Application of FAP protein function inhibitor in preparation of medicine for inhibiting formation of fibrous scar

Abstract

The invention provides application of an FAP protein function inhibitor in preparation of a medicine for inhibiting formation of fibrous scars. The invention finds that the formation of the fibrous scar can be obviously inhibited by knocking out the Fap gene and inhibiting the activity of the FAP protein, so that the FAP protein function inhibitor can be used for preparing the medicine for inhibiting the formation of the fibrous scar. Besides, the invention also finds that the expression levels of the Fap gene and the FAP protein are also increased along with the development of the fibroscar disease course, which indicates that the FAP can be used as a marker of the fibroscar, that is, the FAP can be used as a target for screening products for inhibiting the formation of the fibroscar, and a detection reagent of the FAP can be used for preparing fibroscar diagnosis products and / or prognosis evaluation products.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biomedical technology. More particularly, it relates to the use of a FAP protein function inhibitor in the preparation of a medicament for inhibiting fibrous scar formation. BACKGROUND

[0002] Fibrous scar is a kind of fibrotic tissue formed by a group of proliferative fibroblasts gathering, proliferating and synthesizing and secreting a large amount of fibrous extracellular matrix, which often occurs in the development process of diseases such as central nervous system injury, has the characteristics of mature and irreversibility, and after the formation of fibrous scar, the regenerated axons of the central nervous system cannot penetrate this dense barrier structure, and a large amount of fibrous extracellular matrix will further hinder the regeneration of axons as inhibitory molecules, ultimately leading to the difficulty of recovery of nerve function after central nervous system injury.

[0003] Recent studies have shown that inhibiting the activation of fibroblasts can effectively reduce the formation of fibrous scar, so inflammation, oxidative stress, transforming growth factor β1 (TGF-β1), periostin and Wnt / β-catenin signaling pathway, which are closely related to the activation of fibroblasts, have become research hotspots. At present, TGF-β1 not only can activate fibroblasts and induce them to secrete a large amount of fibrous extracellular matrix, but also can further inhibit the degradation of fibrous extracellular matrix, which is very suitable as a target for inhibiting fibrous scar formation. However, TGF-β1 is also involved in other biological processes such as cell proliferation and differentiation, and blocking its signal transduction will also affect the normal physiological needs of the body, resulting in great side effects and making it difficult to achieve clinical transformation. Therefore, it is imperative to find a safe and effective new target for inhibiting fibrous scar formation in clinical practice. SUMMARY

[0004] The present application aims to provide the use of a fibroblast activation protein (FAP) function inhibitor in the preparation of a medicament for inhibiting fibrous scar formation. The present application found that inhibiting the function of FAP as a target can effectively inhibit the formation of fibrous scar; and FAP has no expression or low expression in normal tissues, and inhibiting its function will not cause significant impact on normal tissues, and has no obvious side effects, so it can be used as a safe and effective new target for inhibiting fibrous scar formation in clinical practice.

[0005] The second object of the present application is to provide the use of a detection reagent for FAP in the preparation of a fibrous scar diagnosis product and / or a prognosis evaluation product.

[0006] A third object of the present application is to provide the use of FAP as a target in screening products for inhibiting fibrous scar formation.

[0007] A fourth object of the present application is to provide a method for screening products for inhibiting fibrous scar formation.

[0008] The above objects of the present application are achieved by the following technical solutions.

[0009] The present application finds that knocking out Fap gene or inhibiting FAP protein activity can significantly inhibit the formation of fibrous scar. Therefore, the use of FAP protein function inhibitors in the preparation of drugs for inhibiting fibrous scar formation should be within the scope of protection of the present application.

[0010] Preferably, the FAP protein function inhibitor is a preparation having one or more of the effects of knocking out Fap gene, inhibiting Fap gene expression, inhibiting FAP protein activity, and inhibiting FAP protein expression.

