Application of chemotactic factor CCL14 in preparation of products for resisting tendon adhesion
Through the human plasma chemokine CCL14 and its promoters, the biocompatibility and side effects of existing drugs in preventing tendon adhesions is solved, and the adhesion between tendons and surrounding tissues is significantly reduced, providing new drug targets and clinical application potential.
Patent Information
- Application Number
- CN202510423966.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-07
- Publication Date
- 2025-08-19
AI Technical Summary
Existing drugs that inhibit tendon adhesions have problems affecting local tendon healing and biocompatibility, and may bring side effects, and lack effective drug targets to prevent tendon adhesions.
Using human plasma chemokine CCL14 and its promoter, its high expression in damaged tendon cells was verified through single-cell sequencing and animal experiments, significantly reducing the adhesion of tendons to surrounding tissues, and using its binding to CCR1, CCR3 and CCR5 chemokine receptors to provide good biocompatibility and low immunogenicity.
It significantly reduces adhesions between tendons and surrounding tissues, improves the safety and effectiveness of drugs, provides new drug targets to prevent adhesions after tendon injury, and meets the clinical potential of autologous biological agents.
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Abstract
Description
Technical Field
[0001] The present invention relates to application of chemokine CCL14 in preparing products for resisting tendon adhesion, and belongs to the technical field of biomedicine. Background Art
[0002] Adhesions formed after tendon injury are a clinically important complication of tendon healing and can lead to functional disability. Hydrogels are a physical barrier used to prevent or limit tendon adhesions. Loading drugs into hydrogels to inhibit the growth of adhesion tissue is a common strategy. Although biological barrier materials have the effect of physical barriers, foreign body reactions can aggravate adhesions. Currently used drugs for the treatment of tendon adhesions, such as corticosteroids, nonsteroidal anti-inflammatory drugs, and decorin, either have unsatisfactory anti-adhesion effects or have side effects, such as poor tendon healing.
[0003] Existing drugs for inhibiting tendon adhesions mainly include anti-inflammatory drugs (such as celecoxib), anti-oxidative stress drugs (such as hydrogen-rich water), and antimetabolites (such as angiotensin-converting enzyme inhibitors and rapamycin). However, these drugs may affect local tendon healing and biocompatibility, as well as other side effects caused by the drugs themselves.
[0004] CCL14 is a human plasma chemokine that belongs to the small cytokine family of CC chemokines. It is a cluster of chemokines located on human chromosome 17 and is expressed in multiple tissues, including the spleen, bone marrow, liver, muscle, and intestine. CCL14 has activity against human monocytes and binds to the CCR1, CCR3, and CCR5 chemokine receptors. Current studies have shown that CCL14 can inhibit the proliferation of hepatocellular carcinoma (HCC) cells by inhibiting cell cycle progression and promoting apoptosis. In vivo animal studies have shown that CCL14 can inhibit HCC tumor growth in nude mice. However, there are currently no studies on the role of CCL14 in preventing and treating tendon adhesions. Summary of the Invention
[0005] The purpose of the present invention is to provide the application of chemokine CCL14 in the preparation of anti-tendon adhesion products to solve the problems existing in the above-mentioned prior art. The present invention found that plasma chemokine CCL14 is highly expressed in damaged tendon pericytes, and confirmed through animal experiments that human plasma chemokine CCL14 can significantly reduce the degree of adhesion between damaged tendons and surrounding tissues, and prevent tendon adhesion. In addition, human-derived plasma chemokine CCL14 has good biocompatibility and low immunogenicity, can reduce the immune response and toxicity of drugs in the body, improve the safety and efficacy of drugs, and has clinical potential for transformation into polypeptide drugs, providing new drug targets and theoretical support for preventing adhesions after tendon injury.
[0006] To achieve the above object, the present invention provides the following solutions:
[0007] The present invention provides use of chemokine CCL14 or a promoter thereof in preparing a product for resisting tendon adhesion.
