低分子量肝素干扰免疫细胞黏附类风湿性关节炎滑膜细胞的体外研究

Low Molecular Weight Heparin's Interference with Immunocytes' Adhesion to Fibroblast-like Synovocyte (FLS) from Patients with Rheumatoid Arthritis(RA):An in Vitro Study

目的:研究低分子量肝素(LMWH)对免疫细胞黏附类风湿性关节炎(RA)滑膜细胞(FLS)的影响和初步机制,探讨LMWH在RA抗炎应用中的可行性。方法:用流式细胞仪定量计数法研究LMWH在体外干扰健康人外周血单核细胞(PBMC)黏附RAFLS的作用。用实时定量聚合酶链式反应(PCR)检测体外培养的RAFLS中血管诱导生成因子61(CYR61)表达格局。结果:在FLS培养系统中加入PBMC时,PBMC明显黏附于FLS。当加入LMWH时,PBMC对于FLS的黏附减少,表现为脱落PBMC数量增加。随着加入LMWH剂量增加,脱落PBMC数量相应增加。结论:RA患者滑膜组织高表达CYR61分子。LMWH能抑制RAFLS和PBMC的黏附,且呈剂量依赖性。LMWH干扰PBMC黏附FLS的作用与竞争结合了CYR61上的硫酸肝素位点有关。

OBJECTIVE： To study the effect and thepossible mechanism of low molecular weight heparin （LMWH） on immunoeytes＇ adhesion to fibroblast-like synovocytes （FLS） isolated from patients with rheumatoid arthritis （RA） and to study LMWH＇ S possible anti- inflammatory effect on RA. METHODS： LMWH＇ s interference on the adhesion of peripheral blood monouelear cells （PBMC） isolated from healthy volunteers to FLS of RA patients was determined by quantitative counting using flow cytometry. The expression of CYR61 in the in vitro cultured FLS of RA patients was detected using real - time PCR technology. RESULTS： When FLS culture system was added with PBMC, PBMC were obviously found to adhere to FLS, but the number of adhered PBMC decreased after LMWH treatment, which manifested as increase of deciduous PBMC, increased more with the increase of LMWH dose. There was a high expression of CYR61 in synovium tissue in RA patients. CONCLUSION： LMWH inhibited the adheresion of PBMC to FLS from RA patients in a dose - dependent manner, which might be attributed to its competitive combination with heparin sulfate sites on CYR61.