Traditional Chinese medicine composition for preventing and treating senile dementia as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for preventing and treating senile dementia and a preparation method and application thereof. The traditional Chinese medicine composition for preventing and treating senile dementia comprises the following components in parts by weight: 10-30 parts of codonopsis pilosula, 10-15 parts of rhizoma polygonati, 10-20 parts of human placenta, 6-12 parts of radix polygoni multiflori preparata, 6-12 parts of semen cuscutae, 6-10 parts of herba epimedii, 10-15 parts of polygonum cuspidatum, 3-10 parts of Chinese polyphaga, 3-6 parts of scorpion, 10-15 parts of stenoloma chusana and 10-15 parts of rhizoma acori graminei. The traditional Chinese medicine composition disclosed by the invention has the effects of reinforcing intelligence and resisting senescence, exerts the effect of treating senile dementia in a multi-component, multi-path and multi-target manner, and can resist oxidative stress of an organism, relieve cognitive impairment and obviously improve nerve cell injury, so that senescence is delayed, senile dementia symptoms are prevented and treated, and the daily living ability of a patient is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for preventing and treating Alzheimer's disease, and a preparation method and use thereof. Background Art

[0002] Alzheimer's disease (AD) is the most common clinical form of senile dementia, with persistent memory loss and progressive cognitive impairment as the main manifestations, and Aβ deposition, Tau protein phosphorylation and neuronal loss as the main pathological features. The pathogenesis of AD is still unclear. The most recognized pathogenesis currently includes: cholinergic damage hypothesis, Aβ cascade hypothesis, abnormal Tau protein phosphorylation hypothesis, oxidative stress hypothesis, neuroinflammation hypothesis, genetic hypothesis, metal ion disorder hypothesis, etc. Senile dementia is more common in the elderly and is gradually becoming younger. Nearly 70%-80% of patients cannot receive timely and effective treatment. Most of the drugs currently used to treat senile dementia are developed based on the pathogenesis, mainly acetylcholinesterase inhibitors, NMDA receptor antagonists, brain-gut axis regulating drugs, targeted drugs, etc., which can only alleviate the disease progression of senile dementia, with limited therapeutic effects and significant adverse reactions. They mainly improve cognitive function and relieve clinical symptoms, and have certain limitations in clinical use.

[0003] Traditional Chinese medicine has unique advantages in the treatment of Alzheimer's disease. Natural medicines represented by traditional Chinese medicine have the characteristics of small toxic and side effects, diverse targets, and good tolerance. They are suitable for patients with sub-health, cognitive decline and Alzheimer's disease. However, the currently reported anti-aging and intelligence-enhancing drugs have different formulations, uncertain main ingredients, and slow clinical effects. Therefore, it is of great significance to research and develop a Chinese medicine composition with good anti-aging effects, a certain formulation, stable ingredients, and small toxic and side effects, which has a good effect in preventing and treating Alzheimer's disease. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a Chinese medicine composition for preventing and treating Alzheimer's disease, and its preparation method and use, in view of the deficiencies in the prior art. The Chinese medicine composition has a reasonable and scientific ratio, few adverse reactions, and good antioxidant capacity, and can significantly improve nerve cell damage, alleviate cognitive dysfunction, delay aging, and prevent and treat symptoms of Alzheimer's disease.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0006] The present invention discloses a Chinese medicine composition for preventing and treating Alzheimer's disease, which comprises the following components in parts by weight:

[0007] 10-30 portions of Codonopsis pilosula,

[0008] 10-15 parts of Polygonatum sibiricum,

[0009] 10-20 portions of placenta,

[0010] 6-12 portions of processed Polygonum multiflorum,

[0011] 6-12 parts of Cuscuta seeds,

[0012] 6-10 portions of Epimedium,

[0013] 10-15 portions of Japanese knotweed,

[0014] 3-10 portions of Eupolyphaga sinensis,

[0015] 3 to 6 servings of whole scorpion,

[0016] 10-15 portions of black fern,

[0017] 10-15 servings of Acorus calamus.

[0018] In some embodiments, preferably, it includes the following components in parts by weight:

[0019] 10-20 portions of Codonopsis pilosula,

[0020] 10-12 parts of Polygonatum sibiricum,

[0021] 10-15 portions of placenta,

[0022] 6-10 portions of processed Polygonum multiflorum,

[0023] 6-10 parts of Cuscuta seeds,

[0024] 8-10 portions of Epimedium,

[0025] 10-12 portions of Japanese knotweed,

[0026] 3-6 portions of Eupolyphaga sinensis,

[0027] 3-5 servings of whole scorpion,

[0028] 10-12 portions of black fern,

[0029] 10-12 portions of Acorus calamus.

