牛磺酸拮抗锰诱导体外培养的皮层神经细胞凋亡
首发时间:2007-05-14
摘要:目的:研究锰诱导体外培养的皮层神经细胞凋亡,同时观察牛磺酸对锰诱导凋亡的拮抗作用。方法:体外分离新生乳大鼠皮层神经细胞,培养至最佳生长状态时进行实验。细胞随机分为7组,即对照组、染锰组(设有低中高3个剂量)、干预组(3组,培养液同时加有锰及牛磺酸)。低中高锰的浓度分别为0.2mmol/L,0.6mmol/L,1.0mmol/L,牛磺酸的浓度为1.5mmol/L。干预24小时后终止培养,用末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)及流式细胞术(FCM)定性、定量检测神经细胞凋亡的变化。结果:⑴TUNEL显示锰干预后神经细胞可呈现典型的凋亡形态学变化:核固缩、碎裂,部分甚至核崩解,出现凋亡小体;正常细胞则染色质均匀,形态完整,且凋亡的神经数量有随着锰浓度升高而增多的趋势;牛磺酸干预组对中低浓度锰有一定的拮抗作用,高浓度未见明显改善;⑵FCM定量研究表明锰诱导神经细胞凋亡与其浓度有剂量-依赖关系,牛磺酸能拮抗中低浓度锰所致的细胞细胞凋亡,但对于过高浓度锰所致细胞急剧的死亡—坏死则无改善作用。结论:锰可致体外培养的皮层神经细胞死亡—坏死或凋亡,牛磺酸对中低锰所致的凋亡有保护作用,但对于过高浓度锰致使神经细胞急剧坏死无保护作用。
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Taurine protect cortex neurons cultured in vitro from apoptosis induced by manganese
Abstract:[Objection]: To study manganese induce cortex neurons apoptosis cultured in vitro and taurine protect neurons from apoptosis induced by manganese. Methods:Cortex neurons were separated from Wistar neonatal rats and cultured in vitro.The neurons were treated up to optimal growth conditions. Groupings: Control group,neurons were cultured in normal media; manganese-added group, neurons were incubated with high,medium and low concentration chloride manganese respectively, intervention group, neurons were treated with chloride manganese and taurine together.Chloride manganese’s dosage was 0.2mmol/L,0.6mmol/L and 1.0mmol/L, Taurine was 1.5mmol/L. All treatments lasted 24h. Terminal -deoxynucleotidyl transferase mediated nick end labeling (TUNEL) was used to observe the morphology of apoptosic neurons, Flow cytometry(FCM) quantitated neuron apoptosic rate. Results:⑴Typical morphologic charateristic was found in Mn-added groups, the number of apopotosic nuerons were increased along with Mn’s dosage increasing.The apoptosic neurons were of nucleus shrinkage, fragmentation or even breakdow and apoptosis body found.While in normal neurons,the chromatin were dyed uniform and the shape was integrity. Taurine intervention could protect the effect of medium and low concentrations Mn on neurons, but high Mn. ⑵Quatity assay showed that cortex neurons apoptosis induced by Mn was dosa-dependent relationship with Mn, Taurin could protect neurons from apoptosis induced by medium and low dose Mn, but it had nothing to do with high neurons necrosis induced bu high Mn.Conclusion: Chloride manganese can induce cortex neurons death in vitro—apotosis or necrosis, Taurine can protect neurons from apoptosis induced by medium and low dose Mn, but no high Mn.
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牛磺酸拮抗锰诱导体外培养的皮层神经细胞凋亡
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