[0008] Furthermore, the chemokine CCL14 is derived from mammals, more preferably from rodents (such as mice and rats), primates and humans, and most preferably is human chemokine CCL14.
[0009] Furthermore, the CCL14 promoter refers to a substance that can increase the activity and / or content of the CCL14 gene or its protein in vivo or in vitro, or a substance that can promote the binding of CCL14 to the CCR1, CCR3 or CCR5 chemokine receptors; the substance can be a synthetic or natural compound, protein, nucleotide, etc.
[0010] Optionally, the CCL14 promoter is selected from the group consisting of: a small molecule compound, a CCL14 gene delivery vector, and a CCL14 protein agonist, or a combination of one or more thereof.
[0011] The gene delivery vector refers to a medium necessary for assembling genes and introducing them into target cells or corresponding host cells to express the genes; the gene delivery vector includes viral vectors or non-viral vectors.
[0012] Optionally, the viral vector is selected from a combination of one or more retroviral vectors, adenoviral vectors, and adeno-associated virus (AAV) vectors.
[0013] Optionally, the non-viral vector comprises mRNA liposome.
[0014] Optionally, the CCL14 gene or protein is derived from mammals, more preferably from rodents (such as mice, rats), primates and humans, and most preferably from humans.
[0015] Optionally, the CCL14 protein agonist includes a full-length CCL14 protein, a recombinant CCL14 protein, or an active fragment of a CCL14 protein.
[0016] In some embodiments of the present invention, the CCL14 protein agonist is a CCL14 recombinant protein, which can be commercially available.
[0017] The recombinant protein refers to first obtaining the target gene (GOI) using gene cloning or chemical synthesis technology, then connecting the target gene to a suitable expression vector, introducing it into a specific host cell, and utilizing the genetic system of the host cell to express a functional protein molecule.
[0018] The present invention also provides a product for resisting tendon adhesion, wherein the active ingredient of the product is chemokine CCL14 or a promoter thereof.
[0019] Optionally, the concentration of chemokine CCL14 in the product is 15-30 ng / mL.
[0020] Optionally, the product further comprises pharmaceutically acceptable excipients.
[0021] Optionally, the dosage forms of the product include tablets, injections and gels.
[0022] Optionally, the product is in the form of an injection and is administered by subcutaneous injection.
[0023] Compared with the prior art, the present invention has the following beneficial effects:
[0024] The present invention discovered through single-cell sequencing technology that plasma chemokine CCL14 is highly expressed in pericytes of injured tendons. Further experiments on mice and primates confirmed that additional administration of human plasma chemokine CCL14 can significantly reduce the degree of adhesion between injured tendons and surrounding tissues, preventing tendon adhesion, revealing the role of human plasma chemokine CCL14 in preventing tendon adhesion; the present invention also proved through molecular simulation and cell experiments that human CCL14 has a high binding ability with CCR1 and is conserved among species, and that human plasma chemokine CCL14 can meet the requirements of autologous sources, has good biocompatibility and low immunogenicity, can reduce the immune response and toxicity of drugs in the body, improve the safety and efficacy of drugs, and has clinical potential for transformation into autologous biological preparations; the present invention provides new drug targets and theoretical support for preventing adhesion formation after tendon injury. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] Figure 1 This is a flowchart for single-cell sequencing;
[0026] Figure 2 Single-cell sequencing results;
[0027] Figure 3 The figure shows the results of immunofluorescence co-staining;
[0028] Figure 4The results of the test of the binding ability of CCL14 factor to CCR1 receptor and its species conservation; A is the result of DiscoveryStudio molecular simulation; B is the result of cell immunofluorescence; C is the result of cell immunoprecipitation;
[0029] Figure 5 The therapeutic effect of CCL14 on tendon adhesion in mice;
[0030] Figure 6 The therapeutic effect of CCL14 in tendon adhesion in cynomolgus monkeys; A is the gross score and HE & MASSON staining results; B is the Col1 / 3 Westein Blot detection result; C is the Col1 / 3 immunohistochemical staining result. DETAILED DESCRIPTION
[0031] To make the present invention more clearly understood, preferred embodiments are described in detail below with reference to the accompanying drawings.