[0011] Preferably, the FAP protein function inhibitor is an expression cassette, a recombinant vector, or a recombinant microorganism having FAP protein function inhibiting effect.

[0012] Preferably, the FAP protein function inhibitor is fibroblast activation protein (FAPi).

[0013] Preferably, the fibrous scar is a fibrous scar formed after central nervous system injury, such as a fibrous scar formed after spinal cord injury.

[0014] In addition, the present application also finds that as the course of fibrous scar develops, the expression level of Fap gene and FAP protein also increases, indicating that FAP can be used as a marker for fibrous scar. Therefore, the use of FAP detection reagents in the preparation of fibrous scar diagnostic products and / or prognosis evaluation products, and the use of FAP as a target in screening products for inhibiting fibrous scar formation should be within the scope of protection of the present application.

[0015] Preferably, the expression amount of FAP in the spinal cord tissue sample and / or cerebrospinal fluid sample of the fibrous scar patient is significantly higher than that in the normal spinal cord tissue sample and / or cerebrospinal fluid sample.

[0016] Preferably, the FAP is one or more of Fap peptide, FAP protein, Fap mRNA, Fap mRNA recombinant vector, and Fap mRNA recombinant cell.

[0017] Preferably, the product is a product having one or more of the effects of knocking out Fap gene, inhibiting Fap gene expression, inhibiting FAP protein activity, and inhibiting FAP protein expression.

[0018] Based on this, the application further provides a method for screening a product for inhibiting fibrous scar formation, specifically: screening by using one or more of the following biological materials:

[0019] (1) a nucleic acid molecule encoding FAP protein;

[0020] (2) an expression cassette containing the nucleic acid molecule in (1);

[0021] (3) a recombinant vector containing the nucleic acid molecule in (1) and / or containing the expression cassette in (2);

[0022] (4) a recombinant microorganism containing the nucleic acid molecule in (1), the expression cassette in (2) and / or the recombinant vector in (3).

[0023] The application has the following beneficial effects:

[0024] 1. The application finds that knocking out Fap gene and inhibiting FAP protein activity can significantly inhibit the formation of fibrous scar, so that FAP protein function inhibitors can be used for preparing a medicine for inhibiting fibrous scar formation; and FAP has no expression or low expression in normal tissues, and inhibiting its function will not cause obvious influence on normal tissues, and has no obvious side effects, so it can be used as a safe and effective new target for inhibiting fibrous scar formation in clinic.

[0025] 2. The application finds that with the development of fibrous scar course, the expression level of Fap gene and FAP protein is also increased, indicating that FAP can be used as a marker of fibrous scar, that is, it can be used as a target for screening a product for inhibiting fibrous scar formation, and its detection reagent can be used for preparing a fibrous scar diagnosis product and / or a prognosis evaluation product. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 A schematic diagram of the method for constructing a fibrous scar model.

[0027] Figure 2 The expression of Col1a1, Col1a2 and Fap gene in Example 1.

[0028] Figure 3 The expression of Collagen I and FAP protein in Example 1.

[0029] Figure 4 A in the above is a result diagram of agarose gel electrophoresis in Example 2; Figure 4 B in the above is the expression of Fap gene in Example 2.

[0030] Figure 5 The expression of FAP protein in Example 2.

[0031] Figure 6In this context, A represents the tissue structure of the mouse spinal cord in Example 2; Figure 6 In this context, B represents the area measurement result of collagen deposition in the tissue structure of Example 2; Figure 6 In the figure, C represents the expression of the fibrous scar-related gene Col1a1 in Example 2; Figure 6 In the figure, D represents the expression of the fibrous scar-related gene Col1a2 in Example 2.

[0032] Figure 7 Example 3: Activity of FAP protein in mouse cerebrospinal fluid.

