Use of histatin-1 polypeptide in the preparation of a formulation for promoting sciatic nerve regeneration or repair

Abstract

The application discloses application of a Histatin-1 polypeptide in preparation of a preparation for promoting sciatic nerve regeneration or repair, and an amino acid sequence of the Histatin-1 polypeptide is shown as SEQ ID NO. 1. The Histatin-1 polypeptide can promote expression of a nerve differentiation index, promote recovery of a functional index after sciatic nerve injury, and promote recovery of a histological index after sciatic nerve injury, for example, recovery of myelin structure and morphology. In addition, in combination with gelatin methacrylate (Gel-MA) as a biological scaffold, slow release of the Histatin-1 polypeptide can be realized, continuous repair support can be provided for a nerve injury site, myelin repair effect is further optimized, a more efficient and more durable solution is provided for treatment of sciatic nerve injury, and the application has a wide application prospect in the field of treatment of myelinated nerve injury.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to the field of sciatic nerve regeneration or repair, and particularly to the application of Histolin-1 peptide in the preparation of agents that promote sciatic nerve regeneration or repair. Background Technology

[0002] The nervous system is a complex neural network, characterized by its microscopic nerve fibers (myelinated and unmyelinated axons, Schwann cells, and myelin sheaths) and macroscopic connective tissue (epidermal, perineurium, and endoneurium). These structures together constitute a highly organized complex organ, enabling it to effectively transmit electrical signals and possess a certain degree of repair capability after injury.

[0003] Nerve injuries range in severity from temporary conduction block to complete nerve rupture. Their clinical prevalence is influenced by various factors, with compressive neuropathy being particularly common. Other injuries encountered include traction injuries, lacerations, avulsions, gunshot wounds, crush injuries, and iatrogenic injuries. However, the self-repair capacity of nerves after injury is limited by several factors. Although the nervous system possesses regenerative potential to some extent, this regeneration is often incomplete, leading to poor functional recovery. The reasons for this include: First, after nerve injury, Wallerian degeneration of distal nerve fibers—the disintegration of axons and myelin sheaths distal to the injury—while helping to clear debris, also impairs the structural integrity of the microenvironment within the distal neural canal, lacking the effective guiding structures needed for axon regeneration. Although Schwann cells proliferate after injury and form bigner bands, providing some guidance for regenerating axons, if the injury distance is too long or the time is too long, the structure of the bigner bands may degenerate, leading to decreased guiding ability, which is a significant limitation on nerve regeneration. Second, the endoneural canal is composed of endoneural connective tissue. In more severe injuries such as nerve transection, the endoneural canal, along with the perineurium and epineurium, is completely severed. This structural damage makes it difficult for regenerating axons to accurately locate and enter the distal endoneural canal, leading to axonal labyrinthine growth, neuroma formation, and severely hindering functional recovery. Third, the regeneration rate of axons is relatively slow, typically about 1 millimeter per day. For longer nerve injuries, axons may take months or even years to reach their distal target organs. During this period, if target organs (such as muscles) lose nerve innervation for an extended period, severe atrophy and fibrosis can occur. Even if axons eventually regenerate and regain innervation, the function of the target organ may not fully recover. This temporal mismatch is a significant reason for poor functional recovery. Fourth, the physical barriers of nerves, including the blood-nerve barrier of the perineurium and the blood-nerve barrier of the endoneurotic vessels, are also crucial for maintaining the stability of the nerve's internal microenvironment. After injury, these barriers may be disrupted, allowing harmful substances, inflammatory cells, and cytokines from the blood to enter the nerve and trigger an inflammatory response. While moderate inflammation is necessary to clear damage debris, excessive or persistent inflammation may inhibit axonal regeneration and adversely affect Schwann cells and the regenerative microenvironment. Furthermore, age is also a significant factor affecting nerve regeneration capacity. The nerve regeneration capacity of older individuals is generally lower than that of younger individuals, which may be related to decreased Schwann cell activity, reduced growth factor levels, and unfavorable changes in the repair microenvironment. Systemic diseases such as diabetes and vascular diseases can also affect the nerve repair process, leading to weakened regenerative capacity and poor functional recovery.

