目的建立体外染锰细胞模型,探讨锰神经毒性作用机制。方法成熟原代皮层与低、中、高不同浓度的锰液,(分别为0.2,0.6,1.0mmol/L,共孵育24h。观察各组神经细胞形态学的变化,单细胞凝胶电泳试验(SCGE)检测神经细胞的DNA损伤,以彗星细胞尾长及彗星样细胞百分率为评价损伤指标。结果光镜下可见不同浓度锰孵育后神经细胞形态学发生改变,SCGE显示神经细胞DNA出现不同程度的损伤,彗星尾长分别为(117.3±30.8),(136.6±29.0),(218.7±22.6)μm;彗星样细胞百分率分别为40%,43%,96%,2指标均较对照组[(84.1±11.8)μm,15%]显著增加(P〈0.01)。尤以高浓度锰损伤组严重,显著高于中、低浓度组(P〈0.01)。结论锰不但能引起体外培养的神经细胞外在的形态学损伤,还可导致神经细胞DNA损伤。
Objective To investigate the mechanisms of Mn neurotoxicity basing on neurons model dealed with Mn in vitro. Methods Mature cortical neurons were incubated with Mn of different dosages (0.2, 0.6, 1.0 tool/L) for 24 hours. The morphological changes in neurons were observed with light microscope and DNA damage was determined through single cells gel electrophoresis (SCGE). The length of comet tail and the percent of comet like neurons were applied to score DNA damage. Results Mn could cause obvious morphological change under light microscope and DNA damage by SCGE. The length of comet tail was (117.3 + 30.8), ( 136.6 + 29.0), (218.7 + 22.6)μm. The percent of comet like neurons was 40%, 43 %, 96 % in different Mn - treated groups respectively. These two indexes in Mn treated groups were higher than that of in control [ ( 184.1 + 11.8) μm, 15 % ] ( P 〈 0.01). DNA damage was specially serious in higher Mn group than that of in lower and medium Mn groups( P 〈 0.01 ). Conclusion Mn could induce morphological change and the damage of DNA in neurons.