[0033] Figure 8 In this context, A represents the tissue structure of the mouse spinal cord in Example 3; Figure 8 In this context, B represents the area measurement result of collagen deposition in the tissue structure of Example 3; Figure 8 In the figure, C represents the expression of the fibrous scar-related gene Col1a1 in Example 3; Figure 8 In the figure, D represents the expression of the fibrous scar-related gene Col1a2 in Example 3. Detailed Implementation

[0034] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0035] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0036] Example 1: FAP can serve as a marker for fibrous scars.

[0037] Eight-week-old female C57 mice were intraperitoneally injected with 20 μL / g 小鼠体重 After anesthetizing with 1.25% (w / v) aphthylazine (Sigma-Aldrich, T48402), the skin on the back of the mouse spinal cord from T9 to T10 was prepared. The skin was then incised, and a 1 mm long lamina of the T9-T10 segment was removed. The corresponding segment of spinal cord tissue was then completely clamped with microforceps for 3 seconds, released after 3 seconds, and sutured to achieve hemostasis. This completed the construction of the fibrous scar model caused by spinal cord injury. A schematic diagram of the method for constructing the fibrous scar model is shown below. Figure 1 As shown in the figure. Mice in which only the lamina was removed but not clamped with microforceps were used as the sham-operated group.

[0038] Post-surgery, the mice were transferred to a 37°C heating pad to maintain stable body temperature and await recovery. Then, 100 μL / 20g of medication was administered subcutaneously to each mouse based on its individual condition. 小鼠体重 physiological saline and / or 5 mg / kg 小鼠体重Ketoprofen (Solarbio, K7980) (for replenishing the loss of body fluid in mice due to modeling and relieving postoperative pain in mice, respectively), and the mice were actively urinated during the subsequent daily feeding process.

[0039] On the 3rd, 7th and 14th day after the completion of the fibrous scar model, the mice were randomly taken, anesthetized and heart perfused, and then the spinal cord tissue of 1 cm long from the 8th to the 11th thoracic segment was taken out. The spinal cord tissue was homogenized and the mRNA and protein were extracted for testing the relationship between Fap gene and fibrous scar, and the relationship between FAP protein and fibrous scar, as follows:

[0040] (1) Relationship between Fap gene and fibrous scar

[0041] The mRNA of the aforementioned extracted spinal cord tissue was reverse transcribed to obtain cDNA, and then the Roche LC480 system was used for fluorescence quantitative PCR detection (the primers are shown in Table 1). The expression of Col1a1, Col1a2 and Fap gene was determined by △△Ct method, and the corresponding best fitting curve was obtained by using a simple linear regression model (the relationship between Fap gene and Col1a1 gene: the equation is Y1=1.049X1+0.09469, R 2 =0.8634; the relationship between Fap gene and Col1a2 gene: the equation is Y2=1.146X1+0.4887, R 2 =0.7511. Wherein, X1 is the expression level of Fap gene, Y1 is the expression level of Col1a1 gene, and Y2 is the expression level of Col1a2 gene).

[0042] The results are shown in Figure 2 . It can be seen that the expression level of Fap gene is positively correlated with the expression level of fibrous scar related genes Col1a1 and Col1a2.

[0043] With the increase of the expression level of fibrous scar related genes Col1a1 and Col1a2, the expression level of Fap gene also increases.

[0044] Table 1

[0045]

[0046] (2) Relationship between FAP protein and fibrous scar

[0047] The extracted spinal cord tissue protein was subjected to SDS-PAGE electrophoresis, PVDF membrane constant current transfer, 5wt% skim milk blocking, TBS / T washing three times, FAP protein primary antibody (Abeam, ab207178) and fibrous scar related protein Collagen I primary antibody (Abeam, ab270993) 4°C incubation for 16h, secondary antibody (Cell Signaling Technology, 7074) 25°C incubation for 1.5h after washing, development, Image J software data analysis, and the corresponding best fitting curve was obtained using a simple linear regression model (FAP protein and Collagen I protein relationship: equation Y3=1.049X2+0.09469, R 2 =0.6424, wherein X2 is the expression level of FAP protein, Y3 is the expression level of Collagen I protein).

