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目的研究鱼藤酮(rotenone)对多巴胺(DA)能神经元的早期毒性作用,并探索一种较理想的组织模型。方法采用界面组织培养法建立SD乳鼠的中脑脑片长期培养体系。加入鱼藤酮作用一定的时间,并用测定培养基中乳酸脱氢酶(LDH)活力、组织中DA含量以及进行酪氨酸羟化酶(TH)染色等技术观察它对整个脑片及脑片上DA能神经元的毒性效应。结果不同浓度的鱼藤酮作用24 h后,组织中DA含量随浓度增加明显下降,TH阳性神经元突起呈串珠样改变,数量减少甚至消失。低浓度鱼藤酮作用14 d后,脑片组织中的DA水平显著降低,但未见明显的DA能神经元形态学变化。结论建立了长期、稳定的中脑脑片培养体系;鱼藤酮对整个脑片以及脑片上DA能神经元的毒性作用具有浓度依赖性;功能性损伤早于形态学变化,突起的变性是形态学变化的早期特征。
Objective To study the early toxic effects of rotenone on dopaminergic neurons and to explore a more ideal tissue model. Methods The long-term culture system of midbrain brain slices of neonatal SD rats was established by interfacial tissue culture. Adding rotenone for a certain period of time, and measuring the activity of lactate dehydrogenase (LDH) in the culture medium, the content of DA in the tissue and tyrosine hydroxylase (TH) staining and other techniques to observe it on the whole brain slices and brain DA energy Toxic effects of neurons. Results After treated with different concentrations of rotenone for 24 h, the content of DA in the tissue decreased significantly with the increase of concentration, and the number of TH positive neurons changed in a beaded pattern. The number decreased or even disappeared. After 14 days of low concentration of rotenone, the level of DA in brain slices decreased significantly, but no obvious change of DA neurons was found. Conclusion The long-term and stable culture system of midbrain brain slices has been established. Rotenone has a concentration-dependent effect on the whole slices and on DA neurons in brain slices. The functional damage is earlier than the morphological changes, and the degeneration of the protrusions is the morphological change Early features.