地塞米松诱导小鼠腹腔巨噬细胞凋亡过程中[Ca~(2 )]i的变化

目的　研究地塞米松诱导凋亡小鼠腹腔巨噬细胞 Ca2 ]i的变化及其对凋亡的影响以及信使分子对凋亡和 Ca2 ]i变化的影响。方法　激光扫描共聚焦显微术、流式细胞术和荧光标记术。结果　 1 凋亡巨噬细胞内fluo 3荧光强度逐渐增强。胞内钙库受体抑制剂 ,尤其是Ca2 内流阻断剂抑制细胞内fluo 3荧光强度变化 ,同时使细胞凋亡率降低 ;2 .staurosporine和DcAMP显著降低巨噬细胞凋亡率并明显抑制fluo 3荧光强度改变。genistein和亚甲蓝稍降低巨噬细胞凋亡率 ,并降低fluo 3荧光强度升高幅度。结论　 1 胞外Ca2 内流和内源性Ca2 释放 ,主要是胞外Ca2 内流使Ca2 ]i逐渐升高并促进巨噬细胞凋亡 ;2 .PKC促进 Ca2 ]i升高和巨噬细胞凋亡。cAMP抑制Ca2 ]i升高和巨噬细胞凋亡。cGMP、TPK降低 Ca2 ]i升高幅度并稍抑制巨噬细胞凋亡。结果提示 ,Ca2 ]i是信使分子调控巨噬细胞凋亡的主要靶点。

Changes of cytoplasmic free Ca~(2 ) during apoptosis of murine peritoneal macrophage induced with dexamethasone

Objective To investigate the changes in Ca 2 ]i and their effects on macrophage apoptosis, the effects of signal molecules on apoptosis and Ca 2 ]i change of apoptotic macrophage induced by dexamethasone. Methods Laser scanning confocal microscopy (LSCM), cytometry and fluorescence labeling. Results 1. The fluo-3 fluorescence in apoptotic macrophage increased gradually. Inhibitors for different receptors of intracellular calcium pool, especially inhibitors of calcium influx inhibited both the increase of fluo-3 fluorescence and apoptosis ratio simultaneously. 2. Staurosporine and DcAMP clearly alleviated macrophage apoptosis and inhibited the changes of fluo-3. Genistein and methyl blue decreased apoptosis ratio slightly and reduced the amplitude of fluo-3 change. Conclusions The results indicated that intracellular Ca 2 release as well as the extracellular Ca 2 influx resulted in the gradual elevation of Ca 2 ]i, which promoted macrophage apoptosis and that PKC accelerated elevation of Ca 2 ]i, then promoted macrophage apoptosis; cAMP restrained elevation of Ca 2 ]i, then in turn inhibited macrophage apoptosis obviously; cGMP, TPK slightly reduced amplitude of Ca 2 ]i change and low-down apoptosis ratio. These results implicated that Ca 2 is one of the main targets at which signal molecules regulate macrophage apoptosis.