[0032] It should be understood that the terms used herein are intended only to describe particular embodiments and are not intended to limit the present invention. Furthermore, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range, within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range are also specifically disclosed. The upper and lower limits of these smaller ranges may independently be included or excluded within the range.
[0033] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.
[0034] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative only.
[0035] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0036] The present invention provides application of chemokine CCL14 or a promoter thereof in preparing a product for resisting tendon adhesion.
[0037] In the present invention, the chemokine CCL14 is derived from mammals, more preferably rodents (such as mice and rats), primates, and humans, and most preferably human-derived chemokine CCL14. Human-derived chemokine CCL14 can meet the requirement of autologous origin, has good biocompatibility, and can reduce the immune response and toxic side effects of the drug in the human body. In some embodiments of the present invention, the human-derived chemokine CCL14 is a commercially available product, which can be purchased from ThermoFisher. In other embodiments of the present invention, it can also be prepared by genetic recombination technology using NCBI reference sequence NM032963.4 (amino acid sequence: NP116739.1) or NCBI reference sequence NM032962.5 (amino acid sequence ID: NP116738.1).
[0038] Optionally, the concentration of the chemokine CCL14 in the product is 15-30 ng / mL; in some specific embodiments of the present invention, the concentration of the chemokine CCL14 in the product is 20 ng / mL.
[0039] Optionally, the product further comprises pharmaceutically acceptable excipients.
[0040] Optionally, the dosage form of the product includes tablets, injections and gels. In some specific embodiments of the present invention, the dosage form of the product is an injection, and the mode of use is subcutaneous injection.
[0041] Unless otherwise specified, the biological materials or reagents used in the following examples of the present invention are all commercially available products and can be purchased through conventional channels.
[0042] Example 1
[0043] 1. Single-cell sequencing
[0044] according to Figure 1 The process shown was used to analyze the results of single-cell sequencing of normal and injured tendon peritendons. Single-cell tissues were obtained from Shanghai Sixth People's Hospital, where normal tendon peritendon tissues were derived from normal tendons of amputees, while injured tendon peritendon tissues were obtained from multiple surgeries after hand trauma. All patients were collected from the Department of Orthopedics of Shanghai Sixth People's Hospital. This study has been approved by the Ethics Committee of Shanghai Sixth People's Hospital, and all patients signed informed consent forms, numbered 2019-KY-047 (K). All specimens were submitted to Beijing Xunyin Biotechnology Co., Ltd. for single-cell sequencing. The tissues were washed with PBS and then cut into 1mm 3The samples were digested with DMEM containing type I collagenase (3 mg / mL) and DNase (0.25 mg / mL) at 37°C for 2 hours, and then filtered through a 100 μm cell strainer. Finally, the cells were centrifuged and collected for scRNA-seq. The scRNA-seq results are shown in Figure 2. Figure 2 As shown, CCL14 is highly expressed in pericytes of injured tendons.
[0045] The results of the single-cell sequencing data were further verified by immunofluorescence co-staining. The process is as follows:
[0046] The obtained samples were fixed with 4% paraformaldehyde for 24 hours, then dehydrated with 30% sucrose solution, embedded in OCT and frozen sectioned. The sections were hydrated in PBS containing 0.3% TritonX-100 (Sigma, 9036-19-5) and then blocked with 5% serum. The primary antibodies CCL14 (NOVUS, NBP2-15760) and CD31 (Abcam, 76533) were incubated overnight at 4°C. After washing with PBS, the secondary antibodies were added and incubated at room temperature for 1 hour. Finally, the sections were covered with anti-fluorescence quenching mounting medium containing DAPI (Beyotime, P0131). The images were taken with a Zeiss 780 confocal microscope, and the positive cells were quantified using ImageJ software. The immunofluorescence co-staining results are shown in Figure 2. Figure 3 As shown, 3D, 14D and over3M represent injured tendon pericytes 3 days, 14 days and more than 3 months after injury, respectively. It can be seen that CCL14 is highly expressed in injured tendon pericytes, and the longer the injury time, the higher the CCL14 expression.
