慢性低氧低糖培养对大鼠海马神经元α-和β-分泌酶活性的影响

Chronic hypoxic and hypoglycemic culture affect activity of α-and β-secretase in rat hippolcampal neuron

目的探讨慢性低氧低糖培养对大鼠海马神经元α-和β-分泌酶活性的影响。方法取新生24h的wistar大鼠海马神经元培养,用慢性低氧低糖进行干预,MTT法检测神经元细胞活力,直接荧光法检测α、β-分泌酶活性。结果低氧低糖组较对照组细胞活力降低,具有显著性差异(P<0.05),低氧低糖培养海马神经元12h的α-分泌酶活性升高(P<0.0001),低氧低糖培养海马神经元12、24、36h的β-分泌酶活性升高(P=0.0032、0.0018、<0.0001)。结论慢性低氧低糖培养对海马神经元具有毒性,可能与其升高β-分泌酶活性使具有神经营养作用的sAPPα降低和具有神经毒性作用的Aβ升高有关。

Objective To investigate the effect of chronic hypoxic and hypoglycemic culture on activity of α- and β-secretase in rat hippolcampal neurons. Methods Primary cultures of 1-day-old wistar rat hippolcampal neurons were preparied for following experiments, cells were exposed to chronic hypoxic and hypoglycemic culture for 12 h, 24 h and 36 h respectively. Cell viability was assessed by the MTT assay, α- secretase and β-secretase cleavage activity were measured by Secretase Activity Kit. Results MTT assay displayed cell viability of chronic hypoxic and hypoglycemic group decreased versus control group （P〈0. 05）. After chronic hypoxic and hypoglycemic culture, α- and β- secretase cleavage activity （12 h） increased dramatically （P〈 0. 05）. Conclusions Chronic hypoxic and hypoglycemic culture is neurotoxic to hippocampus neuron, which may associate with the alteration of cleavage activity of α- and β- secretase, and then result in neurotoxic Aβ increased and neurotrophic SAPPα decreased.