Application of ancient-Chinese health-preserving essence in preparation of nerve repairing medicine

Abstract

The invention provides an application of ancient-Chinese health-preserving essence in preparation of a nerve repairing medicine, and belongs to the technical field of biological medicines. The invention provides an application of ancient-Chinese health-preserving essence in preparation of a nerve repairing medicine. The ancient-Chinese health-preserving essence can improve the plasticity of nerve synapses and improve the communication capability of neurons by promoting the repair of a dendritic structure and enhancing the synaptic function. And secondly, the ancient-Chinese health-preserving essence can further improve the function of VGLUT1 and the stability of MAP2 by regulating NCAM2, so that the health of nerve synapses is promoted, and nerve repair is realized.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to the application of Guhan Yangshengjing in the preparation of nerve repair drugs. Background Technology

[0002] Gu Han Yang Sheng Jing is a traditional Chinese medicine preparation, mainly composed of ginseng, roasted astragalus, wolfberry, processed privet fruit, dodder seed, rosehip, epimedium, white peony root, roasted malt, processed polygonatum, roasted licorice root, and refined honey (excipients). It has the effects of tonifying qi, nourishing the kidneys, and replenishing essence. However, there are currently no reports on the use of Gu Han Yang Sheng Jing for nerve repair. Summary of the Invention

[0003] The purpose of this invention is to provide the application of Guhan Yangshengjing in the preparation of nerve repair drugs, thereby enriching the medicinal uses of Guhan Yangshengjing.

[0004] This invention provides the application of GuHan YangShengJing in the preparation of nerve repair drugs.

[0005] This invention provides the application of GuHan YangShengJing in the preparation of drugs that improve the plasticity of neural synapses.

[0006] This invention provides the application of GuHan YangShengJing in the preparation of drugs that improve the communication ability of neurons and / or enhance signal transmission between neurons.

[0007] This invention provides the application of GuHan YangShengJing in the preparation of drugs that improve and / or enhance the function of nerve synapses.

[0008] This invention provides the application of GuHan YangShengJing in the preparation of drugs that enhance the neurotransmission function of glutamate.

[0009] This invention provides the application of GuHan YangShengJing in the preparation of drugs that improve the regeneration function of synaptic vesicles.

[0010] This invention provides the application of GuHan YangShengJing in the preparation of drugs that promote the repair of dendritic structures.

[0011] This invention provides the application of GuHan YangShengJing in the preparation of drugs that promote the restoration of health and / or function of nerve synapses.

[0012] Preferably, the ancient Chinese herbal essence is the only active ingredient of the drug.

[0013] Preferably, the dosage form of the drug includes an oral dosage form.

[0014] This invention provides the application of GuHan Yangshengjing in the preparation of nerve repair drugs. GuHan Yangshengjing can improve synaptic plasticity and enhance neuronal communication by promoting the repair of dendritic structures and enhancing synaptic function. Furthermore, GuHan Yangshengjing can further improve the function of VGLUT1 and the stability of MAP2 by regulating NCAM2, thereby promoting synaptic health and achieving nerve repair. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 The images show stained sections of vGlut1 in the hippocampus of AD rats treated with GuHan Yangshengjing. (A) Normal group, (B) Model group, (C) Low-dose GuHan Yangshengjing group, (D) Medium-dose GuHan Yangshengjing group, (E) High-dose GuHan Yangshengjing group, and (F) Positive drug group.

[0017] Figure 2 Statistical plot of vGlut1 staining in the hippocampus of AD rats by Guhan Yangshengjing; (*P<0.05, **P<0.01, ***P<0.001, n=3);

[0018] Figure 3 The images show stained sections of the hippocampus MAP2 of AD rats treated with Guhan Yangshengjing; (A) normal group, (B) model group, (C) low-dose Guhan Yangshengjing group, (D) medium-dose Guhan Yangshengjing group, (E) high-dose Guhan Yangshengjing group, and (F) positive drug group.

