 6 parts of diverse wormwood herb, 4 parts of semen katsumadai, 10 parts of malva seed, 4.5 parts of cortex eucalypti radicis and 12 parts of fiveleaf akebia fruit.
The behavioral test experimental result shows that compared with the control group without administration, the model verification group is obviously prolonged in incubation period (P < 0.05) of the Kunming mice, and the incubation period of the Kunming mice in the simulated operation group is not obviously changed (P > 0.05); compared with the model verification group, the Western medicine intervention group, the Chinese medicinal composition high-dose group and the Chinese medicinal composition medium-dose group have obviously shortened incubation period (P is smaller than 0.05), and the Chinese medicinal composition low-dose group has insignificant incubation period change (P is larger than 0.05), which indicates that the Chinese medicinal composition high-dose (12.74 g/kg) and the Chinese medicinal composition medium-dose (6.37 g/kg) can improve the learning and memory capacity of the cleaning-grade Kunming mouse model of the beta-amyloid induced Alzheimer disease.
The ELISA test result shows that compared with a control group without drug administration, the model verifies that the beta-amyloid precursor protein content of the Kunming mice in the group is obviously increased (P is less than 0.05) and the interleukin-1 beta content is obviously increased (P is less than 0.05), and the beta-amyloid precursor protein content and interleukin-1 beta content of the Kunming mice in the simulated operation group are not obviously changed (P is more than 0.05); compared with the model verification group, western medicine intervention group, chinese medicine composition high-dose group and Chinese medicine composition medium-dose group Kunming mice have obviously reduced beta-amyloid precursor protein content and interleukin-1 beta content (P < 0.05), and composition low-dose group Kunming mice have unobvious change of beta-amyloid precursor protein content and interleukin-1 beta content (P > 0.05), which indicates that Chinese medicine composition high dose (12.74 g/kg) and Chinese medicine composition medium dose (6.37 g/kg) can improve beta-amyloid precursor protein content and interleukin-1 beta content of beta-amyloid-induced Alzheimer disease cleaning-grade Kunming mice model, and Chinese medicine composition high dose (12.74 g/kg) and Chinese medicine composition medium dose (6.37 g/kg) can improve beta-amyloid-induced Alzheimer disease cleaning-grade Kunming mice model inflammatory medium level and reduce plaque deposition, which is related to microglial cell and astrocyte activation level.
The results of immunohistochemistry and gene detection show that compared with a control group without administration, the model verifies that the protein and gene expression quantity of the acetylcholine metabolizing enzyme of the Kunming mice in the group are obviously increased (P is less than 0.05), the protein and gene expression quantity of the choline acetylating enzyme is obviously reduced (P is less than 0.05), and the protein and gene expression quantity of the acetylcholine metabolizing enzyme and the choline acetylating enzyme of the Kunming mice in the simulated operation group are not obviously changed (P is more than 0.05); compared with a model verification group, western medicine intervention groups, chinese medicinal composition high-dose groups and Chinese medicinal composition medium-dose groups are used for obviously reducing the expression level of acetylcholine metabolizing enzyme protein and gene of Kunming mice (P < 0.05), obviously increasing the expression level of choline acetyltransferase protein and gene (P < 0.05), and the composition low-dose groups are used for obviously reducing the expression level of the Kunming mice acetylcholine metabolizing enzyme and choline acetyltransferase protein and gene (P > 0.05), which indicates that abnormal expression of the choline acetyltransferase of the brain tissue of the organism and the acetylcholine metabolizing enzyme of the brain tissue of the organism can cause abnormal acetylcholine content of the brain tissue of the organism, the intervention of the choline acetyltransferase of the brain tissue of the organism and the acetylcholine metabolizing enzyme expression of the brain tissue of the organism, and the improvement of beta-amyloid protein induced Alzheimer disease by the Chinese medicinal composition high dose (12.74 g/kg) and the Chinese medicinal composition medium dose (6.37 g/kg).
