南方医科大学学报

南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (04): 504-.

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外源性硫化氢对高糖条件下原代神经元β淀粉样蛋白的影响

朱丽娟，陈筱山，何选丽，戚韵雯，晏勇   

  • 出版日期:2014-04-20 发布日期:2014-04-20

Effect of exogenous hydrogen sulfide on BACE-1 enzyme expression and β-amyloid
peptide metabolism in high-glucose primary neuronal culture

  • Online:2014-04-20 Published:2014-04-20

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摘要： 目的探讨气体信号分子外源性硫化氢（H2S）对高糖条件下培养的原代皮层及海马神经元β位淀粉样前体蛋白裂解酶-1
（BACE-1）和β淀粉样蛋白（Aβ）代谢的影响。方法原代培养大鼠皮层及海马神经元，随机分组为正常培养神经元组、高糖
（60 mmol/L）培养神经元组、高糖（60 mmol/L）加25 μmol/L NaHS组、高糖（60 mmol/L）加50 μmol/L NaHS组和高糖（60 mmol/L）
加100 μmol/L NaHS 组，酶联免疫吸附试验检测Aβ1-42 水平，荧光实时定量检测BACE-1 mRNA表达，Western blot 检测
BACE-1蛋白表达水平。结果高糖培养的神经元组Aβ1-42水平明显高于正常培养神经元组（P<0.05），加入25、50和100 μmol/L
NaHS后，Aβ水平显著降低（P<0.05）；高糖培养的神经元组BACE-1 mRNA表达显著高于正常培养神经元组（P<0.05），加入25、
50及100 μmol/L NaHS后，BACE-1 mRNA水平显著降低（P<0.05）；高糖培养的神经元组BACE-1蛋白显著高于正常培养神经
元组（P<0.05），加入25、50及100 μmol/L NaHS后，BACE-1蛋白表达水平显著降低（P<0.05）。结论在高糖条件下，神经元Aβ
1-42、BACE-1 mRNA和蛋白表达明显升高，NaHS 呈浓度依耐性降低高糖条件下培养神经元中升高的Aβ1-42 水平，降低
BACE-1 mRNA及蛋白表达。

Abstract: Objective To investigate the effects of exogenous hydrogen sulfide (H2S) on β-site APP cleaving enzyme 1 (BACE-1)
and β-amyloid peptide (Aβ) metabolism in primary culture of neurons under high-glucose condition. Methods The cortical
neurons in primary culture under normal and high glucose (60 mmol/L) conditions for 24 h were exposed to 25, 50 and 100
μmol/L NaHS. Aβ1-42 concentration in the cell culture was measured by ELISA, and BACE-1 mRNA and protein levels were
detectedby fluorescent quantitative real-time PCR and Western blotting, respectively. Results Compared with the neurons
cultured in normal glucose, the neurons exposed to high glucose showed significantly increased Aβ1-42 concentration and
BACE-1 mRNA and protein expressions (P<0.05). Exposure to 25, 50 and 100 μmol/L NaHS significantly decreased Aβ1-42
concentration and BACE-1 mRNA and protein expressions in the high-glucose cell culture (P<0.05). Conclusion Neurons
exposed to high glucose exhibit increased Aβ1-42 levels and BACE-1 mRNA and protein expressions, which can be
concentration-dependently decreased by NaHS.