Research On The Effects Of Components Of Chuandilong On The Cartilage Destruction In CIA

Background: Rheumatoid arthritis (RA) is characterized by the presence of an inflammatory synovitis accompanied by destruction of joint cartilage and bone. Destruction of cartilage matrix results predominantly from the action of connective tissue proteinases released by RA synovial tissues, lymphocytic subsets(CD4, CD8), chondrocytes, and pannus tissue. RA synovial tissue produces a variety of cytokines and growth factors including interleukin 1 (IL-1), tumour necrosis factor-a(TNF-a), matrix metalloproteinases (MMPs) etc. Collagen-induced arthritis (CIA) is a generally accepted animal model because of resembling RA in pathology. Good response to treatment of RA has been got by using Chuandilong, a kind of traditional Chinese herbal medicine. However, to our knowledge, its effective components and actional mechanism has not been published yet. Objective: To observe therapeutic effect of aqueous and liposoluble components of Chuandilong on CIA of rats and to elucidate the pharmacologic mechanism in treatment of CIA.Materials and Methods:Type II collagen-induced arthritis model in wistar rats were established. Aqueous and liposoluble components ofChuandilong were extracted and analysised by thin-layer chromatography (TLC). Both of them were respectively used to treat CIA rats for 6 weeks; and(1)The changes of swelling rate of indirect proinflamation ankle joint were observed; (2)The levels of serum WBC, PLT, SA, TNF-a were determined by ELISA; (3)The histopathology and ultrastructure of synovium of joint were respectively observed microscoply and electroscoply; (4)T-cell subsets including CD4, CD8 and TGF-β1, bFGF in synovial and cartilage tissue of CIA rats were detected by Immunohistochemistry techniques; ㏕he action of TNF-a to chondrocyte secreting MMP-1 was observed by means of chondrocyte culture; Diosgenin, hydrolyzate of Chuandilong, was used to intervene the effects. Results:1. The ideal animal model of CIA was successfully established;2. Diosgenin was found in aqueous components of Chuandilong by thin-layer chromatography (TLC), but was not in the liposolublecomponents;3. The joint swelling rate of the left ankle joint was suppressed after taking aqueous components of Chuandilong;4. The levels of WBC, PLT, SA and TNF-a in blood were downregulated after using aqueous components of Chuandilong;5. After taking aqueous components of Chuandilong, the lymphocytesinfiltration in the synovial was ameliorated, and the mitochondrion swelling of synovial cell and vascular endothelial cell and the synovial hyperplasia and pannus formation were relieved;6. The lymophocyte subsets (CD4,CD8) in the synovium of left ankle joint were regulated by using aqueous components of Chuandilong;7. The expression of TGF-β1 and bFGF in the synovial and cartilage was increased and the degree of cartilage destruction was decreased after taking the aqueous components of Chuandilong;8. There was little effect on CIA, but there was the side-effect on gastrointestinal tract after taking the liposoluble components of Chuandilong;9. The amount of MMP-1 secreting by chondrocytes was decreased and the degree of destruction in chondrocytes was reduced after using diosgenin, the effective component of Chuandilong;Conclusion: Chuandilong can efficiently treat collagen-induced arthritis (CIA), whose effective components are mainly aqueous components of Chuandilong. Treatment mechanisms include (1)regulating lyphocytic subsets CD4, CD8 expression in CIA joint synovium, suppressing cellular immunity; (2)suppressing MMP-1 secreting from articular cartilage cells, and relieving the destruction degree of cartilage matrix; (3)increasing the expression of TGF-β1 and bFGF in joint synovium, and protecting joint cartilage, and facilitating thejoint repair; (4)decreasing blood serum inflamation medium sialic acid (SA) and cytokine (TNF-a), thus reducing synovitis and swelling of joint, ameliorating articular pathologic status, relieving articular cartilage destruction. But there was li...