类风湿关节炎成纤维样滑膜细胞诱导分化的体外研究  

In vitro proliferation and differentiation characteristics of fibroblast like synovial cells in patients with rheumatoid arthritis

作　　者：舒强[1] 李兴福[1] 戴国峰[2] 刘花香[1] 

机构地区：[1]山东大学齐鲁医院风湿科,济南250012 [2]山东大学齐鲁医院骨科,济南250012

出　　处：《中华风湿病学杂志》2007年第6期348-351,共4页Chinese Journal of Rheumatology

基　　金：山东省科技厅基金(YZ001C16)

摘　　要：目的研究类风湿关节炎(RA)成纤维样滑膜细胞(FLS)体外培养时的增生分化特点及细胞分化诱导剂全反式维甲酸(ATRA)、骨化三醇[1,25(OH)_2D_3]、地塞米松(DEX)对RA-FLS增殖分化的影响。方法关节镜、关节活检针取RA患者滑膜组织或抽取RA患者关节积液分离培养鉴定滑膜细胞,四甲基偶氮唑蓝(MTT)法和流式细胞仪法分别观察ATRA、骨化三醇和地塞米松对FLS细胞存活分数(SF)和细胞周期的影响。结果RA-FLS在体外无诱导剂作用时呈现良性增殖方式。ATRA、骨化三醇和地塞米松对FLS的细胞标记和免疫染色无明显影响。三种诱导剂对RA-FLS的SF值均有不同程度的抑制(P<0.05),其中地塞米松抑制作用最明显;骨化三醇和地塞米松对RA-FLS的SF抑制作用呈现剂量依赖性曲线(P<0.01)。三种诱导剂均促使RA-FLS细胞周期改变即凋亡峰出现、S期缩短。结论体外培养的RA-FLS生长特性为非恶性无限制增生。ATRA、骨化三醇和地塞米松抑制RA-FLS细胞增殖,促进细胞分化,诱导FLS凋亡,其中地塞米松抗增殖作用最明显;但未发现三者将FLS诱导分化为其他类型细胞。Objective To investigate the proliferation and differentiation characteristics of fibroblast like synovial cells （FLS） in rheumatoid arthritis （RA） in vitro and the mechanism of the immunosuppressive effect of differentiation inducers, such as all trans retinoid acid （ATRA）, calcitriol [ 1,25 （OH）2D3] and dexamethasone （DEX）. Methods FLS of knee synovial tissues from RA patients were cultured and identified in vitro in the presence or absence of ATRA, 1,25（OH）2D3 and DEX respectively. Synoviocyte proliferation in RA were measured by MTY colorimetric assay and the survival fraction （SF） of FLS was evaluated. Cell cycle of FLS was observed using fluorescence-activated cell sorting （FCS） method in RA patients. Results The identified synovial cells in patients with RA were FLS（Vimentin and Fibronectin expression was positive）, and hadn＇t been transformed or differentiated to adipocytes and osteoblasts with the three inducers. The SF of all RA-FLS interfered by ATRA, 1,25（OH）2D3 and DEX was much lower than that without drugs vehicle group in RA-FLS（P〈0.05）. The immunoregulation effects of DEX were the strongest among them. 1,25 （OH）2D3 and DEX at physiological doses demonstrated markedly dose-dependent inhibitory effect on FLS proliferation efficiency. ATRA, 1,25（OH）2D3 and DEX induced cell apoptosis in RA-FLS evidently, because FACS assay had shown that the S phase was shortened and an apoptosis peak appeared markedly. Conclusion RA synovial fi- broblasts do not have malignant unlimited growth pattern without stimulated by proinflammatory cytokines in vitro. The therapeutic effect of ATRA, 1,25（OH）2D3 and DEX can be associated with their abilities to inhibit RA-FLS proliferation. They can markedly suppress abnormal differentiation and induce cell apoptosis effctively at physiological dosage, whereas RA-FLS fails to be transformed to other types of cells by these inducers.

关 键 词：关节炎 类风湿 维甲酸 骨化三醇 地塞米松 成纤维样滑膜细胞 

分 类 号：R593.22[医药卫生—内科学]