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

薄涛; 易璐; 王团美; 里健; 李杏芳; 毛定安

PDF(1103 KB)

中国当代儿科杂志 ›› 2010, Vol. 12 ›› Issue (3) : 211-214.

论著·实验研究

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

薄涛，易璐，王团美，里健，李杏芳，毛定安

作者信息 +

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

BO Tao, YI Lu, WANG Tuan-Mei, LI Jian, LI Xing-Fang, MAO Ding-An

Author information +

文章历史 +

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］

导出引用

薄涛, 易璐, 王团美, 里健, 李杏芳, 毛定安. 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[J]. 中国当代儿科杂志. 2010, 12(3): 211-214

BO Tao, YI Lu, WANG Tuan-Mei, LI Jian, LI Xing-Fang, MAO Ding-An. Effect of magnesium-free on glucocorticoid receptor expression in primary cultured cortical neurons of fetal rats in vitro[J]. Chinese Journal of Contemporary Pediatrics. 2010, 12(3): 211-214

中图分类号： R-33

参考文献

［1］Ferriero DM. Neonatal brain injury ［J］. N Engl J Med, 2004, 351(19): 1985-1995.
［2］Hauwal M, Furon E, Canova C, Griffths M, Neal J, Gasque P. Innate (inherent) control of brain infection, brain inflammation and brain repair: the role of microglia, astrocytes, "protective" glial stem cells and stromal ependymal cells ［J］. Brain Res Rev, 2005, 48 (2):220-233.
［3］Milla R, Riitta P, Helena L, Liisa L, Leena H. Interleukin-6-174 and-572 genotypes and the volume of deep gray matter in preterm infants ［J］. Pediatr Res, 2009, 65(1): 90-96.
［4］Dammon O, Matthews SG. The effects of repeated antenatal glucocorticoid therapy in the developing brain［J］. Pediatr Res, 2001, 50 (5):563-564.
［5］Cao HY, Jiang YW, Liu ZW, Wu XR. Effect of recurrent epileptiform discharges induced by magnesium-free treatment on developing cortical neurons in vitro［J］. Dev Brain Res, 2003, 142 (1):1-6.
［6］Altura BM, Gebrewold A, Zhang A, Altura BT. Low extracellular magnesium ions induce lipid perioxidation and activation of nuclear factor-kappa B in canine cerebral vascular smooth muscle: possible relation to traumatic brain injury and strokes［J］. Neurosci Lett, 2003, 341(3):189-192.
［7］曹海燕，姜玉武，薄涛，吴希如. 无镁诱导的反复惊厥样放电对体外培养发育中神经元损伤的研究［J］. 中华儿科杂志, 2003, 41(1):25-28.
［8］曹海燕，姜玉武，吴希如. 无镁诱导培养的大鼠皮质神经元癫痫样放电［J］. 北京大学学报（医学版）, 2002, 34(3):237-241.
［9］Donley MP, Schμlkin J, Rosen JB. Glucocorticoid receptor antagonism in the basolateral amygdala and ventral hippocampus interferes with long-term memory of contextual fear［J］. Behav Brain Res, 2005, 164(2):197-205.
［10］Velísek L. Prenatal corticosteroid impact on hippocampus: implication for postnatal outcomes ［J］. Epliepsy Behav, 2005, 7(1): 57-67.
［11］Han F, Ozawa H, Matsuda K, Nishi M, Kawata M. Colocafization of mineralocorticoid receptor and glucocorticoid receptor in the hippocampus and hypothalamus ［J］. Neurosci Res, 2005, 51(4): 371-381.
［12］Pryce CR, Feldon J, Fuchs E, Knuesel I, Oertle T, Sengstag C, et al. Postnatal ontogeny of hippocampal expression of the mineralocorticoid and glucocorticoid receptor in the common monkey ［J］. Eur J Neurosci, 2005, 21(6): 1521-1535.
［13］Donley MP, Schulkin J, Rosen JB. Glucocorticoid receptor antagonism in the basolateral amygdala and ventral hippocampus interferes with long-term memory of contextual fear ［J］. Behav Brain Res, 2005, 164(2):197-205.
［14］Nishi M, Kawata M. Dynamics of glucocorticoid receptor and mineralocorticoid receptor: implications from live cell imaging studies ［J］. Neuroendocrinology, 2007, 85(3):186-192.
［15］Marin F, Pozzto C, Andreetta V, Jansson B, Arban R, Domenici E, et al. Single exposure to social defeat increases corticotropin-releasing factor and glucocorticoid receptor on RNA expression in rat hippocampus［J］. Brain Res, 2006, 1067(1):25-35.
［16］Tamura G ,Olson D, Miron J, Clark TG. Tolloid-like I is negatively regulated by stress and glucocorticoid［J］. Brain Res Mol Brain Res, 2005, 142(2):81-90.
［17］Isgor C, Kabbaj M, Akil H, Watson SJ. Delayed effects of chronic variable stress during peripubertal juvenile period on hippocampal morphology and on cognitive and stress axis functions in rats［J］. Hippocampus, 2004, 14(5):636-648.
［18］Yang CH, Huang CC, Hsu KS. Behavioral stress modifies hippocampal synaptic plasticity through corticosterone-induced sustained extracellular signal-regulated Kinase/mitogen-activated protein Kinase activation［J］. J Neurosci, 2004, 24(49):11029-11034.