[0048] The results are shown in Figure 3 . It can be seen that the expression level of FAP protein is positively correlated with the expression level of fibrous scar related protein Collagen I.

[0049] In summary, with the development of fibrous scar course, the expression level of Fap gene and FAP protein also increases, indicating that FAP can be used as a marker of fibrous scar, i.e. it can be used as a target for screening products that inhibit fibrous scar formation, and its detection reagent can be used to prepare fibrous scar diagnostic products and / or prognosis evaluation products.

[0050] Example 2 Inhibition of fibrous scar formation by knocking out Fap gene

[0051] I. Verification of Fap knockout mice and construction of fibrous scar model

[0052] The tail end of 0.4cm of Fap knockout 3-week-old female C57 mice (The Jackson Laboratory, #024288, donated by Professor Yue Rui's research group, School of Life Sciences and Technology, Shanghai Jiao Tong University) was taken, and the DNA of the mouse tail tissue was extracted using a mouse tail identification kit (Yuanjing Biology, YK-MG-100) (using the DNA of the tail tissue of wild-type 3-week-old female C57 mice as a positive control, and DEPC water as a negative control) and subjected to PCR amplification, followed by agarose gel electrophoresis. The results are shown in Figure 4 A of

[0053] Fap knockout and Fap non-knockout 8-week-old female C57 mice were taken, and 20μL / g 小鼠体重After anesthesia with 1.25% (w / v) avertin (Sigma-Aldrich, T48402), the dorsal skin of the mouse at the thoracic 9 to thoracic 10 segment of the spinal cord was prepared, the skin was incised and the lamina of the thoracic 9 to thoracic 10 segment of 1 mm long was removed, then the spinal cord tissue of the mouse at this segment was completely clamped with microforceps for 3 seconds, and then released, fully hemostatic and sutured, that is, the construction of the fibrous scar model was completed.

[0054] After the operation, the mouse was transferred to a heating pad at 37°C to maintain the stability of the body temperature of the mouse and wait for it to wake up, and then 100 μL / 20 g 小鼠体重 of normal saline and / or 5 mg / kg 小鼠体重 of ketoprofen (Solarbio, K7980) (respectively used for supplementing the loss of body fluid of the mouse caused by modeling and relieving the postoperative pain of the mouse) were subcutaneously administered according to the specific circumstances of each mouse, and the mouse was actively urinated during the subsequent daily feeding process.

[0055] On the 14th day after the construction of the fibrous scar model, the mouse was randomly taken, anesthetized and heart perfused, and then the spinal cord tissue of the mouse at the thoracic 8 to thoracic 11 segment of 1 cm long was taken out, the spinal cord tissue was homogenized and the mRNA and protein thereof were extracted, which were respectively used for detecting the expression level of Fap gene and FAP protein, and the specific steps were as follows:

[0056] (1) Expression level of Fap gene

[0057] The mRNA of the spinal cord tissue extracted as described above was subjected to reverse transcription to obtain cDNA, and then fluorescence quantitative PCR detection was performed by using the LC480 system of Roche company (the primers are shown in Table 1), and the expression of Fap gene was determined by △△Ct method.

[0058] The results are shown in B of FIG. Figure 4 It can be seen that, compared with the Fap non-knockout group, the expression level of Fap gene of the Fap knockout group mouse was significantly reduced.

[0059] (2) Expression level of FAP protein

[0060] The protein of the spinal cord tissue extracted as described above was subjected to SDS-PAGE electrophoresis, PVDF membrane constant current transfer, 5wt% skimmed milk blocking, TBS / T washing three times, FAP protein primary antibody (Abeam, ab207178) 4°C incubation for 16h, secondary antibody (Cell Signaling Technology, 7074) 25°C incubation for 1.5h after washing, development, and Image J software data analysis.

