Regulation of Mcl-1 expression in rheumatoid arthritis synovial macrophages
Regulation of Mcl-1 expression in rheumatoid arthritis synovial macrophages
复制标题
DOI：
10.1002/art.22132
复制
发表时间：
2006-10-01
影响因子：
--
通讯作者：
Pope, Richard M.
中科院分区：
文献类型：
--
作者：
Liu, Hongtao;Huang, QiQuan;Pope, Richard M.
目标。对细胞凋亡的抵抗可能是类风湿关节炎（RA）持续存在的一个重要机制。本研究旨在研究抗凋亡Bcl-2家族成员Mcl-1在ra患者关节巨噬细胞中的表达、调控和功能。从RA患者的滑液（SF）中分离出单个核细胞。流式细胞术对CD14+细胞进行细胞内染色，对分离的巨噬细胞进行实时聚合酶链反应或免疫印迹分析，记录Mcl-1的表达。Mcl-1的表达被小干扰RNA （siRNA）或磷脂酰肌醇3-激酶（PI 3-激酶）/Akt-1和转录信号传导和激活因子3 （STAT-3）通路的化学抑制剂抑制。细胞凋亡的定义是线粒体跨膜电位的丧失和DNA的断裂。与正常体外分化的巨噬细胞相比，RA患者SF的CD14+巨噬细胞中Mcl-1的表达升高。抑制PI 3-激酶/Akt-1或STAT-3通路可显著降低SF内CD14+细胞的百分比，导致Mcl-1减少，诱导滑膜巨噬细胞凋亡。Mcl-1 siRNA转染RA滑膜巨噬细胞可导致凋亡细胞死亡。Mcl-1对RA患者关节巨噬细胞的存活至关重要，因此是该疾病的潜在治疗靶点。
Objective. Resistance to apoptosis may be an important mechanism contributing to the persistence of rheumatoid arthritis (RA). This study was undertaken to characterize the expression, regulation, and function of the antiapoptotic Bcl-2 family member Mcl-1 in macrophages isolated from the joints of patients with RA.Methods. Mononuclear cells were isolated from the synovial fluid (SF) of patients with RA. Mcl-1 expression was documented by intracellular staining of CD14+ cells using flow cytometry, and by real-time polymerase chain reaction or immunoblot analysis of isolated macrophages. The expression of Mcl-1 was suppressed with small interfering RNA (siRNA) or chemical inhibitors of the phosphatidylinositol 3-kinase (PI 3-kinase)/Akt-1 and signal transducer and activator of transcription 3 (STAT-3) pathways. Apoptosis was defined by the loss of mitochondrial transmembrane potential and by DNA fragmentation.Results. The expression of Mcl-1 was increased in CD14+ macrophages from the SF of patients with RA compared with normal in vitro-differentiated macrophages. Inhibition of the PI 3-kinase/Akt-1 or STAT-3 pathways significantly reduced the percentage of CD14+ cells within the SF and resulted in the reduction of Mcl-1 and the induction of apoptosis of synovial macrophages. Transfection of RA synovial macrophages with Mcl-1 siRNA resulted in apoptotic cell death.Conclusion. Mcl-1 is critical for the survival of macrophages in the joints of patients with RA, and is therefore a potential therapeutic target in this disease.