无镁对体外原代培养胎鼠皮层神经元糖皮质激素受体表达的影响

Effect of magnesium-free on glucocorticoid receptor expression in primary cultured cortical neurons of fetal rats in vitro

摘要:

目的：研究无镁诱导惊厥后发育中皮层神经元表达糖皮质激素受体（glucocorticoid receptor, GR）的影响。方法：以原代培养的胎鼠大脑皮层神经元无镁诱导的反复惊厥样放电为模型，根据对培养6 d皮层神经元的不同处理分为正常细胞外液组和无镁细胞外液组。神经元在上述2种液体中孵育3 h，然后恢复正常DMEM培养液继续培养，在无镁损害后1 d、7 d及12 d时应用免疫细胞化学技术及实时定量PCR测定发育中大鼠皮层神经元无镁细胞外液处理后GR表达的变化。结果：应用免疫细胞化学方法发现无镁损伤后12 d惊厥组GR蛋白表达较对照组显著下降（P<0.01）；实时定量PCR发现无镁损伤后1 d、7 d惊厥组GR mRNA表达较对照组显著增高（P<0.05）。结论：无镁细胞外液处理可以诱导发育中大鼠皮层神经元GR表达的改变。［中国当代儿科杂志，2010，12（3）：211-214］

Abstract:

OBJECTIVE: To study the changes of glucocorticoid receptor (GR) expression in embryonic rat cortical neurons exposed to transient Mg2+-free treatment. METHODS: Six days after rat cortical neuronal cultures, two groups were created based on the medium to which were transiently exposed. The control group was exposed to a physiological solution (PS), and the Mg2+-free group was exposed to the same medium as the control group except for the removal of magnesium. The expression of GR mRNA and protein was determined by real-time PCR and immunocytochemistry staining 1, 7 and 12 days after transient Mg2+-free treatment. RESULTS: Compared to the control group, the Mg2+-free group displayed the significantly less accumulated optical density (AOD) of GR immunoreactivity 12 days after transient Mg2+-free treatment (P<0.05). On the contrary, GR mRNA expression increased significantly 1 and 7 days after transient Mg2+-free treatment in the Mg2+-free group (P<0.05). CONCLUSIONS: GR expression is modified following Mg-free-induced injury in cultured developing neurons in rats.［Chin J Contemp Pediatr, 2010, 12 (3):211-214］