醋酸铅对原代培养大鼠皮层神经元的毒性效应

目的探讨醋酸铅对体外培养大鼠皮层神经元的毒性效应。方法用不同浓度的醋酸铅(50、100和200μg/ml)染毒后,原代培养乳大鼠皮层神经元,通过中性红染色和PI-Hoechst 33342双染色法观察醋酸铅作用6、12、24和48h对细胞存活的影响;结果PI-Hoechst 33342双染色结果表明,各组醋酸铅染毒12h后,部分细胞出现核固缩,随着醋酸铅浓度的增加和培养时间的延长,出现核固缩细胞增多,呈红色荧光细胞比例增加,凋亡细胞增加。对照组细胞凋亡比例明显低与中浓度和高浓度醋酸铅组P0.05)。中性红染色结果表明,不同浓度醋酸铅作用皮层神经元不同时间后各染铅组细胞活性随时间的延长而降低,但除高醋酸铅染毒组在24和48h较6h差异有统计学意义(P0.05)外,其余差异均无统计学意义(P0.05)。各染铅组细胞活性随染铅剂量的增加而降低,尤其是高铅组与对照组及低铅组相比,差异有统计学意义(P0.05)。结论低、中、高浓度醋酸铅(50、100、200μg/ml)作用皮层神经元12h后细胞存活都明显低于正常对照组,并呈剂量和时间依赖关系。

Toxic Effect of Lead Acetate on Cortical Neurons of Primary Cultured Rats

[Objective] To investigate the toxic effect of lead acetate on in vitro cultured cortical neurons of rats.[Methods]Primary cultured cortical neurons of newborn rats were treated with different concentration of lead acetate （50,100 and 200 μg/ml）. Neutral red stain and PI-Hoechst 33342 double stain were used to compare the viability of cortex neurons, which were cultured in the medium containing different concentration of lead acetate for 6 hours, 12 hours, 24 hours and 48 hours respectively.[Results]PI-Hoechst 33 342 double staining showed pyknosis occurred in some cortical neurons and that increased with the increase of the dose of lead acetate and culture time, the cortical neurons with red fluorescent and apoptosis increased, after 12 hours of lead acetate exposure. Apoptotic rate of cortical neurons of control group was significantly lower than that in both medium and high concentration of lead acetate groups （P0.05）. Neutral red staining showed time, and the difference was significant between cortical neurons treated with high concentration of lead acetate for 24 and 48 hours and cortical neurons treated for 6 hours, the rest was not significantly different （P0.05）. The viability of cortical neurons treated with different concentration of lead acetate decreased with the increase of lead acetate dose, especially for the high and low concentration of lead acetate groups, the difference was significant （P0.05）.[Conclusion]The viability of cortical neurons treated with high, medium and high concentration of lead acetate for 12 hours is significantly lower than that of control group, and shows dose-and time-dependent relationship.