Application of GCA-NAb in preparation of medicine for treating osteoarthritis

Abstract

The invention discloses an application of GCA-NAb in preparation of a medicine for treating osteoarthritis. The prevention and treatment effect of the GCA neutralizing antibody GCA-Nab on osteoarthritis is found for the first time, and a research basis and a theoretical basis are provided for developing medicines for treating osteoarthritis. Experiments prove that the GCA-Nab can effectively block the regulation effect of GCA on chondrocyte metabolism, so that the loss of articular cartilage is obviously improved, and the occurrence and development of osteoarthritis diseases are delayed.

Description

Technical Field

[0001] The invention belongs to the technical field of medicine and biochemistry, and relates to application of GCA-Nab in preparing a medicine for treating osteoarthritis. Background Art

[0002] Osteoarthritis is one of the common chronic degenerative disabling diseases. In 2021, more than 22% of adults over 40 years old suffered from knee arthritis. It is estimated that more than 500 million people worldwide currently suffer from osteoarthritis. The pathological characteristics of osteoarthritis are wear and loss of articular cartilage, formation of osteophytes and bone spurs, hypertrophy of synovial tissue, increase in blood vessels and nerves, and chronic pain. There is currently no cure for osteoarthritis. For a long time, the clinical treatment of osteoarthritis can only improve the symptoms of joint pain, and there is a lack of drugs to slow the progression of the disease. Due to the lack of drugs that effectively control the progression of the disease, patients with end-stage osteoarthritis can only undergo joint replacement surgery. However, the service life of artificial joints is limited, and there is a risk of poor prognosis. In summary, it is of great clinical significance to find drugs to control or alleviate the progression of osteoarthritis.

[0003] The Penta-EF-hand (PEF) protein family is a group of calcium-binding proteins with 5 EF-hand-shaped motifs. Grancalcin (GCA) is specifically expressed in neutrophils and monocytes / macrophages. As a member of the PEF family, it participates in regulating cell migration, apoptosis, and mobilization of immune cells. There is a pro-inflammatory and pro-aging immune cell subset (including macrophages, neutrophils, etc.) in the aging bone marrow microenvironment. Previous studies have found that aging immune cells in the bone marrow can regulate the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells by secreting GCA. Knocking out GCA in mice or injecting GCA neutralizing antibodies into the bone marrow cavity can alleviate bone loss in mice and prevent the occurrence and development of osteoporosis. Osteoporosis is a systemic bone disease characterized by decreased bone mass and destruction of bone microstructure, bone fragility, and increased risk of fractures. The pathogenesis of osteoporosis is mainly that osteoclast-mediated bone resorption is greater than osteoblast-mediated bone formation. The main drugs for the treatment of osteoporosis in clinical practice are calcium agents. and inhibitors such as bisphosphonates, calcitonin, estrogen, and selective estrogen receptor modulators Bone breaking Drugs that promote osteoblastic bone formation, such as osteoblastic bone resorption activity and parathyroid hormone analogs.

[0004] Unlike osteoporosis, osteoarthritis is a type of joint degenerative disease characterized by cartilage loss accompanied by joint surface sclerosis, osteophyte formation, and synovial inflammation. Chondrocytes are the only cell type in cartilage tissue, and they maintain the dynamic balance of cartilage tissue by secreting extracellular matrix. The metabolic imbalance of chondrocytes is an important cause of the occurrence and development of osteoarthritis. The treatment drugs for osteoarthritis are mainly non-steroidal anti-inflammatory drugs with anti-inflammatory and analgesic effects, and there is a lack of drugs that can alleviate the progression of the disease.

[0005] Although both osteoarthritis and osteoporosis are diseases of abnormal bone metabolism, there are huge differences in the clinical symptoms and pathogenesis of the two diseases, which leads to different drug treatments for osteoporosis and osteoarthritis. Teriparatide, a parathyroid hormone analogue, has a strong ability to regulate calcium metabolism, promote new bone synthesis, and reshape bone tissue. It is often used clinically for patients with severe osteoporosis. Clinical studies have found that some patients taking teriparatide have the side effect of joint pain. Teriparatide cannot be used clinically to treat osteoarthritis.

