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Establishment and Evaluztion of Collagen-induced Arthritis Model in Rats and Investigation of Mechanism in CIA

Author: KongZuo
Tutor: LiJun
School: Anhui Medical University,
Course: Pharmacology
Keywords: Collagen-induced arthritis Collagen type Ⅱ Rheumatoid arthritis Regulatory T cells of CD4 ~ CD25 ~ Foxp3
CLC: R593.2
Type: Master's thesis
Year: 2008
Downloads: 231
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Abstract


Objective: chicken sternum xiphoid cartilage extract, purified collagen type Ⅱ (C Ⅱ) and preliminary identification of raw materials for the preparation of collagen-induced arthritis (CIA) rat model; preparation CIA rat model, from morphological immunology and histopathology multi-angle observation the CIA rats model characteristics; further clarify the pathogenesis of CIA rats. Methods and Results: type Ⅱ collagen protein (collagen type Ⅱ, C Ⅱ) the extraction, purification and identification of chicken sternum xiphoid cartilage for raw materials, pepsin digestion method extract C Ⅱ under acidic conditions; using hydrochloric acid guanidine extraction, of NaCl repeated salting purified, DEAE-52 fiber resin processing, dialyzed and other methods to remove proteoglycans, C II degradation fragments and denatured product to purified C II; identified using SDS-PAGE electrophoresis and absorption spectroscopy. The experimental results show that the molecular weights of the extracted C II with standard, the maximum absorption peak of 226nm, to confirm that the extraction and purification of the protein collagen type II. 2 collagen-induced arthritis in rats (collagen-induced arthritis, CIA) model preparation and evaluation of the laboratory from mentioning C Ⅱ plus complete Freund's adjuvant (CFA) intradermal injection rat CIA model. Volumetric method detection rat paw swelling and ratings; HE staining of joint tissue for histological examination; ELISA assay in vivo anti-C Ⅱ antibody content; radioimmunoassay of serum cytokines IL-1β, TNF-α content. The results showed strong the CIA rat hind claw inflammatory response, joint swelling; pathological examination of joint tissue visible to a large number of inflammatory cell infiltration, and associated with tissue fibrosis; serum anti-C Ⅱ antibody levels and IL-1β, TNF-α significantly increased, suggesting that collagen-induced arthritis model in rats success. Part of the mechanism of collagen-induced arthritis in rats in CIA rats models were based on the success, the MTT assay spleen lymphocyte proliferative responses; RT-PCR assay the CIA rat spleen regulatory T cells specific markers - the of Foxp3 surface of TGF-β the 1 expression. The results show that the the CIA rats spleen lymphocyte proliferative response enhancement; Foxp3mRNA with TGF-β 1 mRNA expression decreased. Conclusion: 1. Extracted purified protein collagen type II, the purified C Ⅱ simple and efficient. CIA rat model prepared clinical manifestations, immunological and pathological changes to human RA similar model was successfully. In the CIA model, lymphocytes decrease closely related to the strength of the inflammatory response, regulatory T cells because of their functional defects can not be effectively suppress effector T lymphocyte proliferation activation, resulting in the body's autoimmune reaction occurs, The cause of the occurrence of arthritis.

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CLC: > Medicine, health > Internal Medicine > Systemic disease > Autoimmune diseases > Autoimmune diseases, connective tissue disease
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