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本文研究1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)对体外培养大鼠中脑多巴胺(DA)神经元的毒性作用。取胚胎腹侧中脑,制成细胞悬液,常规体外培养。实验分为:正常对照组、高浓度MPTP(30μmol/ml)组、低浓度MPTP(3μmol/ml)组和胆碱能神经元对照组。培养细胞定期抽出作免疫组化和荧光组化染色。高浓度MPTP组培养10天,荧光组化阳性细胞已基本消失,此后酪氨酸羟化酶(TH)免疫反应的阳性细胞开始减少,至体外培养2周,每孔平均仅剩14.5个,与正常对照组相比有显著性差异。残存的DA神经元突起缩短或消失。低浓度MPTP组培养2周后,大部分荧光组化阳性细胞消失,但对TH免疫反应阳性细胞的数量无明显影响。基底前脑胆碱能神经元体外生长不受MPTP干扰。实验结果提示MPTP对大鼠中脑DA神经元同样具有特异性损伤作用,其损伤程度与MPTP浓度有关。
In this paper, the toxic effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on dopamine (DA) neurons in rat midbrain were studied. Embryonic ventral midbrain, made of cell suspension, routine in vitro culture. The experiment was divided into: normal control group, high concentration MPTP (30μmol / ml) group, low concentration MPTP (3μmol / ml) group and cholinergic neuron control group. The cultured cells are regularly extracted for immunohistochemistry and fluorescent staining. High concentration of MPTP group cultured for 10 days, the fluorescence-positive cells have disappeared, then tyrosine hydroxylase (TH) immunoreactive cells began to decline, to 2 weeks in vitro, an average of only 14.5 per well , Compared with the normal control group, there are significant differences. Remnants of DA neurites shorten or disappear. After two weeks of culture in low concentration MPTP group, most of the fluorescence group positive cells disappeared, but there was no significant effect on the number of TH immunoreactive positive cells. Basal forebrain cholinergic neurons grow in vitro without MPTP interference. Experimental results suggest that MPTP also has specific damage to DA neurons in rat midbrain, and the degree of damage is related to MPTP concentration.