A traditional Chinese medicine composition for treating cognitive decline due to kidney yang deficiency

Abstract

This invention discloses a traditional Chinese medicine composition for treating cognitive decline caused by kidney-yang deficiency. Compared with existing technologies, the traditional Chinese medicine composition of this invention, composed of Eucommia ulmoides and total ginsenosides, has significant effects in preventing or treating cognitive decline, especially cognitive decline caused by kidney-yang deficiency. In comparison with the effects of using Eucommia ulmoides alone or total ginsenosides alone, it can be clearly seen that the combined use of the traditional Chinese medicine composition of this invention has a synergistic effect on the prevention or treatment of cognitive decline.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a traditional Chinese medicine composition for treating cognitive decline due to kidney yang deficiency. Background Technology

[0002] Kidney Yang deficiency is a constitutional disorder in Traditional Chinese Medicine (TCM), referring to the symptoms manifested by a decline in the Yang energy of the kidneys. The earliest record of Kidney Yang deficiency is found in the *Huangdi Neijing* (Yellow Emperor's Inner Classic). With continuous research and development in TCM, the symptoms and treatments for Kidney Yang deficiency have gradually evolved. Clinical studies have shown that patients with Kidney Yang deficiency are more prone to cognitive decline, but treatment methods for Kidney Yang deficiency-related cognitive decline still require further exploration.

[0003] Eucommia bark is the bark of the Eucommia ulmoides tree, a plant in the Eucommiaceae family. It has a sweet and slightly spicy taste, is warm in nature, and enters the liver and kidney meridians. It is mainly used to tonify the liver and kidneys, strengthen muscles and bones, and calm the fetus. It is used to treat lower back pain, weakness in the legs and knees, dribbling urination, dampness and itching in the genital area, threatened miscarriage, fetal restlessness, and high blood pressure.

[0004] Ginsenosides are important active components of ginseng, belonging to the triterpenoid glycoside class of compounds. They can be classified into protopanaxadiol saponins (PPD type saponins), protopanatriol saponins (PPT type saponins), and oleanane type. Ginsenosides possess various therapeutic effects, including antioxidant, anti-inflammatory, vasodilatory, anti-allergic, and anti-diabetic properties. Some ginsenosides exhibit anticancer properties by reducing DNA damage, decreasing host susceptibility to mutations, and increasing immune surveillance and apoptosis. Furthermore, ginsenosides can effectively improve the efficacy of traditional chemotherapy drugs and prevent damage to normal tissues. Summary of the Invention

[0005] In order to solve the technical problems existing in the prior art, the present invention provides the following technical solution:

[0006] The first aspect of the present invention provides a traditional Chinese medicine composition for treating cognitive decline, the composition comprising Eucommia ulmoides and total ginsenosides.

[0007] Furthermore, the mass ratio of Eucommia ulmoides to total ginsenosides is (100 - 400): (20 - 80).

[0008] Furthermore, the mass ratio of Eucommia ulmoides to total ginsenosides is 5:1.

[0009] Furthermore, the individual per kilogram weights of Eucommia ulmoides and total ginsenosides are: 100mg, 200mg, or 400mg of Eucommia ulmoides; and 20mg, 40mg, or 80mg of total ginsenosides.

[0010] Furthermore, the individual mass per kilogram of Eucommia ulmoides and total ginsenosides is: 100 mg of Eucommia ulmoides and 20 mg of total ginsenosides.

[0011] Furthermore, the cognitive decline described is a type of cognitive decline caused by kidney yang deficiency.

[0012] Furthermore, the Eucommia ulmoides includes Eucommia ulmoides raw material, effective components extracted from Eucommia ulmoides raw material, or medicinal salts of effective components; the total ginsenosides include total ginsenosides or their medicinal salts.

[0013] Furthermore, the types of raw materials include dried medicinal materials, fresh medicinal materials, raw medicinal materials, processed medicinal materials, whole plant of medicinal materials, effective parts of whole plant of medicinal materials, or combinations thereof.