[0004] Building upon this foundation, the repair of myelinated nerve injuries, exemplified by the sciatic nerve, faces multiple challenges. These challenges primarily stem from its complex biological mechanisms, challenging microenvironment, and the long-term and uncertain nature of functional recovery. The myelin sheath, a lipid layer surrounding nerve axons, is crucial for the rapid transmission of nerve signals. Therefore, myelinated nerve injury not only signifies physical damage to the axon but also involves the destruction and re-regeneration of the myelin sheath (remyelination), making repair far more complex than for unmyelinated nerve injuries. The combined effect of these factors often results in incomplete functional recovery after sciatic nerve injury, prompting researchers to continuously explore new treatment strategies, such as bioengineered neural conduits and exosome therapy, to overcome these inherent limitations and promote more effective nerve regeneration.

[0005] Histatin-1 (Hst1) is primarily secreted by the parotid and submandibular glands in humans. As one of the most abundant histidine peptides in saliva, it is considered part of the oral defense system. Hst1 possesses multiple biological functions, including promoting cell adhesion and migration, enhancing wound healing, immunomodulation, and promoting bone repair. These properties make it a promising candidate for applications in tissue engineering and regenerative medicine. However, its effects on myelinated nerves remain unexplored and require further investigation. Summary of the Invention

[0006] The purpose of this invention is to address the problems existing in the prior art by providing the application of Histrin-1 peptide and related compositions in the repair of sciatic nerve injury or the promotion of sciatic nerve growth. Specifically, this invention discloses that the composition has one or more of the following uses: promoting changes in neuronal-like morphology of cells, high expression of protein markers such as β-III tubulin, microtubule-associated protein 2, and neurofilament proteins, promoting neurofunctional properties such as the sciatic nerve functional index, restoring the wet weight ratio of target muscles, and promoting neurohistochemical properties such as the restoration of myelin sheath structure and morphology.

[0007] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides the use of Histolin-1 polypeptide in the preparation of agents that promote sciatic nerve regeneration or repair, wherein the amino acid sequence of the Histolin-1 polypeptide is shown in SEQ ID NO.1, specifically as follows: DSHEKRHHGYRRKFHEKHHSHREFPFYGDYGSNYLYDN.

[0008] Furthermore, the Histolin-1 polypeptide is derived from saliva or synthesized artificially.

[0009] Furthermore, the repair specifically involves: promoting the neural differentiation and functional recovery of damaged nerve tissue or promoting the axonal growth of dorsal root ganglion (DRG) cells, and repairing the myelin sheath structure of the sciatic nerve.

[0010] Furthermore, the Histolin-1 polypeptide promotes the expression of neural differentiation indicators, which include one or more of neuron-like morphological changes, protein markers such as β-III tubulin, microtubule-associated protein 2, and neurofilament proteins.

[0011] Furthermore, the Histolin-1 peptide promotes the recovery of functional indicators after sciatic nerve injury, including one or more of the sciatic nerve function index (SFI) and the wet weight ratio of the sciatic nerve target muscle.

[0012] Furthermore, the Histolin-1 peptide promotes the recovery of histological indicators after sciatic nerve injury, including one or more of the following histological recovery indicators: sciatic nerve morphology, myelin sheath structure and morphology, and nerve repair markers such as BDNF, MBP, and NF200.

[0013] Secondly, this invention provides the application of Histolin-1 peptide in the preparation of agents that promote the recovery of nerve myelin sheath structure.

