钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用

畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (3): 360-365.doi:

钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用

陈蕾¹，李志鹏^1,2，李术¹，徐世文¹*

1.东北农业大学动物医学学院，哈尔滨 150030；2.山东省安丘市凌河镇畜牧兽医管理站，安丘 262100

收稿日期:2009-11-06 修回日期:1900-01-01 出版日期:2010-03-24 发布日期:2010-03-24
通讯作者: 徐世文

Effect of the Disequilibrium of Calcium Homeostasis on the Apoptosis of the Neurons in the Chicken Brain Induced by Manganese in Vitro

CHEN Lei ¹, LI Zhipeng^1,2, LI Shu ¹, XU Shiwen¹*

1. College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China;2. Linghe Animal Husbandry and Veterinary Station, Anqiu 262100, China

Received:2009-11-06 Revised:1900-01-01 Online:2010-03-24 Published:2010-03-24
Contact: XU Shiwen

摘要/Abstract

摘要： 旨在探讨钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用。以体外培养鸡胚脑神经元为研究对象，在含终浓度为0、1.5、2、2.5 mmol·L-1 MnCl2的DMEM培养液中培养24 h，应用流式细胞仪检测磷脂酰丝氨酸（PS）和线粒体膜电位（ΔΨm），琼脂糖凝胶电泳法（DNA Ladder）检测细胞染色质的断裂，以Fura3/AM为探针检测细胞内游离钙离子浓度（［Ca2+］i），用实时定量PCR法检测细胞内CaM mRNA的表达水平。结果表明，随着MnCl2浓度的升高，ΔΨm呈降低趋势，PS膜外翻增加，细胞染色质DNA发生损伤，产生明显的梯形条带，细胞内［Ca2+］i呈升高趋势，CaM mRNA的表达水平下降。结果提示，过量锰通过抑制CaM活性，引起神经元内［Ca2+］i升高，膜通透性发生改变，ΔΨm降低，从而导致鸡胚脑神经元发生凋亡。

Abstract: To study the effect of the disequilibrium of calcium homeostasis on the apoptosis of the neurons in the chicken brain induced by manganese in vitro, neurons cultured in vitro were chosen as the research object. Neurons were cultured in the DMEM for 24 h with the final concentration of 0, 1.5, 2 and 2.5 mmol·L-1 MnCl2. The mitochondrial membrane potential (ΔΨm) and phosphatidylserine (PS) were assessed by flow cytometry. DNA fragmentations were detected by DNA Ladder. The calcium ion concentration within neurons (［Ca2+］i) was detected using the Fura3/AM as the probe and expression level of intracellular CaM mRNA was detected by FQPCR. The results showed that ΔΨm tended to decline the expansion of the pore, DNA fragmentations and ［Ca2+］i were increased. Moreover, the expression of CaM mRNA was decreased. This study indicated that MnCl2 could decrease the activity of CaM, ΔΨm and increase ［Ca2+］ result in apoptosis of the chicken neurons.

陈蕾;李志鹏;李术;徐世文. 钙稳态失衡在锰致体外培养鸡胚脑神经细胞凋亡中的作用[J]. 畜牧兽医学报, 2010, 41(3): 360-365.

CHEN Lei;LI Zhipeng;LI Shu;XU Shiwen . Effect of the Disequilibrium of Calcium Homeostasis on the Apoptosis of the Neurons in the Chicken Brain Induced by Manganese in Vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(3): 360-365.

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