[0014] In this invention, "raw material" is synonymous with "active pharmaceutical ingredient," referring to the traditional Chinese medicinal materials that enable the invention to exert its active functions. These materials include various forms, specifically dried, fresh, raw, processed, or prepared medicinal materials, the whole plant of the medicinal material, the effective parts of the whole plant, or combinations thereof, but are not limited to these. In some embodiments, the raw material further comprises the main active ingredients extracted from the raw material, all of which are components proven in the prior art.

[0015] Furthermore, the pharmaceutically usable salts include acid addition salts and alkali addition salts.

[0016] The term "medicinal salt" as used herein includes ammonium phosphates, metal salts, salts formed with amino acids, salts formed with amines, and salts formed with other bases such as piperidine or morpholine. The term "medicinal salt" includes monosalts or disalts. Specifically, it includes ammonium, sodium, calcium, magnesium, cesium, lithium, potassium, barium, zinc, aluminum, lysine, arginine, histidine, methylamine, ethylamine, tert-butylamine, cyclohexylamine, N-methylglucosamine, ethylenediamine, glycine, procaine, benzathine, diethanolamine, triethanolamine, piperidine, and morpholine.

[0017] In some embodiments, the term "medicinal salt" may also include hydrates of the pharmaceutically acceptable salts of the active ingredient of the present invention. Examples of salts include, but are not limited to, sulfates, citrates, acetates, oxalates, chlorides, hydrochlorides, bromides, hydrobromides, iodides, nitrates, bisulfates, phosphates, acid phosphates, isonicotinate, lactate, salicylates, acid citrates, tartrates, oleates, tannates, pantothenates, hydrogen tartrate, ascorbate, gentisinate, gluconate, glucuronate, glycosides, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorsulfonate, naphthalenesulfonate, propionate, succinate, fumarate, maleate, malonate, mandelate, malate, palmitate, aspartate, phthalates, and pyrates. Each of the salts mentioned above may be anhydrous or hydrated. As used herein, the term "medicinal salt" also refers to salts formed by the active ingredients of the medicinal materials of this invention having acidic functional groups, such as carboxylic acid functional groups, and alkalis. Examples of alkalis include, but are not limited to: hydroxides of alkali metals, including sodium, potassium, and lithium; hydroxides of alkaline earth metals, such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia; organic amines, such as unsubstituted or hydroxylated mono-, di-, or trialkylamines, dicyclohexylamine; tributylamine; pyridine; N-methyl, N-ethylamine; diethylamine; triethylamine; mono-, di-, or tri-(2-OH-(C1-C6)-alkylamines), such as N,N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2-hydroxyethyl)amine; N-methyl-D-glucosamine; morpholine; thiomorpholine; piperidine; pyrrolidine; and amino acids such as arginine, lysine, etc.

[0018] In this invention, prevention refers to various means or measures, including medical, physical or chemical methods, used to prevent the occurrence or development of a disease before it is recognized by clinical standards, in order to stop and reduce the occurrence or development of various symptoms of the disease. In a specific embodiment of this invention, the disease is preferably cognitive decline due to kidney yang deficiency.

[0019] In this invention, the treatment refers to the act of preventing and reducing the occurrence or development of a disease, thereby inhibiting, suppressing, alleviating, improving, slowing down, stopping, delaying, or reversing the progression or aggravation of the disease. The various indicators of maintaining and / or using the medication include alleviating or reducing symptoms or complications, or curing or eliminating the disease, disorder, or condition. In a specific embodiment of this invention, the disease is preferably a type of cognitive decline due to kidney yang deficiency.

[0020] A second aspect of the present invention provides a traditional Chinese medicine preparation for treating cognitive decline, the traditional Chinese medicine preparation comprising the traditional Chinese medicine composition provided in the first aspect of the present invention.

[0021] Furthermore, the dosage forms of the traditional Chinese medicine preparations include granules, tablets, capsules, oral liquids, pills, and drop pills.

