Application of human synovial membrane fibroblasts to treating rheumatoid arthritis

A fibroblast, rheumatoid technology, applied in medical preparations containing active ingredients, anti-inflammatory agents, non-central analgesics, etc. , improve the effect of occurrence and progression

Inactive Publication Date: 2017-09-22
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report of related or similar treatment methods at home and abroad.

Method used

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  • Application of human synovial membrane fibroblasts to treating rheumatoid arthritis
  • Application of human synovial membrane fibroblasts to treating rheumatoid arthritis
  • Application of human synovial membrane fibroblasts to treating rheumatoid arthritis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Isolation and culture of fibroblasts derived from human synovial tissue

[0027] 1. Method

[0028] (1) Take the knee joint synovial tissue of osteoarthritis patients under aseptic conditions, and obtain P3 generation synovial fibroblasts through shredding, digestion, filtration, culture, and passage;

[0029] (2) Morphological observation of P3 generation synovial fibroblasts under a microscope;

[0030] (3) P3 generation fibroblasts were cultured in adipogenic differentiation medium and osteogenic differentiation medium for 21 days, stained with Oil Red O and Alizarin Red respectively, and detected and analyzed the multidirectional differentiation potential;

[0031] (4) The method of flow cytometry is used to detect the surface antigen molecules of P3 generation fibroblasts.

[0032] 2. Results

[0033] Biological properties of synovial fibroblasts such as figure 1 shown, where figure 1 (a) shows the fibroblast-like adherent growth of synovial fibrobl...

Embodiment 2

[0034] Example 2: Therapeutic effect of fibroblasts derived from human synovium injected into the knee joint cavity on CIA mice

[0035] 1. Method

[0036] (1) Establishment of type II collagen-induced arthritis (CIA) mouse model: All mice in the model group were subcutaneously injected with 100 μg of bovine type II collagen at the base of the tail on d0 and d21;

[0037] (2) Treat CIA mice with fibroblasts derived from human synovium injected into the right knee joint cavity: use 10 μl containing 1×10 6 PBS of fibroblasts was injected into the right knee joint cavity of CIA mice on d28, d32 and d38 days after the first immunization; 10 μl PBS was injected into the right knee joint cavity of the control group;

[0038] (3) Evaluation of CIA arthritis: score the arthritis of the mice in the model group every other day after the second immunization. The scoring method was double-blind, and the scoring criteria were: 0, normal; 1, redness involving one finger joint; 2, ≥ 2 fing...

Embodiment 3

[0041] Example 3: Detection of mRNA expression levels of cytokines in synovial tissue

[0042] On the 70th day, the mice in the control group and the fibroblast treatment group of Example 2 were sacrificed, the patella and adjacent synovial tissues were separated, and the mRNA levels of cytokines in the synovial tissues were detected by Real time-PCR.

[0043] The result is as image 3 As shown, it can be seen from the results that human synovium-derived fibroblasts can reduce tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ) inflammatory factor IL-17A and transforming growth factor (TGF -β) secretion, and up-regulate the levels of immune regulatory molecules IL-10 and IL-4. It can be seen that the fibroblasts derived from human synovium have immunoregulatory effect and tissue repair ability, and will have broad application prospects in the field of rheumatoid arthritis treatment.

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PUM

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Abstract

The invention discloses application of human synovial membrane fibroblasts to treating rheumatoid arthritis. The application includes building CIA (collagen-induced arthritis) mouse models which are rheumatoid arthritis animal models, injecting fibroblasts from human synovial membranes in knee joint cavities of mice and observing arthritis treatment effects of the fibroblasts from two aspects including the joint swelling degree of the mice and the gene expression level of inflammatory cytokines of synovial membrane tissues of joints of the mice. The application has the advantages that as shown by results, arthritis symptoms of the mice can be obviously improved after the human synovial membrane fibroblasts are injected in the knee joint cavities; immune imbalance states due to the rheumatoid arthritis can be regulated by the aid of the fibroblasts from the human synovial membranes, effective biotherapy means can be provided for treating rheumatoid arthritis patients, and accordingly the application has great social and economic benefit values.

Description

technical field [0001] The invention relates to a novel biological treatment method for rheumatoid arthritis, in particular to a method for treating rheumatoid arthritis by injecting fibroblasts derived from human synovium into the knee joint cavity. The invention belongs to the technical field of biology and new medicine. Background technique [0002] Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease mediated by immunocompetent cells and various cytokines. The prevalence of this disease in my country's population is 0.34%-0.36%, and the incidence rate of women is more than three times higher than that of men. It is one of the main reasons for the loss of labor force and disability in my country's population. The clinical manifestations of RA are multiple symmetrical arthropathy, joint pain, swelling, stiffness, and dyskinesia. The basic pathological changes are synovitis and progressive joint destruction. The disorder and imbalance of the immune system a...

Claims

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Application Information

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IPC IPC(8): A61K35/33A61P29/00A61P19/02
CPCA61K35/33
Inventor 李洋颜明露刘鑫
Owner HARBIN MEDICAL UNIVERSITY
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