[0019] Figure 4 Statistical plot of staining of MAP2 in the hippocampus of AD rats by Guhan Yangshengjing; (*P<0.05, **P<0.01, ***P<0.001, n=3);

[0020] Figure 5This diagram shows the molecular docking results of NCAM2 with the main active ingredients of Guhan Yangshengjing. In the diagram, A represents the 2D interaction between NCAM2 and ginsenoside Rb1; B represents the 2D interaction between NCAM2 and ginsenoside Re; C represents the 2D interaction between NCAM2 and ginsenoside Rg1; D represents the 2D interaction between NCAM2 and astragaloside A; E represents the 2D interaction between NCAM2 and isoflavone glycoside; F represents the 2D interaction between NCAM2 and icariin; and G represents the 2D interaction between NCAM2 and astragaloside A. Force; H is the 2D force of NCAM2-asparagine B; I is the 2D force of NCAM2-asparagine C; J is the 2D force of NCAM2-betaine; K is the 2D force of NCAM2-ligustin; L is the 2D force of NCAM2-rhodioloside; M is the 2D force of NCAM2-hyperoside; N is the 2D force of NCAM2-paeoniflorin; O is the 2D force of NCAM2-glycyrrhizic acid; P is the 2D force of NCAM2-glycyrrhizin.

[0021] Figure 6 A thermal diagram showing the molecular docking between NCAM2 and the main active ingredients of Guhan Yangsheng Jing;

[0022] Figure 7 The diagram shows the 3D docking of NCAM2 with the main active ingredient of Guhan Yangshengjing; (A) 3D docking diagram of NCAM2-icariin; (B) 3D docking diagram of NCAM2-ascorbic acid A; (C) 3D docking diagram of NCAM2-ascorbic acid B; (D) 3D docking diagram of NCAM2-ascorbic acid C.

[0023] Figure 8 This is a schematic diagram of the BLI results; where A is NCAM2-icariin; B is NCAM2-icariin; C is NCAM2-icariin B; and D is NCAM2-icariin C.

[0024] Figure 9 The figure shows the effect of GuHan Yangshengjing on NCAM2 expression; (A) Immunofluorescence detection of siNCAM2 transient transfection results; (B) Relative expression of NCAM2 mRNA; (*P<0.05, **P<0.01, ***P<0.001, n=3). Detailed Implementation

[0025] This invention provides the application of GuHan YangShengJing in the preparation of nerve repair drugs.

[0026] This invention also provides the application of Guhan Yangshengjing in the preparation of drugs that improve neural synaptic plasticity.

[0027] The present invention also provides the application of Guhan Yangshengjing in the preparation of drugs that improve the communication ability of neurons and / or enhance signal transmission between neurons.

[0028] This invention also provides the application of GuHan YangShengJing in the preparation of drugs that improve and / or enhance the function of nerve synapses.

[0029] This invention also provides the application of GuHan YangShengJing in the preparation of drugs that enhance the neurotransmission function of glutamate.

[0030] This invention also provides the application of GuHan YangShengJing in the preparation of drugs that improve the regeneration function of synaptic vesicles.

[0031] This invention also provides the application of Guhan Yangshengjing in the preparation of drugs that promote the repair of dendritic structures.

[0032] This invention also provides the application of GuHan YangShengJing in the preparation of drugs that promote the restoration of health and / or function of nerve synapses.

[0033] In one implementation, the Guhan Yangsheng Jing is the sole active ingredient of the drug.

[0034] As one implementation method, the dosage form of the drug includes an oral dosage form.

[0035] To further illustrate the present invention, the application of the Guhan Yangsheng Essence provided by the present invention in the preparation of nerve repair drugs is described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0036] Example 1

[0037] I. Medicines and Reagents

[0038] Lipofectamine 2000 Reagent (Invitrogen), NCAM2 antibody (proteintech), APP (proteintech), BACE1 antibody (proteintech), vGlut1 antibody (proteintech), MAP2 antibody (proteintech), DAPI staining solution (Beyotime), NCAM2 protein (sinobiolog).

