Study Of The Expression And Clinical Significance Of VEGF, TGF-β1, BMP-2 Of Peripheral Arthritis In Ankylosing Spondylitis Patients

Part i Study of the Expression and Clinical Significance of VEGF ofPeripheral Arthritis in Ankylosing spondylitis OBJECTIVES:(1) To evaluate the role of vascular endothelial growth factor (VEGF) in the pathogenesis of ankylosing spondylitis(AS); and (2) to study whether the serum level of VEGF correlated with disease activity. METHODS :1. Forty-four patients who met the 1984 New York AS classification criteria were recruited from Chinese PLA General Hospital's Rheumatology department. Twenty healthy volunteers with no abnormalities on physical examination and laboratory tests were chosen as controls. The definition of active disease for AS patients with peripheral joint involvement was: (1) presence of active arthritis (joint swelling, tenderness, and limitation of range of motion), (2) physician's global assessment of disease activity > 5 on a 0-10cm VAS scale, (3) ESR > 30mm/hr. For AS patients with axial joint involvement only, the definition of inactive disease was: (1) absence of any sign of active arthritis at the time of clinical examination, (2) physician's global assessment of disease activity <4 on a 0-10cm VAS scale, (3) ESR <30mm/hr.SFMC was isolated from patients' synovial fluid (SF) and was stimulated by IPS. The expression of VEGF mRNA was determined by reverse transcription polymerase chian reaction (RT-PtCR). Paired samples of PBMCwere obtained from 24 AS patients with peripheral arthritis. Sera were collected from 20 AS patients with axial joint involvement only, 24 AS patients with peripheral arthritis and 20 normal controls. The levels of VEGF in sera and SF were measured using enzyme linked immunosorbent assay (ELISA). The degree of correlation between VEGF levels/VEGF mRNA and clinical and laboratory parameters was calculated respectively by regression analysis.2. Synovial samples were collected twenty-two patients (AS, n=12; RA, n=10) during arthroscopic procedures. They all had early knee arthritis with disease duration <24 months. Synovial samples were collected from 12 hips of AS and 13 knees of RA (with late arthritis disease duration >10 years) during joint replacement surgery or arthroscopic procedures.Macroscopic observation during arthroscopy: the vascular morphology of synovial membrane was assessed using a VAS for vascular morphology, tortuous vessels, straight vessels, vascular density, synovial hypertrophy, by 2 blinded observers using a validated methods.Samples were fixed and dehydrated in ethanol, transferred to xylens, and embedded in paraffin. VEGF mRNA and VEGF were detected by both in situ hybridization and immunohistochemistry. Synovial membrane was divided to different regions (synovium lining, perivascular region, sub-lining region) in order to study the relation between VEGF and parameters of disease activity and understand the relation between VEGF and the vascular morphology of synovial membrane. Result:1. Activated SFMC by IPS led to high VEGF mRNA expression. The concentration of SFVEGF and sVEGF in 24 SpA patients with active disease were significantly higher than inactive AS patients and healthy controls. Asignificant correlation was found between SF and serum VEGF concentrations. The concentrations of SFVEGF were significantly higher than those detected in corresponding serum sample. Moreover, the levels of serum VEGF in AS with peripheral Arthritis correlated with BASDAI, number of swollen joint, degree of body fatigue, thrombocytes. The levels of SFVEGF displayed a significant correlation with BASDAI, number of swallen joint, and peripheral WBC count.2. VEGF mRNA and VEGF expression were determined in all synovial membrane samples. The expression of VEGF mRNA showed correlation with VEGF in every regions in synovial membrane. The expression of VEGF demonstrated correlation with vascular morphology (VM), straight vessels (SV) in synovium lining layer. The relation was found between the VEGF and synovials hypertrophy in perivascular region. Others correlation were found between VEGF and CRP in synovium lining layer of early AS and in perivas...