[0030] In some embodiments, it is further preferred that it comprises the following components in parts by weight: 15 parts of Codonopsis pilosula,

[0031] 10 parts of Polygonatum sibiricum,

[0032] 10 portions of placenta,

[0033] 10 portions of processed Polygonum multiflorum,

[0034] 10 parts of Cuscuta seeds,

[0035] 10 portions of Epimedium,

[0036] 10 portions of Japanese knotweed,

[0037] 3 parts of Eupolyphaga sinensis,

[0038] 3 servings of whole scorpion,

[0039] 10 portions of black fern,

[0040] 10 portions of Acorus calamus.

[0041] In some embodiments, the Chinese medicine composition is in the form of an extract, granules, tablets, capsules, pills, pills, powders or liquid preparations.

[0042] In some embodiments, the Chinese medicine composition is combined with one or more pharmaceutically acceptable excipients to prepare a preparation.

[0043] In some embodiments, the excipients include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, and synergists in the pharmaceutical field; the preparation is in the form of an extract, granules, tablets, capsules, pills, pills, powders, or liquid preparations.

[0044] Furthermore, the present invention discloses a method for preparing the above-mentioned traditional Chinese medicine composition, comprising boiling the above-mentioned parts by weight of Codonopsis pilosula, Polygonatum sibiricum, Placenta centipede, Polygonum multiflorum processed, Cuscuta australis, Epimedium, Polygonum cuspidatum, Eupolyphaga sinensis, Scorpio scorpio, Pteris crenata and Acorus calamus with water, filtering, concentrating the filtrate to obtain a concentrated solution.

[0045] In some embodiments, the weight of water added for each decoction is 6 to 10 times the total weight of the Chinese medicinal materials, the decoction is performed 2 to 3 times, and the decoction time for each decoction is 30 to 60 minutes.

[0046] In some embodiments, the concentration temperature is 70°C to 100°C, and the filtrate is concentrated to a relative density of 1.32 to 1.42 after cooling to obtain a concentrated solution.

[0047] In some embodiments, pharmaceutically acceptable excipients are added to the concentrate during the preparation process.

[0048] The use of the above-mentioned traditional Chinese medicine composition in the preparation of drugs for preventing and / or treating Alzheimer's disease is also within the protection scope of the present invention.

[0049] In the present invention, the efficacy and medicinal taste of each component in the Chinese medicine composition are as follows:

[0050] Codonopsis: sweet, neutral in nature, enters the spleen and lung meridians. It has the functions of tonifying the middle qi, strengthening the spleen and lungs, nourishing blood and promoting body fluid, and is suitable for treating symptoms such as insufficient qi and blood, deficiency of lung and spleen qi, lack of appetite, fatigue, cough and asthma.

[0051] Polygonatum: sweet in taste, neutral in nature, enters the spleen, lung and kidney meridians. It has the functions of replenishing qi and nourishing yin, strengthening the spleen, moistening the lungs and benefiting the kidneys. It is suitable for treating symptoms such as spleen and stomach weakness, soreness of waist and knees, premature graying of hair, fatigue, dry mouth and poor appetite, and dry cough due to lung deficiency.

[0052] Placenta: sweet, salty, warm in nature, enters the lung, liver, and kidney meridians. It has the function of nourishing essence, blood, and qi, and is suitable for treating symptoms such as bone steaming, fatigue, soreness of waist and knees, infertility, impotence, spermatorrhea, asthenia, cough, and lack of milk after childbirth.

[0053] Prepared Polygonum multiflorum: bitter, sweet, astringent, slightly warm in nature, enters the liver, heart, and kidney meridians. It has the functions of nourishing yin and blood, replenishing essence and marrow, and is suitable for treating physical weakness after illness, old age, blood deficiency, sallow complexion, dizziness, tinnitus, premature graying of hair, soreness of waist and knees, and numbness of limbs.

[0054] Cuscuta: sweet in taste, warm in nature, enters the kidney, liver, and spleen meridians. It has the functions of nourishing the liver and kidney, consolidating essence and reducing urine, calming the fetus, improving eyesight, and stopping diarrhea. It is suitable for treating kidney deficiency and low back pain, impotence and spermatorrhea, frequent urination, uterine coldness and infertility.

[0055] Epimedium: Spicy and sweet, warm in nature, enters the liver and kidney meridians. It has the functions of tonifying kidney yang, strengthening tendons and bones, benefiting essence and qi, and removing rheumatism. It is suitable for treating impotence, spermatorrhea, tendon and bone weakness, rheumatic pain, numbness and cramps, etc.

[0056] Polygonum cuspidatum: slightly bitter, slightly cold in nature, enters the liver, gallbladder, and lung meridians. It has the functions of dispelling wind and dampness, dispersing blood stasis and relieving pain, relieving cough and reducing phlegm, and is suitable for treating joint pain, amenorrhea, mass, injuries from falls, carbuncle, sore, and other diseases.

[0057] Eupolyphaga sinensis: salty in taste, cold in nature, enters the liver meridian. It has the functions of breaking blood, removing blood stasis, and repairing tendons and bones. It is suitable for treating amenorrhea, postpartum abdominal pain, lumps, fractures and other diseases.