[0022] As a non-limiting example, the preparation of the dosage form of the traditional Chinese medicine preparation is as follows:

[0023] Granules: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of the present invention as the active ingredient, pharmaceutical excipients are added, mixed, granulated, dried, and packaged to obtain traditional Chinese medicine granules containing the traditional Chinese medicine composition for treating cognitive decline.

[0024] Tablets: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of this invention as the active ingredient, the ingredients are dried, pharmaceutical excipients are added, mixed, granulated, dried, compressed, and coated to obtain a traditional Chinese medicine tablet containing the traditional Chinese medicine composition for treating cognitive decline. The tablet can be a plain tablet or a coated tablet, such as a sugar-coated, gastric-coated, or enteric-coated tablet.

[0025] Capsules: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of this invention as the active ingredient, the ingredients are dried, then pharmaceutical excipients are added, mixed, granulated, dried, and encapsulated to obtain a traditional Chinese medicine capsule containing the traditional Chinese medicine composition for treating cognitive decline. The capsule can be a hard capsule or a soft capsule.

[0026] Oral liquid: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of the present invention as the active ingredient, water is added and stirred until homogeneous, then pharmaceutical excipients are added, stirred until homogeneous, filtered, filled, sterilized, and packaged to obtain a traditional Chinese medicine oral liquid containing the traditional Chinese medicine composition for treating cognitive decline.

[0027] Pills: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of this invention as the active ingredient, after drying and pulverizing, pharmaceutical excipients are added, the mixture is placed in a sugar coating pan, a polar solvent is added while rotating the sugar coating machine, the mixture is stirred, formed into pills, dried, coated, and packaged to obtain traditional Chinese medicine pills containing the traditional Chinese medicine composition for treating cognitive decline.

[0028] Droplets: Using the traditional Chinese medicine composition for treating cognitive decline described in the first aspect of this invention as the active ingredient, after drying and pulverizing, pharmaceutical excipients are added, mixed, heated and melted, stirred evenly, made into pills, washed, dried, and packaged to obtain traditional Chinese medicine droplets containing the traditional Chinese medicine composition for treating cognitive decline.

[0029] Furthermore, the traditional Chinese medicine preparation also includes pharmaceutically acceptable excipients.

[0030] Furthermore, the excipients include binders, surfactants, flavoring agents, osmotic pressure regulators, antibacterial agents, solubilizers, antioxidants, and stabilizers.

[0031] Furthermore, the adhesive includes carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, povidone, glycerin, polyvinyl alcohol, polyvinylpyrrolidone, polyalkyl styrene, polymethacrylate and / or polyacrylate.

[0032] Furthermore, the surfactant includes hydroxypropyl-β-cyclodextrin, polyoxyethylene alkyl ether, Tween, sodium lauryl sulfate, sodium lauryl sulfate, poloxamer, polysorbate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate and / or polyethylene glycol alkyl.

[0033] Furthermore, the flavoring agents include sodium saccharin, cyclamate, aspartame, acetylsupan-K, glycerin, sorbitol, mannitol, sucrose, simple syrup, and aromatic syrup.

[0034] Furthermore, the osmotic pressure regulator includes sodium chloride, potassium chloride, mannitol, glycerol, and / or other pharmaceutically or traditional Chinese medicine-acceptable osmotic pressure regulators.

[0035] Furthermore, the antibacterial agents include BAK, benzalkonium chloride, sorbic acid, oxychloride complex, citric acid, chlorobutanol, thimerosal, phenylmercuric acetate, disodium EDTA, phenylmercuric nitrate, perborate, and / or benzyl alcohol.

[0036] Furthermore, the co-solvents include hydrochloric acid, phosphoric acid, propionic acid, acetic acid, lactic acid, citric acid, tartaric acid, boric acid, glucuronic acid, gluconic acid, lactobionic acid, malic acid, threonic acid, glucoheponic acid, 2,5-dihydroxybenzoic acid, and acidic amino acids.

