眼镜蛇毒直接溶解因子对人肝癌细胞株细胞内游离钙动态变化的影响

用荧光染料FIuo-3标记人肝癌细胞株H_(7402)细胞内游离钙,在粘附式细胞仪观察检测单个细胞内游离钙水平的动态变化,细胞在无钙环境中,直接溶解因子(DLF)刺激下细胞内游离钙迅速升高,达到峰值后下降;在细胞培养皿中加入1mmol/L CaCl_2,DLF使胞浆游离钙持续升高;加入10mmol/L CaCl_2,DLF刺激后胞浆游离钙水平无明显变化,表明DLF能引起胞内Ca~(2 )释放和胞外Ca~(2 )内流,细胞外高浓度Ca~(2 )能阻断DLF升高细胞内Ca~(2 )浓度的 作用。

EFFECT OF DIRECT LYTIC FACTOR FROM NAJA NAJA ATRA VENOM ON INTRACELLULAR FREE CALCIUM DYNAMIC CHANGES IN HUMAN HEPATIC CANCER CELL LINE

The free calcium in human hepatic cancer cell line H7402 was labelled with fluorescent dye Fluo-3 and its dynamic changes were observed by using Adherent Cell Analysis and Sorting Cytometer 570 (ACAS 570). When the cells were in a calcium-free environment, the Ievel of intracellular Ca2- elevated rapidly after the stimulation of direct lytic factor (DLF). When it reached the peak, then soon decreased. The Ievel of intracellular Ca2- elevated constantly after the stimulation of DLF when l mmol/L CaCl2 was added into the culture dish. No obvious elevation could be seen in the Ievel of intracellular Ca2- after the stimulation of DLF when 10 mmol/L CaCl2 was added into the culture dish. The results show that DLF can cause intracellular Ca2- release and extracellular Ca2- influx, and the effect of the elevation of intracellular Ca2- caused by DLF can be blocked by high level of extracellular Ca2- concentration.