[0058] Scorpion: Spicy, neutral, enters the liver meridian. It has the functions of calming wind and stopping spasms, detoxifying and dispersing knots, unblocking meridians and relieving pain. It is suitable for treating acute and chronic convulsions, facial paralysis caused by stroke, tetanus, sores, swelling and poison, scrofula and tuberculosis, rheumatism and pain, etc.

[0059] Black fern: bitter, cold in nature, enters the heart, liver, lung, stomach, large intestine, small intestine meridians. It has the functions of clearing away heat and detoxifying, promoting dampness and stopping bleeding. It is suitable for treating wind-heat cold, cough, sore throat, dysentery, damp-heat jaundice, hemoptysis, blood in stool, sores, snake bites and other diseases.

[0060] Acorus calamus: pungent, bitter, warm in nature, enters the heart and stomach meridians. It has the functions of removing dampness and stimulating appetite, opening the orifices and removing phlegm, refreshing the mind and improving intelligence. It is suitable for treating abdominal distension without hunger, coma, epilepsy, forgetfulness, deafness, muteness and dysentery.

[0061] In the Chinese medicine composition of the present invention, Codonopsis pilosula and Polygonatum sibiricum are used as the main medicines to nourish the water and fire of the kidney, improve intelligence and resist aging. Placenta, Radix Polygoni Multiflori, Semen Cuscutae, Epimedium are used to replenish essence and fill marrow, and help generate kidney qi; Polygonum cuspidatum, Scolopendra subspinipes, Scorpio, and Fern are used to promote blood circulation and remove blood stasis, clear away heat and detoxify; Acorus calamus is selected as the assistant medicine to lead the medicine into the brain, open the marrow and open the orifices, calm the mind and improve the will. The above medicines are used together to have good effects such as nourishing yin and tonifying yang, filling marrow and improving intelligence, removing blood stasis and removing toxins, attacking and replenishing at the same time, treating both the symptoms and the root cause, improving cognitive function, and delaying aging. The prescription of the present invention attacks and replenishes at the same time, takes both the symptoms and the root cause into consideration, replenishes without stagnation, attacks without loss, and has a focus; the meridians are mainly kidney, involving the heart, liver and brain; the whole prescription has a mild nature, is not warm or dry, and is not cold, and cooperates with each other, which is very critical to the pathogenesis of senile AD aging.

[0062] The prescription of the present invention nourishes yin and yang, replenishes the marrow and improves intelligence, removes blood stasis and eliminates toxins, aims to achieve balance, improve cognition, and delay the progression of aging. It is mainly aimed at the clinically common AD of kidney deficiency and blood stasis type such as soreness of waist and knees, memory loss, etc., the aging population, the sub-healthy population, and the patients recovering from tumor radiotherapy and chemotherapy.

[0063] Beneficial effects:

[0064] (1) The Chinese medicine composition provided by the present invention is screened according to the theory of traditional Chinese medicine differentiation and treatment, and the ratio is scientific and reasonable, and has good effects of nourishing yin and tonifying yang, replenishing marrow and improving intelligence, and removing blood stasis and detoxifying.

[0065] (2) The Chinese medicine composition provided by the present invention is suitable for patients with Alzheimer's disease, and is also suitable for aging patients with kidney deficiency and blood stasis type. The Chinese medicine composition of the present invention has the effect of improving intelligence and anti-aging, and plays a multi-component, multi-pathway, and multi-target role in treating Alzheimer's disease. It can resist oxidative stress in the body, relieve cognitive dysfunction, and significantly improve nerve cell damage, thereby delaying aging, preventing and treating symptoms of Alzheimer's disease, and improving the patient's daily living ability; at the same time, it has high safety and few adverse reactions.

[0066] (3) The experimental results show that the Chinese medicine composition of the present application can effectively increase the SOD and GSH-Px levels of AD rats and reduce the MDA level. Among them, the high-dose group of the Chinese medicine composition is more effective than the previously patented Chinese medicine composition (the Chinese medicine composition in the Chinese patent with application number 202110408892.9). BRIEF DESCRIPTION OF THE DRAWINGS

[0067] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments, and the above and / or other advantages of the present invention will become more clear.

[0068] Figure 1 HE staining of pathological changes in CA3 region of hippocampus of rats in different treatment groups.

[0069] Figure 2Nissl staining of pathological changes in CA3 region of hippocampus of rats in different treatment groups. DETAILED DESCRIPTION

[0070] The present invention can be better understood according to the following examples. However, it is easy for those skilled in the art to understand that the contents described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims.

[0071] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0072] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents and materials described are commercially available unless otherwise specified.

[0073] Embodiment 1:

[0074] A traditional Chinese medicine composition for preventing and treating Alzheimer's disease is prepared from the following raw materials in parts by weight: 30 parts of Codonopsis pilosula, 15 parts of Polygonatum sibiricum, 20 parts of Placenta centifolia, 10 parts of processed Polygonum multiflorum, 10 parts of Cuscuta seeds, 10 parts of Epimedium, 15 parts of Polygonum cuspidatum, 6 parts of Eupolyphaga sinensis, 6 parts of Scorpio, 15 parts of Pteris crenata and 15 parts of Acorus calamus.

