Application of astragaloside IV in preparation of medicine for treating cognitive impairment after heart failure

Abstract

The invention discloses application of astragaloside IV in preparation of a medicine for treating cognitive impairment after heart failure. The astragaloside IV inhibits the signal channels of Erk-1, Erk-2 and NF-kB through targeted inhibition of the expression of RAGE, and reduces the expression of proinflammatory factors IL-1beta, IL-6 and TNF-alpha so as to inhibit neuroinflammation. The medicine for treating cognitive impairment after heart failure comprises astragaloside IV and pharmaceutically acceptable salts or pharmaceutically acceptable carriers or excipients thereof.

Description

Technical Field

[0001] This invention relates to the field of drugs for cognitive impairment after heart failure, and more particularly to the application of astragaloside IV in the preparation of drugs for cognitive impairment after heart failure. Background Technology

[0002] The 2024 HFSA expert consensus statement indicates that approximately 64.3 million people worldwide currently suffer from heart failure, and up to 78% of heart failure patients experience cognitive impairment. Cognitive impairment is an independent risk factor for increased mortality and readmission rates in heart failure patients, increasing the risk of cardiovascular death. However, even though the use of "new quadruple therapy" drugs provides significant benefits for heart failure symptoms, there is no clinical evidence that these drugs effectively improve cognitive impairment caused by heart failure. Furthermore, heart failure patients often have mild cognitive impairment, not yet meeting the diagnostic criteria for dementia, and therefore cannot use medications for dementia. Using psychiatric medications such as cholinesterase inhibitors may increase adverse drug reactions. Therefore, how to alleviate cognitive impairment while improving heart failure symptoms is an important clinical challenge.

[0003] Traditional Chinese medicine (TCM) has the advantages of multiple targets and multiple components. Astragaloside IV (CAS No. 84687-43-4) is the main active compound of the TCM herb Astragalus membranaceus. Studies have shown that astragaloside IV can cross the blood-brain barrier to enter the brain and exert its effects, exhibiting significant neuroprotective and repair activities in various experimental models. In Parkinson's disease-related models, astragaloside IV can reduce the neuroprotective effects of levodopa (L-DOPA) on dopaminergic substantia nigra neurons. This component exerts an anti-apoptotic effect by inhibiting apoptosis induced by neurotoxins such as DOPA. Furthermore, it can enhance the survival rate of primary cortical neurons in rats and promote axonal elongation; its effect is related to the inhibition of the PTEN / AKT signaling pathway. Notably, recent studies on experimental autoimmune encephalomyelitis (EAE, an animal model of multiple sclerosis) have shown that astragaloside A can regulate Nogo The A / ROCK2 pathway promotes axonal repair and regeneration. To date, there have been no reports on the application of astragaloside IV in the preparation of drugs for cognitive impairment following heart failure. Summary of the Invention

[0004] To overcome the shortcomings of existing technologies, the main objective of this invention is to provide an application of astragaloside IV in the preparation of drugs for cognitive impairment following heart failure. This solves the problem in existing technologies where there are no available medications for heart failure patients with mild cognitive impairment. Furthermore, using drugs for treating dementia, such as cholinesterase inhibitors, does not comply with clinical guidelines and may increase adverse drug reactions.

[0005] To achieve the aforementioned objectives, the technical solution adopted by this invention includes: Application of astragaloside IV in the preparation of drugs for cognitive impairment after heart failure.

[0006] Astragaloside IV inhibits neuroinflammation by targeting and inhibiting the expression of RAGE, thereby suppressing the Erk-1, Erk-2 and NF-κB signaling pathways, and simultaneously reducing the expression of pro-inflammatory factors IL-1β, IL-6 and TNF-α.

[0007] Astragaloside IV can be used alone or in combination with other medicinal ingredients.

[0008] The drug includes astragaloside IV and its pharmaceutically acceptable salts or pharmaceutically acceptable carriers or excipients.

[0009] A drug for post-heart failure cognitive impairment, the drug comprising astragaloside IV.

[0010] Astragaloside IV inhibits neuroinflammation by targeting and inhibiting the expression of RAGE, thereby suppressing the Erk-1, Erk-2 and NF-κB signaling pathways, and simultaneously reducing the expression of pro-inflammatory factors IL-1β, IL-6 and TNF-α.