[0014] Thirdly, the present invention provides a method for promoting the recovery of nerve myelin sheath structure in vitro, comprising: treating in vitro cultured damaged nerve cells or tissues with Histrin-1 peptide in a laboratory environment.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention is the first to apply Histolin-1 peptide to the repair of myelinated nerves (such as the sciatic nerve), demonstrating its significant efficacy in promoting sciatic nerve regeneration and dorsal root ganglion (DRG) axon growth. Specifically, Histolin-1 peptide can be used to prepare preparations that promote changes in cell neuron-like morphology, expression of one or more protein markers such as β-III tubulin, microtubule-associated protein 2, and neurofilament proteins, promote the recovery of sciatic nerve function such as the sciatic nerve function index and target muscle wet weight ratio, and promote the recovery of nerve histology such as myelin sheath structure and morphology.

[0016] This invention not only provides a new improvement strategy for the repair of sciatic nerve injuries, but also provides a reference for the treatment of other myelinated nerve injuries, and offers new treatment methods for nerve injury-related diseases, such as sciatic nerve structural and functional disorders caused by acute contusion or compression, surgical iatrogenic injury, lumbar disc herniation, lumbar spinal stenosis, piriformis syndrome, etc. Attached Figure Description

[0017] Figure 1 This is the overall flowchart of the present invention.

[0018] Figure 2 The results of immunofluorescence staining and synapse number analysis of Hst1 promoting axonal growth in mouse DRG cells are shown in Figure A: Fluorescence confocal microscopy showing the effect of Hst1 intervention on MAP2 protein expression in DRG cells; Figure B: Image J quantitative analysis of MAP2 protein expression and axonal growth in DRG cells of different groups.

[0019] Figure 3The results of Hst1 intervention on the recovery of sciatic nerve function index and target muscle wet weight ratio in mice within 4 weeks after surgery in an experiment on promoting sciatic nerve injury repair in mice were as follows: A: Analysis of the effect of Hst1 intervention on the recovery of sciatic nerve function index in mice; B: Analysis of the effect of Hst1 intervention on the recovery of target muscle wet weight ratio of sciatic nerve in mice (group description: a. no sciatic nerve injury group, b. sciatic nerve compression injury group, c. Hst1 treatment group after sciatic nerve compression injury).

[0020] Figure 4 This study presents the results of immunofluorescence staining and analysis of the sciatic nerve in mice 4 weeks post-surgery in an experiment demonstrating how Hst1 promotes sciatic nerve injury repair in mice. A: Fluorescence confocal microscopy shows the effect of Hst1 intervention on the expression of proteins related to peripheral nerve repair in mice; B: Image J quantitative analysis of the expression of NF200, MBP, and BDNF proteins in different groups (group descriptions: a. normal nerve tissue, b. nerve tissue damaged by compression, c. nerve tissue treated with Hst1 after compression injury).

[0021] Figure 5 This study analyzed the results of TEM examination of the sciatic nerve cross-section and myelin regeneration in mice 4 weeks post-surgery during an experiment on Hst1-induced sciatic nerve injury repair in mice. A: Transmission electron microscopy showing the effect of Hst1 intervention on the repair and regeneration of myelin sheaths in peripheral nerves of mice; B: Image J quantitative analysis of the thickness and area of ​​myelin sheaths in nerve tissues of different groups (group descriptions: a. normal nerve tissue, b. nerve tissue damaged by compression, c. nerve tissue treated with Hst1 after compression injury).

[0022] Figure 6 The results are as follows: KEGG enrichment analysis and cluster heatmap (top 200 genes) of miRNA sequencing; A: KEGG enrichment analysis of the difference in gene expression in the injured nerves in the Hst1 treatment group; B: Cluster heatmap analysis of the difference in gene expression in the injured nerves in the Hst1 treatment group. Legend: H, Hst1: experimental group, nerve tissue treated with Hst1 after crush injury; C, Control: control group, nerve tissue after crush injury. Detailed Implementation

[0023] The present invention will be further described in detail below with reference to the embodiments. It should be noted that the embodiments described below are intended to facilitate the understanding of the present invention and do not limit it in any way.