[0075] The preparation method of the Chinese medicine composition comprises the following steps:

[0076] (1) Weigh the API according to the weight proportions stated in the API formula, add water 10 times the total weight of the API to decoct, filter after the first decoction for 40 minutes to obtain a primary filtrate and a primary filter residue; add water 10 times the total weight of the API to the primary filter residue to decoct for a second time for 40 minutes and filter to obtain a secondary filtrate; combine the primary filtrate and the secondary filtrate, and concentrate the combined filtrate at 100° C. and normal pressure to a relative density of 1.32 to 1.42 after cooling to obtain an extract concentrate.

[0077] (2) The extract obtained in step (1) is dried at 60° C. to 70° C. for 8 h, and crushed into fine powder to obtain a Chinese medicine composition powder.

[0078] Embodiment 2:

[0079] A traditional Chinese medicine composition for preventing and treating Alzheimer's disease is prepared from the following raw materials in parts by weight: 10 parts of Codonopsis pilosula, 10 parts of Polygonatum sibiricum, 10 parts of Placenta centella asiatica, 6 parts of processed Polygonum multiflorum, 6 parts of Cuscuta australis, 8 parts of Epimedium, 10 parts of Polygonum cuspidatum, 3 parts of Eupolyphaga sinensis, 3 parts of Scorpio, 10 parts of Pteris crenata and 10 parts of Acorus calamus.

[0080] The preparation method of the Chinese medicine composition comprises the following steps:

[0081] (1) Weigh the API according to the weight proportions stated in the API formula, add water 10 times the total weight of the API and boil for 40 minutes, filter after the first boiling for 40 minutes to obtain a primary filtrate and a primary filter residue; add water 10 times the total weight of the API to the primary filter residue and boil for 40 minutes for the second time and filter to obtain a secondary filtrate; combine the primary filtrate and the secondary filtrate, and concentrate the combined filtrate under reduced pressure at 70° C. to a relative density of 1.32 to 1.42 after cooling to obtain an extract concentrate.

[0082] (2) The extract obtained in step (1) is dried at 60° C. to 70° C. for 8 h, and crushed into fine powder to obtain a Chinese medicine composition powder.

[0083] Embodiment 3:

[0084] A traditional Chinese medicine composition for preventing and treating Alzheimer's disease is prepared from the following raw materials in parts by weight: 15 parts of Codonopsis pilosula, 10 parts of Polygonatum sibiricum, 10 parts of Placenta centella asiatica, 10 parts of Polygonum multiflorum processed, 10 parts of Cuscuta australis, 10 parts of Epimedium, 10 parts of Polygonum cuspidatum, 3 parts of Eupolyphaga sinensis, 3 parts of Scorpio, 10 parts of Pteris crenata and 10 parts of Acorus calamus.

[0085] The preparation method of the Chinese medicine composition comprises the following steps:

[0086] (1) Weigh the API according to the weight proportions stated in the API formula, add water 10 times the total weight of the API and boil for 40 minutes, filter after the first boiling for 40 minutes to obtain a primary filtrate and a primary filter residue; add water 10 times the total weight of the API to the primary filter residue and boil for 40 minutes for the second time and filter to obtain a secondary filtrate; combine the primary filtrate and the secondary filtrate, and concentrate the combined filtrate under reduced pressure at 70° C. to a relative density of 1.32 to 1.42 after cooling to obtain an extract concentrate.

[0087] (2) The extract obtained in step (1) is dried at 60° C. to 70° C. for 8 h, and crushed into fine powder to obtain a Chinese medicine composition powder.

[0088] Embodiment 4:

[0089] An oral preparation, the dosage form of which is a tablet, and the preparation steps are as follows:

[0090] Prepare extract powder according to the method and raw material drug ratio of Example 3, pass the obtained extract powder of the traditional Chinese medicine composition through a 100-mesh sieve, granulate with 75% ethanol, dry at 65° C. for 10 h, add lubricant magnesium stearate, mix well, calculate the tablet weight, and press into tablets to obtain tablets.

[0091] Embodiment 5:

[0092] An oral preparation, the dosage form of which is a granule, and the preparation steps are as follows:

[0093] The extract powder was prepared according to the method and raw material drug ratio of Example 3, the obtained Chinese medicine composition extract powder was passed through a 100-mesh sieve, granulated with 75% ethanol, dried at 65° C. for 10 h, and then granulated to obtain granules.

[0094] Embodiment 6:

[0095] An oral preparation, the dosage form of which is a capsule, and the preparation steps are as follows:

[0096] The extract powder was prepared according to the method and raw material drug ratio of Example 3, the obtained Chinese medicine composition extract powder was passed through a 100-mesh sieve, granulated with 75% ethanol, dried at 65° C. for 10 h, granulated, and filled into capsules to obtain capsules.