[0011] Astragaloside IV can be used alone or in combination with other medicinal ingredients.

[0012] The drug includes astragaloside IV and its pharmaceutically acceptable salts or pharmaceutically acceptable carriers or excipients.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: Astragaloside IV significantly improved cognitive impairment in heart failure rats, manifested in enhanced learning and memory abilities and reduced cognitive decline. Molecular biology experiments, molecular docking, and molecular dynamics simulations verified that astragaloside IV inhibits the RAGE-Erk1 / 2-NF-κB signaling pathway by targeting and inhibiting RAGE expression, while simultaneously reducing the expression of pro-inflammatory factors IL-1β, IL-6, and TNF-α, thereby suppressing neuroinflammation. These results indicate that astragaloside IV has significant value in the preparation of drugs for cognitive impairment following heart failure, demonstrating both cardiovascular benefits and significant implications for improving cognitive impairment caused by heart failure. Attached Figure Description

[0014] Figure 1 Figure A shows the effect of astragaloside IV on cardiac function improvement in rats with heart failure; Figure B shows the changes in LVEF values ​​in each group of rats; Figure B shows the changes in FS values ​​in each group of rats. Figure 2Figure 1 shows the results of evaluating cognitive impairment in heart failure rats after intervention with astragaloside IV using the Morris water maze test. Figure 2 shows the comparison of latency in each group of rats during the orienteering test. Figure 3 shows the comparison of total swimming distance in each group of rats during the orienteering test. Figure 4 shows the comparison of swimming speed in each group of rats during the orienteering test. Figure 5 shows the comparison of the number of times rats crossed platforms in each group of rats during the platform crossing test. Figure 6 shows the comparison of swimming speed in each group of rats during the platform crossing test. Figure 7 shows the representative trajectories of each group of rats during the platform crossing test. Figure 3 Figure 1 shows the results of evaluating cognitive impairment in heart failure rats after intervention with astragaloside IV using the Y-maze test; Figure 2 shows the comparison of the number of times rats in each group entered the new open arm; Figure 3 shows the comparison of the time it took for rats in each group to enter the new open arm for the first time; Figure 4 shows the comparison of the average speed of rats in each group; Figure 5 shows the representative trajectory of rats in each group. Figure 4 Figure A shows the results of evaluating cognitive impairment in heart failure rats after intervention with astragaloside IV through a novel object recognition experiment; Figure B shows a comparison of cognitive indices among the rat groups; Figure B shows representative trajectory diagrams of rats from each group. Figure 5 Figure 1 shows the regulation of protein expression of AGE, Erk1, and Erk-2 in the hippocampus of rats with cognitive impairment due to heart failure. Figure A shows representative immunoblot bands of AGE in each group of rats. Figure B shows the protein expression level of AGE in each group of rats. Figure C shows representative immunoblot bands of Erk-1 and Erk-2 in each group of rats. Figure D shows the protein expression level of Erk1+Erk2 in each group of rats. Figure 6 Figure 1 shows the regulatory effect of astragaloside IV on the gene expression of Ager, Erk1, Erk-2, and NF-κB in the hippocampus of rats with cognitive impairment due to heart failure. Figure A shows the expression of the Ager gene in each group of rats; Figure B shows the gene expression of Erk-1 in each group of rats; Figure C shows the gene expression of Erk-2 in each group of rats; and Figure D shows the gene expression of NF-κB in each group of rats. Figure 7 Figure 1 shows the inhibitory effect of astragaloside IV on hippocampal inflammation in rats with cognitive impairment due to heart failure; Figure A shows the gene expression of IL-1β in each group of rats; Figure B shows the gene expression of IL-6 in each group of rats; Figure C shows the gene expression of IL-10 in each group of rats; Figure D shows the gene expression of TNf-α in each group of rats. Figure 8Figure 1 shows the molecular docking and molecular dynamics simulation diagrams of key proteins regulated by astragaloside IV. Figure A is a schematic diagram of molecular docking between astragaloside IV and RAGE; Figure B is a schematic diagram of molecular docking between astragaloside IV and Erk-1; Figure C is a schematic diagram of molecular docking between astragaloside IV and Erk-2; Figure D is a schematic diagram of molecular docking between astragaloside IV and NF-κB; Figure E is a schematic diagram of the molecular dynamics simulation diagram of astragaloside IV and RAGE, where E1 is the RMSD value of astragaloside IV binding to RAGE; E2 is the Rg value of astragaloside IV binding to RAGE; E3 is the SASA value of astragaloside IV binding to RAGE; E4 is the number of hydrogen bonds in the binding of astragaloside IV to RAGE; and E5 is the RMSF value of astragaloside IV binding to RAGE. Detailed Implementation

