CD40在大鼠腹膜间皮细胞的表达及其功能研究

Expression and function of CD40 on rat peritoneal mesothelial cells

目的 研究腹膜间皮细胞CD40的表达、调节因素及其活化后与细胞间黏附分子1(ICAM-1)分泌的相关性。方法 分离、培养大鼠腹膜问皮细胞,用IFN-γ、TNF-α、IL-1刺激24 h,通过逆转录-多聚酶链反应(RT-PCR)及流式细胞仪(FACS)检测分析间皮细胞CD40表达。通过CD40单克隆抗体(CD40mAb)活化间皮细胞CD40,用FACS检测分析间皮细胞ICAM-1的表达。结果 间皮细胞结构性表达低水平的CD40。IFN-γ、IL-1可显著增加间皮细胞表面CD40mRNA及其蛋白的表达,而以IFN-γ增幅最大。TNF-α对间皮细胞CD40mRNA及其蛋白的表达无显著增加作用。用IFN-γ增加间皮细胞CD40表达的同时,加入CD40mAb活化CD40受体可显著增强间皮细胞ICAM-1的表达。结论 腹膜间皮细胞功能性表达CD40,其与腹腔中阳性CD40配体(CD40L+)细胞相互作用,可能在腹腔局部防御中发挥重要作用,其结果对于限制炎症的发生和发展具有积极意义。

Objective To determine whether peritoneal mesothelial cells express CD40 and to investigate potential mechanism by which CD40-CD40 ligand(CD40L) interaction may be involved in the inflammation of peritoneal membrane. Methods Rat peritoneal mesothelial cells (MC) were harvested from peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α), interleukin(IL)-1 was examined by RT-PCR and FACS analysis. After activation with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. Results MC cultured in vitro expressed CD40 constitutively. The expression of CD40mRNA and CD40 protein up-regulated markedly following the stimulations with IFN-γ, but not with IL-1, TNF-α. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with IFN-γ and CD40mAb. Conclusions MC functionally expresses CD40. The interaction of CD40 on MC and CD40L+ cells in peritoneal cavity may play an important role in peritoneal local defense and may be involved in the inflammation process of peritoneum.