Evaluation of the Chinese medicinal composition improved the side effects of beta-amyloid induced Alzheimer's disease, which was related to evaluation of body weight and organ index. The results showed that the heart, lung, kidney, liver index and body weight changes were not evident in the model validation group compared to the simulated surgery group in the Kunming mice. After the treatment of the high dosage (12.74 g/kg) of the traditional Chinese medicine composition, the medium dosage (6.37 g/kg) of the traditional Chinese medicine composition and the low dosage (3.19 g/kg) of the traditional Chinese medicine composition, the heart, lung, kidney, liver indexes and body weight of important organs of all groups of Kunming mice are not obvious, which indicates that the treatment of the high dosage (12.74 g/kg) of the traditional Chinese medicine composition, the medium dosage (6.37 g/kg) of the traditional Chinese medicine composition and the low dosage (3.19 g/kg) of the traditional Chinese medicine composition does not have side effects.
In conclusion, the traditional Chinese medicine composition composed of 6 raw medicinal materials of ink can effectively treat beta-amyloid induced Alzheimer disease without side effects.
Furthermore, the invention also provides application of the traditional Chinese medicine composition in preparing medicines for preventing and treating Alzheimer disease.
Preferably, the Alzheimer's disease is induced by beta-amyloid.
Furthermore, the invention also provides a method for preparing the traditional Chinese medicine composition, which comprises the following steps:
(1) Weighing the raw materials according to the weight parts, uniformly mixing the raw materials, adding distilled water which is 20-30 times of the weight of the raw materials, soaking for 2.5-3.5 hours, decocting for 2.5-3.5 hours, and cooling;
(2) Filtering the liquid medicine cooled in the step (1) by using gauze to obtain a first filtrate;
(3) Repeating the steps (1) and (2) to obtain a second filtrate, and combining the second filtrate;
(4) Mixing the first filtrate and the second filtrate, and baking at 60-70deg.C for 6-8 days to obtain Chinese medicinal composition dry extract;
(5) And (3) putting the obtained dry paste into a pulverizer to pulverize, so as to obtain the dry powder of the traditional Chinese medicine composition.
Preferably, the step (2) of filtering with gauze means filtering with 3 layers of gauze, filtering with 5 layers of gauze, filtering with 8 layers of gauze and filtering with 10 layers of gauze in sequence.
Furthermore, the invention also provides a clinically suitable traditional Chinese medicine preparation for treating Alzheimer disease, which is prepared by adding auxiliary materials required by preparation molding into the traditional Chinese medicine composition according to a conventional method for preparing the medicine preparation.
Wherein, the preparation is preferably powder, decoction, capsule, pill, granule, tablet or oral liquid.
Compared with the prior art, the invention has the beneficial effects that:
the invention discloses a traditional Chinese medicine composition for treating Alzheimer disease, in particular to a beta-amyloid induced Alzheimer disease, and a preparation method and application thereof. The basic composition of the traditional Chinese medicine composition comprises 6 traditional Chinese medicines, namely, eclipta, diverse wormwood herb, semen katsumadai, malva seed, cortex acanthopanacis and fructus akebiae. The traditional Chinese medicine composition has the basic effects of tonifying kidney, removing stasis, invigorating stomach, moistening intestines, eliminating dampness and regulating qi. Animal experiment results show that the high dosage (12.74 g/kg) of the traditional Chinese medicine composition and the dosage (6.37 g/kg) of the traditional Chinese medicine composition can improve the learning ability of the Kunming mice with the Alzheimer's disease induced by the beta-amyloid protein, the expression level of the precursor protein of the beta-amyloid protein, the expression level of interleukin-1 beta and the expression level of the acetylcholinergic enzyme, and meanwhile, the traditional Chinese medicine composition has no influence on the main organ index and the body weight of the Kunming mice with the Alzheimer's disease induced by the beta-amyloid protein, so that the traditional Chinese medicine composition is effective in treating the Alzheimer's disease and has no side effect.