[0037] Furthermore, the antioxidants and stabilizers include sulfurous acid, sulfites, bisulfites, metabisulfites, dithionite, thiosulfates, thiourea, glutathione, dimercaprol, mercaptoacetic acid and its salts, thiolactic acid and its salts, thiodipropionic acid and its salts, gallic acid and its salts, caffeic acid or its pharmaceutical salts, ferulic acid or its pharmaceutical salts, di-tert-butyl-p-phenol, 2,5-dihydroxybenzoic acid or its salts, salicylic acid or its salts, ascorbic acid and its salts, isoascorbic acid and its salts, nicotinamide, tartaric acid, phosphates, pharmaceutical salts of acetate, citrates, EDTA and its salts.

[0038] In some embodiments, the excipients may also include substances recognized in the art and including, for example, substances involved in carrying or transporting any subject composition from one organ or part of the body to another organ or part of the body, including but not limited to: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) cocoa butter and suppository wax; (9) Oils, such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer; (21) other non-toxic compatible substances used in traditional Chinese medicine preparations.

[0039] In specific implementation schemes, the choice of which pharmaceutical excipients to combine with the traditional Chinese medicine preparation depends on the preparation to be made. When making ordinary preparations, the choice of which pharmaceutical excipients to use is well known to those skilled in the art.

[0040] In some embodiments, the traditional Chinese medicine preparations of the present invention further include a drug delivery system, specifically including biodegradable polymers, biodegradable grafts, non-biodegradable grafts, and biodegradable microparticles such as biodegradable microspheres and nanoparticles.

[0041] In some implementations, the dosage of the traditional Chinese medicine preparation can be varied based on factors such as formulation method, administration method, patient's age, weight, gender, condition, diet, administration time, route of administration, excretion rate, and responsiveness, and such variations can be easily determined and identified by a skilled physician in the art.

[0042] In some implementation schemes, when administering the traditional Chinese medicine preparations described in this invention to a specific individual, the effective therapeutic dose and specific treatment plan for that individual can be affected by many factors, including the pharmacodynamic activity of the drug used, the age, weight, general condition, gender, diet, administration time, disease susceptibility, disease progression, and the judgment of the attending physician. Furthermore, those skilled in the art understand that the administration method, dosage form, and dosage of a drug are affected by many factors such as the patient's age, weight, gender, pathological condition, diet, administration time, excretion rate, and responsiveness. Therefore, the dosage of the traditional Chinese medicine preparations described in this invention is not limited to the dosage in the embodiments of this invention, and the administration method and dosage form are similar.

[0043] The third aspect of the present invention provides the use of the traditional Chinese medicine composition described in the first aspect of the present invention or the traditional Chinese medicine preparation described in the second aspect of the present invention in the preparation of medicines for the prevention or treatment of cognitive decline.

[0044] The fourth aspect of the present invention provides the use of the traditional Chinese medicine composition described in the first aspect of the present invention or the traditional Chinese medicine preparation described in the second aspect of the present invention in the preparation of a medicine for restoring the density of hippocampal dendritic spines.

[0045] Furthermore, the application described is for non-therapeutic purposes.

[0046] The term "Eucommia" used in this invention refers to the bark of the Eucommia ulmoides tree, a plant in the Eucommiaceae family. It has a sweet and slightly pungent taste, is warm in nature, and enters the liver and kidney meridians. Its main functions are to tonify the liver and kidneys, strengthen tendons and bones, and calm the fetus. It is used to treat lower back pain, weakness in the legs and knees, dribbling urination, itching in the genital area, threatened miscarriage, fetal restlessness, and high blood pressure.

[0047] The term "total ginsenosides" used in this invention refers to important active components of ginseng, belonging to the triterpenoid glycoside compounds. They can be classified into protopanaxadiol saponins (PPD type saponins), protopanatriol saponins (PPT type saponins), and oleanane type, generally appearing as a yellowish-white or pale yellow amorphous powder; slightly odorous, bitter in taste; and hygroscopic. Ginsenosides possess various therapeutic effects, including antioxidant, anti-inflammatory, vasodilatory, anti-allergic, and anti-diabetic properties. Some ginsenosides exhibit anticancer properties by reducing DNA damage, decreasing host susceptibility to mutations, and increasing immune surveillance and apoptosis. Furthermore, ginsenosides can effectively improve the efficacy of traditional chemotherapy drugs and prevent damage to normal tissues.

