Application of rhizoma corydalis extract in medicine for treating myocardial infarction with depression and medicine for preventing myocardial infarction with depression

Abstract

The invention belongs to the field of application of biological extracts, and particularly relates to application of a rhizoma corydalis extract. The rhizoma corydalis extract is used for improving and / or treating myocardial infarction or depression or myocardial infarction with depression; the rhizoma corydalis extract contains alkaloid; the alkaloid comprises dehydrocorydaline and / or tetrahydropalmatine and / or berberine and / or dehydrocorydaline and / or tetrahydropalmatine and / or berberine, and at least comprises dehydrocorydaline; the corydalis tuber extract is used for preparing medicines and / or health foods for improving and / or treating myocardial infarction or depression or myocardial infarction with depression. The effectiveness of the rhizoma corydalis extract in the aspect of treating myocardial infarction with depression and the effective and safe dosage are verified through experiments, and corresponding bases and effective references are provided for guiding and developing application of the rhizoma corydalis extract and application of the rhizoma corydalis extract in the aspect of preparing corresponding medicines and health-care foods.

Description

Technical Field

[0001] The present invention belongs to the application field of biological extracts, and in particular relates to the application of Corydalis yanhusuo extract in drugs for treating myocardial infarction accompanied by depression and drugs for preventing myocardial infarction accompanied by depression. Background Art

[0002] Corydalis yanhusuo, also known as Yuanhu, is the dried tuber of the poppy family plant Corydalis yanhusuo WT Wang. As one of the traditional Chinese medicinal herbs known as the "Eight Herbs of Zhejiang," it boasts benefits such as promoting blood circulation, promoting qi circulation, and relieving pain. Its primary bioactive components are alkaloids, including dehydrocorydalis, tetrahydropalmatine, and berberine. Corydalis yanhusuo extracts are currently widely used clinically to treat a variety of conditions, including cardiovascular diseases such as improving myocardial function, alleviating myocardial ischemia, and dilating coronary arteries, as well as neurological disorders such as pain relief and anxiety relief.

[0003] In addition, Corydalis yanhusuo extract has also shown certain potential in anti-inflammatory, anti-tumor, anti-gastric ulcer, and anti-platelet aggregation. However, although studies have shown that Corydalis yanhusuo extract has a protective effect on myocardial cells and nerve cells, its specific efficacy and mechanism of action in the treatment of myocardial infarction and / or depression still need further in-depth research and verification.

[0004] This invention aims to provide a safe and effective treatment option for patients with myocardial infarction and depression by further studying the pharmacological mechanisms of action of Corydalis yanhusuo extract, specifically its protective effects on cardiomyocytes and neurons. This novel application will leverage the multi-target mechanism of action of Corydalis yanhusuo extract to achieve a synergistic therapeutic effect on myocardial infarction and depression. Summary of the Invention

[0005] In order to solve the problem that the existing application fields of Corydalis yanhusuo extract are limited and more, richer and more practical application fields are urgently needed to be developed, the present invention provides the application of Corydalis yanhusuo extract in drugs for treating myocardial infarction accompanied by depression.

[0006] The main objectives of the present invention are: 1. Provide new application areas for Corydalis yanhusuo extract; 2. Clarify the various effects of Corydalis yanhusuo extract after its use in pharmaceutical production.

[0007] Application of Corydalis yanhusuo extract in the treatment of myocardial infarction-associated depression.

[0008] As a preference, The treatment of myocardial infarction-associated depression includes reducing the anhedonia symptoms in patients with myocardial infarction-associated depression.

[0009] As a preference, The treatment of myocardial infarction with depression includes improving the exercise tolerance and autonomous motor activity of patients with myocardial infarction with depression, and alleviating the despair and anxiety behaviors of patients with myocardial infarction with depression.

[0010] As a preference, The treatment of myocardial infarction-associated depression includes reducing apoptosis or vacuolation of neurons in the prefrontal cortex of patients with myocardial infarction-associated depression.

[0011] As a preference, The treatment of myocardial infarction with depression includes improving myocardial cell pathological damage in patients with myocardial infarction with depression and promoting the neat arrangement of myocardial fibers.