Drawings
FIG. 1 is a graph showing the effect of a Chinese medicinal composition on the learning ability of a beta-amyloid-induced Alzheimer's disease Kunming mouse;
note that: the abscissa is the group; the ordinate is latency(s);
FIG. 2 is a graph showing the effect of a traditional Chinese medicinal composition on the beta-amyloid precursor protein content of a beta-amyloid-induced Alzheimer's disease Kunming mouse;
note that: the abscissa is the group; the ordinate is beta-amyloid precursor protein (ng/L);
FIG. 3 is a graph showing the effect of a traditional Chinese medicine composition on interleukin-1 beta content in a beta-amyloid-induced Alzheimer's disease-infected mouse;
note that: the abscissa is the group; the ordinate is interleukin-1β (pg/mL);
FIG. 4 is a graph showing the effect of a traditional Chinese medicine composition on the content of beta-amyloid-induced Alzheimer's disease Kunming mouse choline acetyltransferase;
note that: the abscissa is the group; the ordinate is choline acetyltransferase (average optical density value);
FIG. 5 is a graph showing the effect of a traditional Chinese medicine composition on beta-amyloid-induced Alzheimer's disease on acetylcholine metabolizing enzyme in Kunming mice;
note that: the abscissa is the group; the ordinate is acetylcholine metabolizing enzyme (average optical density value);
FIG. 6 is a graph showing the effect of a traditional Chinese medicine composition on the beta-amyloid-induced Alzheimer's disease Kunming mouse choline acetyltransferase gene;
note that: the abscissa is the group; the ordinate is choline acetyltransferase (gene expression amount);
FIG. 7 is a graph showing the effect of a traditional Chinese medicine composition on the beta-amyloid-induced Alzheimer's disease Kunming mouse acetylcholine metabolizing enzyme gene;
note that: the abscissa is the group; the ordinate is acetylcholine metabolizing enzyme (gene expression level);
FIG. 8 is a graph showing the effect of a Chinese medicinal composition on the heart index and lung index of a Alzheimer's disease-induced beta-amyloid Kunming mouse;
note that: the abscissa is the group; the ordinate is the heart index, lung index (mg/g);
FIG. 9 is a graph showing the effect of a Chinese medicinal composition on kidney index and liver index of a beta-amyloid-induced Alzheimer's disease-induced Kunming mouse;
note that: the abscissa is the group; the ordinate is kidney index, liver index (mg/g);
FIG. 10 is a graph showing the effect of a traditional Chinese medicinal composition on body weight of a beta-amyloid-induced Alzheimer's disease Kunming mouse;
note that: the abscissa is the group; the ordinate is the body weight (g) of Kunming mice at day 0, day 14, day 28.
Detailed Description
The invention is further described below, the embodiments presented in this description are only exemplary and do not limit the scope of the invention. It will be understood by those skilled in the art that the details and forms of the invention may be modified or substituted without departing from the spirit and scope of the invention.
Example 1 preparation of a Chinese medicinal composition for preventing and treating Alzheimer's disease
The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 12.5g of eclipta, 6g of diverse wormwood herb, 4g of semen katsumadai, 10g of malva seed, 4.5g of cortex caryophylli and 12g of fiveleaf akebia fruit.
Example 2 preparation of a Chinese medicinal composition for preventing and treating Alzheimer's disease
The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10g of eclipta, 8g of diverse wormwood herb, 3g of semen katsumadai, 8g of malva seed, 6g of cortex caryophylli and 15g of fiveleaf akebia fruit.
Example 3 preparation of a Chinese medicinal composition for preventing and treating Alzheimer's disease
The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 15g of eclipta, 5g of diverse wormwood herb, 5g of semen katsumadai, 15g of malva seed, 4g of cortex caryophylli and 10g of fiveleaf akebia fruit.
Example 4 preparation of a Chinese medicinal composition (Dry powder) for preventing and treating Alzheimer's disease
(1) 12.5g of eclipta, 6g of diverse wormwood herb, 4g of cardamom, 10g of malva seed, 4.5g of cortex caryophylli, 12g of fiveleaf akebia fruit, and 5 parts of common akebia fruit are purchased for 49g each part, and the total weight is 245g.