[0006] Therefore, although GCA neutralizing antibodies can be used to prevent and treat osteoporosis, their preventive and therapeutic effects on osteoarthritis are unclear. In addition, there has been no report on the use of GCA neutralizing antibodies in the preparation of drugs for treating osteoarthritis. Summary of the invention

[0007] The technical problem to be solved by the present invention is to overcome the defect of the prior art that the preventive and therapeutic effect of GCA neutralizing antibodies on osteoarthritis is unclear, and provide a use of GCA-NAb in the preparation of drugs for treating osteoarthritis. The present invention creatively discovered a new application of a neutralizing antibody (GCA neutralizing antibody, GCA-NAb) of granular calcitonin (GCA) in the preparation of a preparation for treating osteoarthritis.

[0008] The inventors of the present invention have found that GCA secreted by macrophages in the synovial tissue of the joints of aged mice can regulate the anabolism and catabolism of chondrocytes, and knocking out GCA in macrophages in mice can alleviate the joint pathology of osteoarthritis with destabilization of the medial meniscus (DMM) and age-related joint degeneration. In other words, GCA recombinant protein can regulate the synthesis of human and mouse articular chondrocytes. Metabolism and catabolism, intra-articular injection of GCA neutralizing antibody GCA-NAb can effectively alleviate Jie Lao Joint degeneration in mice aged 2 years and 4 years. It can be seen that GCA neutralizing antibody (GCA-Nab) can block the effect of GCA on chondrocyte metabolism, thereby alleviating the occurrence and development of osteoarthritis.

[0009] The present invention provides an application of a GCA neutralizing antibody (GCA-Nab) in preparing a drug for treating osteoarthritis.

[0010] In the present invention, the GCA neutralizing antibody refers to a neutralizing antibody (GCA-NAb) of Grancalcin (GCA).

[0011] In the present invention, the GCA neutralizing antibody (GCA-nAb) can be prepared with reference to the following literature: Li CJ, Xiao Y, Sun Y, He W, Liu L, Su T, Sun X, Guo Q, Huang Y, Yang M, Liu G, Luo XH*. (2021) Seductive immune cells release grancalcin to promote skeletal aging. Cell Metab. 33(10), 1957-1973. doi: 10.1016 / j.cmet.2021.08.009.

[0012] In the present invention, the GCA neutralizing antibody (GCA-nAb) can be prepared with reference to Examples 1 to 5 of Chinese Patent CN113528456A.

[0013] In the present invention, the GCA neutralizing antibody can be secreted by the hybridoma cell line with a deposit number of CCTCC NO: C2021182.

[0014] In the present invention, the osteoarthritis may be osteoarthritis caused by increased expression of GCA protein.

[0015] In the present invention, the GCA gene sequence may be:

[0016] ATGCATCATCACCATCACCACCACCATCACCATCTGGAAGTCCTGTTTCAGGGACCCATGGCCTACCCGGGATACGGAGGAGGGTTTGGAAATTTTAGCATTCAGGTGCCAGGAATGCAGATGGGACAGCCAGTGCCAGAAACAGGCCCAGCTATACTCCTCGATGGATACTCTGGGCCAGCATATTCAGACACTTATTCCTCAGCTGGTGACTCCGTGTATACTTACTTCAGTGCTGTTGCTGGACAGGATGGTGAAGTGGATGCTGAAGAACTTCAGAGATGTTTGACACAGTCTGGAATTAATGGAACTTACTCTCCCTTCAGTTTGGAAACCTGCAGAATTATGATTGCCATGTTGGATAGAGATCACACAGGAAAAATGGGATTTAATGCATTCAAAGAGCTATGGGCAGCTCTTAATGCCTGGAAGGAAAACTTCATGACTGTTGATCAAGATGGAAGTGGCACAGTAGAACATCATGAGTTGCGTCAAGCCATTGGTCTTATGGGTTATAGGTTGAGTCCTCAAACATTAACTACTATTGTTAAACGTTATAGCAAGAATGGCAGAATTTTCTTTGATGATTATGTTGCTTGCTGTGTGAAGCTTCGAGCATTGACAGATTTCTTTAGGAAAAGAGACCACTTGCAACAAGGGTCTGCGAATTTCATATATGAC GATTTTTTGCAGGGCACTATGGCAATTTAA(SEQ ID NO.1).

[0017] In the present invention, the sequence of the GCA recombinant protein may be:

[0018] MHHHHHHHHHHLEVLFQGPMAYPGYGGGFGNFSIQVPGMQMGQPVPETGPAILLDGYSGPAYSDTYSSAGDSVYTYFSAVAGQDGEVDAEELQRCLTQSGINGTYSPFSLETCRIMIAML DRDHTGKMGFNAFKELWAALNAWKENFMTVDQDGSGTVEHHELRQAIGLMGYRLSPQTLTTIVKRYSKNGRIFFDDYVACCVKLRALTDFRKRDHLQQGSANFIYDDFLQGTMAI(SEQ ID NO.2).