[0039] II. Primer Synthesis and Design

[0040] Mouse NCAM2 gene primers

[0041] Upstream primer: 5'-TGGAGGTGTG CCCATTCAT-3' (SEQ ID NO.1);

[0042] Downstream primer: 5'-GCGTACTATC TTCCAGGTTTCTGAT-3' (SEQ ID NO.2);

[0043] Mouse-derived GAPDH gene primers

[0044] Upstream primer: 5'-CTGGAGAAACCTGCCAAGTATG-3' (SEQ ID NO.3);

[0045] Downstream primer: 5'-GGTGGAAGAATGGGAGTTGCT-3' (SEQ ID NO.4);

[0046] siRNA synthesis

[0047] siNCAM2-Mus-1455:

[0048] Chain of Justice: 5'-GGGAAGGAAAUCCAAUCAATT-3 (SEQ ID NO.5);

[0049] Antonym: 5'-UUGAUUGGAUUUCCUUCCCTT-3' (SEQ ID NO.6);

[0050] Negative control FAM

[0051] Chain of Justice: 5'-UUCUCCGAACGUGUCACGUTT-3 (SEQ ID NO.7);

[0052] Antonym: 5'-ACGUGACACGUUCGGAGA-3' (SEQ ID NO.8);

[0053] III. Experiment Content

[0054] 1. Establishment and treatment of AD rat model

[0055] Seventy-two rats were randomly divided into six groups of 12 rats each: a normal control group, a positive drug group, a high-dose Guhan Yangshengjing group, a medium-dose Guhan Yangshengjing group, a low-dose Guhan Yangshengjing group, and a model group. During the experiment, all rats in each group received the drug via intraperitoneal injection and gavage, prepared with double-distilled water and heated to approximately 37°C. Drug dosage conversion was based on the dosage for animal type A, expressed in kilograms of body weight. The conversion formula is as follows: Required dosage for animal type B (mg) = Required dosage for animal type A (mg·kg) -1 ) × W (conversion factor) × body weight of animal B (kg).

[0056] For the first 8 weeks of the experiment, except for the normal control group, all other groups of rats received intraperitoneal injection of D-galactose and gavage with aluminum trichloride. The normal control group received only double-distilled water gavage once daily. After 8 weeks of modeling, the rats were divided into groups and different treatment interventions were administered to each group. The specific administration methods were as follows: Guhan Yangshengjing was administered by gavage at low, medium, and high doses respectively; the positive control group was administered donepezil hydrochloride tablets by gavage; the normal control group and the model group received an equal volume of double-distilled water by gavage.

[0057] The specific groupings and interventions are as follows:

[0058] 1-8 weeks:

[0059] ① Normal control group: Normal rat group + double-distilled water (double-distilled water of the same volume as the Chinese herbal medicine compound);

[0060] ② Model group: Model rat group + D-galactose (180mg / kg) + aluminum trichloride (15mg / kg); 8-20 weeks:

[0061] ① Normal control group: Normal rat group + double-distilled water (double-distilled water of the same volume as the Chinese herbal medicine compound);

[0062] ② Model group: Model rat group + D-galactose (180mg / kg) + aluminum trichloride (15mg / kg) + double-distilled water (equal volume of double-distilled water to the traditional Chinese medicine compound);

[0063] ③ Low-dose traditional Chinese medicine group: Model rat group + D-galactose (180mg / kg) + aluminum trichloride (15mg / kg) + Guhan Yangshengjing (1mL / kg);

[0064] ④ Medium-dose group of traditional Chinese medicine: Model rat group + D-galactose (180mg / kg) + aluminum trichloride (15mg / kg) + Guhan Yangshengjing (2mL / kg);

[0065] ⑤ High-dose traditional Chinese medicine group: Model rat group + D-galactose (180mg / kg) + aluminum trichloride (15mg / kg) + Guhan Yangshengjing (4mL / kg);

[0066] ⑥ Positive drug group: Model rat group + D-galactose (180mg / kg) + donepezil hydrochloride tablets (1mg / kg).