[0012] As a preference, The treatment of myocardial infarction-associated depression includes reducing ventricular remodeling and / or myocardial fibrosis symptoms in patients with myocardial infarction-associated depression.

[0013] As a preference, The treatment of myocardial infarction with depression includes reducing myocardial cell apoptosis in patients with myocardial infarction with depression.

[0014] As a preference, The treatment of myocardial infarction with depression includes inhibiting the expression of myocardial fibrosis proteins in patients with myocardial infarction with depression.

[0015] As a preference, The pharmaceutical dosage form is tablets and / or capsules and / or granules and / or powders and / or liquid preparations.

[0016] As a preference, The extraction method of the Corydalis yanhusuo extract comprises the following steps: crushing the Corydalis yanhusuo, heating and refluxing with ethanol with a mass fraction of 75-85%, and obtaining the Corydalis yanhusuo extract.

[0017] Application of Corydalis yanhusuo extract in preventing myocardial infarction-associated depression.

[0018] Given the advantages of isoproterenol (ISO)-induced myocardial infarction (MI) models in mice, including ease of use (no complex surgical procedures), controllability (fractionated dosing avoids animal mortality), and high model stability, and the high clinical resemblance and success rate of chronic unpredictable mild stress (CUMS)-induced depression models in mice, this study employed subcutaneous ISO injection combined with CUMS to establish a MI-associated depression model in mice. The efficacy of Corydalis yanhusuo extract against MI in mice was evaluated by hematoxylin-eosin (HE) and Masson staining of mouse heart tissue. Behavioral tests such as sucrose preference, tail suspension, and forced swimming, combined with Nissl staining of the prefrontal cortex, were used to assess the efficacy of Corydalis yanhusuo extract in ameliorating depressive symptoms in mice with MI. In addition, a high-dose administration group of Corydalis yanhusuo extract but without modeling was set up to evaluate its safety; captopril and novitus, commonly used drugs for myocardial infarction or post-myocardial infarction heart failure, were selected as positive control drugs, and by comparing their efficacy with the two, the efficacy of Corydalis yanhusuo extract in preventing myocardial infarction-associated depression in mice was more comprehensively evaluated.

[0019] This study established a mouse model of myocardial infarction-associated depression. Using cardiac histopathological staining and Western blot analysis, the pharmacological effects of Corydalis yanhusuo extract on cardiomyocyte damage, apoptosis, and myocardial fibrosis in mice with myocardial infarction-associated depression were investigated. Results showed that Corydalis yanhusuo extract (466 mg / kg, twice the clinically equivalent dose, high-dose group), captopril, and Entresto all exhibited significant anti-MI and anti-myocardial fibrosis effects. Furthermore, this study evaluated the behavioral performance of mice with myocardial infarction-associated depression and the ameliorative effects of Corydalis yanhusuo extract on their depressive symptoms using behavioral tests (sucrose preference, tail suspension, and forced swimming) combined with Nissl staining of the prefrontal cortex. Results showed that high-dose Corydalis yanhusuo extract significantly alleviated anhedonia and significantly reduced levels of despair and anxiety in mice with myocardial infarction-associated depression. However, captopril and Entresto, commonly used cardiovascular drugs, had no significant effect on the mice's depression. These results suggest that simply administering cardioprotective drugs is ineffective in alleviating depressive symptoms in mice with myocardial infarction-associated depression. In addition, compared with the clinical equivalent dose group, the twice clinical equivalent dose of Corydalis yanhusuo extract is more effective in preventing myocardial infarction-associated depression, and the twice clinical equivalent dose of Corydalis yanhusuo extract has no obvious damaging effects on the behavior, myocardium and nerves of normal mice.