[0061] The results are shown in FIG. Figure 5As shown in the figure, the FAP protein expression level in the Fap knockout group mice was significantly reduced compared to the non-knockout group.

[0062] In conclusion, the genotype of the Fap knockout mice is indeed Fap. - / - .

[0063] II. Inhibitory effect of Fap gene knockout on fibrous scar formation

[0064] (1) Pathological condition of fibrous scar tissue

[0065] On day 14 after the fibrous scar model was constructed, mice were randomly selected, euthanized, and their spinal cords were perfused with 4% (w / v) paraformaldehyde. Tissue sections were prepared, and the tissue structure of the mouse spinal cords was observed by Masson staining. The area of ​​collagen deposition in the tissue structure was measured using ImageJ software.

[0066] The histological structure of the mouse spinal cord is as follows Figure 6 As shown in Figure A, the area measurement results of collagen deposition in the tissue structure are as follows: Figure 6 As shown in B, compared to the non-Fap knockout group, the collagen deposition area in the tissue structure of the Fap knockout group mice was significantly reduced, indicating that Fap knockout can significantly inhibit the formation of fibrous scars.

[0067] (2) Expression levels of genes related to fibrous scars

[0068] On day 14 after the fibrous scar model was constructed, mice were randomly selected. After anesthesia and cardiac perfusion, a 1 cm long section of spinal cord tissue from T8 to T11 was taken from the mice. The spinal cord tissue was homogenized and its mRNA was extracted. cDNA was obtained by reverse transcription and then detected by real-time PCR using the Roche LC480 system (primers are shown in Table 1). The expression of fibrous scar-related genes Col1a1 and Col1a2 was determined by ΔΔCt method.

[0069] The expression level of the fibrous scar-associated gene Col1a1 is as follows: Figure 6 As shown in C, the expression level of the fibrous scar-related gene Col1a2 is as follows: Figure 6 As shown in D in the figure, compared with the non-Fap knockout group, the expression level of fibrous scar-related genes in the Fap knockout group mice was significantly reduced, indicating that Fap knockout can significantly inhibit the formation of fibrous scars.

[0070] In summary, knocking out the Fap gene can significantly inhibit the formation of fibrous scars. Therefore, FAP protein function inhibitors can be used to prepare drugs that inhibit the formation of fibrous scars. Moreover, FAP is not expressed or has low expression levels in normal tissues, and inhibiting its function will not have a significant impact on normal tissues and will not have obvious side effects. Therefore, it can serve as a safe and effective new target for inhibiting the formation of fibrous scars in clinical practice.

[0071] Example 3: Inhibition of FAP protein activity on fibrous scar formation

[0072] Eight-week-old female C57 mice were intraperitoneally injected with 20 μL / g 小鼠体重 After anesthetizing with 1.25% (w / v) aphthylazine (Sigma-Aldrich, T48402), the dorsal skin of the spinal cord segment from T9 to T10 in mice was prepared. The skin was then incised, and a 1 mm long lamina of the T9-T10 segment was removed. The spinal cord tissue of this segment was then completely clamped with microforceps for 3 seconds, released after 3 seconds, and sutured to achieve hemostasis, thus completing the construction of the fibrous scar model. Postoperatively, the mice were transferred to a 37°C heating pad to maintain stable body temperature and await awakening. Then, 100 μL / 20g of aphthylazine was administered subcutaneously to each mouse according to its specific condition. 小鼠体重 physiological saline and / or 5 mg / kg 小鼠体重 Ketoprofen (Solarbio, K7980) was used to replenish fluid loss in mice caused by modeling and to relieve postoperative pain in mice, and active urination was induced in mice during subsequent daily feeding.