[0067] 2. Serum Sample Collection and Preparation

[0068] Rat serum samples were collected after the experiment using the abdominal vein method. After collection, the blood samples were allowed to stand at room temperature for 30 minutes to promote coagulation, and then centrifuged to separate the serum. Centrifugation was performed at 3000 rpm for 10 minutes at 4°C. The supernatant was collected as the serum sample and stored frozen at -80°C for later use.

[0069] 3. Hippocampal sample collection and preparation

[0070] Rats collected hippocampal samples after the experiment. The rat brain was removed via abdominal dissection and quickly placed on ice. The hippocampus was carefully separated with a scalpel, and after removing the surrounding brain tissue, the hippocampal sample was rapidly frozen in liquid nitrogen. The sample was stored at -80°C for later analysis.

[0071] 4. Histopathological analysis

[0072] In this embodiment, tissue embedding was performed using a conventional paraffin embedding method. First, relevant tissues from the experimental rats were extracted, quickly washed with physiological saline to remove blood impurities, and fixed in 10% neutral buffered formalin for 24 hours. The fixed tissue specimens underwent dehydration, clearing, and paraffin embedding. The tissues were sequentially dehydrated using different volume concentrations of ethanol (70%, 80%, 90%, 100%), cleared with xylene, and then embedded in molten paraffin. After embedding, the tissues were cooled to room temperature and prepared into paraffin blocks for subsequent sectioning.

[0073] Sectioning was performed using a conventional microtome. The paraffin-embedded tissue blocks were placed in the microtome, adjusted to an appropriate thickness (generally 4–5 μm), and the section quality was checked under a microscope. After sectioning, dewaxing was performed using xylene, followed by hydration with ethanol of varying concentrations, and finally rinsing with distilled water to remove any residual ethanol.

[0074] In immunofluorescence staining, tissue sections were routinely dewaxed and hydrated, then treated with 0.3% Triton X-100 to increase cell membrane permeability. Subsequently, the tissue sections were incubated with a specific primary antibody according to the manufacturer's recommended time and temperature conditions for antigen-antibody binding. After washing away unbound primary antibody, fluorescently labeled secondary antibody was added, and incubation continued until excess secondary antibody was washed away. Finally, observation and imaging were performed using a fluorescence microscope to analyze the localization and expression of the target protein.

[0075] In immunohistochemical staining, sections undergo routine dewaxing and hydration, and endogenous peroxidase activity is blocked using hydrogen peroxide. Subsequently, the sections are incubated with the corresponding primary antibody. After the antigen binds to the primary antibody, a secondary antibody binds to horseradish peroxidase (HRP), forming a complex. Finally, by adding a DAB chromogenic substrate, an HRP-catalyzed reaction occurs, producing a brown precipitate that indicates the distribution of the target antigen. This method allows for qualitative or quantitative assessment of target antigen expression by observing the staining reaction of tissue sections under a microscope.

[0076] 5. RT-qPCR (Real-time quantitative reverse transcription polymerase chain reaction)

[0077] This experiment uses Total RNA was extracted from rat tissues using the Reagent method to ensure its quality met the requirements for subsequent analysis. After RNA extraction, reverse transcriptase was used to convert the total RNA into complementary DNA (cDNA). Random primers or oligo(dT) primers were added to the reverse transcription reaction system to ensure comprehensive synthesis covering all mRNAs. After the reverse transcription reaction was completed, a heat inactivation step was performed to ensure the completeness of the reverse transcription process.

[0078] Subsequently, real-time quantitative PCR (qPCR) was performed using a kit to detect the expression level of the target gene. Primer design was based on the NCBI Primer-BLAST tool, with appropriate primers selected according to the mRNA sequence of the target gene, ensuring that the primers spanned exon linker regions to avoid interference from genomic DNA. The amplification specificity of all primers was verified by melting curve analysis, and the amplification efficiency was evaluated using the standard curve method to ensure the accuracy and reliability of the reaction.