[0047] 2. Binding ability of CCL14 factor to CCR1 receptor and its species conservation test
[0048] 2.1 The amino acid sequences and structural models of CCL14 (NCBI gene ID: 6358) and CCR1 (NCBI gene ID: 1230) were downloaded from NCBI. Molecular simulations were performed using Discovery Studio to verify the high binding ability of human CCL14 and CCR1.
[0049] The results are as follows Figure 4 As shown in A, it can be seen that human CCL14 and CCR1 have a higher binding ability.
[0050] 2.2 FITC fluorescent label was coupled to CCL14, and the conserved binding between human CCL14 and CCR1 was demonstrated through cell immunofluorescence and cell immunoprecipitation. The process is as follows:
[0051] Dissolve FITC powder in dimethyl sulfoxide (DMSO). Mix the FITC solution with CCL14 protein and bind FITC to CCL14 via isocyanation. Stop the reaction with glycine and remove unreacted FITC using column chromatography. Fluorescence microscopy is used to determine if FITC binds to CCL14.
[0052] The macrophages used in immunofluorescence and immunoprecipitation were derived from primary mouse giant cells, obtained by induction with ascites starch injection, and cultured in DMEM supplemented with 5% fetal bovine serum at 37°C and 5% CO2. For immunofluorescence, 10 μL of FITC-conjugated CCL14 protein was added to a 24-well plate at 80% cell density at a concentration of 10 ng / well. After 5 minutes, the cells were fixed with 4% paraformaldehyde and blocked with 5% serum. The primary antibody, CCR1 (NOVUS, NB100-56334), was incubated overnight at 4°C. Subsequently, the secondary antibody was added to the 24-well plate and incubated at room temperature for 1 hour. Finally, the sections were covered with anti-fluorescence quenching mounting medium containing DAPI (Beyotime, P0131), and images were captured using a Zeiss 780 confocal microscope.
[0053] Cell immunoprecipitation: CCL14 protein was added to a 24-well plate at a concentration of 10 ng / mL at 10 μL per well with a cell density of 80%. After 5 minutes, the cells were washed with pre-cooled PBS and pre-cooled RIPA Bufer (1 ml / 10 μL) was added. 7 Cells were scraped from the culture dish with a cell scraper and centrifuged for 15 minutes. The supernatant was transferred to a new centrifuge tube, and the cell lysate was mixed with agarose beads bound to CCR1 antibody (Abcam, 276049). After incubation for a period of time, the immune complex was collected by centrifugation, and the immune complex was subjected to SDS-PAGE electrophoresis and then transferred to a PVDF membrane. The membrane was blocked with BSA for 1 hour, and then the primary antibody CCL14 (NOVUS, NBP2-15760) and the primary antibody CCR1 were added and incubated at 4°C overnight. After washing, the secondary antibody was incubated at room temperature for 30 minutes, and the membrane was imaged using ECL (Beyotime, P0018S) developer.
[0054] The results are as follows Figure 4 As shown in Figures B and C, it can be seen that the binding between human CCL14 and CCR1 is conserved among species.