[0020] The beneficial effects of the present invention are: The present invention verifies the effectiveness of Corydalis yanhusuo extract in treating myocardial infarction with depression, as well as the effective and safe dosage, through experiments. This provides a corresponding basis and effective reference for guiding and developing the application of Corydalis yanhusuo extract, as well as for using Corydalis yanhusuo extract in preparing corresponding medicines and health foods. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Schematic diagram of the establishment, drug administration, and behavioral testing of the mouse myocardial infarction-combined depression model in Example 1; Figure 2 Representative images of HE staining of heart tissues of mice in each drug-treated group in Example 2; Figure 3 These are representative images of Masson staining of the hearts of mice in each drug-treated group in Example 2; Figure 4 The protein expression changes of α-SMA, TGF-β1 and Bax in the heart tissue of mice in each drug-treated group in Example 1; Figure 5 The effect of each drug-administered group in Example 1 on the sucrose preference experiment in mice with myocardial infarction and depression; Figure 6 The effects of each drug-administered group in Example 1 on the forced swimming and tail suspension tests in mice with myocardial infarction and depression; Figure 7 The effect of each drug-administered group intervention in Example 2 on Nissl staining of the prefrontal cortex tissue of mice with myocardial infarction and depression. DETAILED DESCRIPTION

[0022] The present invention is further described in detail below with reference to specific embodiments and the accompanying drawings. Based on these descriptions, those skilled in the art will be able to implement the present invention. Furthermore, the embodiments of the present invention described below are generally only a portion of the embodiments of the present invention, rather than all of the embodiments. Therefore, all other embodiments derived by those skilled in the art based on the embodiments of the present invention without inventive effort should fall within the scope of protection of the present invention.

[0023] Unless otherwise specified, the raw materials used in the examples of the present invention are all commercially available or available to those skilled in the art; unless otherwise specified, the methods used in the examples of the present invention are all methods known to those skilled in the art.

[0024] Example 1: Construction of a Myocardial Infarction-Combined Depression Model and Drug Efficacy Verification Animal grouping: C57BL / 6 mice were divided into blank control group, model group, low-dose group (233 mg / kg), and high-dose group (466 mg / kg); Experimental solution preparation: 1% sodium carboxymethylcellulose solution: Accurately weigh 5.0 g of sodium carboxymethylcellulose powder, add 500 mL of phosphate buffered saline (PBS), stir at 25 °C for 8 h, and store at 4 °C until use.

[0025] ISO injection: Weigh an appropriate amount of ISO powder and add phosphate buffered saline (PBS) solution to prepare an ISO injection solution with a final concentration of 5 mg / mL. Prepare and use immediately.

[0026] Corydalis yanhusuo extract suspension: Accurately weigh an appropriate amount of Corydalis yanhusuo extract and use 1% sodium carboxymethylcellulose solution to prepare low-dose (23.3 mg / mL) and high-dose (46.6 mg / mL) Corydalis yanhusuo extract suspensions (dosing volume 0.1 mL / 10 g, prepared and used immediately).

[0027] Captopril suspension: Based on the clinical dosage of captopril (100 mg / 60 kg), calculate the daily dose for mice using a body surface area conversion factor of 12.3 between humans and mice. Accurately weigh captopril powder at a dosage volume of 0.1 mL / 10 g mouse and add 1% sodium carboxymethylcellulose solution to prepare a captopril suspension with a density of 2 mg / mL. Prepare and use immediately.

[0028] Entresto suspension: Based on the clinical dosage of Entresto (100 mg / 60 kg), calculate the daily dosage for mice according to the human-to-mouse body surface area conversion coefficient of 12.3. Based on a dosage volume of 0.1 mL / 10 g mouse, accurately weigh sacubitril / valsartan sodium powder and add 1% sodium carboxymethylcellulose solution to prepare a Entresto suspension with a density of 2 mg / mL. Prepare and use immediately.

[0029] Protein lysis buffer: Accurately measure 2 mL of 1 M tris(hydroxymethyl)aminomethane hydrochloride solution (Tris-HCl), 4 mL of 5 M sodium chloride solution, 0.4 mL of 0.5 M ethylenediaminetetraacetic acid solution (EDTA), 10 mL of 0.5% Triton X-100 solution (Triton X-100 solution), 20 mL of 10% glycerol solution, and 163.6 mL of sterile ultrapure water to prepare 200 mL of working solution. Mix well and store at 4°C. Mix 10 μL of 0.5 M sodium fluoride solution, 10 μL of 0.1 M sodium vanadate (Na3VO4) solution, 10 μL of phenylmethylsulfonyl fluoride (PMSF) solution, and 970 μL of working solution to prepare protein lysis buffer.