[0073] I. Inhibitory effect of FAPi on FAP protein activity

[0074] (1) FAPi treatment and cerebrospinal fluid extraction

[0075] Fibrous scar model mice were randomly divided into 7 groups, with mice that underwent only laminectomy but were not clamped with microforceps serving as the sham surgery group. The treatment of the 7 groups of fibrous scar model mice was as follows: (1) 3-day group: Cerebrospinal fluid was extracted from mice on the 3rd day after the fibrous scar model was established. (2) 7-day group: Cerebrospinal fluid was extracted from mice on the 7th day after the fibrous scar model was established. (3) 14-day group: Cerebrospinal fluid was extracted from mice on the 14th day after the fibrous scar model was established. (4) Vehicle group: 100 μL of physiological saline was injected once a day from the 1st to the 14th day after the fibrous scar model was established, and cerebrospinal fluid was extracted from mice on the 14th day. (5) FAPi 0.15 mg / kg group: 0.15 mg / kg was injected once a day from the 1st to the 14th day after the fibrous scar model was established. 小鼠体重FAPi (100 μL in volume) and the cerebrospinal fluid of the mice was extracted on day 14. (7) FAPi 1.50 mg / kg group: 1.50 mg / kg of FAPi was injected once a day from day 1 to day 14 after the construction of the fibrous scar model was completed. 小鼠体重 FAPi (100 μL in volume) and the cerebrospinal fluid of the mice was extracted on day 14. (7) FAPi 1.50 mg / kg group: 1.50 mg / kg of FAPi was injected once a day from day 1 to day 14 after the construction of the fibrous scar model was completed. 小鼠体重 FAPi (100 μL in volume) and the cerebrospinal fluid of the mice was extracted on day 14.

[0076] The method for extracting the cerebrospinal fluid of the mice is as follows: after the mice are anesthetized, the skin and the first layer of muscle on the back of the neck of the mice are cut and pulled to the two sides to move away from the occipital region, then the last thin layer of muscle outside the skull is peeled off and moved to the side under a dissecting microscope to expose the dura mater, then under a micro-manipulator, a glass capillary (which is pre-sucked by a syringe to ensure that the glass capillary is under negative pressure and is not contaminated) is aimed at the back of the head of the mouse, the tip is tilted at an angle of 30° to 45°, and is close to the surface of the dura mater until resistance is felt, and finally the glass capillary is tapped to penetrate the dura mater, and the cerebrospinal fluid is sucked into the glass capillary.

[0077] (2) Activity of FAP protein in the cerebrospinal fluid

[0078] 5 μL of the cerebrospinal fluid is diluted with 45 μL of PBS buffer solution, mixed with 50 μL of Z-glycyl proline-4-methyl-7-coumarin (Z-Gly-Pro-AMC) (Bachem, 4002518.0250) with a concentration of 50 μM to form a reaction system of 100 μL, and then incubated at 25°C for 10 min. After excitation by a 380 nm laser, the intensity of the emitted light is detected at 460 nm to determine the activity of the FAP protein.

[0079] The results are shown in Table 1. Figure 7 As can be seen, the activity of the FAP protein in the cerebrospinal fluid of the fibrous scar model mice is higher than that of the sham operation group, and gradually increases with the development of the fibrous scar, while the injection of FAPi can effectively inhibit the increase of the activity of the FAP protein, and the effect of the FAPi 0.5 mg / kg group is the most significant. It is indicated that FAPi can inhibit the formation of fibrous scar by inhibiting the activity of FAP protein.

[0080] II. Inhibitory effect of FAPi on the formation of fibrous scar

[0081] (1) Pathological condition of fibrous scar tissue

[0082] The fibrous scar model mice were randomly divided into two groups, and 0.50 mg / kg of FAPi was injected once a day from day 1 to day 14 after the construction of the fibrous scar model was completed.小鼠体重 The mice were euthanized on day 14, the spinal cord of the mice was perfused with 4% (w / v) paraformaldehyde, and the mice spinal cord was sectioned and observed for tissue structure by Masson staining, and the area of collagen deposition in the tissue structure was measured by Image J software.