[0048] The term "cognitive function" as used in this invention refers to functions such as thinking, reasoning, memory, impression, and learning. In some embodiments, cognition refers to mental processes including attention, memory, language production and comprehension, learning, reasoning, problem-solving, and decision-making.

[0049] Advantages and beneficial effects of the present invention:

[0050] This invention provides a traditional Chinese medicine composition for treating cognitive decline caused by kidney yang deficiency. Compared with the prior art, its beneficial effects are that the traditional Chinese medicine composition of this invention is composed of Eucommia ulmoides and total ginsenosides, and has significant effects in treating and preventing cognitive decline, especially cognitive decline caused by kidney yang deficiency. In the comparison of the effects of using Eucommia ulmoides alone and using total ginsenosides alone, it can be clearly seen that the combined use of the traditional Chinese medicine composition of this invention has a synergistic effect on the treatment and prevention of cognitive decline caused by kidney yang deficiency.

[0051] Unless otherwise stated, all figures used in this specification and claims to represent volume, weight, temperature, time, density, parts by weight, technical effect, etc., should in any case be understood to be modified by the terms "about" or "approximately". Therefore, unless indicated to the contrary, the numerical parameters listed in the specification and appended claims are approximate values. Unless otherwise stated, the terminology used herein has its common meaning as understood by one of ordinary skill in the art, and each numerical parameter should be interpreted according to the number of significant figures and conventional rounding methods or in a manner understood by one of those skilled in the art. Attached Figure Description

[0052] Figure 1 The figure shows the experimental results of the combined use of Eucommia ulmoides and total ginsenosides on the cognitive function of rats with kidney-yang deficiency; where A is the behavioral path diagram of the water maze; B is the total path quantification diagram; and C is the escape latency quantification diagram.

[0053] Figure 2 The figure shows the experimental results of the combined use of Eucommia ulmoides and total ginsenosides on the dendritic spines of the hippocampus in rats with kidney-yang deficiency. Detailed Implementation

[0054] The present invention will be further illustrated below with reference to specific embodiments. These specific embodiments are for illustrative purposes only and should not be construed as limiting the invention. Those skilled in the art will understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the invention. The scope of the invention is defined by the claims and their equivalents. The experimental consumables, reagents, and raw materials used in this invention are readily available to those skilled in the art and, unless otherwise specified, can be obtained commercially. Experimental methods in this invention that do not specify specific conditions are generally performed under conventional conditions or according to the manufacturer's recommended conditions.

[0055] The terminology used in this application is for the purpose of describing particular embodiments only and is not to be construed as limiting. As used herein, the singular forms “an” and “a” are also intended to include the plural forms unless the context clearly indicates otherwise. As used herein, the term “and / or” includes any and all combinations of one or more of the associated enumerated items. Expressions such as “at least one of…” modify the entire list of elements when preceding or following it, without modifying any individual element in the list.

[0056] Example 1: Water maze test to assess cognitive function in rats with kidney-yang deficiency

[0057] I. Experimental Methods

[0058] 1. Medicinal materials: Eucommia ulmoides (Shanghai Yuanye Biotechnology Co., Ltd., B26137); Ginsenosides (Shanghai Yuanye Biotechnology Co., Ltd., S31132).

[0059] 2. Experimental animals: SD rats, male, 10-12 weeks old, 8 rats per group.

[0060] Rat model of kidney yang deficiency: Hydrocortisone (HC) 15 mg / kg, intramuscular injection in the hind leg (alternating between left and right legs), for 21 consecutive days.

[0061] 3. Experimental Grouping

[0062] 1) Normal group: intramuscular injection of physiological saline;

[0063] 2) HC group: HC 15 mg / kg / day was administered intramuscularly for 21 consecutive days;

[0064] 3) HC + Eucommia ulmoides group: HC and Eucommia ulmoides (100, 200, 400 mg / kg / d) were administered by gavage for 21 days;

[0065] 4) HC + Ginsenosides group: HC and ginsenosides (20, 40, 80 mg / kg / d) were administered by gavage for 21 days;

[0066] 5) HC + Eucommia ulmoides + Ginseng total saponins group: Eucommia ulmoides 100 mg / kg + Ginseng total saponins 20 mg / kg.