[0079] Data analysis employed the 2-ΔΔCt relative quantification method, using the internal reference gene GAPDH as a standardized internal reference. Three technical replicates were set up for each sample, and three biological replicates were performed for each independent experiment. Ct values ​​were acquired and data processed using Bio-Rad CFX Manager 3.1 software.

[0080] 6. Molecular docking experiments

[0081] Based on the network pharmacology results of GuHan Yangshengjing, the core target and active ingredient were identified by selecting proteins and active ingredients with the highest concentration in the component-target-pathway network. The 3D structure files of the active ingredient were obtained from PubChem, while the crystal structures of the protein target were obtained from the RCSB PDB database (http: / / www.rcsb.org / ). The protein structures were processed using Pymol, including solvent removal, equilibrium ion removal, and hydrogenation. Finally, molecular docking between the protein and component was performed using Autodock, and 2D force field maps and 3D spatial environment maps were generated using Discovery Studio 2016.

[0082] 7. Biolayer Interference (BLI) Experiment

[0083] In the experiment, the NCAM2 protein was first immobilized on the surface of a BLI sensor, and ligands such as icariin or astragaloside ABC were injected to monitor their binding and dissociation reactions with NCAM2 in real time. The experimental steps included: first, activating the sensor surface and immobilizing the NCAM2 protein; then, adding icariin or astragaloside ABC and monitoring their binding kinetics with NCAM2; finally, treating with elution buffer and observing the molecular dissociation process. Experimental data were collected in real time using a BLI instrument, and the binding constant (Kbinding constant) of intermolecular interactions was finally analyzed. D Dynamic parameters such as )

[0084] 8. Transient transfection experiment of siNCAM2

[0085] Transfection procedure: Seed cells in six-well plates (30%–50% density) → Wash with PBS → Prepare transfection complex (Solution A: 4 μl Lipo2000 + serum-free medium; Solution B: 4 μl 50 nM siRNA + serum-free medium) → Mix solutions A and B and let stand for 15 min → Add to cell culture system → Incubate at 37°C for 48 h → Collect cells for downstream analysis.

[0086] 9. Cell Experiment Design

[0087] In this experiment, mouse hippocampal neurons HT22 cells were first introduced at a rate of 5 × 10⁻⁶. 5 Cells were seeded at a density of [number] cells / well. When cells reached 80% confluence, they were treated with 30 μM Aβ for 24 hours to induce cell damage. After treatment, fresh culture medium was added, and different concentrations of drug-containing serum were added for further treatment: 5%, 10%, and 20%. Serum volumes for all treatment groups were adjusted to the appropriate total volume using blank serum to ensure accurate final concentrations. Three replicates were set up for each group to ensure experimental reproducibility and data reliability. The treatment time was 24 hours.

[0088] 10. Statistical Analysis

[0089] Unless otherwise stated, data are presented as mean ± standard error. Analyses were performed using GraphPad Prism 10 with a critical α level set at P < 0.05, and comparisons between multiple groups were conducted using one-way ANOVA.

[0090] 11. Results

[0091] (1) This embodiment evaluated the regulatory effect of GuHan Yangshengjing on NCAM2 and its related synaptic markers. The effect of GuHan Yangshengjing on improving neural synaptic function was evaluated by detecting changes in the expression of glutamatergic synaptic marker VGLUT1 and neuronal cytoskeletal protein MAP2.

[0092] First, the immunohistochemical results for VGLUT1 showed that ( Figure 1 and Figure 2 In comparison with the model group, VGLUT1 expression was significantly upregulated in the normal, medium-dose, and high-dose groups. VGLUT1, as a glutamate transporter, participates in the formation and release of presynaptic vesicles. Its upregulated expression suggests that GuHan Yangshengjing may enhance glutamatergic neurotransmission, improve synaptic vesicle reuptake function, thereby enhancing signal transmission between neurons and promoting synaptic function recovery. This result further supports the effect of GuHan Yangshengjing on improving synaptic plasticity and cognitive function.

