原钒酸钠的细胞毒理学实验研究

目的　探讨原钒酸钠 (SOV)对心肌细胞的毒性作用及其机制。方法　采用循环灌流的方法急性分离成年豚鼠心肌细胞 ,使其在原钒酸钠浓度为 1 ,1 0 ,1 0 0 μmol·L- 1 ,1mmol·L- 1 作用后 ,分别检测 30 ,60 ,1 2 0 ,1 80min 4个时相心肌细胞保存液中LDH、CK含量以及心肌细胞蛋白含量和Na+ ,K+ ATP酶活性 ,检测心肌细胞凋亡率和细胞活力 ;电镜下观察死亡和凋亡细胞的细胞器和细胞核形态学变化。结果　在原钒酸钠 1 0 0 μmol·L- 1 和 1mmol·L- 1 两个组的各时相保存液中LDH、CK以及心肌细胞中蛋白含量增加 ;心肌细胞中Na+ ,K+ ATP酶活性和细胞活力降低 ,与对照组相比差异有显著性 (P <0 0 1 )。流式细胞仪显示 :在 1 80min时 1 0 0 μmol·L- 1 和 1mmol·L- 1 两组的细胞凋亡率与对照组相比增加 (P <0 0 5)。结论　在原钒酸钠浓度高于 1 0 0μmol·L- 1 且作用时间在 30min以上时 ,心肌细胞出现不可逆性损害

Toxicity of sodium orthovanadate on guinea pig myocardial cells

AIM To study toxicity and mechanism of SOV on ventricular myocardial cells. METHODS Cyclical perfusion was used to acutely detach the mature guinea pig myocytes into single cells. The CK,LDH activity in a medium of myocardial cells was measured for 30,60,120 and 180 min after the effect of SOV at 1,10, 100 μmol·L -1 and 1 mmol·L -1 . Rates of cell viability and apoptosis were also measured. The protein content and Na +,K + ATP enzyme activity of the myocardial cells were measured as well. An electron microscope was used to investigate cell nucleus and organelle morphology. RESULTS The release of CK, LDH and protein from the myocardial cell gradually increased after administering SOV at 100 μmol·L -1 and 1 mmol·L -1 to the cells. While ,the cell activity decreased. CK release, which indicated the extent of myocardial cell injury, was more sensitively than that of LDH and cell viability. Meanwhile ,the activity of Na +,K + ATP enzyme decreased gradually. The rate of apoptosis of myocardial cells significantly changed for 180 min after treated with SOV at 100 μmol·L -1 and 1 mmol·L -1 . CONCLUSION Guinea pig ventricular myocardial cells were only injured when exposed for 30 min to 100 μmol·L -1 and 1 mmol·L -1 concentrations of SOV higher than the effective concentration of the drug.