(2) 245g of the traditional Chinese medicine composition of the Chinese medicinal materials of the eclipta, the diverse wormwood herb, the semen katsumadai, the malva seed, the cortex acanthopanacis and the fiveleaf akebia fruit are put into a 20000mL high-capacity round bottom flask (6000 mL of distilled water) to be soaked for 3 hours, then decocted for 3 hours, and filtered by 3 layers of gauze, 5 layers of gauze, 8 layers of gauze and 10 layers of gauze to obtain a first filtrate.
(3) The drug residue was put into 20000mL high capacity round bottom flask (6000 mL of distilled water) and soaked for 3 hours, then decocted for 3 hours, 3 layers of gauze filtration, 5 layers of gauze filtration, 8 layers of gauze filtration, 10 layers of gauze filtration, and a second filtrate was obtained.
(4) Mixing the first filtrate and the second filtrate, and baking in a drying oven (65deg.C) for 7 days to obtain 68.35g of Chinese medicinal composition dry extract.
(5) 68.35g of the dry paste of the traditional Chinese medicine composition is put into a pulverizer to be pulverized, and 65.24g of dry powder of the traditional Chinese medicine composition is obtained.
(6) 65.24g of the traditional Chinese medicine composition dry powder is packaged separately, 10g of each bag and 5.24g of the last bag. Preserving in a refrigerator at-80 ℃.
EXAMPLE 5 use of the Chinese medicinal composition of the present invention in the treatment of Alzheimer's disease
1 Experimental materials
1.1 laboratory animals and instruments therefor
Kunming mice 56 (SCXK 2012-0001), body weight (19.6857 + -0.6887) g, clean grade, male, beijing Vetong Lihua, routinely raised in the university of traditional Chinese medicine laboratory animal center in Heilongjiang. Incubator, model BC-J160S, shanghai five-time company; electronic balance, qingdao dishing company, model JC-TP; the brain stereotactic apparatus, hebei Huiya company, model ZH-C; a water maze, model MWM101, of north hua corporation; color analysis system, model HMIAS, wuhan Qian Screen company; centrifuges, shanghai Anpavilion, model TGL-16G, other instruments are provided by the Zhang Yingbo pair of professor.
1.2 Experimental drugs and reagents therefor
12.5g of Chinese ink-early lotus, 6g of diverse wormwood herb, 4g of cardamom, 10g of malva seed, 4.5g of cortex caryophylli and 12g of fiveleaf akebia fruit are purchased in a first affiliated hospital of traditional Chinese medicine university in black dragon river, 12.5g of Chinese ink-early lotus, 6g of diverse wormwood herb, 4g of cardamom seed, 10g of malva seed, 4.5g of cortex caryophylli and 12g of fiveleaf akebia fruit are identified, 12.5g of Chinese ink-early lotus, 6g of diverse wormwood herb, 4g of cardamom seed, 10g of malva seed, 4.5g of cortex caryophylli and 12g of fiveleaf akebia fruit are compatible. Donepezil hydrochloride tablet, a company of the health and medicine industry; beta-amyloid precursor protein, interleukin-1 beta kit, nanjing build company; acetylcholine metabolizing enzymes, choline acetylase, santa Inc. of USA; other reagents are provided by the Zhang Yingbo pair of teachings.
2 Experimental methods
2.1 Experimental animals grouping and model preparation thereof
The number of Kunming mice was 56, and the mice were randomly divided into a control group without administration, a simulated operation group, a model verification group, a western medicine intervention group, a composition low dose group, a composition medium dose group and a composition high dose group, and 8 mice each. Use of beta-amyloid 1 Preparation of beta-amyloid induced Alzheimer's disease model by injection of Hippocampus and cat rat scaring method of 42 Hippocampus, and beta-amyloid is carried out on model validation group, western medicine intervention group, composition low dose group, composition medium dose group, and composition high dose group every day 1 -42 hippocampus (80 pmol/. Mu.L) was injected, the simulated surgical group was injected with sterile physiological saline, and the control group was not administered without intervention for 20 days. The model verification group, western medicine intervention group, composition low-dose group, composition medium-dose group and composition high-dose group are interfered by adopting a cat rat-scaring method every afternoon, and the non-administration control group and the simulated operation group are not interfered by adopting a cat rat-scaring methodFor 20 days.
