Smith R A, Orr D J, Haetzman M L, MacPherson N, Storey N D
Department of Anatomy, University of Glasgow, Scotland.
Virchows Arch B Cell Pathol Incl Mol Pathol. 1990;58(5):323-30. doi: 10.1007/BF02890088.
Primary cultures of adult mouse sensory neurons maintained for 8 days in vitro (8 div), in both the presence of non-neuronal cell (NNC) outgrowth and in NNC-reduced cultures, were exposed to doses of ethanol, propanol, acetaldehyde and acrolein. The effects on cell viability were monitored: LD50's of 600 microM acrolein and 100 mM propanol were obtained after 24 h exposures and after 48 h with 1 mM acetaldehyde and 500 mM ethanol. Morphological effects were evident by scanning electron microscopy with sub-acute doses for each agent, using both lower concentrations and shorter exposures. Membrane pitting of the perikaryon and a reduction in the proportion of neurons bearing neurites were common signs of toxic insult. The neurites of treated cells were thicker and more irregular than those of untreated cells; this proved a good indicator of specific neurotoxicity rather than merely a cytotoxic response. Fetal calf serum in the medium lessened the response of neurons to ethanol treatments. Comparison with other in vitro studies suggests these primary cultures are a more sensitive system than established cell lines of neuronal origin for use in neurotoxicity testing.
将成年小鼠感觉神经元的原代培养物在体外培养8天(8日龄),分别置于有非神经元细胞(NNC)生长和NNC减少的培养环境中,然后分别用不同剂量的乙醇、丙醇、乙醛和丙烯醛进行处理。监测对细胞活力的影响:暴露24小时后,丙烯醛的半数致死剂量(LD50)为600微摩尔,丙醇为100毫摩尔;暴露48小时后,乙醛为1毫摩尔,乙醇为500毫摩尔。通过扫描电子显微镜观察各试剂亚急性剂量(使用较低浓度和较短暴露时间)的形态学效应。胞体膜凹陷以及有神经突的神经元比例降低是毒性损伤的常见迹象。处理后细胞的神经突比未处理细胞的更粗且更不规则;这证明是特异性神经毒性的良好指标,而不仅仅是细胞毒性反应。培养基中的胎牛血清减轻了神经元对乙醇处理的反应。与其他体外研究相比,这些原代培养物在神经毒性测试中是比已建立的神经元起源细胞系更敏感的系统。