[0097] Example 7: Experiment on the protective effect of Chinese medicine composition on nerve cells

[0098] (1) Preparation of medicines

[0099] The powder of the Chinese medicine composition prepared in Example 3 was dissolved in sterile PBS buffer (pH = 7.3-7.5, ion concentration of 0.01 mol / L), and diluted to the concentrations of a low-dose solution of the Chinese medicine composition (0.3 mg / mL), a medium-dose solution of the Chinese medicine composition (0.6 mg / mL), and a high-dose solution of the Chinese medicine composition (1.2 mg / mL).

[0100] (2) Cell culture

[0101] The mouse hippocampal neural cell line HT22 cells were thawed in a 37°C water bath, added to a 10 mL centrifuge tube, and then 4 times the volume of DMEM culture medium containing 10% fetal bovine serum was added, centrifuged at 1000 rpm for 5 min, the supernatant was discarded and DMEM complete culture medium containing 10% FBS was added, and the cells were resuspended and inoculated into a culture dish and cultured in an incubator at 37°C and 5% CO2.

[0102] When the cells were 80% to 90% full, the culture medium was discarded, the cells were washed twice with PBS (pH = 7.3-7.5, concentration of 0.01 mol / L), 1 mL of 0.25% trypsin digestion solution was added, and digestion was carried out at 37°C for 1 min; immediately after digestion, 2 volumes of DMEM complete culture solution containing 10% FBS were added, and the cells were transferred to a 10 mL centrifuge tube and centrifuged at 1000 rpm for 5 min; the supernatant was discarded, fresh complete culture solution was added, the cells were transferred to a culture dish, and cultured in an incubator at 37°C and 5% CO2.

[0103] HT22 cells in the logarithmic growth phase were inoculated in a 96-well culture plate at a density of 6000 cells per well, with 100 μL complete culture medium per well, and blank group, control group, model group, low-dose group of Chinese medicine composition, medium-dose group of Chinese medicine composition, and high-dose group of Chinese medicine composition were set up, with 4 replicates in each group. There were no HT22 cells in the blank group.

[0104] After culturing for 24 hours, the culture medium was discarded, and 100 μL of complete culture medium was added to the blank group, control group and model group, respectively; 100 μL of low-dose solution of the Chinese medicine composition (0.3 mg / mL) was added to the low-dose group of the Chinese medicine composition; 100 μL of medium-dose solution of the Chinese medicine composition (0.6 mg / mL) was added to the medium-dose group of the Chinese medicine composition; 100 μL of high-dose solution of the Chinese medicine composition (1.2 mg / mL) was added to the high-dose group of the Chinese medicine composition; after culturing for 24 hours in the above groups, the culture medium was discarded, 100 μL of complete culture medium was added to the blank group and control group, and 40 μM Aβ was added to the model group and the low-dose group of the Chinese medicine composition, the medium-dose group of the Chinese medicine composition, and the high-dose group of the Chinese medicine composition, respectively. 25-35 After culturing for 24 hours, the culture medium was discarded, and 10% CCK8 solution prepared in DMEM was added to each well. After incubation in the incubator for 1 hour, the absorbance value at 450nm was measured using an enzyme marker to calculate the cell survival rate. The results are shown in Table 1.

[0105] Cell survival rate = (OD value of low-dose Chinese medicine composition group / medium-dose Chinese medicine composition group / high-dose Chinese medicine composition group / model group - OD value of blank group)) / (OD value of control group - OD value of blank group) × 100%.

[0106] Table 1 Cell survival rate in different treatment groups

[0107] Group Cell survival rate (%) Control group 100±4.61 Model Group 61.05±1.27 Low dose group of Chinese medicine combination 72.92±3.76 Chinese medicine composition medium dose group 80.15±7.09 High dose group of Chinese medicine combination 86.52±5.74

[0108] As can be seen from Table 1, after the HT22 cells in the low-dose group of the Chinese medicine composition / the medium-dose group of the Chinese medicine composition / the high-dose group of the Chinese medicine composition were intervened with the Chinese medicine composition for preventing and treating Alzheimer's disease, the Aβ 25-35 The damaged HT22 cells had a certain protective effect, among which the high-dose group of the Chinese medicine composition had the most significant protective effect.

[0109] Example 8: Learning and memory behavior test

[0110] (1) The Chinese medicine composition powder prepared in Example 3 was diluted with physiological saline to obtain a Chinese medicine composition solution.

[0111] (2) 36 male SPF SD rats with a body weight range of 180-220 g were selected and provided by the Experimental Animal Center of Hangzhou Medical College. The rats were randomly divided into 6 groups, 6 rats in each group, namely, blank control group, model group, positive control group, previously patented Chinese medicine composition group (202110408892.9 group), low-dose group of Chinese medicine composition of this application, and high-dose group of Chinese medicine composition.

