共聚焦镜观察凋亡巨噬细胞内pH的变化

DETECTION OF THE CHANGES IN INTRACELLULAR PH OF THE APOPTOTIC MACROPHAGES BY LASER SCANNING CONFOCAL MICROSCOPE

用透射电镜观察巨噬细胞的形态学改变，结果显示，地塞米松处理８小时后，大部分巨噬细胞发生凋亡特征变化：胞突缩短、减少，胞膜完整。胞体皱缩，胞质密度增加，其中出现大量空泡。胞核染色质边聚、浓缩。另外用激光扫描共聚焦显微镜（ＡＣＡＳ５７０）和ｐＨ荧光探针ＳＮＡＲＦ┐１／ＡＭ实时检测地塞米松处理巨噬细胞胞浆ｐＨ的动态变化。加入地塞米松，多数巨噬细胞胞浆马上发生快速和短期的碱化。随后，胞浆ｐＨ缓慢降低，胞浆酸化。结果表明，胞浆酸化是细胞凋亡发展的必然过程，胞浆碱化则很可能与细胞凋亡的发生相关，也可能与细胞种类。

The changes of morphology and intracellular pH of apoptotic macrophages induced by dexamethasone were studied. Using TEM, laser scanning confocal microscope (ACAS570) and pH sensitive fluorescence probes SNARF 1/AM. Eight hours after dexamethasone treatment, the macrophages undrwent apoptotic changes characterized by shortening and decrease of processes, cell shrinkage, compact of cytoplasm with emergence of many vacuoles, nuclear condensation, etc. A quick and short time alkaliztion immediately took place. Then, the intracellular pH decreased slowly, and the cytoplasm was acidificated. The results indicated that the cytoplasm acidification was the inevitable process in apoptosis. However, the cytoplasm alkalization might be related with cell types, and have a direct bearing on the induction of apoptosis.