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目的 研究重组人白细胞介素 1β(IL 1β)对骨髓基质细胞 (BMSC)膜电位的影响及其与Cl-通道的关系 ,探讨离子通道是否参与细胞因子IL 1的生物信号转导。方法 用荧光染料DiBAC4 (3)标记原代培养的BMSC ,在激光扫描共聚焦显微镜下直接监测IL 1β刺激下细胞膜电位的实时动态变化以及Cl-通道阻断剂呋喃苯胺酸 (furosemide)对IL 1β膜电位效应的影响。结果 IL 1β加入测定体系后浓度依赖性地引起BMSC膜电位的迅速改变。低浓度时 (8～ 10 0kU·L-1)发生去极化反应 ,高浓度时 (2 0 0～ 5 0 0kU·L-1)发生超极化反应。非受体方式作用的IL 1β肽段 16 3～ 171对膜电位无影响。Furosemide(2 0～ 5 0 0 μmol·L-1)浓度依赖性地抑制IL 1β的膜电位效应。结论 膜电位的变化为IL 1受体后早期信号事件 ,Cl-通道的活化参与了IL 1的生物信号转导。
Objective To investigate the effect of recombinant human interleukin-1β (IL-1β) on the membrane potential of bone marrow stromal cells (BMSCs) and its relationship with Cl-channels and to explore whether ion channels participate in the biological signal transduction of cytokine IL1. Methods Primary cultured BMSCs were labeled with fluorescent dye DiBAC4 (3). The real-time dynamic changes of cell membrane potential under IL-1β stimulation were monitored by laser scanning confocal microscopy and the effects of Cl-channel blocker furosemide on IL 1β Effect of membrane potential effect. Results After IL 1β was added to the assay system, the BMSC membrane potential was rapidly changed in a concentration-dependent manner. The depolarization reaction occurred at low concentration (8-10 kU · L-1) and hyperpolarization occurred at high concentration (200-2500 kU · L-1). Non-receptor IL 1β peptide 16 3 ~ 171 had no effect on membrane potential. Furosemide (20 ~ 500 μmol·L -1) inhibited the membrane potential of IL 1β in a concentration-dependent manner. Conclusion The change of membrane potential is early signal event after IL 1 receptor, and the activation of Cl-channel is involved in the biological signal transduction of IL 1.