[0024] like Figure 1 As shown, this invention provides the application of Histolin-1 polypeptide in the preparation of agents that promote sciatic nerve regeneration or repair, which promotes the neural differentiation and functional recovery of damaged nerve tissue, promotes the axonal growth of dorsal root ganglion cells, and repairs the myelin sheath structure of the sciatic nerve.

[0025] In this embodiment of the invention, to achieve more stable drug delivery, the Histolin-1 peptide is combined with a biological scaffold material, which includes gelatin, collagen, hyaluronic acid, chitosan, sodium alginate, heparin, polyvinyl alcohol, dextran, carboxymethyl cellulose, ethylene glycol chitosan, propylene glycol chitosan, chitosan lactate, carboxymethyl chitosan, chitosan quaternary ammonium salt, hydrophilic or water-soluble animal and plant proteins, collagen, serum proteins, bi- or multi-armed polyethylene glycol, polyethyleneimine, dendrites, synthetic peptides, polylysine or (meth)acrylate or (meth)acrylamide, and one or more of the above-mentioned components as modifications.

[0026] Biological scaffold materials are generally reagents that carry active ingredients. These reagents are safe for biological tissues and can maintain the release and slow release of active ingredients, thereby allowing the active ingredients to exert their greater or better physiological functions. Further, the biological scaffold material described in this invention is gelatin methacrylate (Gel-MA). Gelatin methacrylate has a similar chemical composition to collagen and exhibits good biocompatibility. Unlike collagen, Gel-MA does not pose a risk of pathogen transmission. Moreover, Gel-MA contains a large number of adhesion ligands, such as the arginine-glycine-aspartic acid sequence, thereby promoting cell adhesion and migration. Before curing, Gel-MA has good fluidity, allowing it to flexibly adapt to nerve lesions of different regions and sizes. After photocuring, Gel-MA transforms from a liquid state to a hydrogel state, which is non-irritating to biological tissues while facilitating the slow release of Hst-1 in vivo.

[0027] Using Gel-MA as a biological scaffold and loading Histolin-1 peptide as a bioactive agent, the repair of peripheral nerve injury can be effectively promoted. In the above composition, the mass-to-volume ratio of Histolin-1 peptide to gelatin methacrylate is 100 μg:(20-25) μL.

[0028] The present invention will be further described below.

[0029] Example 1: Preparation of Gel-MA hydrogel prepolymer solution and Hst1

[0030] The Gel-MA hydrogel used in this embodiment was purchased from EFL-Tech (Suzhou Yongqinquan Intelligent Equipment Co., Ltd., Suzhou, China). 1. Prepare a standard solution of photoinitiator (0.25% (w / v), i.e., 2.5 mg / ml): Take 20 ml of PBS and add it to a brown bottle containing the initiator phenyl-2,4,6-trimethylbenzoyl lithium phosphine (LAP) (containing 0.05 g LAP); heat in a water bath at 40-50 °C for 15 minutes, shaking several times during the process.

[0031] 2. Prepare GelMA solution (GelMA concentration is 5% (w / v), i.e., 50 mg / ml): Place the required mass of GelMA into a centrifuge tube; add the initiator standard solution to the centrifuge tube and shake to fully wet the GelMA; heat in a 60-70℃ water bath in the dark for 30 minutes, shaking several times during the process; remove air bubbles from the system by centrifugation (3000 rpm, 2 min); immediately sterilize the GelMA solution using a 0.22 μm sterile syringe filter (to prevent low-temperature gelation) and store it in a sterile centrifuge tube.

[0032] 3. The lyophilized linear Hst1 peptide was obtained from the University of Amsterdam (Amsterdam, Netherlands, amino acid sequence: DSHEEKRHHGYRRKFHEKHHSHREFPFYGDYGSNYLYDN, SEQ ID NO.1) and stored at -20°C. 100 μg of the lyophilized Hst1 peptide was dissolved in 21.1 μL of Gel-MA prepolymer solution to prepare the implantation reagent.