[0015] The embodiments described below are exemplary descriptions of key experimental evidence and are not intended to limit the core content and application scope of this invention due to the amount of evidence. It should be noted that all the accompanying drawings and corresponding descriptions merely illustrate the concept, principles, and representative experimental evidence of the disclosed embodiments of this invention. Where the chain of evidence is complete, it is unnecessary to show all the specific details and extended details of the various embodiments listed in this invention.

[0016] Unless otherwise defined, the technical terms used in the following embodiments have the same meaning as commonly understood by those skilled in the art to which this invention pertains.

[0017] Unless otherwise specified, the experimental materials used in the examples below can be purchased from conventional chemical reagent companies.

[0018] The total RNA extraction kit was purchased from Guangzhou Meiji Biotechnology Co., Ltd., model R4310. The reverse transcription kit was purchased from Tiangen Biotech Co., Ltd., model KR116. The real-time PCR kit was purchased from Tiangen Biotech Co., Ltd., model FP217. All primers were designed and synthesized by Sangon Biotech Co., Ltd.

[0019] The present invention will now be described in detail with reference to the accompanying drawings and embodiments.

[0020] This invention primarily focuses on animal experiments. Male SD rats (220g-240g) were used as research subjects. A heart failure model following myocardial infarction was established by ligating the left anterior descending coronary artery. The drug groups were administered low-dose (30mg / kg / d) and high-dose (60mg / kg / d) astragaloside IV via gavage on the second day after surgery. After 8 weeks, cognitive function was evaluated using behavioral tests (Morris water maze, Y maze, and new object recognition). Western blotting and qPCR were used to observe the effects of astragaloside IV on the expression of AGE-RAGE and downstream signaling pathway proteins Erk-1 / 2 and NF-κB. Furthermore, the inhibitory effects of astragaloside IV on pro-inflammatory factors IL-1, IL-6, and TNF-α, and the activating effect on the anti-inflammatory factor IL-10 were observed. To clarify the specific role of astragaloside IV as a "drug-target", molecular docking was performed on proteins with differentials in the AGE-RAGE signaling pathway and inflammation-related proteins, and further molecular dynamics simulations were conducted with the core protein to verify the specific target of astragaloside IV.

[0021] Specific technical solution: 1. Astragaloside IV Source: Purchased from Chengdu Pufeed Biotechnology Co., Ltd., molecular formula C41H68O14, structure as follows:

[0022] 2. Preparation of a heart failure model after myocardial infarction: This invention comprises four animal groups: A: sham-operated group; B: model group; C: low-dose astragaloside IV group; D: high-dose astragaloside IV group. The modeling and gavage protocols for each group are as follows: A: Sham surgery group: After thoracotomy, only sutures were threaded, without ligation. Distilled water was administered by gavage at a rate of 1 ml / 100g for 2 months starting on the second postoperative day.

[0023] B: Model group: The heart failure model after myocardial infarction was prepared by ligation of the left coronary artery. The specific method is described in the following text. The model was prepared by gavage with distilled water at a volume of 1 ml / 100 g for 2 months on the second day after the operation. C; Low-dose astragaloside IV group: The surgical procedure was the same as that of the model group. Starting from the second day after surgery, low-dose (30mg / kg / d) astragaloside IV was administered by gavage at a volume of 1ml / 100g for 2 months.

[0024] D: High-dose astragaloside IV group: The surgical procedure was the same as the model group. Starting from the second day after surgery, high-dose (60mg / kg / d) astragaloside IV was administered by gavage at a volume of 1ml / 100g for 2 months.

[0025] Methods for preparing a heart failure model after myocardial infarction: (1) Preoperative preparation: The rats were fasted but allowed to drink water the day before the operation. The rats were anesthetized with 1% sodium pentobarbital (40 mg / kg) before the operation. They were fixed in a supine position on the rat board. After wiping the hair of the left chest area with alcohol, the blade was prepared in the left chest area with the needle holder and disinfected with iodine.

