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Pharmaceutical application of CYR61 protein

A protein and drug technology, applied in the application field of CYR61 protein in pharmaceuticals, can solve the problem of no report on the effect of rheumatoid arthritis, and achieve the effect of promoting proliferation

Inactive Publication Date: 2010-05-19
SHANGHAI INST OF IMMUNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on CYR61 is mostly focused on tumors, but its role in rheumatoid arthritis has not been reported yet

Method used

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  • Pharmaceutical application of CYR61 protein
  • Pharmaceutical application of CYR61 protein
  • Pharmaceutical application of CYR61 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Expression Pattern of CYR61 in Synovial Tissue of Rheumatoid Arthritis

[0038] 1. Materials and methods

[0039] 1 Experimental materials

[0040] 1.1 Clinical samples:

[0041] All samples in the study were from RA patients and OA patients undergoing knee replacement or synovectomy in clinical orthopedics department, and the diagnosis of all cases met the international diagnostic criteria. Two normal controls were from trauma patients. The patient's synovial tissue was used for cell culture in vitro, part of the tissue was fixed with 4% paraformaldehyde and reserved for immunohistochemistry, serum and synovial fluid (SF) were centrifuged at high speed to obtain supernatant, and stored at -80°C. All clinical samples used in this study were informed to the patients.

[0042] 2 Experimental methods

[0043] 2.1 Cell preparation

[0044] 2.1.1 Preparation of peripheral blood mononuclear cells:

[0045] Mononuclear cells were separated from heparin-antico...

Embodiment 2

[0070] Example 2: CYR61 in rheumatoid arthritis synovial fluid promotes the proliferation of FLS

[0071] 1. Materials and methods

[0072] 1.1 Synovial fluid stimulation experiment: Mix 3-5 parts of synovial fluid and add to FLS in logarithmic growth phase to continue culturing for 24 hours, then detect CYR61 mRNA and protein expression levels, or FLS proliferation (see below for details).

[0073] 1.2 Cell proliferation and antibody blocking experiments: take FLS in the logarithmic growth phase, digest with 0.25% trypsin and adjust the cell concentration to 1×10 4 cells / ml, inoculate in 96-well cell culture plate, add different concentrations of SF (the ratio of SF to culture medium is 1:32, 1:16, 1:8, 1:4, 1:2) or cytokines (IL-10, IL12, IFNγ, TNFa and IL-17), or pure CYR61 protein was co-cultured with FLS. 1 μCi of 3H (per well) was added 16 hours before the end of the culture, the cells were collected, and the proliferation was detected by a β liquid scintillation instr...

Embodiment 3

[0087] Example 3: Study on the mechanism of CYR61 promoting the proliferation of rheumatoid arthritis FLS

[0088] 1. Materials and methods

[0089] 1. Clinical samples, CYR61 gene and protein expression detection are the same as those described in the materials and methods of "Example 1: CYR61 expression pattern in rheumatoid arthritis synovial tissue".

[0090] 2. FLS culture and proliferation stimulation experiments are the same as those described in the materials and methods of "Example 2: CYR61 promotes the proliferation of FLS in rheumatoid arthritis synovial fluid".

[0091] 3. siRNA interference experiment:

[0092] 3.1: siRNA sequence and synthesis: 3 siRNAs were synthesized according to the siRNA design principle, the sequence is as follows:

[0093] Table 2 Three siRNA sequences

[0094]

[0095] 3.2 siRNA transfection: transfection reagent liposome (geneporter) and serum-free DMEM solution containing antibiotics were used, and the transfection system (take 24...

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Abstract

The invention relates to a purpose of CYR61 protein, in particular to an application of CYR61 protein in the pharmaceutical field. The adopted technical proposal is an application of the CYR61 protein in the preparation of medicaments for rheumatic diseases, and the rheumatic diseases refer to chronic infectious arthritis. The invention has the advantages of developing the medical purpose of the CYR61 and opening a new application field. Results of the invention indicate that the CYR61 has the functions of promoting the proliferation of fibroblast-like synoviocytes (FLS) and participating in the damages to articular cartilages and bone tissues. One method for realizing the functions is to promote the fibroblast-like synoviocytes to proliferate by inhibiting the apoptosis of synoviocytes, and the other method is to participate in the damages to articular cartilages and bone tissues by up-regulating metalloproteinase. The result provides test basis for finding biological therapeutic targets and therapeutic targets of other small-molecule medicaments, thereby having the clinical application values.

Description

【Technical field】 [0001] The present invention relates to the use of CYR61 protein, in particular to the application of CYR61 protein in the pharmaceutical field. 【Background technique】 [0002] Rheumatoid arthritis (RA) is one of the most common rheumatic diseases. Prevalence varies slightly across countries and regions. The global prevalence of RA averages 0.5-1%. In my country, the prevalence of RA is 0.45%, and the total number of patients is about 4 million. RA is not a benign disease, compared with the same age, the average life expectancy of RA patients can be shortened by 10-15 years. RA is also one of the arthritis with the highest morbidity rate. For untreated patients, the 2-year and 3-year morbidity rates can reach 50% and 70%. RA is a systemic chronic autoimmune disease characterized by inflammation of the joint synovium, clinically characterized by erosive synovitis involving peripheral joints, with a proliferative and destructive pattern resembling "locali...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P29/00A61P19/02A61P19/04A61P37/06
CPCC07K16/18A61K38/18C07K2317/76A61K31/713C07K14/705C12N15/00A61P19/02A61P19/04A61P29/00A61P37/06
Inventor 李宁丽王利沈佰华肖涟波欧阳桂林
Owner SHANGHAI INST OF IMMUNOLOGY
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