CAJ | 학술논문

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.
목적 연구류풍습관절염(RA)환자관절활막단핵거서세포Rho격매(ROK)활화정황,병탐토해격매억제제대활막단핵거서세포분비염증세포인자적영향.방법 단핵거서세포분리채용Ficoll밀도제도리심법화점부법;ROK활성용린산화MYPTl단백표체래표시,린산화MYPTl단백검측채용면역인적법,단핵거서세포활성검측채용MTT법,세포인자수평채용ELISA검측.결과 신선분리적RA환자관절활액단핵거서세포ROK활성현저고우자신배대적화정상대조조외주혈단핵세포;신선분리적RA환자활액단핵거서세포린산화MYPTl표체량여RA환자DAS28평분정정상관관계(r=0.69,P<0.05),ROK억제제Y27632재억제RA활막단핵거서세포ROK활성적동시,대종류배사인자(TNr)α、백세포개소(IL)-1β、IL-6등염증세포인자분비야유현저억제작용,단대항염세포인자IL-10분비몰유영향.결론 RA활액거서세포중ROK처우이상활화상태,병여RA질병활동유관.ROK특이성억제제대RA활액거서세포분비다충염증세포인자균유억제작용,제시ROK가능시치료RA활막염증적잠재파점.
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.