[0026] (2) Preoperative electrocardiogram and endotracheal intubation: A 12-lead small animal electrocardiograph was used to detect the preoperative electrocardiogram of rats. The chest and limb leads of the electrocardiogram were inserted subcutaneously into the corresponding sites, and the paper output speed of the electrocardiograph was set to 25 mm / s. The rat was placed on a triangular board, and the neck blood vessels were directly illuminated by a desk lamp. The rat's tongue was gently lifted by holding gauze. At this time, a clear bright spot could be seen along the direction of the neck blood vessels. The endotracheal tube was placed on the indwelling needle of the artery and vein. The tube was inserted into the trachea through the pharynx along the bright spot. The indwelling needle of the artery and vein was immediately removed to keep the trachea patent. The success of the endotracheal intubation was observed outside the endotracheal tube with a small cotton ball. The tube was adjusted to a suitable position and fixed with tape.

[0027] (3) Ligation of the left anterior descending coronary artery to prepare a myocardial infarction model: The rat was fixed supine on a rat board. The manubrium of the sternum was positioned at the level of the 2nd rib, and the two intercostal spaces below it were the 3rd and 4th intercostal spaces. A small incision was made in the 3rd and 4th intercostal spaces parallel to the ribs using small scissors. The muscles were bluntly separated with ophthalmic forceps. After the ribs were clearly seen, the ventilator was connected and the ventilator parameters were adjusted (respiratory rate: 80 breaths / min, tidal volume: 7ml, inspiratory-to-expiratory ratio: 1:2). Using ophthalmic scissors, cut the intercostal muscles between the 3rd and 4th ribs. Use a chest expander to open the ribs. Use ophthalmic forceps to tear open the pericardium to fully expose the heart. Support the rat from behind with your left hand until you can see the left atrial appendage and the beating heart. Hold a 12.5 cm needle holder in your right hand, clamping a suture needle (5-0 suture). Insert the needle 2-3 mm below the edge of the left atrial appendage (to the left of the blood vessel). Following the vertical direction of the rat's body, exit the needle approximately 2-3 mm to the right of the anterior descending limb. Gently lift the ligation suture and observe the insertion and exit points to ensure they are appropriate. Preliminary judgment of ligation success is based on whether the apex of the heart turns white after ligation. Remove the cotton ball from the pleural cavity and clean away any blood. Gently squeeze the pleural cavity to expel air and prevent pneumothorax. Use 2-0 suture and suture the ribs, muscles, and skin sequentially using a figure-eight suture technique. Remove the ventilator connection tube and squeeze the pleura to restore spontaneous breathing in the rat. After surgery, rats were injected intraperitoneally with 0.1 mL lidocaine to prevent ventricular fibrillation; 0.1 mL furosemide was injected intraperitoneally for diuresis; and 400,000 U penicillin was injected intraperitoneally to prevent infection.

[0028] (4) Model evaluation: The success criteria for model establishment were determined based on the number of pathological Q waves on the electrocardiogram on the second day after surgery and the value of LVEF by echocardiography at 60 days. If there were 6-8 pathological Q waves and LVEF < 40%, heart failure could be diagnosed (according to the literature published by the research group, Wang Lei, Zhao Mingjing, Yang Tao, et al. Early evaluation and screening method of heart failure model after myocardial infarction from the correlation analysis of electrocardiogram and echocardiogram [J]. Journal of Integrated Traditional and Western Medicine on Cardiovascular and Cerebrovascular Diseases, 2017, 15(22): 2816-20.). The number of pathological Q waves on the electrocardiogram 24 hours after model establishment was significantly negatively correlated with the LVEF value at 8 weeks. When the number of pathological Q waves at 24 hours was 6-8, the specificity of diagnosing heart failure based on the LVEF value detected by echocardiography at 8 weeks was 100%, the sensitivity was 95%, and the accuracy was 96.7%. Eight weeks later, the cognitive function of rats with heart failure was evaluated using the Morris water maze, Y-maze, and novel object recognition tests. The success rate of the heart failure model after myocardial infarction was 85%, and the incidence of cognitive impairment after heart failure was 80%.