[0093] In addition, in MAP2 immunofluorescence analysis ( Figure 3 and Figure 4 Significant upregulation of MAP2 expression was also observed in both the normal and high-dose groups. MAP2 is an important marker of dendritic microtubules, involved in the stability of dendritic structure and the maintenance of synaptic function. The upregulation of its expression suggests that Guhan Yangshengjing may improve neuronal communication by promoting the repair of dendritic structure and enhancing synaptic function, thereby restoring synaptic function and cognitive ability in AD model rats.

[0094] These results indicate that NCAM2, as an important regulatory molecule, is closely related to the expression of synaptic function-related molecules such as VGLUT1 and MAP2. GuHan Yangshengjing, by regulating NCAM2, further improved the function of VGLUT1 and the stability of MAP2, thereby promoting the health of neural synapses.

[0095] (2) The key role of NCAM2 in regulating Aβ deposition in ancient Chinese herbal medicine

[0096] The expression level of NCAM2 in the differentially expressed protein network closely related to synaptic function showed significant changes (P<0.05). A multi-stage experimental strategy was adopted: first, molecular docking technology was used to screen components in the compound with potential binding activity to NCAM2; then, BLI was used to quantitatively determine the affinity of candidate components for NCAM2; finally, qPCR and IF were used to detect the regulatory effect of the effective components on the expression level of NCAM2.

[0097] Molecular docking results of NCAM2 with the main chemical components in Guhan Yangshengjing showed that ( Figure 5 , Figure 6Among the flavonoids, icariin exhibited the strongest theoretical binding affinity (-6.7 kcal / mol), followed by ginsenoside A (-6.4 kcal / mol) and ginsenoside Re (-6.4 kcal / mol). Notably, saponins (ginsenoside Rb1, ginsenoside Re, ginsenoside Rg1, etc.) generally showed high binding energies (≤-6.0 kcal / mol), while among flavonoids, icariin and its derivatives (ginsenoside A, ginsenoside B, ginsenoside C) showed superior binding affinity compared to other flavonoids (such as hyperoside -5.6 kcal / mol).

[0098] In the molecular docking analysis of NCAM2 and icariin ( Figure 7 The results showed that multiple amino acid residues formed stable bonds with icariin. Specifically, LEUA:7, THRA:21, and ARGA:57 formed conventional hydrogen bonds with icariin, indicating a strong intermolecular affinity, especially near the protein active site. Furthermore, SERA:6 formed a carbon-hydrogen bond with icariin, which further promoted the stability of the molecular binding. However, the interaction between ARGA:55 and icariin exhibited an unfavorable donor-donor interaction (red dashed line), which could lead to unstable molecular binding or functional limitations in vivo.

[0099] The interaction characteristics between the main active ingredients in Guhan Yangshengjing and NCAM2 were characterized using BLI. Experimental data revealed significant differences in the binding abilities of each component. Figure 8 ): Asparagine C exhibits the strongest binding properties (K) D =238 μM), its binding response (0.0361-0.0777 nm) showed good concentration dependence and reached 88.1% binding saturation; icariin was the second best (K D =1.18 mM), showing moderate binding saturation (67.9%) and a clear dose-response effect. In contrast, the binding abilities of ascorbic acid A and B were weaker: the response values ​​of ascorbic acid A at all concentrations (156.3-1250 μM) were all below 0.1 nm and showed no concentration dependence (R0). 2 =0.3659), its abnormal dynamic parameter (k on =4.32×10^861 / Ms) suggests that only weak binding may exist; although astragalus B shows a certain concentration dependence (R 2 =0.9373), but the binding signal was weak (highest response 0.1351 nm) and not saturated (Req / Rmax ≤ 39.9%). It is noteworthy that the experimental data for both icariin C and epimedium glycoside showed excellent goodness of fit (R = 0.9373), but the binding signal was weak (highest response 0.1351 nm) and not saturated (Req / Rmax ≤ 39.9%). 2>0.92), confirming the reliability of the results.