[0030] Runnig Buffer (10×): Weigh 100.00 g of sodium dodecyl sulfate, 151.00 g of Tris-base, and 720.00 g of glycine, add ultrapure water to 5 L, and mix thoroughly. Runnig Buffer (1×): Take 200 mL of Running Buffer (10×) solution, add ultrapure water to 2 L, and mix thoroughly; Transfer Buffer (10×): Weigh 100.00 g of sodium dodecyl sulfate, 150.00 g of Tris, and 720.00 g of glycine, add ultrapure water to 5 L, and mix thoroughly. Transfer Buffer (1×): Take 200 mL of Transfer Buffer (10×) solution and 400 mL of methanol solution, add ultrapure water to 2 L, and mix thoroughly. TBS buffer (10×): Take 100 mL of Tris-HCl solution (1 M), take 80.00 g of sodium chloride, add ultrapure water to 1 L, and dissolve and mix thoroughly; TBST buffer (1×): Take 100 mL of TBS buffer (10×), add 1 mL of Tween-20 solution, add ultrapure water to 1 L, and mix thoroughly; 5% skim milk powder blocking solution: Weigh 10.00 g of skim milk powder, add 200 mL of TBST buffer, and dissolve and mix thoroughly.

[0031] Model building: Myocardial infarction model induction: Mice were housed in individually ventilated cages at an ambient temperature of 25°C, a relative humidity of 50 ± 10%, and a 12-h light-dark cycle. Water and food were available ad libitum. After 7 days of adaptive feeding, the mice were subcutaneously injected with ISO solution at a dose of 50 mg / kg daily for 7 days based on body weight to establish a mouse myocardial infarction model. After the injection, hearts from three mice were randomly sampled for HE staining to confirm myocardial vacuolation and fibrosis.

[0032] Depression Model Induction: After the myocardial infarction model was established, mice were exposed to a random, unpredictable mild stimulus daily for 6 weeks. These mild stimulations included: empty cage for 24 hours, moist bedding for 16 hours (150 mL of water per 100 g of bedding), 24 hours of fasting, 24 hours of water deprivation, horizontal shaking for 10 minutes, restraint in a 50 mL centrifuge tube for 4 hours, a dark cycle with light for 12 hours, and a 45-degree cage tilt for 24 hours.

[0033] The experimental animals were randomly divided into the following 7 groups according to their body weight, with 10 animals in each group. They were blank control group (Control), blank administration group (Control+YHS-high), model group (Model), low-dose Corydalis extract administration group (YHS-low, clinical equivalent dose), high-dose Corydalis extract administration group (YHS-high, twice the clinical equivalent dose), captopril positive drug control group (Captopril), and Entresto positive drug control group (Entresto). The blank control group and the blank administration group were subcutaneously injected with PBS in the first week, and were raised normally for the next 6 weeks. They were gavaged with 1% sodium carboxymethyl cellulose in the 5th and 6th weeks. The remaining five groups were subcutaneously injected with ISO injection in the first week, and then received 6 weeks of CUMS stimulation. They were gavaged in the 5th and 6th weeks of CUMS stimulation, and then underwent 4 days of behavioral experiments. The specific dosage and experimental procedures are shown in the following table and Figure 1 shown. Figure 1 Middle: Control: blank control group; Con+YHS-high: blank administration group; no model was established but high-dose of Corydalis yanhusuo extract was administered; Model: myocardial infarction with depression model group; YHS-low: low-dose Corydalis yanhusuo extract administration group; YHS-high: high-dose Corydalis yanhusuo extract administration group; Captopril: captopril positive drug control group; Entresto: Entresto positive drug control group; d: day;

[0034] The following drug efficacy tests were performed 6 weeks after oral administration.