[0019] In the present invention, the dosage of the GCA neutralizing antibody can be converted according to body weight, for example, 0.1 μg / g-1 μg / g, and for example 0.15 μg / g.

[0020] In the present invention, the GCA neutralizing antibody can be administered by intra-articular injection.

[0021] In the present invention, the osteoarthritis may be osteoarthritis with downregulated expression of COL-2.

[0022] In the present invention, the osteoarthritis may be osteoarthritis with upregulated expression of MMP13.

[0023] The present invention also provides a use of a GCA neutralizing antibody (GCA-Nab) in preparing a drug for improving the down-regulated expression of COL-2 caused by osteoarthritis.

[0024] In the present invention, the COL-2 is type II collagen, which is secreted by chondrocytes and is the main structural protein of cartilage tissue. It forms a network structure of extracellular matrix (ECM) together with proteoglycans and other proteoglycans. On the surface of cartilage, the collagen fibers composed of type II collagen are parallel to the joint surface and participate in alleviating the shear stress borne by the joint. Studies have shown that with the increase of age or the progress of osteoarthritis, the synthesis of type II collagen is relatively reduced, the decomposition is relatively increased, and it is no longer limited to the surrounding parts of chondrocytes. In the early stage of the disease, the collagen in the joint surface and its vicinity is damaged and lost. As the disease becomes more serious, the loss of type II collagen accelerates and penetrates deep into the cartilage tissue until the cartilage tissue is completely lost.

[0025] The present invention also provides a use of a GCA neutralizing antibody (GCA-Nab) in preparing a drug for reducing up-regulation of MMP13 expression caused by osteoarthritis.

[0026] In the present invention, the MMP13 refers to matrix metalloproteinase 13, which has the ability to specifically decompose type II collagen and is the main decomposition factor involved in the degradation of cartilage tissue. Chondrocytes secrete extracellular matrix through anabolism and degrade extracellular matrix through catabolism. Under physiological conditions, the slow and relatively balanced anabolism and catabolism of chondrocytes maintain the normal structure of cartilage tissue. In the process of the occurrence and development of osteoarthritis, the anabolism of chondrocytes and the catabolism of chondrocytes are unbalanced, and the synthesis and secretion of MMP13 by chondrocytes increase, resulting in increased degradation of the extracellular matrix and accelerated loss of cartilage tissue.

[0027] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0028] The reagents and raw materials used in the present invention are commercially available.

[0029] The positive and progressive effects of the present invention are:

[0030] The present invention discovers for the first time the preventive and therapeutic effect of GCA neutralizing antibody GCA-Nab on osteoarthritis, providing a research basis and theoretical basis for the development of drugs for treating osteoarthritis.

[0031] Experiments have shown that the GCA-Nab of the present invention can effectively block the regulatory effect of GCA on chondrocyte metabolism, thereby significantly improving the loss of articular cartilage and delaying the occurrence and development of osteoarthritis.

[0032] Biomaterial Deposit Information

[0033] The hybridoma cell line of the present invention was deposited in the China Center for Type Culture Collection (CCTCC) on August 3, 2021. The deposit address is Wuhan University, Luojia Mountain, Wuchang, Wuhan City, Hubei Province, China, Postal Code 430072, the deposit number is CCTCCNO: C2021182, and the culture name is hybridoma cell line GCA-NAB4. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1A This is a WB schematic diagram of the GCA recombinant protein inhibiting the anabolism (COL-2) and promoting the catabolism (MMP13) of mouse primary chondrocytes in Example 1;

[0035] Figure 1B This is a WB schematic diagram of the GCA recombinant protein inhibiting the anabolism (COL-2) of human primary chondrocytes and promoting their catabolism (MMP13) in Example 1;

[0036] Figure 2AThis is a schematic diagram of the qPCR results of GCA recombinant protein inhibiting the anabolism (COL-2) and promoting the catabolism (MMP13) of primary chondrocytes in mice in Example 1;

[0037] Figure 3A This is a schematic diagram showing the results of knocking out GCA in macrophages to alleviate DMM-induced osteoarthritis pain in mice in Example 2;

