Expression analysis of Notch-related molecules in peripheral blood T helper cells of patients with rheumatoid arthritis
Expression analysis of Notch-related molecules in peripheral blood T helper cells of patients with rheumatoid arthritis
复制标题
类风湿关节炎患者外周血T辅助细胞中Notch相关分子的表达分析
DOI：
10.3109/03009740903124424
复制
发表时间：
2010-01-01
影响因子：
2.1
通讯作者：
Li, J.
中科院分区：
文献类型：
--
作者：
Jiao, Z.;Wang, W.;Li, J.
目的：Notch同源物在类风湿关节炎（RA）和培养的滑膜细胞局部组织炎症中的表达已有报道，但Notch相关分子在RA外周血淋巴细胞中的表达谱尚不清楚。在本研究中，我们检测了RA患者外周血淋巴细胞中Notch受体及其下游分子的表达。方法：采用流式细胞术和实时聚合酶链反应（PCR）检测RA患者外周血淋巴细胞中Notch受体的表达。real-time PCR检测RA患者T辅助细胞中代表性Notch靶基因HES-1和调控基因NUMB的表达，免疫印迹法检测Notch胞内结构域（ICD）的表达。结果：活动期RA患者T辅助细胞中Notch 2、Notch 3、Notch 4表达升高，其中Notch 3主要通过活化的T细胞表达升高。值得注意的是，在不活跃的RA患者中，T细胞中Notch 3的表达下降，其水平与健康对照组相似（HC）。B细胞上很少观察到Notch受体，RA患者和HC患者在表达上没有差异。RA患者的T辅助细胞靶基因HES-1表达增加，Notch信号负调控基因NUMB表达减少。在活动期RA疾病的T辅助细胞中，Notch-ICD的核易位也增加。结论：本研究表明，与HC相比，RA患者的T辅助细胞的Notch受体表达谱显著改变，Notch信号激活增强。
Objective: Expression of Notch homologues in local tissue inflammation in rheumatoid arthritis (RA) and cultured synoviocytes has been reported, but the expression profile of Notch-related molecules in peripheral lymphocytes in RA remains unclear. In this study, we measured the expression of Notch receptors and downstream molecules in peripheral lymphocytes from RA patients. Methods: Expression of Notch receptors in peripheral lymphocytes of RA patients was assessed by both flow cytometry and real-time polymerase chain reaction (PCR). Expression of the representative Notch target gene HES-1 and the regulatory gene NUMB in purified T helper cells from RA patients was determined by real-time PCR, and expression of Notch intracellular domain (ICD) was determined by immunoblot analysis. Results: There was an increased expression of Notch 2, Notch 3, and Notch 4 in T helper cells from active RA patients, among which increased expression of Notch 3 was mainly by activated T cells. Notably, expression of Notch 3 in T cells decreased in inactive RA patients and the level was similar to that of healthy controls (HC). Notch receptors were rarely observed on B cells and no difference in expression was found between RA patients and HC. T helper cells from RA patients exhibited increased expression of the target gene HES-1 but decreased expression of the negative modulation gene NUMB of Notch signalling. There was also an increased nuclear translocation of Notch-ICD in T helper cells from active RA disease. Conclusion: The present study demonstrated that T helper cells from RA patients display a significantly altered expression profile of Notch receptors and enhanced activation of Notch signalling compared with HC.