[0035] Cardioprotective effect assessment: Histopathological Staining: Heart paraffin blocks were cut into 4-μm-thick slices using a microtome. The slices were flattened with warm water, placed on glass slides, dewaxed, and hydrated. Sections for hematoxylin and eosin staining were sequentially stained with hematoxylin and eosin and mounted with neutral gum. Sections for Masson staining were first stained with hematoxylin and ponceau red, then treated with phosphomolybdic acid, counterstained with aniline blue, and finally treated with 1% glacial acetic acid and mounted with neutral gum. Cardiac histopathological changes were observed and images were taken using a NIKON inverted microscope. Three samples were included in each group.

[0036] Western blotting: Proteins from mouse hearts were extracted using protein lysis buffer and determined using the Bradford assay. Protein content was then denatured by heating at 100°C for 10 minutes in a loading buffer containing 10% β-mercaptoethanol. After cooling, the denatured proteins were stored at -20°C. A 10% polyacrylamide gel was prepared according to the manufacturer's instructions. After adding 1× Running Buffer, protein samples were loaded, electrophoresed, and transferred to the membrane. After transferring the proteins from the gel to a PVDF membrane, the membrane was sheared according to the molecular weight of the target protein and placed in an antibody incubation chamber. The membrane was blocked with 5% skim milk powder for 2 hours. For antibody incubation, diluted primary antibodies targeting GAPDH, α-SMA, Bax, and TGF-β were added and incubated overnight at 4°C. After incubation, the primary antibodies were aspirated, and diluted horseradish peroxidase-conjugated secondary antibodies were added and incubated at room temperature for 1 hour. After incubation, the secondary antibodies were washed with TBST buffer, and the exposed protein bands were quantified using EvolutionCapt edge software.

[0037] The results are as follows Figure 4 As shown, Figure 4 Middle: Control: blank control group; Con+YHS-high: blank control group; high-dose Corydalis extract was given without modeling; Model: myocardial infarction-associated depression model group; YHS-low: low-dose Corydalis extract group; YHS-high: high-dose Corydalis extract group. *P<0.05, **P<0.01 compared with the control group. #P<0.05 compared with the model group.

[0038] Evaluation of antidepressant effect: Sucrose preference test (SPT): Before the sucrose preference test, mice were housed in individual cages. A bottle of 1.5% sucrose water and a bottle of pure water were placed in each cage, and the mice were allowed to acclimate for 12 hours to familiarize themselves with the taste of the two liquids. After the acclimatization period, the mice were deprived of food and water for 12 hours to ensure that all mice were in the same physiological state at the start of the experiment. At the start of the sucrose preference test, each mouse was re-equipped with a bottle of 1.5% sucrose water and a bottle of pure water. The bottles were accurately weighed and the initial weight of each bottle was recorded. During the experiment, the positions of the two water bottles were swapped every 6 hours to prevent positional preference from interfering with the experimental results. After 24 hours, all water bottles were collected and weighed again, and the sucrose preference of the mice was calculated according to the following formula.

[0039] Forced swimming test (FST): Before the forced swimming test, mice were placed in the experimental room for 2 hours of pre-acclimatization. A transparent cylindrical container 40 cm high and 20 cm in diameter was prepared and filled with 25°C distilled water to approximately three-quarters of the container's height. Mice were then placed in the container and forced to swim for 6 minutes. The duration of buoyancy during the last 4 minutes was recorded.

[0040] Tail suspension test (TST): Before the experiment, mice were placed in the experimental room for 2 hours to acclimate to the environment. During the test, the mice were tied with tape 1 cm from the tip of their tails and suspended from a hanging rod in an inverted headstand position with the tip of their tail approximately 50 cm above the ground. The total suspension time was 6 minutes, and the time the mice remained motionless within 4 minutes was recorded.

[0041] The results of FST and TST are as follows Figure 6 As shown, Figure 6 Middle: (A) Forced swimming test; (B) Tail suspension test. Control: blank control group; Con+YHS-high: blank administration group; no model was established but high-dose Corydalis extract was administered; Model: myocardial infarction with depression model group; YHS-low: low-dose Corydalis extract administration group; YHS-high: high-dose Corydalis extract administration group; Captopril: captopril positive control group; Entresto: Entresto positive control group. Compared with the Control group, * P <0.05; **** P <0.0001. Compared with the Model group, ### P <0.001; #### P <0.0001. n=10.