2.2 Chinese medicinal composition Dry powder
Same as in example 4.
2.3 calculation of dosing and animal dosing
According to 0.0026:1, calculating an administration dosage of 12.5g of Chinese medicinal composition, 6g of herba artemisiae anomalae, 4g of semen katsumadai, 10g of semen Abutili, 4.5g of cortex caryophylli and 12g of fructus akebiae, wherein the total dosage of 49g is 49g by 0.0026 by 50=6.37 g/kg, the dosage of the Chinese medicinal composition is 6.37g/kg, the high dosage of the Chinese medicinal composition is 12.74g/kg, and the low dosage of the Chinese medicinal composition is 3.19g/kg. Donepezil is administered at a dose of 0.001g/kg. The control group is not administered, the simulated operation group and the model verification group are administered with the same amount of physiological saline to irrigate the stomach; the western medicine intervention group is administrated with 0.001g/kg donepezil for gastric lavage; the composition was administered in low dose groups at 3.19g/kg for lavage; 6.37g/kg of the composition was administered in a dosage group for gastric lavage; the high dose group of the composition was administered 12.74g/kg for gastric lavage. The administration duration was 28 days.
2.4 assessment of learning ability of animals
The learning ability of each group of Kunming mice was evaluated using a water maze, the Kunming mice stained the head with ink, the internal and external references of the water maze were kept consistent throughout, training time was maintained for 4 days in quadrants I, II, and III of the water maze, each time for 60 seconds, incubation period was calculated for each group of Kunming mice on day 5, and animal learning ability was evaluated.
2.5 evaluation of beta-amyloid precursor protein and Interleukin-1 beta expression levels
After the water maze evaluation of learning ability of each group of Kunming mice, the Kunming mice were grasped, whole brains were taken aseptically, hippocampus were peeled off, homogenized and centrifuged, supernatant was taken, absorbance values (OD) were analyzed at a wavelength of 450nm using a microplate reader, and the beta-amyloid precursor protein and interleukin-1 beta expression levels were further evaluated.
2.6 evaluation of acetylcholine Metabolic enzyme and Choline Acetylase protein levels
After the experiment personnel wear gloves and the water maze evaluates the learning ability of each group of Kunming mice, the Kunming mice are grabbed, after physiological saline is infused into hearts of the Kunming mice, about 50mL of 4% paraformaldehyde is infused at a constant speed, and after conventional paraffin section treatment, the contents of acetylcholine metabolizing enzyme and choline acetyltransferase are measured according to a basic method of a kit.
2.7 evaluation of acetylcholine Metabolic enzyme and Choline Acetylase Gene levels
After the experiment personnel wear gloves and the water maze evaluates the learning ability of each group of Kunming mice, the Kunming mice are grabbed, the whole brain and the sea horse thereof are obtained, the acetylcholine metabolizing enzyme and the choline acetylase mRNA are extracted, the cDNA is reversely transcribed, the primers aiming at the acetylcholine metabolizing enzyme and the choline acetylase are designed, the reaction parameters aiming at the acetylcholine metabolizing enzyme and the choline acetylase are set, the reaction parameters comprise 2 minutes after 50 ℃ and 10 minutes after reaction at 95 ℃, 15s after reaction at 95 ℃ and 1 minute after reaction at 60 ℃, 42 basic cycles are obtained by a manager, and the gene expression level of the acetylcholine metabolizing enzyme and the choline acetylase is measured by using an instrument.