[0100] Based on the combined results of molecular docking and biolayer interference experiments, icariin exhibits a significant characteristic of stable binding with NCAM2. Therefore, this study has established it as a core candidate molecule and focuses on elucidating its neuroprotective mechanism regulated by the NCAM2 target.

[0101] In NCAM2 knockdown experiments ( Figure 9 The role of icariin in Aβ clearance was investigated by knocking down NCAM2 expression using siRNA. 23-25 In a HT22 mouse hippocampal neuronal model treated with 30 μM icariin, Aβ deposition was significantly reduced (8.761 ± 1.511 s) after treatment. 23-25 Compared with the 30 μM group, the Aβ fluorescence intensity decreased significantly (P<0.01). This result indicates that icariin can effectively clear Aβ deposits and has a neuroprotective effect.

[0102] Notably, NCAM2 knockdown significantly inhibited the Aβ scavenging effect of icariin, while Aβ deposition increased markedly, indicating that NCAM2 downregulation suppressed icariin's Aβ scavenging ability. This result further confirms the important role of NCAM2 in the regulation of Aβ accumulation and scavenging by icariin. Further mRNA analysis showed that NCAM2 expression was significantly upregulated in the hippocampus of rats in the normal group and the high-dose GuHan Yangshengjing group, while the model group showed lower NCAM2 expression. This finding suggests that NCAM2 may be involved in regulating the icariin-mediated Aβ scavenging process, providing a new molecular target for elucidating the neuroprotective mechanism of icariin.

[0103] In summary, GuHan Yangshengjing significantly modulates the synaptic function-related protein network, and the hypothesis that NCAM2 is a key molecule simultaneously linked to Aβ metabolism and synaptic plasticity is proposed. Furthermore, pharmacological validation results further confirmed the expression changes of APP and BACE1, demonstrating the regulatory effect of GuHan Yangshengjing on these key molecules. Knockdown of NCAM2 expression using siRNA revealed that NCAM2 plays a crucial role in the GuHan Yangshengjing-mediated Aβ clearance process, and icariin may be the core active ingredient. In addition, analysis of synaptic markers VGLUT1 and MAP2 indicates that GuHan Yangshengjing can also significantly promote synaptic function recovery. These findings collectively construct a mechanism by which GuHan Yangshengjing intervenes in the amyloid cascade reaction through a dual pathway of "BACE1-APP-Aβ generation" and "NCAM2-Aβ clearance," while simultaneously exerting multi-target neuroprotective effects by regulating synaptic plasticity and brain energy metabolism.

[0104] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Application of Guhan Yangshengjing in the preparation of nerve repair drugs.

2. Application of Guhan Yangshengjing in the preparation of drugs that improve the plasticity of neural synapses.

3. Application of Guhan Yangshengjing in the preparation of drugs that improve the communication ability of neurons and / or enhance signal transmission between neurons.

4. Application of Guhan Yangshengjing in the preparation of drugs that improve and / or enhance the function of nerve synapses.

5. Application of Guhan Yangshengjing in the preparation of drugs that enhance the neurotransmission function of glutamate.

6. Application of Guhan Yangshengjing in the preparation of drugs that improve the regeneration function of synaptic vesicles.

7. Application of Guhan Yangshengjing in the preparation of drugs that promote the repair of dendritic structures.

8. Application of Guhan Yangshengjing in the preparation of drugs that promote the recovery of the health and / or function of nerve synapses.

9. The application according to any one of claims 1 to 8, characterized in that, The ancient Chinese health essence is the only active ingredient in the drug.

10. The application according to any one of claims 1 to 8, characterized in that, The dosage form of the drug includes oral dosage forms.

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