[0033] Example 2: Cellular evaluation of Hst-1 in promoting nerve repair

[0034] The fetal rat DRG cells used in this example were extracted from c57 female rats at 15-17 days of gestation. All animal experimental protocols were approved by the ethics review of the Animal Experiment Center of Zhejiang University of Traditional Chinese Medicine (IACUC-20240401-02).

[0035] 1. Extraction of fetal rat DRG cells: Female rats were euthanized by cervical dislocation, and the fetuses were removed after soaking in 75% alcohol. The fetal rat dorsal root ganglia were carefully isolated in PBS (Biosharp, China) at 4°C and temporarily stored in DMEM / F12 medium (Gibco, USA) containing 1% P / S (Biosharp, China). All the above operations were performed on ice. Digestion was performed at 37°C for 30 min using 0.1% type IV collagenase (BioFroxx, Germany) + 0.25% trypsin (Senrui Biotechnology, China), with gentle pipetting every 10 min. Digestion was stopped by adding DMEM / F12 medium containing 10% FBS (Neuzerum, New Zealand). After centrifugation at 500g for 5 min, the cells were resuspended in neurobasal-A medium (Gibco, USA) + 2% B27 supplement (Gibco, USA) + 1% GlutaMax (Gibco, USA) + 1% P / S + 50 ng / ml NGF (PeproTech, USA). The cell suspension was filtered through a 40 μm cell sieve and seeded into cell culture plates pre-coated with poly-L-lysine (Biosharp, China) at 37°C for 2 h. The cells were then cultured routinely at 37°C and 5% CO2. Change the medium 24 hours after inoculation. Add 10 μM Cytarabine (Sigma, Germany) to the nerve culture medium to inhibit the growth of non-neuronal cells. Change the medium once every 2 days.

[0036] 2. Verification of Hst-1's role in promoting neural differentiation: Conventionally cultured DRG cells were divided into a control group and an experimental group. The control group was cultured normally, while the experimental group was treated with 10 μM Hst1 on the 4th day of normal culture. After 3 days of treatment, the cell status was observed under a microscope.

[0037] Calcium ion signal expression in the two groups of cells was observed by live cell staining. Three days after treatment, DRG cells were stained with Fluo-4 AM calcium indicator (AbMole, USA) according to the reagent instructions, and calcium ion expression was observed using a laser confocal microscope.

[0038] The expression of the neuronal marker MAP2 was detected by immunofluorescence staining. After 5 days of group treatment, DRG cells were washed three times with PBS, fixed in 4% paraformaldehyde for 20 minutes, permeabilized in 0.1% Triton X-100 for 20 minutes, and blocked with 5% BSA (MedChemexpress, USA) + 0.3% Triton X-100 for 1 hour. Primary antibody was diluted with 1% BSA + 0.3% Triton X-100 + 5% goat serum, and the cell culture plates containing primary antibody were incubated overnight at 4°C in the dark. The next day, the culture plates were warmed, washed three times with PBS, and the secondary antibody was diluted using the same method and added to the cell culture plates, incubated in the dark for 1 hour. After washing three times with PBS, the cells were stained with DAPI. Finally, the cells in the wells were observed using a fluorescence microscope. The primary antibody used was rabbit anti-MAP2 (Proteintech, USA), and the corresponding secondary antibody was goat anti-rabbit IgG (Bioss, China). ImageJ software was used to analyze MAP2 fluorescence expression and cell synapse occurrence.

[0039] All data are expressed as mean ± standard deviation (SD). Data analysis was performed using GraphPad Prism 10, with Ordinary one-way ANOVA and One-sample t-test used for data comparison. P < 0.05 (95% confidence level) was used as the significance threshold.