[0067] 4. Water maze test to assess cognitive function

[0068] The water maze consisted of a circular pool with a diameter of 150 cm and an automated camera and computer analysis system that simultaneously recorded the rats' movement trajectories. The water depth was 40 cm, and the water temperature was 24 ± 2℃. The experimental training phase lasted for 4 consecutive days, with 4 training sessions per day. During training, the animals were submerged headfirst into the water, and the time it took for the rats to find and stand on the underwater platform was recorded as the escape latency, expressed in seconds (s). Rats that failed to find the platform within 60 seconds were led to it and placed there for at least 10 seconds to encourage learning and memorization. The fifth day was the formal testing period, during which the escape latency of each group of experimental animals was recorded.

[0069] II. Experimental Results

[0070] Compared with the normal control group, the escape latency and total distance traveled in the HC group were significantly increased; compared with the HC group, Eucommia ulmoides (EU) and total saponins of Panax ginseng (TSPG) significantly reduced the escape latency and total distance traveled. Figure 1 A, Figure 1 B); The combined use of low doses of EU and TSPG has an efficacy comparable to that of single-dose administration. Figure 1 C).

[0071] Table 1. Cognitive function test data for the water maze.

[0072]

[0073] **p<0.01 vs Con, #p<0.05 vs HC

[0074] The q-value for combined use of Eucommia ulmoides and ginseng total saponin antibodies was calculated according to the King's formula commonly used in this field. A q-value > 1 indicates that the two have a synergistic effect. The data are shown in Table 1.

[0075] The escape latency of 100 mg / kg Eucommia ulmoides was reduced by 16% (n=8) (EA);

[0076] The evasion latency of 20 mg / kg total ginsenosides was reduced by 20.5% (n=8) (EB);

[0077] The combined use of the two methods reduced the escape latency by 26.2% (n=8) (E(A+B));

[0078] Substituting into the King's formula: q=E(A+B) / (EA+EB-EA×EB)=52.4% / ( 16% + 20.5% - 16%×20.5%) = 1.577>1.

[0079] The overall pathway reduction rate of 100 mg / kg Eucommia ulmoides was 21.2% (n=8) (EA);

[0080] The overall pathway reduction rate of 20 mg / kg ginsenosides was 26% (n=8) (EB);

[0081] The combined path reduction rate is 64% (n=8) (E(A+B));

[0082] Substituting into King's formula: q=E(A+B) / (EA+EB-EA×EB)=64% / ( 21.2% + 26% - 21.2%×26%) = 1.535>1.

[0083] Based on the above calculations, the q values ​​of both batches of data are greater than 1, indicating that Eucommia ulmoides and total ginsenosides produce a synergistic effect when used in combination. The combined treatment of Eucommia ulmoides and total ginsenosides with cognitive decline of kidney yang deficiency has a synergistic effect.

[0084] Example 2: Golgi staining to detect the density of hippocampal dendritic spines in rats with kidney-yang deficiency.

[0085] I. Experimental Methods

[0086] Golgi staining:

