First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, and Chongqing Eye Institute, Chongqing 400016, China.
Rheumatology (Oxford). 2014 May;53(5):810-20. doi: 10.1093/rheumatology/ket438. Epub 2014 Jan 20.
Behçet's disease (BD) is a refractory inflammatory disorder with unknown causes. Since the Notch pathway is critically involved in the immune response, the present study was undertaken to investigate the role of this pathway in BD.
Hes-1, Notch 1-4, Jagged-1, DLL-1 and DLL-4 expression, frequency of IFN-γ and IL-17 expressing Th cells, Notch intracellular domain (NICD), phosphorylation of signal transducer and activator of transcription 3 (STAT3) and the production of IFN-γ and IL-17 were examined by real-time PCR, flow cytometry and ELISA. Notch blockade was performed using the γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-1-alanyl]-S-phenylglycine t-butyl ester (DAPT). Transfection with miR-23b mimics and inhibitor was used to examine the effect of miR-23b on Notch pathway activation.
Active BD patients showed an increased activation of the Notch pathway in association with a higher Th17 response. Notch blockade preferentially inhibited Th17 responses. The effect of Notch blockade on the Th17 response was associated with a lower level of STAT3 phosphorylation. miR-23b was significantly decreased in CD4(+) T cells from active BD patients. CD4(+) T cells transfected with miR-23b showed a reduced expression of NICD and a reduced frequency of IL-17- and IFN-γ-expressing T cells.
The present study suggests that an increased activation of the Notch pathway may contribute to the pathogenesis of BD. Decreased expression of miR-23b may be involved in activation of the Notch pathway in BD. Manipulation of the Notch pathway may offer a novel therapeutic approach for BD.
贝切特病(BD)是一种病因不明的难治性炎症性疾病。由于 Notch 通路在免疫反应中起着至关重要的作用,本研究旨在探讨该通路在 BD 中的作用。
通过实时 PCR、流式细胞术和 ELISA 检测 Hes-1、Notch1-4、Jagged-1、DLL-1 和 DLL-4 的表达、IFN-γ和 IL-17 表达 Th 细胞的频率、Notch 细胞内结构域(NICD)、信号转导和转录激活因子 3(STAT3)的磷酸化以及 IFN-γ和 IL-17 的产生。用γ-分泌酶抑制剂 N-[N-(3,5-二氟苯乙酰基)-1-丙氨酸]-S-苯甘氨酸叔丁酯(DAPT)阻断 Notch 通路。用 miR-23b 模拟物和抑制剂转染来检测 miR-23b 对 Notch 通路激活的影响。
活动期 BD 患者 Notch 通路的激活增加,与 Th17 反应增强相关。Notch 阻断优先抑制 Th17 反应。Notch 阻断对 Th17 反应的影响与 STAT3 磷酸化水平降低有关。来自活动期 BD 患者的 CD4+T 细胞中 miR-23b 显著降低。转染 miR-23b 的 CD4+T 细胞显示 NICD 表达降低,IL-17 和 IFN-γ表达 T 细胞的频率降低。
本研究表明,Notch 通路的过度激活可能有助于 BD 的发病机制。miR-23b 的表达下调可能参与了 BD 中 Notch 通路的激活。Notch 通路的调控可能为 BD 提供一种新的治疗方法。