[0040] The results are as follows Figure 2 As shown, by Figure 2 It is evident that cells in the Hst1 intervention group expressed more MAP2 fluorescence signals. Analysis of synaptic development in individual cells revealed that cells in the Hst1 intervention group exhibited a greater number of synapses, indicating that Hst1 intervention promoted axonal growth in mouse DRG cells. Example 3: Functional and histological evaluation of Hst-1 in promoting nerve repair This study used 6-8 week old c57 / BL6 mice. All animal experimental protocols were approved by the Animal Experiment Center of Zhejiang University of Traditional Chinese Medicine (IACUC-20240401-02).

[0041] Six mice were randomly assigned to three groups: a control group, a nerve injury group, and a nerve injury + Hst1 treatment group. The left sciatic nerve of each mouse was surgically exposed, while in the control group, only the nerve was exposed and the wound was sutured. In the nerve injury group, the mid-sciatic nerve was clamped with toothless forceps to induce nerve compression injury. In the Hst1 treatment group, Hst1-loaded GelMA-60 hydrogel (Engineering For Life, China) was placed in the injured nerve segment, the subcutaneous tissue was closed, and the skin was sutured. Functional assessments were performed on the mice 1 to 4 weeks post-surgery. The mice were euthanized in batches at 2 and 4 weeks post-surgery, and the sciatic nerve tissue was fixed or cryopreserved according to subsequent evaluation goals.

[0042] One to four weeks post-surgery, mice had their hind limbs stained with ink up to the ankle joints and placed at one end of a 60cm long, 10cm wide, and 10cm high wooden trough with openings at both ends. A 70g sheet of white paper of equal length and width was placed at the bottom of the trough, allowing the mice to walk to the other side of the trough. Five to six footprints were left on each hind limb. Three indices were measured on the normal (N) and injured (E) paw prints: A) PL (footprint length); B) TS (toe width); and C) IT (middle toe width). These indices were then substituted into the Bain formula to calculate the sciatic nerve function index.

[0043] Bain formula: SFI = 109.5(ETS-NTS) / NTS-38.3(EPL-NPL) / NPL+13.3(EIT-NIT) / NIT-8.8; where ETS represents the toe width (TS) of the injured foot (E), NTS represents the toe width (TS) of the normal foot (N), and other parameters can be deduced similarly. A sciatic nerve function index SFI = 0 indicates normal function, and SFI = -100 indicates complete injury.

[0044] Two and four weeks post-surgery, mice in each group were sacrificed, and tissue samples from the sciatic nerve injury segment were collected to prepare 2 μm thick paraffin sections. Immunofluorescence staining was performed on target proteins including CGRP, Gap43, MBP, β3-tubulin, NF200, and BDNF. ImageJ software was used to analyze the expression of fluorescent markers in different groups. Paraffin sections were dewaxed twice with xylene (10 minutes each time), rehydrated with graded ethanol to PBS, and then microwaved for 10 minutes with 0.01 M sodium citrate buffer to restore antigenic epitopes. All sections were then permeabilized with 1% Triton X-100 at room temperature for 20 minutes. After blocking with 5% goat serum at room temperature for 1 hour, the corresponding primary antibody was added and incubated overnight at 4°C in a humidified chamber. The next day, the sections were warmed, washed three times with PBS, incubated with secondary antibody for 1 hour in the dark, stained with DAPI for 5 minutes, rinsed with PBS, and mounted with glycerol gelatin. Finally, the sections were observed under a confocal microscope. Data acquisition was to be completed within 3 hours to avoid fluorescence quenching.

[0045] Four weeks post-surgery, a batch of mice in each group were sacrificed and perfused for fixation. Tissue from the target muscles (including the gastrocnemius and tibialis anterior) of the sciatic nerve on both the healthy and affected sides of each mouse was collected and weighed. The wet weight ratio of the target muscles on the affected / healthy side of each group of mice was calculated and analyzed. Tissue from the injured segment of the sciatic nerve was collected, and the cross-sectional morphology of the nerve was photographed using transmission electron microscopy to analyze the morphology and regeneration status of the myelin sheath.

