氟化物对原代培养大鼠肝细胞酶活力及超微结构的影响

Effect of fluoride on activities of enzyme and ultrastructure in primary cultured rat hepatocytes

目的研究不同剂量的氟化物对原代培养大鼠肝细胞存活率、酶活力及超微结构的影响。方法采用半原位胶原酶消化法分离大鼠肝细胞;MTT法检测细胞存活率;赖氏法检测培养液中谷草转氨酶(AST)和谷丙转氨酶(ALT)活性;透射电镜观察细胞超微结构改变。结果氟化钠染毒24小时后肝细胞存活率下降,且呈现明显的剂量-效应关系;2mmolL和4mmolL染氟组肝细胞培养液中ALT和AST的活性显著升高(P<005);透射电镜下染氟组肝细胞线粒体肿胀,内质网排列紊乱或断裂。结论过量氟化物对原代培养大鼠肝细胞有明显的毒作用,其主要作用方式是引起细胞膜和细胞器质膜损伤。

Objective To study the cell viability, activities of enzyme and ultrastructure changes induced by sodium fluoride in primary cultured rat hepatocytes. Methods Hepatocytes were isolated using half-in situ collagenase digestion method. Cellular viability was determined by MTT method. The activities of ALT and AST were determined by spectrophotography method. The ultrastructure changes of hepatocyte were observed under transmission electron microscope. Results After cultured with various concentrations of fluoride for 24 hours, a dose-dependent decrease of cell viability was detected in the hepatocytes. The activities of AST and ALT in the 2 mmol/L and 4 mmol/L groups were significantly higher than the control group(P<0.05). Transmission electron microscope study showed that in fluoride treated hepatocytes the changes included swollen mitochondria and disordered, disrupted endoplasm reticulum. Conclusion Excessive fluoride induced significant toxicity in primary cultured hepatocytes which manifested the injuries of membrane and organell plasma membrane.