[0042] Neuroprotective effect assessment: Nissl staining of the prefrontal cortex: paraffin sections were prepared first, followed by Nissl staining, and finally the number of Nissl body-positive cells was counted under a microscope using SlideViewer software.

[0043] Analysis of the results in the attached figures showed that the cardiovascular and neurobehavioral indicators of the myocardial infarction-related depression model mice were significantly improved under the intervention of a high dose of Corydalis yanhusuo extract (466 mg / kg / d), as follows: Histopathological Improvement: HE staining revealed severe structural damage to the myocardial tissue of the model group mice, manifested by fiber breakage and vacuolation. In stark contrast, the myocardial cells in the high-dose group gradually became more regular and orderly, and the pathological damage was significantly alleviated. Further analysis by Masson staining revealed abnormal and extensive collagen fiber deposition in the myocardial tissue of the model group, while collagen fiber deposition was significantly reduced in the high-dose group, directly reflecting a significant reduction in the degree of fibrosis.

[0044] Molecular Mechanism Verification: Western Blot analysis revealed that in the high-dose group, the expression of the pro-apoptotic protein Bax, as well as the fibrosis-related markers α-SMA and TGF-β, were significantly downregulated in myocardial tissue. High BAX expression accelerated myocardial loss, while elevated TGF-β levels promoted scar formation and exacerbated interstitial fibrosis. Elevated fibrosis markers led to increased fibrosis. These results strongly suggest that Corydalis yanhusuo extract can exert its protective effects on the heart by effectively inhibiting myocardial cell apoptosis and fibrosis pathways.

[0045] Sucrose preference experiment: Results as follows Figure 5 As shown, in the sucrose preference experiment, the model group mice showed typical anhedonia symptoms; while the sucrose preference rate of the high-dose group mice increased significantly, approaching normal levels. This change fully demonstrates the positive improvement effect of high-dose intervention on depression-related behaviors in mice, and has a positive effect on improving the anhedonia symptoms of patients with myocardial infarction and depression. Figure 5 Middle: Control: blank control group; Con+YHS-high: blank administration group; no model was established but high-dose Corydalis extract was administered; Model: myocardial infarction with depression model group; YHS-low: low-dose Corydalis extract administration group; YHS-high: high-dose Corydalis extract administration group; Captopril: captopril positive drug control group; Entresto: Entresto positive drug control group. Compared with the Control group, ** P <0.01. Compared with the Model group, ## P <0.01. n=10.

[0046] Tail suspension test: During the tail suspension and forced swim tests, the immobility time of mice in the high-dose group was significantly shorter than that in the model group. This data clearly demonstrates that high-dose Corydalis extract can effectively alleviate the despair and anxiety behaviors of mice, improve their behavioral performance in specific experimental situations, and also has a positive effect on improving patients' exercise tolerance and locomotor activity.

[0047] Furthermore, Nissl staining and analysis of the prefrontal cortex revealed that neurons in the model group's prefrontal cortex suffered severe damage, with a sharp decrease in the number of Nissl bodies. In contrast, the number of Nissl-positive cells in this region of the high-dose group significantly rebounded, returning to near-normal levels, and the structural integrity of neurons was significantly improved. This demonstrates that Corydalis extract has a potent repairing effect on neuronal damage. Neuronal damage is a key pathological mechanism of depression, and its bidirectional relationship with depression—both a cause and a symptom—suggests that Corydalis extract can be used for both treatment and prevention.

[0048] Example 2: Safety Evaluation Male C57BL / 6 mice, 6–8 weeks old and weighing 20–22 g, were selected and housed in an SPF-grade environment. Mice were acclimated for 7 days before use in the experiment. The group design was designed with n = 10 per group: Blank control group (Control): No modeling treatment was performed, and 1% sodium carboxymethylcellulose solution was administered orally at a dose of 0.1 mL / 10 g.

[0049] High-dose group (YHS-high): The dosage was 466 mg / kg / day.