[0055] 3. Test the therapeutic effect of CCL14 in tendon adhesions in mice by subcutaneous injection:
[0056] In this experiment, C57BL / 6J mice were purchased from the Animal Laboratory of Shanghai Sixth People's Hospital and used as wild-type mice (WT); LysM-Cre mice and Rosa26-CCR1-Loxp mice were purchased from Jicui Pharmaceutical Company and used to construct CCL14 receptor CCR1 knockout mice (LysM-Cre mice). CCRI ) and was constructed by the company to obtain homozygous LysM-Cre: Rosa26-CCR1-Loxp mice, which are CCL14 factor receptor CCR1 knockout mice (abbreviated as LysM CCRI This experiment was approved by the Ethics Committee of Shanghai Sixth People's Hospital.
[0057] CCL14 receptor CCR1 knockout mice (LySM CCRI ) and C57BL / 6J mice were divided into the following groups: WT+Veh group, WT+CCL14 group and LySM CCR1 +CCL14 group. Among them, C57BL6J mice in the WT+Veh group and the WT+CCL14 group underwent transection and Kessel suture of the deep flexor tendon of the digitorum to establish a tendon adhesion model. Then, the WT+CCL14 group was injected with CCL14 drug (Human HCC-1 (CCL14) Recombinant Ptotein, purchased from ThermoFisher, 300-38-100UG, dissolved in sterile PBS when used, ready for use). The injection method was local injection at the modeling site, the drug concentration was 20ng / mL, and 10μL was injected each time; the WT+Veh group mice were injected with 10μL of Vehicle (i.e., sterile PBS) to serve as the control group. LySM CCR1 +CCL14 group selected gene mice (LySM CCR1 ) The deep flexor tendon of the digitorum was cut and Kessel sutured to construct a tendon adhesion model. Then, CCL14 drug was injected locally at the modeling site with a drug concentration of 20 ng / mL and 10 μL was injected each time.
[0058] The flexor digitorum profundus tendon was sectioned and Kessel sutured as follows: After anesthetizing the mouse, the skin of the right hind paw was disinfected with 75% ethanol. The flexor digitorum profundus tendon was exposed through a posterior midline incision and transected laterally. The tendon was then repaired using a modified Kessel pattern to create a tendon adhesion model. The wound was closed with 5-0 sutures.
[0059] After modeling, all mice received the same treatment and were allowed to move freely in their cages. 14 days after drug injection, mice were euthanized by carbon dioxide inhalation and then perfused with 4% paraformaldehyde through the femoral artery for 5 minutes. The right foot was then removed and immersed in 4% paraformaldehyde for 24 hours and decalcified in 0.5M ethylenediaminetetraacetic acid (VWR, 0105) (pH 7.5) for one week. Finally, the specimens were dehydrated with 30% sucrose (Sigma-Aldrich, S9378) at 4°C and embedded in Optimal Cutting Temperature Compound (OCT) at -80°C. Gross scoring, HE & Masson staining, and immunofluorescence staining of αSMA were performed on mice in each group to verify their phenotypes.
[0060] Gross Movement Scoring (ROM): The hind limbs of mice in each group were amputated to the knee, and the skin of the ankles was removed. First, the tibia was fixed with a clamp, and then the FDL (Flexor Digitorum Longus) was pulled with a 19-gram weight to measure the change in the metatarsophalangeal joint angle.
[0061] HE & Masson staining: Tendons were stained using a hematoxylin and eosin (HE) staining kit (Beyotime, C0105S) and a Masson trichrome staining kit (Beyotime, C0189s).
[0062] Immunofluorescence staining of αSMA: Before blocking nonspecific sites with 5% serum, the sections were hydrated with PBS (Solarbio, SL050), and then incubated with the primary antibody αSMA (Cell Signaling, 19245) at 4°C overnight. The secondary antibody was then incubated with the sections at room temperature for 1 hour. Finally, the sections were mounted with anti-fluorescence quenching mounting medium containing DAPI (Beyotime, P0131). Images were taken with a Zeiss 780 confocal microscope, and positive cells were quantified using ImageJ software.