[0112] Among them, rats in the model group, positive control group, low-dose group of Chinese medicine composition, high-dose group of Chinese medicine composition, and 202110408892.9 group were treated with intraperitoneal injection of D-gal (D-galactose, 60 mg / kg / d) and oral gavage of AlCl3 (200 mg / kg / d) for 10 weeks, once a day, and the blank control group was intervened with an equal volume of normal saline. Starting from the date of modeling, drug intervention was performed 2 weeks after modeling. The low-dose group of Chinese medicine composition and the high-dose group of Chinese medicine composition of this application were gavaged with the Chinese medicine composition solution prepared in step (1) at a dose of 7.2 mg / kg / d and 14.4 mg / kg / d, respectively, once a day; the 202110408892.9 group was gavaged according to the Chinese medicine composition of the previous patent application (using the Chinese medicine decoction formula recorded in paragraph 0062 of the Chinese patent specification with application number 202110408892.9: ginseng: 15 g; Polygonatum: 15g; Epimedium: 10g; Radix Rehmanniae Preparata: 10g; Radix Polygoni Multiflori Preparata: 10g; Cuscuta Semen: 10g; Pteris Piper: 15g; Polygonum Cuspidatum: 15g; Alpinia Oxyphylla: 12g; Acorus Tatarinowii: 10g, the decoction method is the same as Example 3 of the present application, the prepared Chinese medicine composition powder is dissolved with physiological saline, and then the rats are gavaged), the dosage is 14.4mg / kg / d, 1 time / d; the positive control group is gavaged with 0.45mg / kg / d donepezil hydrochloride tablet suspension, 1 time / d; each group is continuously administered for 8 weeks. Among them, the dosage unit of the Chinese medicine composition administration group is mg / kg / d, mg is the weight of the Chinese medicinal material, and kg is the weight of the rat.

[0113] OLT and NORT tests were used. One week before the experiment, each rat was touched for 1 min every day; on the day of the experiment, the rats were adapted to the environment 1 h in advance. The experiment used a 72×72 cm square behavior box and was conducted in 4 stages over 2 days. After each experiment, the behavior box was thoroughly cleaned with 75% alcohol. The first stage (adaptation period): the rats were placed one by one from a fixed position facing the wall of the behavior box into an open behavior box to explore freely for 5 min. After the exploration, the rats were taken out and the behavior box was cleaned with 75% alcohol. This stage was repeated three times on the first day. The second stage (familiarization period): after placing two different objects (objects A and B) in relative positions on the same side of the behavior box, the rats were placed from the original fixed position facing the wall of the behavior box into the behavior box to explore for 10 min. Phase III (test period - OLT): After an interval of 20 minutes in the second phase, object A remained unchanged, and object B was moved to the diagonal relative position of object A as object B'. The rat was then placed in the behavior box from its original fixed position facing the wall of the behavior box to explore for 10 minutes, and the SuperMaze animal behavior video analysis software was used to analyze and record the time the rat explored the two objects. Phase IV (test period - NORT): After an interval of 20 minutes in the third phase, object A was replaced with a completely different object C, and the rat was then placed in the behavior box from its original fixed position facing the wall of the behavior box to explore for 10 minutes, and the software was used to analyze and record the time the rat explored the two objects. It was considered that the rat facing the object and the distance between the nose and the object was ≤2cm was exploratory behavior.

[0114] Discrimination index = [(new location or new object exploration time - old location or old object exploration time) /

[0115] (exploration time of new location or new object + exploration time of old location or old object)]

[0116] If the index is positive, it means that more time is spent exploring the new location or new object; if the index is zero, it means that the time spent exploring the new and old locations or the new and old objects is equal. The novel object recognition test is a sensitive behavioral method that uses the natural curiosity of rodents for novel objects to test their recognition memory. If the mouse has poor cognitive ability, there will be no difference in the exploration time of new / old objects; if the cognitive ability is normal, the exploration time of new things will be longer than the exploration time of old things (the discrimination index is a positive value).

[0117] The discrimination index of each group was statistically analyzed and the results are shown in Table 2.

[0118] Table 2: Comparison of the discrimination index of the object location task and novel object recognition task in rats of different treatment groups

[0119]

[0120] Note: OLT: object localization task, NORT: novel object recognition task; compared with the blank control group, *P<0.05; compared with the model group, #P<0.05.

[0121] It can be seen from Table 2 that compared with the blank control group, the discrimination index of the rats in the model group for new positions and new objects was significantly reduced (P < 0.05); compared with the model group, the discrimination index of the rats in the positive control group, the high-dose group of the Chinese medicine composition and the 202110408892.9 group for new positions was significantly increased (P < 0.05), indicating that the Chinese medicine composition of the present application can effectively improve the spatial learning and memory ability of rats, and improve the recognition and memory ability of rats to a certain extent. At the same time, the therapeutic effect of the high-dose group of the Chinese medicine composition is significantly improved compared with the previously patented Chinese medicine composition (202110408892.9 group).

[0122] Example 9: Determination of related indicators

[0123] After the rat learning and memory behavior test experiment in Example 8 was completed, the rats were anesthetized and blood was collected from the abdominal aorta. The serum was collected after centrifugation and stored at -80°C. After blood collection, the rats were killed and the brain tissues were taken out and placed in 4% paraformaldehyde for future use.