[0050] The dosing cycle was then set to six weeks of continuous gavage, with daily dosing at a volume of 0.1 mL / 10 g based on mouse body weight. The Corydalis extract was prepared with 1% sodium carboxymethylcellulose solution to a suspension concentration of 46.6 mg / mL for administration.

[0051] The safety test group was then tested as follows.

[0052] Body weight changes: The body weight of mice was measured weekly, and the body weight gain rate was calculated.

[0053] Behavioral observation: Closely record the mice's autonomous activities, food intake, and mental state.

[0054] Blood biochemical analysis: After the last dose, mice were fasted for 12 hours, anesthetized, and blood was collected from the orbital vein. Liver function markers (ALT, AST), renal function markers (BUN, Cr), and myocardial enzymes (CK-MB, LDH) were measured.

[0055] Histopathological examination: The heart and prefrontal cortex tissues of mice were obtained, fixed with 10% formalin, embedded in paraffin, and sectioned. They were then stained with HE to observe pathological changes.

[0056] Characterization results such as Figure 2 and Figure 7 shown.

[0057] Figure 2 Middle: Control: blank control group; Con+YHS-high: blank control group; high-dose Corydalis extract was given without modeling; Model: myocardial infarction-associated depression model group; YHS-low: low-dose Corydalis extract group; YHS-high: high-dose Corydalis extract group; Captopril: captopril positive control group; Entresto: Entresto positive control group. n = 3. Scale bar, 100 μm.

[0058] Figure 7 Middle: (A) Representative image of Nissl staining results of prefrontal cortex tissue; (B) Statistical analysis of the number of Nissl body-positive cells in prefrontal cortex tissue; Control: blank control group; Con+YHS-high: blank administration group; no modeling but administration of high-dose Corydalis extract; Model: myocardial infarction with depression model group; YHS-low: low-dose Corydalis extract administration group; YHS-high: high-dose Corydalis extract administration group; Captopril: captopril positive control group; Entresto: Entresto positive control group. Compared with the Control group, ** P <0.01. Compared with the Model group, ### P <0.001. n = 3. Scale bar, 100 μm.

[0059] Specific results showed no significant difference in weight gain between the two groups (control group: 18.5±2.1%; high-dose group: 19.3±1.8%, P>0.05). The high-dose group showed similar behavior to the control group in terms of locomotor activity, food intake, and mental state.

[0060] Blood biochemical parameters: liver function tests showed no significant differences between the two groups in ALT (control group: 32±5 U / L, high-dose group: 35±6 U / L) and AST (control group: 48±7 U / L, high-dose group: 51±8 U / L) (P>0.05). Renal function tests showed no significant differences in BUN (control group: 8.2±1.1 mmol / L, high-dose group: 8.5±1.3 mmol / L) and Cr (control group: 25±3 μmol / L, high-dose group: 26±4 μmol / L) (P>0.05). Myocardial enzymes showed no abnormally elevated levels of CK-MB (control group: 125±15 U / L, high-dose group: 130±18 U / L) and LDH (control group: 280±25 U / L, high-dose group: 290±30 U / L) (P>0.05).

[0061] Furthermore, the high-dose group showed regular myocardial fiber arrangement, with no evidence of edema, necrosis, or inflammatory infiltration. Prefrontal cortical neurons showed normal morphology, with evenly distributed Nissl bodies and no evidence of apoptosis or vacuolation.

[0062] The present invention discloses a synergistic intervention strategy for the heart and brain based on Corydalis yanhusuo extract, whose mechanism of action covers three core pathways: in terms of cardiovascular system regulation, by targeted inhibition of the cell death signal cascade mediated by the pro-apoptotic protein Bax, the incidence of programmed apoptosis of cardiomyocytes is significantly reduced; at the same time, the fibroblast activation pathway mediated by α-smooth muscle actin (α-SMA) and transforming growth factor β (TGF-β) is downregulated, effectively blocking the process of myocardial fibrosis. In terms of central nervous system repair, by activating the protein synthesis mechanism of prefrontal cortex neurons, the density of Nissl bodies is significantly increased, synaptic plasticity reconstruction is promoted, and dual repair of neural structure and function is achieved. In terms of cross-system synergy, a bidirectional protective effect is established based on the oxidative stress regulatory nodes shared by heart and brain tissues.

