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采用电镜和粘附式细胞仪(ACAS570)观察了胶体金标记的刀豆素A(ConA-Au)与小鼠腹腔巨噬细胞(Mφ)表面ConA受体的结合、内吞及其在Mφ中转运的形态学改变,同时分析ConA刺激后Mφ内游离钙(Ca2+)的变化。ConA孵育1min后,配体进入内吞小泡,第2次内吞约需20min。粘附式细胞仪分析表明:ConA刺激后50s,Ca2+急剧上升,平均住值达0.557U;约100s后,恢复到刺激前水平;18min后,Ca2+重新回升,持续约18min后开始下降。本实验揭示配体再内吞与Ca2+重新上升的时间相吻合,其肯定性和机理有待于进一步研究证实。
The binding of colloidal gold-labeled ConA-Au to the ConA receptor on the surface of mouse peritoneal macrophage (Mφ) was observed by electron microscopy and adherent cytometry (ACAS570) The morphological changes of Ca2 + transport and the changes of Ca2 + in Mφ after ConA stimulation were analyzed. ConA incubated 1min, the ligand into the endocytosis, the second endocytosis takes about 20min. Adherent cytometry analysis showed that Ca2 + increased sharply at 50s after ConA stimulation, with an average of 0.557U. After about 100s, Ca2 + returned to the pre-stimulation level. Ca2 + began to rise again after 18min and continued to decrease after about 18min. This experiment revealed that the endocytosis of ligand and Ca2 + coincide with the time of re-rise, its affirmation and mechanism needs further study confirmed.