[0083] The tissue structure of the mouse spinal cord is shown in A of Figure 8 The area of collagen deposition in the tissue structure is shown in B of Figure 8 It can be seen that the area of collagen deposition in the tissue structure of the FAPi group mice is significantly reduced compared with the Vehicle group, indicating that FAPi can inhibit the formation of fibrous scar by inhibiting the activity of FAP protein.

[0084] (2) Expression level of fibrous scar related genes

[0085] On day 14 after the fibrous scar model was constructed, the mice were randomly selected, anesthetized and heart perfused, and the spinal cord tissue of 1 cm long from the 8th to 11th thoracic segment of the mice was taken out, the spinal cord tissue was homogenized and the mRNA was extracted, the cDNA was obtained by reverse transcription, and the fluorescence quantitative PCR detection was performed by using the LC480 system of Roche company (the primers are shown in Table 1), and the expression of fibrous scar related genes Col1a1 and Col1a2 was determined by △△Ct method.

[0086] The expression level of fibrous scar related gene Col1a1 is shown in C of Figure 8 The expression level of fibrous scar related gene Col1a2 of the mice is shown in D of Figure 8 It can be seen that the expression level of fibrous scar related genes of the FAPi group mice is significantly reduced compared with the Vehicle group, indicating that FAPi can inhibit the formation of fibrous scar by inhibiting the activity of FAP protein.

[0087] In summary, inhibiting the activity of FAP protein can significantly inhibit the formation of fibrous scar, and therefore the FAP protein function inhibitor can be used for preparing a drug for inhibiting the formation of fibrous scar; and FAP has no expression or low expression in normal tissues, and inhibiting its function will not cause obvious impact on normal tissues, and has no obvious side effects, and therefore it can be used as a safe and effective new target for inhibiting the formation of fibrous scar in clinic.

[0088] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application should be equivalent replacement methods, and all are included in the protection scope of the present application.

Claims

1. Use of a FAP protein function inhibitor in the preparation of a drug for inhibiting fibrous scar formation.

2. Use according to claim 1, characterized in that, The FAP protein function inhibitor is a preparation with one or several effects of knocking out Fap gene, inhibiting Fap gene expression, inhibiting FAP protein activity, and inhibiting FAP protein expression.

3. Use according to claim 1, characterized in that, The FAP protein function inhibitor is an expression cassette, a recombinant vector, or a recombinant microorganism with FAP protein function inhibiting effect.

4. The use according to claim 1, characterized in that, The FAP protein function inhibitor is FAPi.

5. The use according to claim 1, characterized in that, The fibrous scar is a fibrous scar formed after central nervous system injury.

6. Use of a detection reagent for FAP in the preparation of a fibrous scar diagnosis product and / or a prognosis evaluation product.

7. Use according to claim 6, characterized in that, The expression amount of FAP in the spinal cord tissue sample and / or the cerebrospinal fluid sample of the fibrous scar patient is significantly higher than that in the normal spinal cord tissue sample and / or the cerebrospinal fluid sample.

8. Use according to claim 6, characterized in that, The FAP is one or several of Fap peptide, FAP protein, Fap mRNA, Fap mRNA recombinant vector, and Fap mRNA recombinant cell.

9. Use of FAP as a target in screening a product for inhibiting fibrous scar formation.

10. A method of screening for a product that inhibits fibrotic scarring, characterized by, The screening is performed using one or several of the following biomaterials: (1) a nucleic acid molecule encoding FAP protein; (2) an expression cassette containing the nucleic acid molecule of (1); (3) a recombinant vector containing the nucleic acid molecule of (1) and / or containing the expression cassette of (2); (4) a recombinant microorganism containing the nucleic acid molecule of (1), the expression cassette of (2), and / or the recombinant vector of (3).

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