[0029] 3. Echocardiography Eight weeks after modeling, cardiac function and structure in rats were evaluated by echocardiography. Rats were anesthetized with 1% sodium pentobarbital via intraperitoneal injection. After skin preparation of the anterior chest area, the rats were fixed in a supine position on the operating table. A blind procedure was performed by a professional technician using a Vevo2100 ultra-high resolution small animal color Doppler imaging system with an MS 200 probe at 15 MHz to perform two-dimensional M-mode ultrasound on the short-axis and long-axis sections of the left ventricular papillary muscles. The following parameters were measured: left ventricular ejection fraction (LVEF) and fractional shortening (FS). Three cardiac cycles were measured for each animal, and the average value was used for analysis.

[0030] like Figure 1 As shown, the LVEF values ​​of rats in the low-dose and high-dose groups of astragaloside IV were significantly increased, the FS value of rats in the high-dose group was significantly increased, while the FS value of rats in the low-dose group showed no significant change. This indicates that the heart failure model was successfully established.

[0031] 4. Morris test for cognitive function: (1) Experimental preparation: The water maze setup consisted of a circular water tank with a diameter of 1.6 m, filled with opaque water containing ink, at a depth of 27 cm and a temperature of 22 ± 2 ℃. The tank was divided into four quadrants (Ⅰ, Ⅱ, Ⅲ, Ⅳ). A platform with a diameter of 12 cm was located in the pool, 1.5 cm below the water surface. The entire pool was surrounded by a blue curtain with four irregular shapes as distal cues. Before the formal experiment, the experimental rats were placed in the water maze room to acclimatize to the environment. The water maze test included a navigation experiment and a platform-crossing experiment.

[0032] (2) Orientation and navigation experiment: The orientation and navigation experiment was conducted from the first to the third day of the experiment, with the platform placed in area III. On the first day, before the rat entered the water for the first time, it was placed on the platform for 5 seconds. Each day, rats were randomly placed into the water from areas I, II, and IV, facing the wall, and allowed to swim freely for 90 seconds. If the rat found the platform and stood stably on it for 5 seconds during this period, the recording was automatically paused. Conversely, if the rat did not find the platform within 90 seconds after its first entry into the water, it was gently guided to the platform with a stick and stood there for 5 seconds. The time it took for the rat to find the platform (latency period), swimming speed, and total swimming distance were recorded.

[0033] (3) Platform crossing test: The platform crossing test was conducted on the fourth day. The platform placed in area III was removed, and the rats were placed in the water from area I facing the wall. The rats were allowed to swim freely in the water for 120 seconds. The number of times the rats crossed the original platform position, the swimming trajectory, and the swimming speed were recorded.

[0034] like Figure 2 As shown, in the Morris water maze, there were no significant differences in various indicators between low-dose and high-dose astragaloside IV in the orienteering experiment; low-dose and high-dose astragaloside IV significantly increased the number of times heart failure rats crossed platforms, indicating an improvement in learning and memory abilities.

[0035] 5. Y-maze test for cognitive function (1) Experimental preparation: The Y-maze apparatus consists of three arms of equal length (a, the initial arm; b, the newly opened arm; c, the open arm), with the arms positioned such that the angle between any two adjacent arms is 120°. Each arm has a movable baffle at the midpoint between it and the apparatus. Before the formal experiment, the experimental rats were placed in a water maze room to acclimatize to the experimental environment. The Y-maze includes a training phase and a testing phase.

[0036] (2) Training phase: During the training phase, arm b was closed, and the rat was placed into the device from the starting segment of arm a, facing the wall, and allowed to explore the maze freely for 3 minutes. After each rat experiment, its feces were cleaned up, and the device was wiped with 75% alcohol to prevent residual odor from affecting subsequent experiments.

[0037] (3) Testing phase: One hour after the end of the training phase, open arm b and place the rat into the device from the starting segment of arm a, facing the wall, allowing it to freely explore the maze for 3 minutes. Recorded indicators include the number of times the rat entered arm b, the time of its first entry into arm b, and its movement speed. Clean the feces of each rat after the experiment and wipe the device with 75% alcohol to avoid affecting subsequent experiments.

[0038] like Figure 3 As shown, in the Y-maze, low-dose astragaloside IV increased the number of times new open arms were entered, while high-dose astragaloside IV showed no significant difference. In comparing the time to first entry into a new open arm, the low-dose and high-dose groups of astragaloside IV only showed a decrease in the mean time, with no significant difference.