[0038] Figure 3B This is a schematic diagram of safranin fast green staining showing that knocking out GCA in macrophages alleviates DMM-induced cartilage degeneration in osteoarthritis mice in Example 2;

[0039] Figure 3C This is a schematic diagram of the safranin fast green staining scoring for the alleviation of cartilage degeneration in osteoarthritis mice induced by DMM by knocking out GCA in macrophages in Example 2;

[0040] Figure 4A This is a schematic diagram of Safranin Fast Green staining showing that knocking out GCA in macrophages alleviates age-related articular cartilage degeneration in mice in Example 3;

[0041] Figure 4B This is a schematic diagram of the safranin fast green staining score for alleviating age-related articular cartilage degeneration in mice by knocking out GCA in macrophages in Example 3;

[0042] Figure 5A This is a schematic diagram of the qPCR results that the neutralizing antibody against GCA alleviates the decrease in anabolism (col-2) and increase in catabolism (MMP13) of primary chondrocytes induced by the supernatant of bone marrow macrophages of aged mice in Example 4;

[0043] Fig. 6A This is a schematic diagram of the WB results of the GCA neutralizing antibody in Example 4 alleviating the decrease in anabolism (col-2) and increase in catabolism (MMP13) of primary chondrocytes induced by the supernatant of bone marrow macrophages of aged mice;

[0044] Figure 6B This is a schematic diagram of the quantitative analysis of WB results showing that the GCA neutralizing antibody (GCA-NAb) alleviates the decrease in anabolism (col-2) and increase in catabolism (MMP13) of primary chondrocytes induced by the supernatant of bone marrow macrophages of 18-month-old mice in Example 4;

[0045] Fig. 7A This is a schematic diagram of Safranin Fast Green staining showing that GCA-NAb alleviates articular cartilage degeneration in 21-month-old mice in Example 5;

[0046] Figure 7B This is a schematic diagram of the Safranin Fast Green staining score for GCA-NAb alleviating articular cartilage degeneration in 21-month-old mice in Example 5;

[0047] Fig. 8A This is a schematic diagram of COL-2 and MMP13 staining after intra-articular injection of GCA-NAb in elderly mice in Example 5;

[0048] Figure 8B This is a schematic diagram of the quantitative analysis of COL-2 and MMP13 staining after intra-articular injection of GCA-NAb in elderly mice in Example 5. DETAILED DESCRIPTION

[0049] The technical solution of the present invention is further described in detail below in conjunction with the accompanying drawings and specific implementation plans.

[0050] Example 1 GCA recombinant protein inhibits chondrocyte anabolism and promotes its catabolism

[0051] First, human or mouse primary chondrocytes were inoculated in 12-well plates and cultured in DMEM-F12 medium containing 10% FBS and 1% penicillin / streptomycin. The next day, the experimental group was intervened by GCA recombinant protein (rGCA) at a final concentration of 100nmol / L, and the control group was added with an equal volume of PBS. After 3 days of continuous culture, cell protein or RNA was extracted for WB or qPCR experiments to evaluate the effect of GCA recombinant protein on the metabolism of human or mouse primary chondrocytes.

[0052] The GCA protein sequence is:

[0053] MHHHHHHHHHHLEVLFQGPMAYPGYGGGFGNFSIQVPGMQMGQPVPETGPAILLDGYSGPAYSDTYSSAGDSVYTYFSAVAGQDGEVDAEELQRCLTQSGINGTYSPFSLETCRIMIAML DRDHTGKMGFNAFKELWAALNAWKENFMTVDQDGSGTVEHHELRQAIGLMGYRLSPQTLTTIVKRYSKNGRIFFDDYVACCVKLRALTDFRKRDHLQQGSANFIYDDFLQGTMAI(SEQ ID NO.2)

[0054] from Figure 1A It can be seen that compared with the control group, the anabolism of mouse primary chondrocytes decreased and the catabolism increased significantly after the addition of GCA recombinant protein. Figure 1B It can be seen that compared with the control group, the anabolism of human primary chondrocytes (COL-2) was significantly decreased and the catabolism (MMP13) was significantly increased after treatment with GCA recombinant protein.

[0055] See also Figure 2A, Each value is expressed as the mean ± SD of three tests. *, compared with the control group cells treated with PBS, P < 0.05; **, compared with the control group cells treated with PBS, P < 0.01.