2.8 evaluation of organ index and body weight
After the learning ability of each group of Kunming mice is evaluated by the water maze, the Kunming mice are grabbed, the body weight of each group of Kunming mice is weighed for 0 day, 14 days and 28 days, the Kunming mice are killed by broken ends, hearts, lungs, kidneys and livers of the Kunming mice are obtained, and liver indexes, kidney indexes, heart indexes and lung indexes are calculated according to a formula. Liver index, kidney index, heart index, lung index calculation formula: organ index (mg/g) of heart, lung, kidney, liver = organ mass (mg) of heart, lung, kidney, liver/body weight (g) of kunming mice.
2.9 statistical analysis of data
The experiment uses SPSS19.0 software to analyze the data (expressed as x+ -s) related to the experiment, and the comparison of the data among multiple groups uses single-factor analysis of variance.
3 results of experiments
3.1 Effect of Chinese medicinal composition on learning ability of beta-amyloid-induced Alzheimer's disease Kunming mice
The experimental result shows that compared with the control group without administration, the model verifies that the incubation period of the Kunming mice in the group is obviously prolonged (P is less than 0.05), and the incubation period of the Kunming mice in the group with simulated operation is not obviously changed (P is more than 0.05); compared with the model verification group, the latency of the Kunming mice in the western medicine intervention group, the composition high-dose group and the composition medium-dose group is obviously shortened (P is smaller than 0.05), and the latency of the Kunming mice in the composition low-dose group is not obviously changed (P is larger than 0.05). See table 1. See fig. 1.
TABLE 1 influence of Chinese medicinal composition on learning ability of beta-amyloid induced Alzheimer's disease Kunming mice
/>
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.2 Effect of the Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse beta-amyloid precursor protein
The experimental result shows that compared with a control group without administration, the model verifies that the beta-amyloid precursor protein content of the Kunming mice in the group is obviously increased (P is less than 0.05), and the beta-amyloid precursor protein content of the Kunming mice in the simulated operation group is not obviously changed (P is more than 0.05); compared with the model verification group, the western medicine intervention group, the composition high-dose group and the composition low-dose group have obviously reduced beta-amyloid precursor protein content (P < 0.05) of Kunming mice, and the composition low-dose group has not obviously changed beta-amyloid precursor protein content (P > 0.05). See table 2. See fig. 2.
TABLE 2 Effect of Chinese medicinal composition on beta-amyloid precursor protein content in beta-amyloid-induced Alzheimer's disease Kunming mice
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.3 Effect of Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse interleukin-1 beta
The experimental result shows that compared with a control group without administration, the interleukin-1 beta content of the Kunming mice in the model verification group is obviously increased (P is less than 0.05), and the interleukin-1 beta content of the Kunming mice in the simulation operation group is not obviously changed (P is more than 0.05); compared with the model verification group, the interleukin-1 beta content of the western medicine intervention group, the composition high-dose group and the composition low-dose group is obviously reduced (P is less than 0.05), and the interleukin-1 beta content of the composition low-dose group is not obviously changed (P is more than 0.05). See table 3. See fig. 3.
TABLE 3 Effect of Chinese medicinal composition on beta-amyloid induced interleukin-1 beta content in Kunming mice with Alzheimer's disease
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.4 Effect of Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse choline acetyltransferase
The results show that compared with a control group without administration, the average optical density value of the Kunming mouse choline acetylase in a model verification group is obviously reduced (P is less than 0.05), and the average optical density value of the Kunming mouse choline acetylase in a simulated operation group is not obviously changed (P is more than 0.05); compared with the model verification group, the average optical density value of the Kunming mouse choline acetyltransferase in the western medicine intervention group, the composition high-dose group and the composition medium-dose group is obviously increased (P is less than 0.05), and the average optical density value of the Kunming mouse choline acetyltransferase in the composition low-dose group is not obviously changed (P is more than 0.05). See table 4. See fig. 4.