[0039] 6. New object recognition experiments test cognitive function and exploratory ability. (1) Experimental preparation: The device for new object recognition was an open box with a side length of 1 meter. There were two objects symmetrically positioned in the open space. The day before the formal experiment, the experimental rats were placed in the open box for 5 minutes to adapt to the environment. The new object recognition experiment included a training phase and a testing phase.

[0040] (2) Training phase: Two identical objects a were placed symmetrically on both sides of the open field, 25 cm away from the edge of the open field. The experimental rats were placed in the box facing the wall in the middle position on opposite sides of the two objects a, and allowed to explore freely in the device for 5 min. After removal, the feces were cleaned and wiped with 75% alcohol before the next rat was tested.

[0041] (3) Testing phase: The testing phase experiment was conducted 1 hour after the end of the training phase. Object a was replaced with a completely different object b, and the rat was placed in the device from its original position and allowed to explore freely for 5 minutes. The time taken for the rat to explore object a and object b was recorded, and the cognitive index (CI) was calculated.

[0042] The formula is CI = b / (a + b).

[0043] Where a represents the time spent exploring the old object a, and b represents the time spent exploring the new object b.

[0044] At the end of the experiment, the feces of each rat were cleaned and the equipment was wiped with 75% alcohol to prevent residual odor from affecting subsequent experiments.

[0045] like Figure 4 As shown, in the recognition of new objects, both doses of astragaloside IV can improve the cognitive index of rats with heart failure (cognitive index = new object time / (new object time + old object time)), indicating that astragaloside IV can increase the rats' ability to explore and remember novel things.

[0046] 7. Western Blot Tissue was lysed using RIPA lysis buffer, and phosphatase and protease inhibitors were added. After centrifugation, total rat hippocampal protein was extracted. Loading buffer was then added to the protein sample, and the mixture was boiled at 99°C for 20 min. Samples (30 µg) were subjected to SDS-PAGE electrophoresis, wet-transferred to a 0.22 μm NC membrane, blocked with 5% skim milk powder, and incubated overnight at 4°C with primary antibody. The membrane was washed with Tris-buffered saline (TBST) containing 0.1% Tween 20, incubated with HRP-labeled secondary antibody (1:8000), and then subjected to enhanced chemiluminescence (ECL) detection using an enhanced chemiluminescence (ECL) substrate. All blocking buffers, secondary antibodies, and skim milk powder were prepared fresh for use. Finally, optical density was measured using ImageJ software, and statistical analysis was performed.

[0047] like Figure 5 As shown, Western blotting revealed that both low and high doses of astragaloside IV reduced the protein expression of Erk-1 and Erk-2. However, there was no significant difference in protein expression at AGE levels among the different groups.

[0048] 8. Real-time quantitative polymerase chain reaction (qPCR) We previously performed transcriptomic analysis on the hippocampus of rats with heart failure, and at the same time, we analyzed the drug targets of astragaloside A through network pharmacology. After screening, we found a common differential pathway, the AGE-RAGE signaling pathway.

[0049] RNA was extracted from the hippocampus of rats with heart failure using a total RNA extraction kit. The concentration (ng / μL) and purity (A260 / A280) of the obtained RNA were determined using a UV spectrophotometer. The expression levels of the following indicators in rat hippocampus were detected using reverse transcription and quantitative real-time PCR kits. Primers are listed in Table 1.

[0050] Table 1

[0051] like Figure 6-7 As shown, low and high doses of astragaloside IV can reduce the expression of the Age gene in RAGE, reduce the expression of Erk-1, Erk-2 and Nf-κb genes, and reduce the expression of pro-inflammatory factors IL-1, IL-6 and Tnf-α genes, thereby inhibiting neuroinflammation. However, it has no significant effect on the expression of the pro-inflammatory factor IL-10.

[0052] 9. Molecular docking and molecular dynamics simulation The binding potential of AS-IV to differentially expressed proteins was determined by molecular docking. The structure of AS-IV was downloaded from TCMSP (http: / / www.tcmspw.com / tcmsp.php), and the molecular structures of RAGE, Erk-1, Erk-2, and NF-κB were downloaded from the PDB database (https: / / www.rcsb.org / ). The binding affinity of AS-IV to target proteins was determined using CB-Dock2 assay. Finally, the docking results were visualized and analyzed using PyMOL (Version 2.2.0) and Ligplus software.

