Rho激酶抑制剂对类风湿关节炎患者滑膜单核巨噬细胞分泌炎症因子的影响

目的 研究类风湿关节炎(RA)患者关节滑膜单核巨噬细胞Rho激酶(ROK)活化情况,并探讨该激酶抑制剂对滑膜单核巨噬细胞分泌炎症细胞因子的影响.方法 单核巨噬细胞分离采用Ficoll密度梯度离心法和黏附法;ROK活性用磷酸化MYPTl蛋白表达来表示,磷酸化MYPTl蛋白检测采用免疫印迹法,单核巨噬细胞活性检测采用MTT法,细胞因子水平采用ELISA检测.结果 新鲜分离的RA患者关节滑液单核巨噬细胞ROK活性显著高于自身配对的和正常对照组外周血单核细胞;新鲜分离的RA患者滑液单核巨噬细胞磷酸化MYPTl表达量与RA患者DAS28评分呈正相关关系(r=0.69,P<0.05),ROK抑制剂Y27632在抑制RA滑膜单核巨噬细胞ROK活性的同时,对肿瘤坏死因子(TNr)α、白细胞介素(IL)-1β、IL-6等炎症细胞因子分泌也有显著抑制作用,但对抗炎细胞因子IL-10分泌没有影响.结论 RA滑液巨噬细胞中ROK处于异常活化状态,并与RA疾病活动有关.ROK特异性抑制剂对RA滑液巨噬细胞分泌多种炎症细胞因子均有抑制作用,提示ROK可能是治疗RA滑膜炎症的潜在靶点.

Cytokine in synovial monocytic cells from rheumatoid arthritis patients

Objective To evaluate the inhibitory effect of blockade of Rho kinase upon mediating the secretion of proinflammatory cytokine in monocytic cells from rheumatoid arthritis (RA)patients. Methods Synovial fluid (SF) monocytic cells and peripheral blood monocytes (PB) from active RA patients were treated with TNFα or LPS respectively in the presence or absence of a specific ROK inhibitor, Y27632. ROK activity was assessed by Western blot and cytokine secretion measured by ELISA. Results Elevated ROK activity was found in synovial fluid monocytic cells from active RA patients.ROK activity was correlated with DAS, an index of disease activity of RA patients. ROK inhibitor Y27632 reduced the secretion of TNFα, IL-1β and IL-6 in RA SF monocytic cells, but had no effect upon the secretion of IL-10, an anti-inflammatory cytokine. Conclusion The present study provides novel evidence that ROK mediates the secretion of proinflammatory cytokines in monocytic cells from RA synovial fluids,suggesting a critical role of ROK in macrophage-mediated synovial inflammation of RA. Thus inhibition of ROK may be a new therapeutic target for RA.