[0056] from Figure 2A It can be seen that compared with the control group, the mRNA level of col-2a1, a gene related to anabolism, in primary chondrocytes of mice was significantly decreased, and the mRNA level of MMP13, a gene related to catabolism, was significantly increased after treatment with GCA recombinant protein.

[0057] Figure 1A , Figure 1B , Figure 2A The results showed that GCA recombinant protein inhibited chondrocyte anabolism and promoted its catabolism.

[0058] Example 2 Knockout of GCA alleviates the progression of osteoarthritis induced by DMM in mice

[0059] The pathological characteristics of osteoarthritis include wear and loss of articular cartilage, accompanied by chronic pain. Previous research results showed that GCA can inhibit chondrocyte anabolism and promote its catabolism, so it is speculated whether knocking out GCA in vivo can alleviate the progression of osteoarthritis. Based on this, osteoarthritis models (medial meniscus destabilization surgery, DMM) were established in 3-month-old male macrophages knocked out GCA mice (GCA-CKO) and wild-type mice (WT). Samples were collected 2 months after surgery and sections were stained.

[0060] from Figure 3A As shown in Table 1, the pain symptoms of GCA-CKO mice were alleviated compared with the control group. Figure 3B , Figure 3C It can be seen that the severity of articular cartilage loss in GCA-CKO mice was reduced compared with that in the control mice.

[0061] Table 1

[0062]

[0063]

[0064] Example 3 Knockout of GCA alleviates age-related articular cartilage degeneration

[0065] To further clarify the role of GCA in age-related articular cartilage degeneration, 23-month-old male GCA-CKO mice and WT mice were sampled, sectioned, and stained.

[0066] from Figure 4A , Figure 4BIt can be seen that the severity of articular cartilage loss in 23-month-old GCA-CKO mice was less severe than that in WT mice.

[0067] Figure 3A , Figure 3B , Figure 3C , Figure 4A , Figure 4B The results showed that specific knockout of GCA in mouse macrophages can alleviate the joint pathology of osteoarthritis with medial meniscus instability (DMM) and age-related articular cartilage degeneration.

[0068] Example 4 GCA neutralizing antibodies alleviate chondrocyte metabolic disorders induced by elderly macrophages

[0069] First, human or mouse primary chondrocytes were inoculated in 12-well plates and cultured in DMEM-F12 medium containing 10% FBS and 1% penicillin / streptomycin. The next day, the supernatant of macrophages from aged mice was added for intervention, and the experimental group was given 5 μg / ml of GCA neutralizing antibody, and the control group was added with an equal volume and concentration of neutralizing antibody isotype control (Isotype). After 3 days of continuous culture, cell protein or RNA was extracted for WB or qPCR experiments to evaluate the therapeutic effect of GCA neutralizing antibody on metabolic disorders of primary chondrocytes in mice.

[0070] GCA neutralizing antibody (GCA-nAb) was prepared with reference to Example 1 to Example 5 of Chinese Patent CN113528456A:

[0071] The GCA gene sequence is:

[0072] ATGCATCATCACCATCACCACCACCATCACCATCTGGAAGTCCTGTTTCAGGGACCCATGGCCTACCCGGGATACGGAGGAGGGTTTGGAAATTTAGCATTCAGGTGCCAGGAATGCAGATGGGACAGCCAGTGCCAGAAACAGGCCCAGCTATACTCCTCGATGGATA CTTCTGGGCCAGCATATTCAGACACTTATTCCTCAGCTGGTGACTCCGTGTATACTTACTTCAGTGCTGTTGCTGGACAGGATGGTGAAGTGGATGCTGAAGAACTTCAGAGATGTTTGACACAGTCTGGAATTAATGGAACTTACTCTCCCTTCAGTTTGGAAACCTGCA GAATTATGATTGCCATGTTGGATAGAGATCACACAGGAAAAAATGGGATTTAATGCATTCAAAGAGCTATGGGCAGCTCTTAATGCCTGGAAGGAAAACTTCATGACTGTTGATCAAGATGGAAGTGGCACAGTAGAACATCATGAGTTGCGTCAAGCCATTGGTCTTATG GGTTATAGGTTGAGTCCTCAAACATTAACTACTATTGTTAAACGTTATAGCAAGAATGGCAGAATTTTCTTTGATGATTATGTTGCTTGCTGTGTGAAGCTTCGAGCATTGACAGATTTCTTTAGGAAAAGAGACCACTTGCAACAAGGGTCTGCGAATTTCATATATGAC GATTTTTTGCAGGGCACTATGGCAATTTAA (SEQ ID NO. 1).