TABLE 4 Effect of Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse choline acetyltransferase
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.5 Effect of the Chinese medicinal composition on beta-amyloid-induced Acetylcholinergic enzymes in Kunming mice with Alzheimer's disease
The results show that compared with the control group without administration, the average optical density value of the acetylcholine metabolizing enzyme of the Kunming mice in the model verification group is obviously increased (P is less than 0.05), and the average optical density value of the acetylcholine metabolizing enzyme of the Kunming mice in the simulated operation group is not obviously changed (P is more than 0.05); compared with the model verification group, the average optical density value of the acetylcholine metabolizing enzyme of the Kunming mice in the western medicine intervention group, the composition high-dose group and the composition medium-dose group is obviously reduced (P is less than 0.05), and the average optical density value of the acetylcholine metabolizing enzyme of the Kunming mice in the composition low-dose group is not obviously changed (P is more than 0.05). See table 5. See fig. 5.
Table 5 Effect of the Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse acetylcholine Metabolic enzyme
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.6 Effect of Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse choline acetyltransferase Gene
The result shows that compared with a control group without administration, the gene expression level of the Kunming mouse choline acetylase is obviously reduced (P is less than 0.05) in the model verification group, and the gene expression level of the Kunming mouse choline acetylase is not obviously changed (P is more than 0.05) in the simulated operation group; compared with the model verification group, the western medicine intervention group, the composition high-dose group and the composition medium-dose group have obviously increased gene expression quantity of the Kunming mouse choline acetyltransferase (P is less than 0.05), and the composition low-dose group has unobvious gene expression quantity change of the Kunming mouse choline acetyltransferase (P is more than 0.05). See table 6. See fig. 6.
TABLE 6 influence of Chinese medicinal composition on beta-amyloid induced Alzheimer's disease Kunming mouse choline acetyltransferase gene
2 -ΔΔCt Method of
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.7 Effect of the Chinese medicinal composition on beta-amyloid-induced Alzheimer's disease Kunming mouse acetylcholine Metabolic enzyme Gene
The result shows that compared with a control group without administration, the model verifies that the gene expression level of the acetylcholine metabolizing enzyme of the Kunming mice in the group is obviously increased (P is less than 0.05), and the gene expression level of the acetylcholine metabolizing enzyme of the Kunming mice in the simulated operation group is not obviously changed (P is more than 0.05); compared with the model verification group, the Western medicine intervention group, the composition high-dose group and the composition medium-dose group have obviously reduced gene expression quantity of the acetylcholine metabolizing enzyme of the Kunming mice (P is less than 0.05), and the composition low-dose group has unobvious gene expression quantity change of the acetylcholine metabolizing enzyme of the Kunming mice (P is more than 0.05). See table 7. See fig. 7.
TABLE 7 influence of the Chinese medicinal composition on the beta-amyloid induced Alzheimer's disease Kunming mouse acetylcholine Metabolic enzyme Gene
2 -ΔΔCt Method of
Note that: comparison to the non-dosed control group: # P＜0.05， ## p is more than 0.05; comparison to model validation set: * P＜ 0.05， ** P＞0.05
3.8 Effect of Chinese medicinal composition on the heart index and the lung index of Kunming mice with beta-amyloid induced Alzheimer's disease
The results show that compared with a control group without administration, the heart index and the lung index of the Kunming mice in a model verification group are not obviously changed (P is more than 0.05), and the heart index and the lung index of the Kunming mice in a simulation operation group are not obviously changed (P is more than 0.05); compared with the model verification group, western medicine intervention group, composition high dose group, composition medium dose group, composition low dose group, kunming mouse heart index and lung index change is not obvious (P > 0.05). See table 8. See fig. 8.
TABLE 8 influence of Chinese medicinal composition on heart index and lung index of Kunming mice with beta-amyloid induced Alzheimer's disease
n＝8)/>
Note that: comparison to the non-dosed control group: # p is more than 0.05; comparison with model verification group： * P＞0.05
3.9 Effect of the Chinese medicinal composition on renal index and hepatic index of Kunming mice with beta-amyloid induced Alzheimer's disease
The results show that compared with a control group without administration, the kidney index and liver index of the Kunming mice in the model verification group are not obviously changed (P is more than 0.05), and the kidney index and liver index of the Kunming mice in the simulation operation group are not obviously changed (P is more than 0.05); compared with the model verification group, the kidney index and liver index of Kunming mice in western medicine intervention group, composition high-dose group, composition medium-dose group and composition low-dose group are not obviously changed (P is more than 0.05). See table 9. See fig. 9.