[0087] 1.1 Preparation of tissue sections (1) According to the instructions of the Rapid Golgi Stain Kit (FD Rapid GolgiStain Kit PK-401), prepare solution A and solution B at least 24 hours before the start of the experiment, mix them in equal volumes to form an infiltration solution, and store it at room temperature for later use; (2) After the rat is deeply anesthetized by injection of 3% sodium pentobarbital, the rat is decapitated on a decapitation device, the skull is quickly opened and fresh brain tissue is taken out (Note: 4% paraformaldehyde cannot be used for fixation), and the brain tissue should be protected from damage during the operation; (3) After obtaining fresh rat brain tissue, firstly, the hippocampus is cut open with a scalpel blade along the coronal plane to obtain the hippocampal brain tissue, and the blood on the surface of the tissue is quickly rinsed off with pure water; (4) The hippocampal brain tissue is soaked in the mixed infiltration solution at room temperature in the dark for 2 weeks. Note: After the rat brain tissue is soaked in the infiltration solution for the first time for 6 hours or after the tissue is soaked overnight, a new infiltration solution should be replaced; after changing the solution, the container containing the brain tissue should be gently rotated left and right several times, twice a week. (5) Gently and carefully transfer the tissue to solution C and keep it at room temperature in the dark for at least 72 h; during this period, change solution C at least once after 24 h or the next day; (6) Before sectioning, turn on the cryostat in advance and set the temperature to -22℃. After the cryostat has stabilized at this temperature for 1 h, section the tissue; (7) Before freezing and sectioning rat brain tissue, fix the tissue on the sample tray with OCT embedding agent and freeze it quickly in liquid nitrogen. Then immediately install the sample tray with the fixed sample on the cryostat, and section it after acclimatizing to the temperature for 5 min. The section thickness is 100 μm; (8) Use a sample recovery device (provided by the kit) to transfer the sample to a microscope slide coated with gelatin containing solution C; (9) Experimental materials and methods; the prepared sections can be stored in a slide box and can be stored for up to 3 days at room temperature in the dark.

[0088] 1.2 Staining (1) Preparation of working solution: The staining working solution consists of solution D, solution E and pure water, prepared in a volume ratio of 1:1:2 and used immediately. (2) Staining: Rinse the tissue sections twice with a slow water flow for 4 min each time; then, place the tissue sections in the working solution and immerse for 10 min. (3) Dehydration: Insert the tissue sections into the slide holder, place them in the antigen retrieval box, add pure water to rinse the sections three times for 3 min each time; then, place the tissue sections in 50%, 75% and 95% ethanol for 4 min each for gradient dehydration; finally, dehydrate the sections in anhydrous ethanol for 5 min, repeating twice. (4) Clearing and mounting: Place the dehydrated sections in xylene and immerse for 10 min for clearing treatment, repeating twice; cover the cleared sections with coverslips and mount with resin mounting medium. Observe and photograph under a regular optical microscope.

[0089] II. Experimental Results

[0090] Compared with the normal control group, the dendritic spine density in the HC group was sparse; compared with the HC group, the dendritic spine density was significantly increased in the EU and TSPG medium-dose groups; the efficacy of the combined low-dose EU and TSPG was comparable to that of the medium-dose single herb, showing a synergistic effect. Figure 2 ).

[0091] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. A traditional Chinese medicine composition for treating cognitive decline, characterized in that, The traditional Chinese medicine composition consists of Eucommia ulmoides and total ginsenosides; the cognitive decline is of the kidney-yang deficiency type; the mass ratio of Eucommia ulmoides to total ginsenosides is 5:1; the Eucommia ulmoides is the raw material of Eucommia ulmoides.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The types of active pharmaceutical ingredients include dried medicinal materials, fresh medicinal materials, or combinations thereof.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The types of active pharmaceutical ingredients include raw medicinal materials, processed medicinal materials, or combinations thereof.

4. A traditional Chinese medicine preparation for treating cognitive decline, characterized in that, The traditional Chinese medicine preparation comprises the traditional Chinese medicine composition according to any one of claims 1-3; The cognitive decline described is a type of cognitive decline caused by kidney yang deficiency.

5. The traditional Chinese medicine preparation according to claim 4, characterized in that, The dosage forms of the traditional Chinese medicine preparations include granules, tablets, capsules, oral liquids, pills, and drop pills.

6. The use of the traditional Chinese medicine composition according to any one of claims 1-3 in the preparation of a medicament for the prevention or treatment of cognitive decline; The cognitive decline described is a type of cognitive decline caused by kidney yang deficiency.

7. The use of the traditional Chinese medicine preparation according to any one of claims 4 or 5 in the preparation of a medicine for the prevention or treatment of cognitive decline; The cognitive decline described is a type of cognitive decline caused by kidney yang deficiency.

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