[0124] (I) Observation of pathological changes in hippocampal tissue by HE staining and Nissl staining

[0125] The fixed rat brain tissue was taken out and placed in an embedding box. After washing away the excess paraformaldehyde with running water, it was soaked in 50% ethanol solution (2h), 60% ethanol solution (2h), 75% ethanol solution (2h), 85% ethanol solution (2h), 95% ethanol solution (1h, 2 times), and 100% ethanol solution (30min, 2 times) to remove the moisture of the tissue. The dehydrated brain tissue was soaked in xylene solution (10min, 2 times). Then, the transparent brain tissue was placed in soft wax (1h) and hard wax (2h) for waxing, and then made into brain tissue wax blocks with an embedding machine. At room temperature, the required part of the tissue wax block was cut into thin slices of about 4μm thickness using a paraffin slicer, and the slices were spread and pasted on a glass slide. After baking, the paraffin slices were preserved for later use.

[0126] (1) HE staining

[0127] Take the prepared paraffin sections and soak them in xylene (10min, 2 times) for dewaxing, then soak them in anhydrous ethanol (5min), 95% ethanol solution (5min), 90% ethanol solution (5min), 85% ethanol solution (5min), 75% ethanol solution (5min), and finally soak them in pure water for 5min. Then, stain the sections with hematoxylin for 7min and rinse them gently with pure water for 1min; after hydrochloric acid ethanol differentiation, rinse with pure water for anti-blue; then stain with eosin for 5min and rinse gently with pure water for 15s. After staining, soak the sections in 75% ethanol solution (5min), 90% ethanol solution (5min), and anhydrous ethanol (5min) for dehydration, then soak them in xylene (5min, 2 times) to make them transparent, air-dry the sections, and finally seal them with neutral gum. The neuronal structure, number of arranged layers, cell morphology, etc. in the CA3 region of the hippocampal tissue of each group of rats were observed under a microscope. The results are shown in Figure 1 (×400, HE staining). The low-dose group of Shenhuang Heji in the figure is the low-dose group of the Chinese medicine composition, and the high-dose group of Shenhuang Heji is the high-dose group of the Chinese medicine composition.

[0128] Figure 1 It shows that the blank control group has a regular and neat structure, abundant neurons, normal morphology, neat arrangement, and a clear and centered nucleolus. The model group has a disordered structure, a reduced number of neurons, abnormal morphology, uneven arrangement, and nuclear condensation. Compared with the model group, the positive control group, the 202110408892.9 group, and the low- and high-dose groups of the Chinese medicine composition restored regular and neat structures, an increased number of neurons, and neat arrangement, and the cell morphology returned to normal, and the nuclear condensation improved. It shows that the Chinese medicine composition of this application can improve the number, morphology, and abnormal arrangement of neurons, and is better than the Chinese medicine composition previously applied for a patent (i.e., the Chinese medicine composition of the 202110408892.9 group).

[0129] (2) Nissl staining

[0130] Take the prepared paraffin sections and dry them at 40℃ for 6 minutes, then soak them in xylene solution (5 minutes, 2 times) for dewaxing, and then soak them in anhydrous ethanol (5 minutes), 95% ethanol solution (5 minutes), 80% ethanol solution (5 minutes), and 75% ethanol solution (5 minutes) for dehydration. After dehydration, place the sections in 50℃ toluidine blue dye for 10 minutes, and then put them in a 60℃ constant temperature box for staining for 25 minutes. After staining, rinse the sections gently with pure water, then differentiate with 95% ethanol and dehydrate with gradient ethanol, then soak them in xylene solution (5 minutes, 3 times), air-dry the sections, and finally seal them with neutral gum. The changes of Nissl bodies in the neurons of the CA3 region of the hippocampal tissue of each group of rats were observed under a microscope. The results are shown in Figure 2(×400, Nissl staining). The low-dose group of Shenhuang Heji in the figure is the low-dose group of the Chinese medicine composition, and the high-dose group of Shenhuang Heji is the high-dose group of the Chinese medicine composition.

[0131] Figure 2 It shows that the neurons in the blank control group are complete in morphology, the cell morphology is clear and normal, and neatly arranged blue-purple Nissl bodies can be seen. The neurons in the model group are scattered, the morphology is incomplete, some cells are wrinkled, the number of Nissl bodies is reduced, and the staining is uneven. Compared with the model group, the neurons in the positive control group, the 202110408892.9 group, and the low- and high-dose groups of the Chinese medicine composition are better in arrangement and morphology, and the number and morphology of Nissl bodies are also improved. It shows that the Chinese medicine composition of this application can effectively improve neuronal damage in the hippocampal tissue of the brain, and is better than the Chinese medicine composition previously applied for a patent (i.e., the Chinese medicine composition of the 202110408892.9 group).