[0046] The results are as follows Figure 3-5 As shown; by Figure 3 It is evident that the sciatic nerve function index of mice in the Hst1 intervention group recovered more rapidly, and the wet weight ratio of the target muscle was also significantly higher than that in the nerve injury group, indicating that Hst1 intervention effectively promoted the recovery of sciatic nerve function in mice.

[0047] Depend on Figure 4 It can be seen that the nerve tissue in the Hst1 intervention group expressed more NF200, MBP and BDNF fluorescence signals, indicating that Hst1 intervention promoted the repair of sciatic nerve injury in mice.

[0048] Depend on Figure 5 It is evident that the myelin sheath structure of nerve tissue subjected to compression injury is damaged, and the thickness and area of ​​the myelin sheath are reduced. In contrast, the nerve tissue in the Hst1 intervention group showed a clearer typical myelin sheath structure, and its thickness and area were also significantly increased, indicating that Hst1 intervention promoted the repair and regeneration of the myelin sheath of the sciatic nerve in mice. Example 4: Molecular biological evaluation of Hst-1's role in promoting neural repair

[0049] This embodiment follows the animal experiment protocol in Example 3 and has been approved by the ethics review of the Animal Experiment Center of Zhejiang University of Traditional Chinese Medicine (IACUC-20240401-02).

[0050] Sciatic nerve samples were taken from the experimental side of three groups of experimental animals on ice and temporarily stored in liquid nitrogen. miRNA sequencing was performed by Qingke Biotechnology Co., Ltd. (Beijing, China) to further analyze the potential mechanism by which Hst1 promotes nerve repair.

[0051] The results are as follows Figure 6 As shown; by Figure 6 It is evident that the Hst-1 intervention group showed significantly high expression of cytokines and cytokine receptor interactions, calcium signaling pathways, etc. in the nerve tissue, which is consistent with the main process and mechanism of nerve repair after injury described above, indicating that Hst1 intervention promoted the repair and regeneration of the sciatic nerve in mice.

[0052] The above description is merely a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited thereto. Any simple changes or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the scope of the technology disclosed in the present invention shall fall within the scope of protection of the present invention.

Claims

1. The application of Histolin-1 polypeptide in the preparation of agents that promote sciatic nerve regeneration or repair, characterized in that, The amino acid sequence of the Histolin-1 polypeptide is shown in SEQ ID NO.

1.

2. The application according to claim 1, characterized in that, The repair specifically involves: promoting the neural differentiation and functional recovery of damaged nerve tissue and / or promoting the axonal growth of dorsal root ganglion cells.

3. The application according to claim 2, characterized in that, Histatin-1 peptide repairs the myelin sheath structure of the sciatic nerve.

4. The application according to claim 1, characterized in that, The Histolin-1 peptide promotes the expression of neural differentiation indicators; these indicators include one or more of neuron-like morphological changes, protein markers such as β-III tubulin, microtubule-associated protein 2, and neurofilament proteins.

5. The application according to claim 1, characterized in that, The Histolin-1 peptide promotes the recovery of functional indicators after sciatic nerve injury, including one or both of the sciatic nerve function index and the wet weight ratio of the sciatic nerve target muscle.

6. The application according to claim 1, characterized in that, The Histolin-1 peptide promotes the recovery of histological indicators after sciatic nerve injury, including one or more of the following: sciatic nerve morphology, myelin sheath structure and morphology, and nerve repair markers.

7. The application of Histolin-1 peptide in the preparation of agents that promote the recovery of nerve myelin sheath structure, wherein the amino acid sequence of Histolin-1 peptide is shown in SEQ ID NO.

1.

8. A method for promoting the recovery of nerve myelin sheath structure in vitro, characterized in that, The method includes: treating in vitro cultured damaged nerve cells or tissues with Histolin-1 peptide in a laboratory environment; the amino acid sequence of the Histolin-1 peptide is shown in SEQ ID NO.1.

Images