[0063] The treatment regimen employed a 6-week continuous dosing schedule, with a dose-response relationship based on pharmacodynamic studies. The baseline dose was converted to a clinically equivalent dose of 233 mg / kg / day, while the double-dose bolus (466 mg / kg / day) was designed to enhance bioavailability by increasing blood-brain barrier penetration. A dual-system independent modeling strategy was employed for model construction: Myocardial ischemia was induced by subcutaneous injection of isoproterenol (ISO) at 50 mg / kg / day for 7 consecutive days, successfully establishing characteristic pathological changes such as cardiomyocyte vacuolation and mitochondrial damage. The depression model employed a modified chronic unpredictable mild stress (CUMS) paradigm, in which a combination of stressful stimulation, including empty cage isolation, bedding humidity adjustment, and intermittent food restriction, was administered for 6 weeks to establish a stable neurobehavioral phenotype.

[0064] Pharmacodynamic verification showed that HE staining of myocardial tissue in the treatment group showed remodeling of muscle fiber arrangement, such as Figure 3 The quantification of Masson staining as shown confirmed that the area of ​​collagen deposition was reduced to a significant level. Figure 3Middle: Control: blank control group; Con+YHS-high: blank control group; high-dose Corydalis extract was administered without modeling; Model: myocardial infarction-associated depression model group; YHS-low: low-dose Corydalis extract group; YHS-high: high-dose Corydalis extract group; Captopril: captopril positive control group; Entresto: Entresto positive control group. n = 3. Scale bar, 100 μm. Western blot analysis revealed a significant downregulation of the expression levels of key apoptosis-regulating proteins and fibrosis signaling molecules, confirming its multidimensional regulatory effects on the cardiomyocyte survival microenvironment. Neurobehavioral assessments revealed significant improvements in behavioral indicators such as sucrose preference, locomotor activity, and stress response threshold in the treatment group. The mechanism may be related to functional remodeling of limbic system-prefrontal cortex neural circuits. Safety assessments confirmed that the high-dose group demonstrated a dose-dependent improvement in cardiac ejection fraction and behavioral reversal. No abnormalities in liver and kidney function or histotoxic pathological changes were detected, highlighting its safety within the therapeutic window.

Claims

1. Application of Corydalis yanhusuo extract in the treatment of myocardial infarction-associated depression.

2. The use according to claim 1, characterized in that The treatment of myocardial infarction-associated depression includes reducing the anhedonia symptoms in patients with myocardial infarction-associated depression.

3. The use according to claim 1, characterized in that The treatment of myocardial infarction with depression includes improving the exercise tolerance and autonomous motor activity of patients with myocardial infarction with depression, and alleviating the despair and anxiety behaviors of patients with myocardial infarction with depression.

4. The use according to claim 1, characterized in that The treatment of myocardial infarction-associated depression includes reducing apoptosis or vacuolation of neurons in the prefrontal cortex of patients with myocardial infarction-associated depression.

5. The use according to claim 1, characterized in that The treatment of myocardial infarction with depression includes improving myocardial cell pathological damage in patients with myocardial infarction with depression and promoting the neat arrangement of myocardial fibers.

6. The use according to claim 1, characterized in that The treatment of myocardial infarction with depression includes reducing ventricular remodeling and / or myocardial fibrosis symptoms in patients with myocardial infarction with depression.

7. The use according to claim 1, characterized in that The treatment of myocardial infarction with depression includes reducing myocardial cell apoptosis in patients with myocardial infarction with depression.

8. The use according to claim 1, characterized in that The treatment of myocardial infarction with depression includes inhibiting the expression of myocardial fibrosis proteins in patients with myocardial infarction with depression.

9. The use according to claim 1, characterized in that The extraction method of the Corydalis yanhusuo extract comprises the following steps: crushing the Corydalis yanhusuo, heating and refluxing with ethanol with a mass fraction of 75-85%, to obtain the Corydalis yanhusuo extract.

10. Application of Corydalis yanhusuo extract in the prevention of myocardial infarction-associated depression.

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