[0073] from Figure 5A It can be seen that compared with the complete blank control, the addition of supernatant of bone marrow macrophages from aged mice will cause metabolic disorders in primary chondrocytes of mice - the mRNA level of the anabolism-related gene col-2a1 is significantly decreased, and the mRNA level of the catabolism-related gene MMP13 is significantly increased, but GCA neutralizing antibodies can partially rescue the metabolic disorders of chondrocytes.

[0074] See also Figure 5A , Each value is expressed as the mean ± SD of three tests. **, P < 0.01 compared with the control group cells treated with isotype.

[0075] from Fig. 6A , Figure 6B It can be seen that compared with the control group, the addition of GCA neutralizing antibody in the experimental group can partially rescue the metabolic disorder of chondrocytes.

[0076] See also Fig. 6A , Figure 6B , Each value is expressed as the mean ± SD of three tests. *, compared with the control group cells treated with isotype, P < 0.05, **, compared with the control group cells treated with isotype, P < 0.01.

[0077] Example 5 GCA neutralizing antibody GCA-NAb can alleviate joint degeneration in elderly mice

[0078] To further clarify whether GCA neutralizing antibodies can alleviate age-related joint degeneration, 19-month-old C57 mice were injected with GCA neutralizing antibodies or Isotype at 1.5 μg / 10 g once a week for 2 months. Samples were collected 2 months after administration and sectioned for staining.

[0079] from Fig. 7A , Figure 7B It can be seen that the severity of articular cartilage degeneration in mice injected with GCA neutralizing antibodies was reduced compared with that in control mice. The results of the study showed that GCA neutralizing antibodies GCA-NAb can alleviate joint degeneration in elderly mice. GCA neutralizing antibodies are the same as those in Example 4.

[0080] from Fig. 8A , Figure 8B It can be seen that compared with control mice, mice injected with GCA neutralizing antibodies had less loss of COL-2 in the extracellular matrix of articular cartilage and decreased expression of MMP13, which decomposes extracellular matrix proteins. The results show that GCA neutralizing antibodies GCA-NAb can alleviate joint degeneration in elderly mice.

[0081] Figure 5A , Fig. 6A , Figure 6B , Fig. 7A , Figure 7B , Fig. 8A , Figure 8B This shows that GCA neutralizing antibodies can alleviate articular cartilage degeneration and delay the occurrence and development of osteoarthritis.

[0082] All the above results indicate that GCA plays an important role in osteoarthritis, and GCA neutralizing antibodies can alleviate the occurrence and development of osteoarthritis.

[0083] The above is only a preferred specific implementation manner of the present invention, and the protection scope of the present invention is not limited thereto. Any simple change or equivalent replacement of the technical solution that can be obviously obtained by any technician familiar with the technical field within the technical scope disclosed in the present invention falls within the protection scope of the present invention.

Claims

1. Use of a GCA neutralizing antibody in the preparation of a drug for treating osteoarthritis.

2. The use according to claim 1, characterized in that The GCA neutralizing antibody is secreted by a hybridoma cell line with a deposit number of CCTCC NO: C2021182.

3. The use according to claim 1, characterized in that The osteoarthritis is osteoarthritis caused by increased expression of GCA protein, and the gene sequence of the GCA protein is SEQ ID NO.1; And / or, the sequence of the GCA protein is shown as SEQ ID NO.

2.

4. The use according to claim 1, characterized in that The osteoarthritis is osteoarthritis with downregulated expression of COL-2.

5. The use according to claim 1, characterized in that The osteoarthritis is osteoarthritis with upregulated expression of MMP13.

6. The use according to any one of claims 1 to 5, characterized in that The dosage of the GCA neutralizing antibody is 0.1 μg / g-1 μg / g; And / or, the GCA neutralizing antibody is administered by intra-articular injection.

7. Use of a GCA neutralizing antibody in the preparation of a drug for improving the down-regulated expression of COL-2 caused by osteoarthritis.

8. The use according to claim 7, characterized in that The GCA neutralizing antibody is secreted by a hybridoma cell line with a deposit number of CCTCC NO: C2021182.

9. Use of a GCA neutralizing antibody in the preparation of a drug for reducing upregulated MMP13 expression caused by osteoarthritis.

10. The use according to claim 9, characterized in that The GCA neutralizing antibody is secreted by a hybridoma cell line with a deposit number of CCTCC NO: C2021182.

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