[0132] (II) Determination of serum superoxide dismutase, malondialdehyde and glutathione peroxidase levels in rats

[0133] According to the instructions of ELISA kit (Nanjing Jiancheng Bioengineering Institute Co., Ltd.), the levels of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in rat serum were measured. The results are shown in Table 3.

[0134] Table 3: Levels of serum SOD, MDA and GSH-Px in rats in different treatment groups

[0135]

[0136]

[0137] Note: Compared with the blank control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0138] Table 3 shows that compared with the blank control group, the activity of antioxidant enzymes SOD and GSH-Px in the serum of rats in the model group was significantly reduced (P<0.05), and the content of oxidation product MDA was significantly increased (P<0.01). Compared with the model group, the MDA content in the serum of rats in each drug intervention group was significantly reduced (P<0.01); the SOD activity in the serum of rats in the positive control group, the high-dose group of the Chinese medicine composition, and the 202110408892.9 group was significantly increased (P<0.05); the GSH-Px activity in the serum of rats in the positive control group, the high-dose group of the Chinese medicine composition, and the 202110408892.9 group was significantly increased compared with the model group. The results show that the Chinese medicine composition of the present application can effectively improve the levels of SOD and GSH-Px in AD rats and reduce the level of MDA. Among them, the high-dose group of the Chinese medicine composition is better than the Chinese medicine composition previously applied for patents (i.e., the Chinese medicine composition of the 202110408892.9 group).

[0139] The present invention provides a Chinese medicine composition for preventing and treating Alzheimer's disease, and a method for preparing and using the composition. There are many methods and approaches to realize the technical solution. The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the protection scope of the present invention. All components not specified in this embodiment can be realized by existing technologies.

Claims

1. A Chinese medicine composition for preventing and treating Alzheimer's disease, characterized in that: The invention comprises the following components by weight: 10 to 30 parts of Codonopsis pilosula, 10-15 parts of Polygonatum sibiricum, 10-20 portions of placenta, 6-12 portions of processed Polygonum multiflorum, 6-12 parts of Cuscuta seeds, 6-10 portions of Epimedium, 10-15 portions of Japanese knotweed, 3-10 portions of Eupolyphaga sinensis, 3 to 6 whole scorpions, 10-15 portions of black fern, 10-15 servings of Acorus calamus.

2. The Chinese medicine composition according to claim 1, characterized in that: The invention comprises the following components by weight: 10 to 20 parts of codonopsis pilosula, 10-12 parts of Polygonatum sibiricum, 10-15 portions of placenta, 6-10 portions of processed Polygonum multiflorum, 6-10 parts of Cuscuta seeds, 8-10 portions of Epimedium, 10-12 portions of Japanese knotweed, 3-6 portions of Eupolyphaga sinensis, 3-5 servings of whole scorpion, 10-12 portions of black fern, 10-12 portions of Acorus calamus.

3. The Chinese medicine composition according to claim 1, characterized in that The invention comprises the following components by weight: 15 parts of Codonopsis pilosula, 10 parts of Polygonatum sibiricum, 10 portions of placenta, 10 portions of processed Polygonum multiflorum, 10 parts of Cuscuta seeds, 10 portions of Epimedium, 10 portions of Japanese knotweed, 3 parts of Eupolyphaga sinensis, 3 servings of whole scorpion, 10 portions of black fern, 10 portions of Acorus calamus.

4. The Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The Chinese medicine composition is in the form of extract, granules, tablets, capsules, pills, pills, powders or liquid preparations.

5. The Chinese medicine composition according to claim 1, characterized in that: The Chinese medicine composition is combined with one or more pharmaceutically acceptable excipients to prepare a preparation.

6. The Chinese medicine composition according to claim 5, characterized in that: The auxiliary materials include conventional diluents, excipients, fillers, adhesives, wetting agents, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, and synergists in the pharmaceutical field; the dosage form of the preparation is extract, granules, tablets, capsules, pills, pills, powders or liquid preparations.

7. The method for preparing the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The above weight proportions of Codonopsis pilosula, Polygonatum sibiricum, Placenta centella asiatica, Radix Polygoni multiflori, Semen Cuscutae, Epimedium, Polygonum cuspidatum, Eupolyphaga sinensis, Scorpio scorpio, Pteris cretica and Acorus calamus are decocted in water, filtered, and the filtrate is concentrated to obtain a concentrated solution.

8. The preparation method according to claim 7, characterized in that: The water-adding and decocting step is as follows: the weight of water added each time is 6 to 10 times of the total weight of the Chinese medicinal materials; the decocting times are 2 to 3 times; and the decocting time each time is 30 to 60 minutes; the concentration step is as follows: the concentration temperature is 70° C. to 100° C., and the filtrate is concentrated to a relative density of 1.32 to 1.42 after cooling to obtain a concentrated solution.

9. The preparation method according to claim 7, characterized in that: During the preparation process, pharmaceutically acceptable excipients are added to the concentrate.

10. Use of the Chinese medicine composition according to any one of claims 1 to 6 in the preparation of a drug for preventing and / or treating Alzheimer's disease.

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