TABLE 9 influence of the Chinese medicinal composition on renal index and hepatic index of Kunming mice with beta-amyloid induced Alzheimer's disease
n＝8)
Note that: comparison to the non-dosed control group: # p is more than 0.05; comparison to model validation set: * P＞0.05
3.10 Effect of Chinese medicinal composition on body weight of beta-amyloid-induced Alzheimer's disease Kunming mice
The results show that the body weight of the Kunming mice in the model verification group is not obviously changed (P > 0.05) in 0 days, 14 days and 28 days compared with the body weight of the Kunming mice in the non-administration control group, and the body weight of the Kunming mice in the model verification group is not obviously changed (P > 0.05) in 0 days, 14 days and 28 days in the simulation operation group; compared with the weights of the Kunming mice in the model verification groups of 0 day, 14 day and 28 day, the weights of the Kunming mice in the western medicine intervention group, the composition high-dose group, the composition medium-dose group and the composition low-dose group are not obviously changed (P is more than 0.05) in the model verification groups of 0 day, 14 day and 28 day. See table 10. See fig. 10.
TABLE 10 influence of Chinese medicinal composition on body weight of clean-class Kunming mice with beta-amyloid induced Alzheimer's disease
n＝8)
Note that: comparison to the non-dosed control group: # p is more than 0.05; comparison to model validation set: * P＞0.05。
Claims (8)
1. the traditional Chinese medicine composition for treating the Alzheimer disease is characterized by being prepared from the following raw materials in parts by weight: 10-15 parts of eclipta, 3-8 parts of diverse wormwood herb, 2-6 parts of semen katsumadai, 5-15 parts of malva seed, 2-6 parts of cortex caryophylli and 10-15 parts of fiveleaf akebia fruit.
2. The traditional Chinese medicine composition for treating Alzheimer's disease according to claim 1, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 12.5 parts of eclipta, 6 parts of diverse wormwood herb, 4 parts of semen katsumadai, 10 parts of malva seed, 4.5 parts of cortex eucalypti radicis and 12 parts of fiveleaf akebia fruit.
3. The use of the Chinese medicinal composition according to claim 1 or 2 for preparing a medicament for preventing and treating alzheimer's disease.
4. The use according to claim 3, wherein the alzheimer's disease is induced by β -amyloid.
5. A method of preparing the traditional Chinese medicine composition of claim 1 or 2, characterized by comprising the steps of:
(1) Weighing the raw materials according to the weight parts of claim 1 or 2, uniformly mixing the raw materials, adding distilled water with the weight being 20-30 times of the weight of the raw materials, soaking for 2.5-3.5 hours, decocting for 2.5-3.5 hours, and cooling;
(2) Filtering the liquid medicine cooled in the step (1) by using gauze to obtain a first filtrate;
(3) Repeating the steps (1) and (2) to obtain a second filtrate;
(4) Mixing the first filtrate and the second filtrate, and baking at 60-70deg.C for 6-8 days to obtain Chinese medicinal composition dry extract;
(5) And (3) putting the obtained dry paste into a pulverizer to pulverize, so as to obtain the dry powder of the traditional Chinese medicine composition.
6. The method of claim 5, wherein the step (2) of filtering with gauze comprises filtering with 3 layers of gauze, filtering with 5 layers of gauze, filtering with 8 layers of gauze, and filtering with 10 layers of gauze.
7. A clinically suitable Chinese medicinal preparation for treating Alzheimer's disease is characterized in that the Chinese medicinal preparation is prepared by adding auxiliary materials required by preparation molding into the Chinese medicinal composition according to claim 1 or 2 according to a conventional method for preparing the medicinal preparation.
8. The Chinese medicinal preparation according to claim 7, wherein the preparation is a powder, a decoction, a capsule, a pill, a granule, a tablet or an oral liquid.
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