[0053] Furthermore, this study utilized Gromacs 2022 software for molecular dynamics (MD) simulations. Force field parameters were obtained using the pdb2gmx tool in Gromacs combined with the AutoFF webpage. The CHARMM36 force field was selected for the receptor protein, and the CGenff force field was selected for the ligand. During solvation, the system was encapsulated in a 1 nm thick TIP3P cubic water box. Ions were added to the system using the gmx genion tool to achieve electroneutrality. Long-range electrostatic interactions were handled using the Particle Mesh Ewald (PME) method, with the cutoff distances for both van der Waals and electrostatic interactions set to 1 nm. All critical constraints were implemented using the SHAKE algorithm, with the Verlet frog-jump algorithm used to set the integration step size to 1 fs. Energy optimization was performed before molecular dynamics simulations. The energy minimization process involved first performing 3000 steps of the steepest descent method optimization, followed by 2000 steps of the conjugate gradient method optimization. The specific optimization steps are as follows: constrain the solute to minimize the energy of water molecules; constrain the reaction force to minimize energy; finally, minimize the energy of the entire system under unconstrained conditions. The simulation environment was set as an NPT constant-pressure system at 310K, with a simulation duration of 100 ns. The simulation trajectory analysis was completed using the GROMACS tool: gmx rms was used to calculate the root mean square deviation (RMSD), gmx rmsf to calculate the root mean square fluctuation (RMSF), gmx hbond to count the number of hydrogen bonds (HBonds), gmx gyrate to calculate the radius of gyration (Rg), and gmx sasa to analyze the solvent accessible surface area (SASA).

[0054] like Figure 8 As shown, molecular docking and molecular dynamics simulation results indicate that astragaloside IV has a good binding effect with RAGE, suggesting that RAGE-astragaloside IV targets key cognitive impairment after heart failure.

[0055] The demonstrated pharmacological effects reveal the role of astragaloside IV in improving cognitive impairment caused by heart failure, suggesting its potential for developing drugs to treat cognitive impairment following heart failure. Astragaloside IV can be used alone or in combination with other pharmaceutically acceptable components, or formulated with pharmaceutically acceptable salts or carriers or excipients into tablets, capsules, granules, drops, or injections.

[0056] The above embodiments only describe a portion of the specific implementation methods of the present invention in detail, and are not limited to the embodiments disclosed herein. Furthermore, the substantive content protected by the present invention is not limited thereto. Any other modifications, equivalent substitutions, improvements, etc., made based on the principles and techniques of the present invention without departing from its design scope are all within the protection scope of the present invention.

Claims

1. Use of Astragaloside IV in the preparation of a drug for cognitive impairment after heart failure.

2. The use of astragaloside IV according to claim 1 in the preparation of a drug for cognitive impairment after heart failure, characterized in that, The Astragaloside IV inhibits neuroinflammation by targeting the inhibition of the expression of RAGE, thereby inhibiting the Erk-1, Erk-2 and NF-κB signaling pathways, while reducing the expression of pro-inflammatory factors IL-1β, IL-6 and TNF-α.

3. The use of astragaloside IV according to claim 2 in the preparation of a drug for cognitive impairment after heart failure, characterized in that, The Astragaloside IV is used alone or in combination with other medicinal ingredients.

4. The use of astragaloside IV according to claim 2 in the preparation of a drug for cognitive impairment after heart failure, characterized in that, The drug comprises Astragaloside IV and pharmaceutically acceptable salts or carriers or excipients.

5. A drug for post-heart failure cognitive impairment, characterized by comprising the compound or salt according to claim 1. The drug comprises Astragaloside IV.

6. The medicament for cognitive impairment after heart failure according to claim 5, characterized by, The Astragaloside IV inhibits neuroinflammation by targeting the inhibition of the expression of RAGE, thereby inhibiting the Erk-1, Erk-2 and NF-κB signaling pathways, while reducing the expression of pro-inflammatory factors IL-1β, IL-6 and TNF-α.

7. The medicament for cognitive impairment after heart failure according to claim 5, wherein The Astragaloside IV is used alone or in combination with other medicinal ingredients.

8. The medicament for cognitive impairment after heart failure according to claim 5, characterized by, The drug comprises Astragaloside IV and pharmaceutically acceptable salts or carriers or excipients.

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