摘要
本试验旨在研究低剂量铅对原代培养的大鼠大脑皮质神经细胞的存活及凋亡的影响。用新生24 h内SD大鼠,建立了大脑皮质神经细胞原代培养方法,通过不同浓度醋酸铅染毒后,用MTT法测定神经细胞的存活率;应用Hoechst 33258荧光染色和流式细胞仪检测铅对神经细胞凋亡的影响。结果表明,低剂量铅（50μmol/L）在3～6 h对神经细胞生长无明显作用,12 h后则表现抑制作用,其他各染毒组随剂量增加和作用时间的延长,细胞存活率逐渐下降,与对照组相比,作用12 h以上差异显著（P〈0.05）;Hoechst 33258荧光染色随染铅剂量的增加,细胞核呈新月形,染色质浓缩,出现凋亡小体,甚至核碎裂;细胞凋亡率随染毒剂量的增加而逐渐升高,对照组为0.7%,染铅组依次为1.1%、1.5%、1.9%、2.2%,而各染铅组细胞坏死率均为0.2%,与对照组比较只增加了0.1%。表明本试验所选择的铅浓度引起神经细胞死亡是以凋亡为主,低剂量铅具有抑制神经细胞存活和促进其凋亡的毒性作用。
To study the impact of low-level lead on the survival and apoptosis of primary cultured cortical neurons, develop a method of primary culture cortical neurons of newborn SD rats and establish experiment cell model of the neurons. Being treated with different dosage of lead acetate, the survival of the cultured cells were observed at different time-point by measurement of optical density （D） value in MTT method, The apoptosis of neurons was investigated by Hoechst 33258 fluorescent staining and flow cytometry. The results showed that low level of 50 μmol/L Pb^2＋ could promote the neuronst growth in a short time （100.27%±1.61% ） ,however,depressant effect was displayed after 6 h （≤99.20%±0.13 %）. Cell survival rate was gradually decreased when increased Pb^2＋ and prolonged time,in comparison with control group, significant decrease was observed after 12 h（P〈0.05）. Cell nucleus developed crescentiform,chromatin condense,apoptotic body and nuclear fragmentation. Apoptotic rate was gradually increased with of increasing concentration Pb^2＋ （0.7 %, 1.1%, 1.5 %, 1.9 %, 2.2 % ）. The data indicated that low-level lead could inhibit the neurons＇ survival and promote neuron apoptosis.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2008年第5期576-580,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30440050,305713647)
关键词
铅
皮质神经元
存活率
细胞凋亡
毒性效应
lead
cortical neuron
survival rate
cell apoptosis
toxic effect