[0063] Test results such as Figure 5As shown, gross scoring results showed that, compared with the WT+Veh group, CCL14 injection increased joint mobility in mice; however, CCL14 receptor knockout reduced joint mobility in mice compared with the WT+CCL14 group. HE and MASSON staining results showed that, compared with the WT+Veh group, CCL14 injection reduced adhesion between tendons and surrounding tissues in mice; however, CCL14 receptor knockout increased adhesion between tendons and surrounding tissues in mice compared with the WT+CCL14 group. (T represents tendon, M represents muscle, black dashed lines represent tendon adhesions, and yellow dashed lines represent tendon non-adhesions.) αSMA staining results showed that, compared with the WT+Veh group, CCL14 injection reduced myofibroblast activation in mice; however, CCL14 receptor knockout increased myofibroblast activation in mice compared with the WT+CCL14 group.
[0064] 4. Test the therapeutic effect of CCL14 in tendon adhesions in a primate model by subcutaneous injection:
[0065] This study used cynomolgus macaques as a primate model. Cynomolgus macaques were obtained from WuXi AppTec (Chengdu). This study was reviewed and approved by the Animal Ethics and Use Committee of Chengdu WuXi AppTec Pharmaceutical Development Co., Ltd., number CD01-0005-2022.
[0066] Cynomolgus monkeys were randomly divided into the following groups: Normal, Vehicle, and CCL14. The Normal group received no treatment. Both the Vehicle and CCL14 groups underwent transection of the flexor digitorum profundus tendon and Kessel suture to establish a tendon adhesion model. Sterile PBS (50 μL per injection) and CCL14 (20 ng / L, 50 μL per injection, prepared in sterile PBS, used immediately) were subcutaneously injected into the modeling site on the day of surgery and every 7 days after surgery in both the Vehicle and CCL14 groups.
[0067] The specific operation of the flexor digitorum profundus tendon cutting and Kessel suture is as follows: after general anesthesia, the right paw is disinfected in a standardized manner, the flexor digitorum profundus tendons of the middle and ring fingers of the right paw are exposed, transversely cut, and then the severed tendons are sutured using the modified Kessler suture method. After suturing the wound, the right paw is immobilized to the upper arm with plaster.
[0068] The macaques were housed normally during the experiment. 21 days after the surgery, all macaques underwent amputation of the middle and ring fingers and stump repair under general anesthesia. After sample processing, gross scoring, tissue section HE & MASSON staining, Col1 / 3 immunohistochemical staining, and Col1 / 3 Westein Blot were performed.
[0069] For the detailed procedures of gross scoring and HE & MASSON staining, please refer to "3. Testing the efficacy of CCL14 in tendon adhesions in mice by subcutaneous injection";
[0070] Immunohistochemical staining of Col1 / 3: The middle and ring fingers were immersed in 4% paraformaldehyde for 24 hours and decalcified in 0.5 M ethylenediaminetetraacetic acid (VWR, 0105) (pH 7.5) for three weeks. Finally, the specimens were dehydrated with 30% sucrose (Sigma-Aldrich, S9378) at 4°C, embedded in optimal cutting temperature compound (OCT) at -80°C, and sectioned. The sections were hydrated in PBS containing 0.3% Triton X-100 (Sigma, 9036-19-5) to permeabilize the membrane, and then 3% hydrogen peroxide was added to block endogenous peroxides. The sections were then blocked with 5% serum and incubated overnight at 4°C with the primary antibodies Collagen 1 (Abcam, 138492) and Collagen 3 (Abcam, 184993). After washing with PBS, the secondary antibodies were added and incubated at room temperature for 1 hour. Finally, DAB (ThermoFisher, 34002) was used for color development. After hematoxylin nuclear staining, the sections were dehydrated with xylene and mounted with neutral resin. Images were captured with a Leica DM6 microscope, and the positive areas were quantified using ImageJ software.
[0071] Col1 / 3 Westein Blot: Peritendon tissue was collected, pre-cooled RIPA buffer was added, and the mixture was shaken and ground. The tube was centrifuged for 15 minutes. The supernatant was transferred to a new centrifuge tube, and loading buffer was added and boiled. SDS-PAGE electrophoresis was then performed, and the membrane was transferred to a PVDF membrane. The membrane was blocked with BSA for 1 hour, and then the primary antibodies Collagen 1 (Abcam, 138492) and Collagen 3 (Abcam, 184993) were added and incubated at 4°C overnight. After washing, the membrane was incubated with the secondary antibody at room temperature for 30 minutes. The membrane was imaged using ECL (Beyotime, P0018S) developer, and the grayscale values of the bands were calculated using ImageJ software.
[0072] The results are as follows Figure 6 As shown, it can be seen that the overall score test results ( Figure 6 A) shows that, compared with the Vehicle group, the active gap of the deep flexor tendon of the cynomolgus monkey increased after injection of CCL14, approaching that of the Normal group; HE & MASSON staining results ( Figure 6Figure A) shows that the adhesion between the tendon and surrounding tissues of cynomolgus monkeys was reduced after CCL14 injection compared with the vehicle group. T represents tendon, M represents muscle, S represents skin, and B represents bone. The black dotted line represents the part where the tendon is adhered to the surrounding tissues, and the yellow dotted line represents the part where the tendon is not adhered to the surrounding tissues. Results of Westein Blot ( Figure 6 B) shows that the protein expression of Col1 / 3 in the local adhesion tissue of cynomolgus monkeys was significantly reduced after CCL14 injection compared with the Vehicle group. Figure 6 In Figure C), T represents tendon, AD represents adhesion tissue, the black dotted line represents the part where the tendon is adhered to the surrounding tissue, and the yellow dotted line represents the part where the tendon is not adhered to the surrounding tissue. This result indicates that compared with the Vehicle group, after injection of CCL14, the degree of adhesion between the tendon and the surrounding tissue of the cynomolgus monkey was reduced, and the protein expression of Col1 / 3 was decreased.
[0073] The above experimental results indicate that plasma chemokine CCL14 can prevent and treat tendon adhesions and can be used to prepare related products for preventing and treating tendon adhesions after tendon injury.
[0074] The above description is only a preferred embodiment of the present invention and does not constitute any formal or substantial limitation to the present invention. It should be noted that ordinary technicians in this technical field can make several improvements and supplements without departing from the present invention, and these improvements and supplements should also be regarded as the scope of protection of the present invention.
Claims
1. Application of chemokine CCL14 or its promoter in the preparation of products against tendon adhesion.
2. The use according to claim 1, characterized in that The chemokine CCL14 is a human chemokine CCL14; The CCL14 promoter refers to a substance that can increase the activity and / or content of the CCL14 gene or its protein in vivo or in vitro, or a substance that can promote the binding of CCL14 to the CCR1, CCR3 or CCR5 chemokine receptors; the substance is a synthetic or natural compound, protein, or nucleotide.
3. The use according to claim 1, characterized in that The CCL14 promoter is selected from the group consisting of: a small molecule compound, a CCL14 gene delivery vector, and a CCL14 protein agonist, or a combination thereof.
4. The use according to claim 3, characterized in that The CCL14 protein agonist includes a full-length CCL14 protein, a recombinant CCL14 protein or an active fragment of a CCL14 protein.
5. A product for resisting tendon adhesion, characterized in that: The active ingredient in the product is chemokine CCL14 or its promoter.
6. The product according to claim 5, characterized in that The concentration of chemokine CCL14 in the product is 15-30 ng / mL.
7. The product according to claim 5, characterized in that The product also includes pharmaceutically acceptable excipients.
8. The product according to claim 5, characterized in that The dosage forms of the product include tablets, injections and gels.
9. The product according to claim 8, characterized in that The dosage form of the product is injection and the method of use is subcutaneous injection.